Profiling of Virulence Determinants in Cronobacter sakazakii Isolates from Different Plant and Environmental Commodities

2017 ◽  
Vol 74 (5) ◽  
pp. 560-565 ◽  
Author(s):  
Niharika Singh ◽  
Mamta Raghav ◽  
Shifa Narula ◽  
Simran Tandon ◽  
Gunjan Goel
Keyword(s):  
Cronobacter Sakazakii ◽  
Virulence Determinants ◽  
Environmental Commodities

scholarly journals Outer Membrane Proteins A (OmpA) and X (OmpX) Are Essential for Basolateral Invasion of Cronobacter sakazakii

2010 ◽  
Vol 76 (15) ◽  
pp. 5188-5198 ◽  
Author(s):  
Kyumson Kim ◽  
Kwang-Pyo Kim ◽  
Jeongjoon Choi ◽  
Jeong-A Lim ◽  
Junghyun Lee ◽  
...  
Keyword(s):  
Outer Membrane ◽  
Outer Membrane Proteins ◽  
Opportunistic Pathogen ◽  
Target Cells ◽  
Deletion Mutants ◽  
Cronobacter Sakazakii ◽  
Virulence Determinants ◽  
Rat Pups ◽  
Recombination System ◽  
Human Enterocyte

ABSTRACT Cronobacter sakazakii is an opportunistic pathogen that actively invades host eukaryotic cells. To identify invasion factors responsible for the intestinal translocation of C. sakazakii, we constructed for the first time outer membrane protein X (OmpX) and A (OmpA) deletion mutants using the lambda Red recombination system. The ompX and ompA deletion mutants showed significantly reduced invasion of human enterocyte-like epithelial Caco-2 and human intestinal epithelial INT-407 cells, and significantly fewer mutant cells were recovered from the livers and spleens of rat pups. Furthermore, compared with intact target cells, the invasion and initial association potentials of the mutants increased at a rate similar to that of the wild type in tight-junction-disrupted target cells, suggesting that OmpX and OmpA are involved in basolateral invasion by C. sakazakii. This is the first report of C. sakazakii virulence determinants that are essential for basolateral invasion and that may be critical for the virulence of C. sakazakii.

Enterococcal Virulence Determinants In Urinary Tract Infection Patients

2012 ◽  
Vol 6 (1) ◽  
pp. 14-17
Author(s):  
Jabin Akhter ◽  
Shaheda Anwar ◽  
Sharmeen Ahmed
Keyword(s):  
Urinary Tract Infection ◽  
Urinary Tract ◽  
Tract Infection ◽  
Virulence Factors ◽  
Virulence Determinants ◽  
Microtitre Plate ◽  
Microbiological Method ◽  
Enterococcal Infection ◽  
Microtitre Plate Assay ◽  
Positive Frequency

Urinary tract infection caused by Enterococci has become frequent occurrences in health care settings. Currently they emerged with increasing resistance to multiple antibiotics.  Haemolysin, gelatinase and biofilm production are some markers that have been proposed as possible Enterococcal virulence factors. In view of the increasing importance of Enterococcal infection, the present study was designed to isolate and identify the Enterococci to the species level from urine of urinary tract infection patients and to investigate their possible virulence factors. Biofilm was detected on polystyrene microtitre plate to see the adherence of microorganism. Haemolysin production and gelatin hydrolysis detected by standard microbiological method. Fifty nine enterococcal isolates were speciated by conventional microbiological method and examined for their ability to form biofilm by microtitre plate assay. In this study, biofilm formations by Enterococci were found in 83.33% isolates from catheterized and 56.09% from non-catheterized patients. Aong them, E.faecalis & 50% E.faecium produced biofilm. About 43.63% E.faecalis & 10% E.faecium produced haemolysin and only one isolate were found to be gelatinase positive. Frequency of virulence factors (VFs) in combination was observed in this study. Two VFs (haemolysin and biofilm) were observed in 27.11% in combination and 3 VFs ( haemolysinm biofilm and gelatinase) were present in 1.69% isolates. These results suggest that although there may not be an absolute role for individual virulence determinants in infectivity, combinations of factors may play a role in allowing a biofilm infection to be more resistant to therapy.DOI: http://dx.doi.org/10.3329/bjmm.v6i1.19361 Bangladesh J Med Microbiol 2012; 06(01): 14-17

Investigate of the Ability of Cronobacter sakazakii Isolated from Clinical Samples of Children Under Two Years to Induce Swimming, Swarming and Biofilm

2018 ◽  
pp. 123
Author(s):  
Luma Abdal Hady Zwein ◽  
Tharieyt Abdulrahman Motlag ◽  
Mohamed Mousa
Keyword(s):  
Cerebrospinal Fluid ◽  
Congo Red ◽  
Biofilm Formation ◽  
Microtiter Plate ◽  
Clinical Samples ◽  
Cronobacter Sakazakii ◽  
Yellow Color ◽  
Golden Yellow ◽  
Vitek 2 ◽  
Biochemical Examination

