scholarly journals Characterization and Analysis of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPRs) in Pandemic and Non-Pandemic Vibrio parahaemolyticus Isolates from Seafood Sources

2021 ◽  
Vol 9 (6) ◽  
pp. 1220
Author(s):  
Nawaporn Jingjit ◽  
Sutima Preeprem ◽  
Komwit Surachat ◽  
Pimonsri Mittraparp-arthorn
Keyword(s):  
Antibiotic Resistance ◽  
Antimicrobial Susceptibility ◽  
Hemolytic Activity ◽  
Vibrio Parahaemolyticus ◽  
Sequence Type ◽  
Multiple Antibiotic Resistance ◽  
Hemolysin Gene ◽  
Shared Ancestry ◽  
Thermostable Direct Hemolysin ◽  
Mar Index

Vibrio parahaemolyticus is one of the significant seafood-borne pathogens causing gastroenteritis in humans. Clustered regularly interspaced short palindromic repeats (CRISPR) are commonly detected in the genomes of V. parahaemolyticus and the polymorphism of CRISPR patterns has been applied as a genetic marker for tracking its evolution. In this work, a total of 15 pandemic and 36 non-pandemic V. parahaemolyticus isolates obtained from seafood between 2000 and 2012 were characterized based on hemolytic activity, antimicrobial susceptibility, and CRISPR elements. The results showed that 15/17 of the V. parahaemolyticus seafood isolates carrying the thermostable direct hemolysin gene (tdh+) were Kanagawa phenomenon (KP) positive. The Multiple Antibiotic Resistance (MAR) index ranged between 0.1 and 0.4, and 45% of the isolates have an MAR index ≥ 0.2. A total of 19 isolates were positive for CRISPR detection, including all tdh+ trh− isolates, two of tdh− trh+, and each of tdh+ trh+ and tdh− trh−. Four spacer types (Sp1 to Sp4) were identified, and CRISPR-positive isolates had at least one type of spacer homolog to the region of Vibrio alginolyticus megaplasmid. It is of interest that a specific CRISPR profile and spacer sequence type was observed with correlations to the hemolysin genotype (tdh/trh). Thus, these provide essential data on the exposure of foreign genetic elements and indicate shared ancestry within different genotypes of V. parahaemolyticus isolates.

scholarly journals Antibiotic resistance pattern and its correlation to the presence of tdh gene and CRISPR-Cas system in Vibrio parahaemolyticus strains isolated from seafood

2019 ◽  
Vol 66 (2) ◽  
Author(s):  
Pallavi Baliga ◽  
Malathi Shekar ◽  
Shaik Thahur Ahamed ◽  
M. N. Venugopal
Keyword(s):  
Antibiotic Resistance ◽  
Negative Correlation ◽  
Antimicrobial Susceptibility ◽  
Vibrio Parahaemolyticus ◽  
Multidrug Resistant ◽  
Resistance Pattern ◽  
Susceptibility Pattern ◽  
Antimicrobial Susceptibility Pattern ◽  
Thermostable Direct Hemolysin ◽  
Tdh Gene

Vibrio parahaemolyticus is a pathogen native to the aquatic environment. In this study, 46 environmental V. parahaemolyticus isolates were subjected to a correlational analysis to find the association between their antimicrobial susceptibility pattern, prevalence of CRISPR-Cas system and thermostable direct hemolysin (tdh) gene. Antibiotic resistance profiling against eleven antibiotics revealed the isolates to be multidrug resistant. Isolates exhibited highest resistance to vancomycin (97.8%) followed by ampicillin (91.3%), cefotaxime (69.6%), ceftazidime/clavulanic acid (54.4%), ceftazidime (45.7%) and gentamicin (39.1%). CRISPR loci and tdh gene were detected in 47.83 and 58.7% of strains respectively. No significant correlation was observed between antibiotic resistance to presence of CRISPR, except in the case of gentamicin wherein, a negative correlation was seen (r=-0.272, p<0.10). Similarly, tdh did not correlate to antibiotic resistance. Seventeen strains in this study harboured the CRISPR loci as well as tdh gene, the association of which was found to be statistically significant.

