scholarly journals IDENTIFICATION TARGETED ON toxR GENE AND DETECTION OF VIRULENT FACTOR ENCODING GENES ON Vibrio parahaemolyticus ISOLATED FROM RAW CHICKEN MEAT

2011 ◽  
Vol 2 (1) ◽  
pp. 7-11
Author(s):  
Eka Fitrianda
Keyword(s):  
Virulence Factors ◽  
Chicken Meat ◽  
Specific Gene ◽  
Gene Encoding ◽  
Genes Encoding ◽  
Thermostable Direct Hemolysin ◽  
Pcr Method ◽  
Toxr Gene ◽  
Meat Samples ◽  
Encoding Genes

A number of V. parahaemolyticus isolates has been isolated from raw chicken meat samples collected in Pasar raya Padang. Isolation was done using ChromagarTM Vibrio media. Identification of toxR gene was then done to these isolates. toxR is a very specific gene in V. parahaemolyticus species. Detection for the presence of genes encoding virulence factors, in this case were the gene encoding thermostable direct hemolysin (tdh) and TDH related hemolysin (trh) was performed on toxR positive V. parahaemolyticus isolates. Identification of toxR gene and detection of tdh and trh genes were done trough amplification using polymerase chain reaction PCR (method). The results showed that all of the tested V. parahaemolyticus isolates (22 isolates) had toxR gene, but none of isolates has gene encoding the production of virulence factors both tdh and trh.

scholarly journals An n-Alkane-Responsive Promoter Element Found in the Gene Encoding the Peroxisomal Protein of Candida tropicalis Does Not Contain a C6 Zinc Cluster DNA-Binding Motif

2000 ◽  
Vol 182 (9) ◽  
pp. 2492-2497 ◽  
Author(s):  
Tamotsu Kanai ◽  
Akihiro Hara ◽  
Naoki Kanayama ◽  
Mitsuyoshi Ueda ◽  
Atsuo Tanaka
Keyword(s):  
Candida Tropicalis ◽  
Promoter Region ◽  
Binding Motif ◽  
Gene Encoding ◽  
Peroxisomal Enzyme ◽  
Zinc Cluster ◽  
Genes Encoding ◽  
Tata Sequence ◽  
Encoding Genes ◽  
Activation Sequence

ABSTRACT When an asporogenic diploid yeast, Candida tropicalis, is cultivated on n-alkane, the expression of the genes encoding enzymes of the peroxisomal β-oxidation pathway is highly induced. An upstream activation sequence (UAS) which can induce transcription in response to n-alkane (UASALK) was identified on the promoter region of the peroxisomal 3-ketoacyl coenzyme A (CoA) thiolase gene of C. tropicalis(CT-T3A). The 29-bp region (from −289 to −261) present upstream of the TATA sequence was sufficient to inducen-alkane-dependent expression of a reporter gene. Besidesn-alkane, UASALK-dependent gene expression also occurred in the cells grown on oleic acid. Several kinds of mutant UASALK were constructed and tested for their UAS activity. It was clarified that the important nucleotides for UASALKactivity were located within 10-bp region from −273 to −264 (5′-TCCTGCACAC-3′). This region did not contain a CGG triplet and therefore differed from the sequence of the oleate-response element (ORE), which is a UAS found on the promoter region of 3-ketoacyl-CoA thiolase gene of Saccharomyces cerevisiae. Similar sequences to UASALK were also found on several peroxisomal enzyme-encoding genes of C. tropicalis.

scholarly journals Identification and Characterization of Hemolysin-Like Proteins Similar to RTX Toxin in Pasteurella pneumotropica

2009 ◽  
Vol 191 (11) ◽  
pp. 3698-3705 ◽  
Author(s):  
Hiraku Sasaki ◽  
Eiichi Kawamoto ◽  
Yoshikazu Tanaka ◽  
Takuo Sawada ◽  
Satoshi Kunita ◽  
...  
Keyword(s):  
Virulence Factors ◽  
Opportunistic Pathogen ◽  
Pcr Analysis ◽  
Type I ◽  
Secretion Systems ◽  
Rtx Toxin ◽  
Rtx Toxins ◽  
Genes Encoding ◽  
Encoding Genes