      The study included 200 samples were collected   from   children  under two   years included (50 samples from each of Cerebrospinal fluid, Blood, Stool and Urine) from, Central Children Hospital and Children's Protections Educational Hospital. Isolates bacterial were obtained cultural, microscopic and biochemical examination and diagnosed to the species by using vitek2 system. The results showed there were contamination in 6.5% of clinical samples. The diagnosed colonies which gave pink color on the MacConkey agar , golden yellow color on the Trypton Soy agar and green color on the Birillent Enterobacter sakazakii agar and gave  a probability of 99% in the vitek 2 and were identified as Cronobacter sakazakii. The identification revealed of thirteen isolates: 6(46.16%) isolated from Cerebrospinal fluid samples, 7(53.84%) isolated from blood samples and not isolated bacteria from stool and urine samples. The results of the investigation of some virulence factors showed that all bacteria isolates were able to swimming with a diameter ranging (1-9 mm) and swarming with a diameter ranging (1-40 mm) and their  ability to biofilm formation  by using three methods. The results show the ability  of  isolates to form biofilm by using  Congo red media  methods where it is 12 (92.30 %) out of 13 isolated bacteria belonging to C. sakazakii  able to form biofilm on the Congo red media  which is 3 (23.07%) were  strong production  biofilm ,   8 (61.53%)  were intermediate  production  biofilm and  1 (7.69% ) were weak  biofilm formation , while the 1 (7.69%)  unable to form biofilm.  Tubes method were all isolates were able to form biofilm, it were found that 3 (23.07%)  isolates strong, and 8 (61.53%) intermediate  and 2( 15.38%)  weak biofilm formation. Microtiter plate method  gave 5 (38.46 %) isolates strong, 6 (46.15%) intermediate and 1 (7.69%) weak biofilm formation.  

First reported case of Weissella confusa abundance in the lung microbiota of a critically ill trauma patient with acute respiratory distress syndrome - caution warranted in the use of the Weissella genus as a probiotic

2020 ◽  
Author(s):  
Sandeep Chakraborty
Keyword(s):  
Acute Respiratory Distress Syndrome ◽  
Respiratory Distress Syndrome ◽  
Respiratory Distress ◽  
Critically Ill ◽  
Distress Syndrome ◽  
Joint Infection ◽  
Starter Cultures ◽  
Trauma Patients ◽  
Virulence Determinants ◽  
Prosthetic Joint

Weissella strains are currently being used for biotechnological and probiotic purposes [1]. While, Weissella hellenica found in flounder intestine had probiotic effects [2], certain species from this genus are opportunistic pathogens in humans. Apart from being implicated in disease in farmed rainbow trout [3], Weissella has been found to cause the following disease in humans.1. endocarditis [4,5]2. bacteraemia [6]3. prosthetic joint infection [7]Whole genome sequences ‘find several virulence determinants such as collagen adhesins, aggregation sub- stances, mucus-binding proteins, and hemolysins in some species’, as well as antibiotic resistance-encoding genes [8]. Caution is warranted in selecting of Weissella strains as starter cultures or probiotics, if at all, since the other option, Lactobacillus, are rarely involved in human disease.Here, the analysis of the lung microbiota in critically ill trauma patients suffering from acute respiratory distress syndrome [9] shows (Accid:ERR1992912) shows complete colonization of Weissella (Fig 1). While, the study mentions ‘significant enrichment of potential pathogens including Streptococcus, Fusobacterium, Prevotella, Haemophilus and Treponema’, there is no reference to the Weissella genus. The percentages of Weissella strains are :confusa=81, soli=7 ,hellenica=5 ,diestrammenae=2. I believe this is the first reported case of Weissella causing ARDS in humans.

scholarly journals Transfer of Plasmid DNA to Clinical Coagulase-Negative Staphylococcal Pathogens by Using a Unique Bacteriophage

2015 ◽  
Vol 81 (7) ◽  
pp. 2481-2488 ◽  
Author(s):  
Volker Winstel ◽  
Petra Kühner ◽  
Bernhard Krismer ◽  
Andreas Peschel ◽  
Holger Rohde
Keyword(s):  
Plasmid Dna ◽  
Genetic Manipulation ◽  
Current Knowledge ◽  
Virulence Determinants ◽  
Coagulase Negative Staphylococci ◽  
Reliable Technique ◽  
Content Type ◽  
Research Activities ◽  
Wide Range ◽  
Genetic Barriers

ABSTRACTGenetic manipulation of emerging bacterial pathogens, such as coagulase-negative staphylococci (CoNS), is a major hurdle in clinical and basic microbiological research. Strong genetic barriers, such as restriction modification systems or clustered regularly interspaced short palindromic repeats (CRISPR), usually interfere with available techniques for DNA transformation and therefore complicate manipulation of CoNS or render it impossible. Thus, current knowledge of pathogenicity and virulence determinants of CoNS is very limited. Here, a rapid, efficient, and highly reliable technique is presented to transfer plasmid DNA essential for genetic engineering to important CoNS pathogens from a uniqueStaphylococcus aureusstrain via a specificS. aureusbacteriophage, Φ187. Even strains refractory to electroporation can be transduced by this technique once donor and recipient strains share similar Φ187 receptor properties. As a proof of principle, this technique was used to delete the alternative transcription factor sigma B (SigB) via allelic replacement in nasal and clinicalStaphylococcus epidermidisisolates at high efficiencies. The described approach will allow the genetic manipulation of a wide range of CoNS pathogens and might inspire research activities to manipulate other important pathogens in a similar fashion.