scholarly journals MULTIPLE ANTIBIOTIC RESISTANCE INDEXING OF NON- FERMENTING GRAM NEGATIVE BACILLI

2017 ◽  
Vol 10 (6) ◽  
pp. 78
Author(s):  
Bhuvaneshwari Gunasekar
Keyword(s):  
Antibiotic Resistance ◽  
Tertiary Care ◽  
Multidrug Resistant ◽  
Diffusion Method ◽  
Care Hospital ◽  
Multiple Antibiotic Resistance ◽  
Antibiotic Resistant ◽  
Gram Negative ◽  
Mar Index ◽  
Surgery Department

Objective: The multiple antibiotic resistance (MAR) indexing and finding Multidrug resistant (MDR) bacteria will help to indicate the origin from high risk of contamination where the antibiotics are often used. Hence this study was carried out to give the MAR index of non-fermenting Gram negative bacilli in a tertiary care hospital which would help our infection control team also.Methods: Drug resistance was tested by Kirby bauer’s disc diffusion method. MAR index was calculated using the formula, a/b (were a= number of antibiotics to which the organism was resistant and b= total number of antibiotics to which the organism was tested).Results: Out of 240 Gram negative non-fermenters isolated, 117 (49%) strains were greater than 0.2 of MAR index, 95(81%) was from in-patient department. 73(62%) were hospitalized for more than 3 days, 44 (38%) was from surgery department. 49(42%) was wound specimen. Out of 117 multiple antibiotic resistant isolates 99 (85%) were MDR isolates.Conclusion: 51% prevalence of isolates >0.2 MAR index shows that the source of contamination can still be brought up down by proper surveillance and management with proper usage of  surface and skin disinfectants especially in surgery ward where the MAR index has indicated more usage of antibiotics

Increased Sensitivity in PCR Detection of tdh-Positive Vibrio parahaemolyticus in Seafood with Purified Template DNA

2003 ◽  
Vol 66 (9) ◽  
pp. 1675-1680 ◽  
Author(s):  
Y. HARA-KUDO ◽  
Y. KASUGA ◽  
A. KIUCHI ◽  
T. HORISAKA ◽  
T. KAWASUMI ◽  
...  
Keyword(s):  
Glass Fiber ◽  
Vibrio Parahaemolyticus ◽  
Detection Sensitivity ◽  
Sea Bream ◽  
Silica Membrane ◽  
Membrane Method ◽  
Hemolysin Gene ◽  
Thermostable Direct Hemolysin ◽  
Increased Sensitivity ◽  
Template Dna

PCR is an important method for the detection of thermostable direct hemolysin gene (tdh)–positive (pathogenichemolysin-producing) strains of Vibrio parahaemolyticus in seafood because tdh-negative (nonpathogenic) V. parahaemolyticus strains often contaminate seafood and interfere with the direct isolation of tdh-positive V. parahaemolyticus. In this study, the use of PCR to detect the tdh gene of V. parahaemolyticus in various seafoods artificially contaminated with tdh-positive V. parahaemolyticus was examined. PCR was inhibited by substances in oysters, squid, mackerel, and yellowtail but not by cod, sea bream, scallop, short-necked clam, and shrimp. To improve detection, DNA was purified by either the silica membrane method, the glass fiber method, or the magnetic separation method, and the purified DNA was used as the PCR primer template. For all samples, the use of the silica membrane method and the glass fiber method increased detection sensitivity. The results of this study demonstrate that the use of properly purified template DNA for PCR markedly increases the effectiveness of the method in detecting pathogenic tdh-positive V. parahaemolyticus in contaminated seafood.

scholarly journals Association of Vibrio parahaemolyticus Thermostable Direct Hemolysin with Lipid Rafts Is Essential for Cytotoxicity but Not Hemolytic Activity

2009 ◽  
Vol 78 (2) ◽  
pp. 603-610 ◽  
Author(s):  
Shigeaki Matsuda ◽  
Toshio Kodama ◽  
Natsumi Okada ◽  
Kanna Okayama ◽  
Takeshi Honda ◽  
...  
Keyword(s):  
Lipid Rafts ◽  
Hemolytic Activity ◽  
Vibrio Parahaemolyticus ◽  
Cultured Cells ◽  
Direct Interaction ◽  
Caveolin 1 ◽  
Marker Proteins ◽  
Thermostable Direct Hemolysin ◽  
Target Molecules