ABSTRACT Pasteurella pneumotropica is an opportunistic pathogen that causes lethal pneumonia in immunodeficient rodents. The virulence factors of this bacterium remain unknown. In this study, we identified the genes encoding two RTX toxins, designated as pnxI and pnxII, from the genomic DNA of P. pneumotropica ATCC 35149 and characterized with respect to hemolysis. The pnxI operon was organized according to the manner in which the genes encoded the structural RTX toxin (pnxIA), the type I secretion systems (pnxIB and pnxID), and the unknown orf. The pnxII gene was involved only with the pnxIIA that coded for a structural RTX toxin. Both the structural RTX toxins of deduced PnxIA and PnxIIA were involved in seven of the RTX repeat and repeat-like sequences. By quantitative PCR analysis of the structural RTX toxin-encoding genes in P. pneumotropica ATCC 35149, the gene expression of pnxIA was found to have increased from the early log phase, while that of pnxIIA increased from the late log to the early stationary phase. As expressed in Escherichia coli, both the recombinant proteins of PnxIA and PnxIIA showed weak hemolytic activity in both sheep and murine erythrocytes. On the basis of the results of the Southern blotting analysis, the pnxIA gene was detected in 82% of the isolates, while the pnxIIA gene was detected in 39%. These results indicate that the products of both pnxIA and pnxIIA were putative associations of virulence factors in the rodent pathogen P. pneumotropica.

scholarly journals MOLECULAR DETECTION OF PROTEIN ENCODING GENES Virb11 ON BRUCELLA ABORTUS LOCAL ISOLATES ORIGIN OF PINRANG, NTT AND VACCINE STRAINS

2020 ◽  
Vol 21 (4) ◽  
pp. 503-511
Author(s):  
Maria Gladis Bupu Maze ◽  
Didik Handijatno ◽  
Wiwik Tyasningsih ◽  
Suwarno Suwarno ◽  
Agnes Theresia Soelih Estoepangestie ◽  
...  
Keyword(s):  
Virulence Factors ◽  
Virulence Factor ◽  
Brucella Abortus ◽  
Molecular Detection ◽  
Livestock Population ◽  
Gram Staining ◽  
Protein Encoding ◽  
Important Virulence Factor ◽  
Pcr Method ◽  
Encoding Genes

Brucellosis in cattle is a disease caused by Brucella abortus due to the reduction in livestock population caused by abortion, stillbirth, weak birth, infertility and sterility. Brucella abortus has several potential virulence factors, i.e. virB11 gene that encodes VirB11 protein is an important virulence factor acts as an ATPase for assembling organelles when the bacteria replicate, helping to complete the bacterial cycle and agress to another cells. The aim of this study are to re-identification Brucella abortus and detect virB11 gene as encoding of B. abortus VirB11 protein in local isolates from Pinrang, NTT, strain vaccines S19 and RB51. The isolates Brucella abortus were re-cultured in Brucella agar base and re-identification is followed by microscopic with Gram staining and biochemical tested with urease, citrat, indol and TSIA test. virB11gene was detected with PCR method. The PCR result showed virB11 gene have DNA band 720 bp. virB11 gene are present in local isolates from Pinrang, NTT, strain vaccines S19 and RB51.

scholarly journals Gas Gangrene of Different Origin Associated with Clostridium perfringens Type A in Three Patients Simultaneously Hospitalized in a Single Department of Orthopedics and Traumatology in Poland

2016 ◽  
Vol 65 (4) ◽  
pp. 399-406
Author(s):  
Monika Brzychczy-Włoch ◽  
Dorota Ochońska ◽  
Anna Piotrowska ◽  
Małgorzata Bulanda
Keyword(s):  
Clostridium Perfringens ◽  
Gas Gangrene ◽  
Pulsed Field Gel Electrophoresis ◽  
Gene Encoding ◽  
The Third ◽  
Resistance Patterns ◽  
Biochemical Profiles ◽  
Pcr Method ◽  
Encoding Genes

The objective of the study was to perform a comparative analysis of phenotypic and genetic similarity, determination of resistance profiles, detection of toxin-encoding genes and molecular typing of Clostridium perfringens isolates originating from patients with gas gangrene. The study encompassed three patients with a clinical and microbiological diagnosis of gas gangrene who were hospitalized in one of the hospitals of the Kujawsko-Pomorskie province in the same period of time between 8th April 2015 and 20th April 2015. The three C. perfringens isolates studied had identical biochemical profiles. Two isolates had identical resistance patterns, while the third presented a different profile. Using the multiplex PCR method, all isolates showed the presence of cpa gene encoding α-toxin; furthermore, the presence of the cpb2 gene encoding β2-toxin was confirmed in two isolates. Genotyping with the use of pulsed field gel electrophoresis (PFGE) indicated that the isolates originating from the three studied patients represent three genetically different restrictive patterns which corresponded to three different clones – clone A, clone B and clone C. As a result of the study, it is possible to conclude that the studied patients simultaneously hospitalized in a single Department of Orthopedics and Traumatology developed three different endogenous infections.

