scholarly journals Characteristics of bactericidal activity of healthy donors' peripheral blood against Escherichia coli

2020 ◽  
pp. 48-51
Author(s):  
A Godovalov ◽  
I Boev
Keyword(s):  
Escherichia Coli ◽  
Immunoglobulin G ◽  
Peripheral Blood ◽  
Clinical Manifestations ◽  
Absorption Capacity ◽  
Antibiofilm Activity ◽  
Microbicidal Activity ◽  
E Coli ◽  
Healthy Donors ◽  
Wide Range

Recently, the role of opportunistic pathogenic microorganisms in the development of inflammatory diseases of various localization has been relatively widely discussed. It is known that Escherichia coli are the etiological agent of such diseases, especially extraintestinal localization, a distinctive feature of which is considered to be a wide range of clinical manifestations. The aim of investigation was to assess the contribution of some components to the total microbicidal activity of healthy donors peripheral blood. Materials and methods. The bactericidal capacity of peripheral blood was assessed by the number of viable E. coli after incubation with blood samples by plating on Endo medium. The antibiofilm activity of whole blood and blood plasma was assessed by direct contact with the biofilm and its subsequent visualization using O'Toole technique. Phagocytic activity of neutrophils and monocytes was assessed separately in a microscopic version of the test. The production of reactive oxygen species was assessed using the stimulated and spontaneous luminol-dependent chemiluminescence reaction. An immunoglobulin G was used to opsonize E. coli. Results and discussion. It was shown that the peripheral blood of healthy volunteers has pronounced bactericidal properties against E. coli, reducing the number of viable bacteria by 2 times. Opsonization of E. coli reduces the number of bacteria by 4 times. Among leukocytes, the predominant place is occupied by neutrophils, which activate their absorption capacity and production of radicals faster than monocytes. The number of actively phagocytic neutrophils after E. coli treatment with an immunoglobulin G increased to 4.1±1.0% (without opsonization - 1.7±0.3%; p<0.05). Conclusion. In general, the microbicidal activity of the blood of healthy people is a multi-component system, in which a special place should be given to the factors of opsonization.

scholarly journals Staphylococcus aureus as a target of peripheral blood microbicidal factors from healthy donors

2021 ◽  
Vol 38 (2) ◽  
pp. 6-13
Author(s):  
Anatoliy P. Godovalov ◽  
Iosif A. Boev
Keyword(s):  
Staphylococcus Aureus ◽  
Hydroxyl Radicals ◽  
Immunoglobulin G ◽  
Peripheral Blood ◽  
Phagocytic Activity ◽  
Clinical Manifestations ◽  
Blood Monocytes ◽  
Complement Components ◽  
Bacteriolytic Activity ◽  
Healthy Donors

Objective. The aim of investigation was to evaluate the bacteriolytic activity of the peripheral blood of healthy donors against S. aureus. Currently, much attention is paid to the involvement of opportunistic microorganisms in the development of infectious and inflammatory diseases, among which one of the leading places is occupied by the processes of staphylococcal etiology. Staphylococcus aureus possesses a unique spectrum of pathogenic factors, which, together with intracellular persistence, allow staphylococci to avoid exposure to immune factors and other agents. Materials and methods. The bacteriolytic activity of the peripheral blood of 32 healthy donors, as well as the ability of the whole blood and serum to destroy biofilms, were evaluated. The phagocytic activity of peripheral blood monocytes and neutrophils and the ability to produce hydroxyl radicals were analyzed. For opsonization of S. aureus, a commercial immunoglobulin G or donor serum was used. Results. It was shown that the whole peripheral blood practically does not have a significant effect on the number of viable cells of S. aureus. However, freshly obtained blood serum significantly destroys the biofilm. It has been established that a fifth part of peripheral blood leukocytes is absorbed by S. aureus. After opsonization of microbial cells with immunoglobulin G, the indices of phagocytic activity of monocytes and neutrophils did not change significantly. When using freshly obtained serum for opsonization of objects, a stimulating effect on the production of hydroxyl radicals by leukocytes was revealed (2758.7 725.3 and 870.6 197.4 related light units, respectively; p 0.05). After heating the serum at 56 C, the stimulating effect was leveled (1091.1 234.7 related light units; p 0.05 for samples with non-opsonized objects). In general, the obtained data indicate that the complement components can be recognized as the most effective system for the elimination of S. aureus. Conclusions. Thus, S. aureus uniquely adapted to the human body that allows staphylococci to persist for a long time without clinical manifestations. It can be assumed that among the factors of the immune system, the proteins of the complement system, which destroy both S. aureus cells and the biofilm matrix, probably have the most effective bactericidal action. However, the effectiveness of this system depends on the protein-synthesizing function of the liver, the availability of microorganisms to the action of complement.

scholarly journals Nanoemulsions of Satureja montana Essential Oil: Antimicrobial and Antibiofilm Activity against Avian Escherichia coli Strains

Pharmaceutics ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 134
Author(s):  
Federica Rinaldi ◽  
Linda Maurizi ◽  
Antonietta Lucia Conte ◽  
Massimiliano Marazzato ◽  
Alessandro Maccelli ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Essential Oil ◽  
Ternary Phase Diagram ◽  
Biological Activities ◽  
Minimal Bactericidal Concentration ◽  
Antibiofilm Activity ◽  
E Coli ◽  
Biofilm Production ◽  
Wide Range ◽  
Satureja Montana