ABSTRACT Thermostable direct hemolysin (TDH), a major virulence factor of Vibrio parahaemolyticus, induces cytotoxicity in cultured cells. However, the mechanism of TDH's cytotoxic effect including its target molecules on the plasma membrane of eukaryotic cells remains unclear. In this study, we identified the role of lipid rafts, cholesterol- and sphingolipid-enriched microdomains, in TDH cytotoxicity. Treatment of cells with methyl-β-cyclodextrin (MβCD), a raft-disrupting agent, inhibited TDH cytotoxicity. TDH was associated with detergent-resistant membranes (DRMs), and MβCD eliminated this association. In contrast, there was no such association between a nontoxic TDH mutant and DRMs. The disruption of lipid rafts neither affected hemolysis nor inhibited Ca2+ influx into HeLa cells induced by TDH. These findings indicate that the cytotoxicity but not the hemolytic activity of TDH is dependent on lipid rafts. The exogenous and endogenous depletion of cellular sphingomyelin also prevented TDH cytotoxicity, but a direct interaction between TDH and sphingomyelin was not detected with either a lipid overlay assay or a liposome absorption test. Treatment with sphingomyelinase (SMase) at 100 mU/ml disrupted the association of TDH with DRMs but did not affect the localization of lipid raft marker proteins (caveolin-1 and flotillin-1) with DRMs. These results suggest that sphingomyelin is important for the association of TDH with lipid rafts but is not a molecular target of TDH. We hypothesize that TDH may target a certain group of rafts that are sensitive to SMase at a certain concentration, which does not affect other types of rafts.

scholarly journals Hfq regulates the expression of the thermostable direct hemolysin gene in Vibrio parahaemolyticus

2008 ◽  
Vol 8 (1) ◽  
pp. 155 ◽  
Author(s):  
Masayuki Nakano ◽  
Akira Takahashi ◽  
Zehong Su ◽  
Nagakatsu Harada ◽  
Kazuaki Mawatari ◽  
...  
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Hemolysin Gene ◽  
Thermostable Direct Hemolysin

scholarly journals Extended Spectrum Beta Lactamases Detection and Multiple Antibiotic Resistance Indexing of Escherichia Coli from Urine Samples of Patients from a Referral Hospital of Eastern Nepal

2015 ◽  
Vol 3 (3) ◽  
pp. 423-426 ◽  
Author(s):  
A. Chakrawarti ◽  
P. Dongol ◽  
H. Khanal ◽  
P. Subba ◽  
J.J. Benerjee
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Urine Samples ◽  
Multiple Antibiotic Resistance ◽  
Beta Lactamases ◽  
E Coli ◽  
Extended Spectrum Beta Lactamases ◽  
Mar Index ◽  
Esbl Producers ◽  
Disc Assay

Background: Escherichia coli is the most common causative agent of urinary tract infection. Antibiotic resistance among uropathogens has become a prominent public health problem. Multidrug resistance bacteria have limited the therapeutic possibilities by producing Extended Spectrum Beta Lactamases (ESBL). Objective: Since routine monitoring of ESBL producers are not conducted in clinical laboratories their true prevalence is still unknown. So the objective of this research was to assess multiple antibiotic resistance (MAR) indices and determine ESBL production among Escherichia coli isolated from urine samples. Methods: Standard microbiological techniques and antibiotic sensitivity test were performed by Kirby Bauer disc diffusion method to identify E. coli. ESBL screening was done by using Ceftriaxone, Aztreonam, Cefotaxime, Ceftazidime and Cefpodoxime whereas confirmation by combined disc assay. SPSS 16 software was used to analyze data. Results: 86.95% E. coli isolates were MDR strains. 27 isolates had multiple antibiotic resistance (MAR) index of 0.2 and 5 isolates had MAR index of 0.7. E. coli isolates showed higher degree of resistance towards Amoxicillin (100%) while 100% were sensitive towards Gentamicin followed by Nitrofurantoin (62.31%). The reliable screening agent for ESBL detection with sensitivity 100% and positive predictive value of 80% was Cefotaxime. Combined disc assay detected 12/69 (17.31%) of E. coli isolates as confirmed ESBL producers. Conclusion: The ubiquity of ESBL-producing E. coli was observed emphasizing the necessity of regular surveillance of ESBL producing clinical isolates in clinical samples to minimize multi-drug resistance strains and avert the ineffectiveness of antimicrobial agent for good health practices.\Int J Appl Sci Biotechnol, Vol 3(3): 423-426

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