Development of a Rapid On-Site Method for the Detection of Chicken Meat in Processed Ground Meat Products by Using a Direct Ultrafast PCR System

2020 ◽  
Vol 83 (6) ◽  
pp. 984-990 ◽  
Author(s):  
SUYEON SUL ◽  
MI-JU KIM ◽  
JUNG-MIN LEE ◽  
SUNG-YEON KIM ◽  
HAE-YEONG KIM
Keyword(s):  
Limit Of Detection ◽  
Meat Products ◽  
Accurate Method ◽  
Chicken Meat ◽  
Rrna Gene ◽  
Target Sequence ◽  
Ground Meat ◽  
Specific Pcr ◽  
Pcr Method ◽  
Meat Samples

ABSTRACT In this study, we developed a rapid on-site detection method by using direct ultrafast PCR coupled with a microfluidic chip to identify the presence of chicken meat in processed ground meat products. Chicken-specific PCR primer targeting mitochondrial 16S rRNA gene was newly designed, and its specificity was confirmed against 17 other animal species and 4 different chicken meat samples from different countries of origin. The sensitivity of the chicken-specific ultrafast PCR was 0.1 pg of chicken DNA. To evaluate the limit of detection of the direct ultrafast PCR method, different percentages of chicken meat mixed with pork or beef were prepared. The limit of detection of the direct ultrafast PCR method for the chicken meat–pork and chicken meat–beef mixtures was 0.1% for both raw meat and autoclaved meat. This method was used for 15 commercialized processed ground meat products. In this method, the target sequence was successfully amplified, and the presence of chicken meat in processed ground meat products was identified within approximately 25 min, including the time for sample preparation. Thus, our study shows that this developed direct ultrafast PCR method is a rapid and accurate method for on-site detection of chicken DNA in commercial food products. HIGHLIGHTS

scholarly journals The katX Gene, Which Codes for the Catalase in Spores of Bacillus subtilis, Is a Forespore-Specific Gene Controlled by ςF, and KatX Is Essential for Hydrogen Peroxide Resistance of the Germinating Spore

1998 ◽  
Vol 180 (8) ◽  
pp. 2057-2062 ◽  
Author(s):  
Irina Bagyan ◽  
Lilliam Casillas-Martinez ◽  
Peter Setlow
Keyword(s):  
Hydrogen Peroxide ◽  
Bacillus Subtilis ◽  
Mature Spore ◽  
Specific Gene ◽  
Gene Encoding ◽  
Major Function ◽  
Gene Coding ◽  
Genes Encoding ◽  
Dormant Spore

ABSTRACT Previous work has shown that the katX gene encodes the major catalase in dormant spores of Bacillus subtilis but that this enzyme has no role in dormant spore resistance to hydrogen peroxide. Expression of a katX-lacZ fusion began at approximately h 2 of sporulation, and >75% of thekatX-driven β-galactosidase was packaged into the mature spore. A mutation in the gene coding for the sporulation-specific RNA polymerase sigma factor ςF abolishedkatX-lacZ expression, while mutations in genes encoding ςE, ςG, and ςK did not. Induction of ςF synthesis in vegetative cells also resulted in katX-lacZ expression, while induction of ςG expression did not; the katX-lacZ fusion was also not induced by hydrogen peroxide. Upstream of the in vivokatX transcription start site there are sequences with good homology to those upstream of known ςF-dependent start sites. These data indicate that katX is an additional member of the forespore-specific ςF regulon. A mutant in the katA gene, encoding the major catalase in growing cells, was sensitive to hydrogen peroxide during sporulation, while akatX mutant was not. However, outgrowth of katXspores, but not katA spores, was sensitive to hydrogen peroxide. Consequently, a major function for KatX is to protect germinating spores from hydrogen peroxide.

scholarly journals Screening of Antimicrobial Resistance Genes and Epidemiological Features in Hospital and Community-Associated Carbapenem Resistant Pseudomonas aeruginosa Infections