scholarly journals The Prevalence of Virulence Determinants and Antibiotic Resistance Patterns in Methicillin—Resistant Staphylococcus aureus in a Nursing Home in Poland

Pathogens ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 427
Author(s):  
Martyna Kasela ◽  
Agnieszka Grzegorczyk ◽  
Bożena Nowakowicz-Dębek ◽  
Anna Malm
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Variable Number Tandem Repeat ◽  
Multidrug Resistant ◽  
Variable Number ◽  
Pulse Field ◽  
Virulence Determinants ◽  
Gene Encoding ◽  
Methicillin Resistant ◽  
Pulse Field Gel Electrophoresis

Nursing homes (NH) contribute to the regional spread of methicillin-resistant Staphylococcus aureus (MRSA). Moreover, residents are vulnerable to the colonization and subsequent infection of MRSA etiology. We aimed at investigating the molecular and phenotypic characteristics of 21 MRSA collected from the residents and personnel in an NH (Lublin, Poland) during 2018. All MRSA were screened for 20 genes encoding virulence determinants (sea-see, eta, etb, tst, lukS-F-PV, eno, cna, ebpS, fib, bbp, fnbA, fnbB, icaADBC) and for resistance to 18 antimicrobials. To establish the relatedness and clonal complexes of MRSA in NH we applied multiple-locus variable-number tandem-repeat fingerprinting (MLVF), pulse field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and staphylococcal cassette chromosome mec (SCCmec) typing. We identified four sequence types (ST) among two clonal complexes (CC): ST (CC22) known as EMRSA-15 as well as three novel STs—ST6295 (CC8), ST6293 (CC8) and ST6294. All tested MRSA were negative for sec, eta, etb, lukS-F-PV, bbp and ebpS genes. The most prevalent gene encoding toxin was sed (52.4%; n = 11/21), and adhesins were eno and fnbA (100%). Only 9.5% (n = 2/21) of MRSA were classified as multidrug-resistant. The emergence of novel MRSA with a unique virulence and the presence of epidemic clone EMRSA-15 creates challenges for controlling the spread of MRSA in NH.

scholarly journals The Distribution of Several Genomic Virulence Determinants Does Not Corroborate the Established Serotyping Classification of Bacillus thuringiensis

2021 ◽  
Vol 22 (5) ◽  
pp. 2244
Author(s):  
Anton E. Shikov ◽  
Yury V. Malovichko ◽  
Arseniy A. Lobov ◽  
Maria E. Belousova ◽  
Anton A. Nizhnikov ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Protein Identification ◽  
Core Gene ◽  
Comparative Genomic ◽  
Virulence Determinants ◽  
Strain Characterization ◽  
Proteomic Approach ◽  
Liquid Chromatography Tandem Mass ◽  
Identity Threshold ◽  
Genome Assemblies

Bacillus thuringiensis, commonly referred to as Bt, is an object of the lasting interest of microbiologists due to its highly effective insecticidal properties, which make Bt a prominent source of biologicals. To categorize the exuberance of Bt strains discovered, serotyping assays are utilized in which flagellin serves as a primary seroreactive molecule. Despite its convenience, this approach is not indicative of Bt strains’ phenotypes, neither it reflects actual phylogenetic relationships within the species. In this respect, comparative genomic and proteomic techniques appear more informative, but their use in Bt strain classification remains limited. In the present work, we used a bottom-up proteomic approach based on fluorescent two-dimensional difference gel electrophoresis (2D-DIGE) coupled with liquid chromatography/tandem mass spectrometry(LC-MS/MS) protein identification to assess which stage of Bt culture, vegetative or spore, would be more informative for strain characterization. To this end, the proteomic differences for the israelensis-attributed strains were assessed to compare sporulating cultures of the virulent derivative to the avirulent one as well as to the vegetative stage virulent bacteria. Using the same approach, virulent spores of the israelensis strain were also compared to the spores of strains belonging to two other major Bt serovars, namely darmstadiensis and thuringiensis. The identified proteins were analyzed regarding the presence of the respective genes in the 104 Bt genome assemblies available at open access with serovar attributions specified. Of 21 proteins identified, 15 were found to be encoded in all the present assemblies at 67% identity threshold, including several virulence factors. Notable, individual phylogenies of these core genes conferred neither the serotyping nor the flagellin-based phylogeny but corroborated the reconstruction based on phylogenomics approaches in terms of tree topology similarity. In its turn, the distribution of accessory protein genes was not confined to the existing serovars. The obtained results indicate that neither gene presence nor the core gene sequence may serve as distinctive bases for the serovar attribution, undermining the notion that the serotyping system reflects strains’ phenotypic or genetic similarity. We also provide a set of loci, which fit in with the phylogenomics data plausibly and thus may serve for draft phylogeny estimation of the novel strains.

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