2020 ◽  
Author(s):  
ayse erturk ◽  
Ayşegül Çopur ÇİÇEK ◽  
Nebahat EJDER ◽  
Uğur KOSTAKOĞLU ◽  
İlknur Esen YILDIZ ◽  
...  
Keyword(s):  
Resistance Genes ◽  
Multidrug Resistant ◽  
Gene Encoding ◽  
Pfge Typing ◽  
Carbapenem Resistant ◽  
Antimicrobial Resistance Genes ◽  
Genes Encoding ◽  
Pcr Method ◽  
Vitek 2 ◽  
Hospital Acquired

Abstract Background: Researching carbapenem-resistant isolates and the use of antibiotics and following infection control policies enable the identification of carbapenemase-producing bacteria and prevent their spread.Methods: P. aeruginosa isolates were recovered from Medicine Faculty of Recep Tayyip Erdoğan University between April 2015 and October 2016 and identified by conventional methods and the automated Vitek 2 Compact (BioMerieux, France) system. Antimicrobial susceptibility experiments were performed in accordance with CLSI criteria and the automated Vitek 2 Compact system. The PCR method was investigated for the presence of β-lactamase resistance genes. PFGE typing was performed to show clonal relation among samples.Results: Seventy P. aeruginosa strains were isolated from seventy patients. The median age of 70 cases was found 66 with minimum 17 and maximum 92 years old. 67.1% of the patients had contact with the health service in the last 90 days and 75.7% of the patients had received antimicrobial therapy in the previous 90 days. The most common comorbidity was cardiovascular diseases. Twenty-four (34.3%) strains were carbapenem resistant, 2 strains were multidrug-resistant except colistin, and none of the samples had colistin resistance. The gene encoding β-lactamase or metallo-β-lactamase was found in a total of 36 strains. The bla VEB gene was identified in only 1 strain alone, but in combination with other resistance genes in a total of 17 strains. While the bla PER gene was detected in 5 samples alone, it was found in 13 samples in combination with other genes. Among the genes encoding metallo-β-lactamase, the most bla NDM positive was detected (n=22), followed by 14 positive samples of bla KPC . bla IMP and bla VIM were detected in 5 and 1 samples, respectively. Also, the association of bla VEB - bla PER and bla VEB - bla KPC - bla NDM was found to be very high. Much more resistance genes and associations were detected in hospital-acquired samples than community-acquired samples, both proportionally and in terms of co-occurrence. Most of the community-associated strains were collected in the F2 clade, while most of the hospital-associated strains were collected in the G1 clade. However, no difference was found between the community and hospital-associated strains according to PFGE results. Simultaneously, other microorganisms were also isolated from patients from which these 6 P. aeruginosa strains were isolated. Of these patients, 5 patients died, except the number 70.Conclusions: The median length of stay (days) was found to be significantly higher in the group with HAI than in the group with CAI. Compared to sample 28 and 37, which carried 5 β-lactamase coding genes, the death of these 5 patients with fewer or no resistance genes showed that the coexistence of other factors - especially other microorganisms in addition to resistance genes, was important.

scholarly journals Diversity of Virulence Factors Associated with West Australian Methicillin-SensitiveStaphylococcus aureusIsolates of Human Origin

2016 ◽  
Vol 2016 ◽  
pp. 1-10 ◽  
Author(s):  
Charlene Babra Waryah ◽  
Jully Gogoi-Tiwari ◽  
Kelsi Wells ◽  
Karina Yui Eto ◽  
Elnaz Masoumi ◽  
...  
Keyword(s):  
Virulence Factors ◽  
Genetic Relatedness ◽  
Teichoic Acid ◽  
Relative Distribution ◽  
Capsular Polysaccharides ◽  
Human Origin ◽  
Gene Encoding ◽  
Factors Associated ◽  
Rapd Pcr ◽  
Genes Encoding

An extensive array of virulence factors associated withS. aureushas contributed significantly to its success as a major nosocomial pathogen in hospitals and community causing variety of infections in affected patients. Virulence factors include immune evading capsular polysaccharides, poly-N-acetyl glucosamine, and teichoic acid in addition to damaging toxins including hemolytic toxins, enterotoxins, cytotoxins, exfoliative toxin, and microbial surface components recognizing adhesive matrix molecules (MSCRAMM). In this investigation, 31 West AustralianS. aureusisolates of human origin and 6 controls were analyzed for relative distribution of virulence-associated genes using PCR and/or an immunoassay kit and MSCRAMM by PCR-based typing. Genes encoding MSCRAMM, namely, Spa, ClfA, ClfB, SdrE, SdrD, IsdA, and IsdB, were detected in >90% of isolates. Gene encodingα-toxin was detected in >90% of isolates whereas genes encodingβ-toxin and SEG were detectable in 50–60% of isolates. Genes encoding toxin proteins, namely, SEA, SEB, SEC, SED, SEE, SEH, SEI, SEJ, TSST, PVL, ETA, and ETB, were detectable in >50% of isolates. Use of RAPD-PCR for determining the virulence factor-based genetic relatedness among the isolates revealed five cluster groups confirming genetic diversity among the MSSA isolates, with the greatest majority of the clinicalS. aureus(84%) isolates clustering in group IIIa.

scholarly journals Occurrence of extended spectrum β-lactamaseand AmpC-producing Escherichia coli in meat samples

2013 ◽  
Vol 57 (4) ◽  
pp. 513-517 ◽  
Author(s):  
Bernard Wasiński ◽  
Hanna Różańska ◽  
Jacek Osek
Keyword(s):  
Escherichia Coli ◽  
Inhibitory Concentration ◽  
E Coli ◽  
Extended Spectrum ◽  
Genes Encoding ◽  
Meat Samples ◽  
Phenotypic Tests ◽  
Encoding Genes ◽  
Pork Samples

Abstract The aim of the study was a preliminary determination of occurrence of extended spectrum β-lactamases (ESBL)- and AmpC-producing Escherichia coli (E.coli) in raw meat samples collected from slaughter-houses located in different regions of Poland. A total of 141 samples were tested, comprising 78 pork samples, 44 beef samples, and 19 chicken meat samples. Isolated and identified E. coli strains were examined for the ESBL and/or AmpC β-lactamases production by the use of four disc diffusion and minimum inhibitory concentration tests. All strains positive in one or both tests were examined by PCR for the presence of the blaCTX, blaTEM, blaSHV, and blaCMY-2 group genes. During the study, 154 E. coli strains were isolated from 95 samples. Among these, 18 (11.7%) strains were identified in phenotypic tests as ESBL-producing and seven (4.5%) strains as AmpC-positive. The presence of the genes encoding selected ESBL-s (TEM, CTX, SHV) was identified in 14 of the strains recognised as ESBLpositive in phenotypic tests. All AmpC-positive isolates showed the presence of the CMY-2 group encoding genes. One of these strains had also the CTX-M and TEM genes, and four of them expressed the TEM marker.

scholarly journals Identification of Three Genes Encoding PII-Like Proteins in Gluconacetobacter diazotrophicus: Studies of Their Role(s) in the Control of Nitrogen Fixation

2003 ◽  
Vol 185 (19) ◽  
pp. 5854-5861 ◽  
Author(s):  
Olena Perlova ◽  
Alejandro Ureta ◽  
Stefan Nordlund ◽  
Dietmar Meletzus
Keyword(s):  
Gene Expression ◽  
Nitrogen Fixation ◽  
Gluconacetobacter Diazotrophicus ◽  
Triple Mutant ◽  
Gene Encoding ◽  
Protein Encoding ◽  
Genes Encoding ◽  
Encoding Gene ◽  
Encoding Genes ◽  
Expression And Activity

ABSTRACT In our studies on the regulation of nitrogen metabolism in Gluconacetobacter diazotrophicus, an endophytic diazotroph of sugarcane, three glnB-like genes were identified and their role(s) in the control of nitrogen fixation was studied. Sequence analysis revealed that one PII protein-encoding gene, glnB, was adjacent to a glnA gene (encoding glutamine synthetase) and that two other PII protein-encoding genes, identified as glnK1 and glnK2, were located upstream of amtB1 and amtB2, respectively, genes which in other organisms encode ammonium (or methylammonium) transporters. Single and double mutants and a triple mutant with respect to the three PII protein-encoding genes were constructed, and the effects of the mutations on nitrogenase expression and activity in the presence of either ammonium starvation or ammonium sufficiency were studied. Based on the results presented here, it is suggested that none of the three PII homologs is required for nif gene expression, that the GlnK2 protein acts primarily as an inhibitor of nif gene expression, and that GlnB and GlnK1 control the expression of nif genes in response to ammonium availability, both directly and by relieving the inhibition by GlnK2. This model includes novel regulatory features of PII proteins.

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