Satureja montana essential oil (SEO) presents a wide range of biological activities due to its high content of active phytochemicals. In order to improve the essential oil’s (EO) properties, oil in water nanoemulsions (NEs) composed of SEO and Tween-80 were prepared, characterized, and their antimicrobial and antibiofilm properties assayed against Escherichia coli strains isolated from healthy chicken. Since surfactant and oil composition can strongly influence NE features and their application field, a ternary phase diagram was constructed and evaluated to select a suitable surfactant/oil/water ratio. Minimal inhibitory concentration and minimal bactericidal concentration of NEs, evaluated by the microdilution method, showed that the SEO NE formulation exhibited higher inhibitory effects against planktonic E. coli than SEO alone. The quantification of biofilm production in the presence of NEs, assessed by crystal violet staining and scanning electron microscopy, evidenced that sub-MIC concentrations of SEO NEs enable an efficient reduction of biofilm production by the strong producer strains. The optimized nanoemulsion formulation could ensure food safety quality, and counteract the antibiotic resistance of poultry associated E. coli, if applied/aerosolized in poultry farms.

An Improved Membrane Filtration Method for Enumeration of Faecal Coliforms and E. Coli by a Fluorogenic β-Glucuronidase Assay

1993 ◽  
Vol 27 (3-4) ◽  
pp. 267-270 ◽  
Author(s):  
M. T. Augoustinos ◽  
N. A. Grabow ◽  
B. Genthe ◽  
R. Kfir
Keyword(s):  
Escherichia Coli ◽  
Water Samples ◽  
Membrane Filtration ◽  
Faecal Coliforms ◽  
Environmental Waters ◽  
Filtration Method ◽  
E Coli ◽  
Wide Range ◽  
Pure Cultures ◽  
Glucuronidase Assay

A fluorogenic β-glucuronidase assay comprising membrane filtration followed by selective enumeration on m-FC agar at 44.5°C and further confirmation using tlie 4-metliylumbelliferyl-β-D-glucuronide (MUG) containing medium was evaluated for the detection of Escherichia coli in water. A total of 200 typical blue and non-typical blue colonies were isolated from sea and fresh water samples using initial selective enumeration on m-FC agar. Pure cultures of the selected colonies were further tested using the MUG assay and identified using the API 20E method. Of the colonies tested which were shown to be positive using the MUG assay 99.4% were Escherichia coli. The results of this study indicate the combination of the m-FC method followed by the MUG assay to be highly efficient for the selection and confirmation of E. coli from a wide range of environmental waters.

scholarly journals Vibrio cholerae Triggers SOS and Mutagenesis in Response to a Wide Range of Antibiotics: a Route towards Multiresistance

2011 ◽  
Vol 55 (5) ◽  
pp. 2438-2441 ◽  
Author(s):  
Zeynep Baharoglu ◽  
Didier Mazel
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Vibrio Cholerae ◽  
Fluorescent Protein ◽  
Resistance Development ◽  
Content Type ◽  
E Coli ◽  
Wide Range ◽  
Green Fluorescent ◽  
Regulated Promoters

ABSTRACTAntibiotic resistance development has been linked to the bacterial SOS stress response. InEscherichia coli, fluoroquinolones are known to induce SOS, whereas other antibiotics, such as aminoglycosides, tetracycline, and chloramphenicol, do not. Here we address whether various antibiotics induce SOS inVibrio cholerae. Reporter green fluorescent protein (GFP) fusions were used to measure the response of SOS-regulated promoters to subinhibitory concentrations of antibiotics. We show that unlike the situation withE. coli, all these antibiotics induce SOS inV. cholerae.

scholarly journals The Repeat-In-Toxin Family Member TosA Mediates Adherence of Uropathogenic Escherichia coli and Survival during Bacteremia

2011 ◽  
Vol 80 (2) ◽  
pp. 493-505 ◽  
Author(s):  
Patrick D. Vigil ◽  
Travis J. Wiles ◽  
Michael D. Engstrom ◽  
Lev Prasov ◽  
Matthew A. Mulvey ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract ◽  
Virulence Factors ◽  
Upper Urinary Tract ◽  
Host Cells ◽  
Content Type ◽  
E Coli ◽  
Wide Range ◽  
Novel Target ◽  
Tract Infections

ABSTRACTUropathogenicEscherichia coli(UPEC) is responsible for the majority of uncomplicated urinary tract infections (UTI) and represents the most common bacterial infection in adults. UPEC utilizes a wide range of virulence factors to colonize the host, including the novel repeat-in-toxin (RTX) protein TosA, which is specifically expressed in the host urinary tract and contributes significantly to the virulence and survival of UPEC.tosA, found in strains within the B2 phylogenetic subgroup ofE. coli, serves as a marker for strains that also contain a large number of well-characterized UPEC virulence factors. The presence oftosAin anE. coliisolate predicts successful colonization of the murine model of ascending UTI, regardless of the source of the isolate. Here, a detailed analysis of the function oftosArevealed that this gene is transcriptionally linked to genes encoding a conserved type 1 secretion system similar to other RTX family members. TosA localized to the cell surface and was found to mediate (i) adherence to host cells derived from the upper urinary tract and (ii) survival in disseminated infections and (iii) to enhance lethality during sepsis (as assessed in two different animal models of infection). An experimental vaccine, using purified TosA, protected vaccinated animals against urosepsis. From this work, it was concluded that TosA belongs to a novel group of RTX proteins that mediate adherence and host damage during UTI and urosepsis and could be a novel target for the development of therapeutics to treat ascending UTIs.

scholarly journals Multi-omic surveillance of Escherichia coli and Klebsiella spp. in hospital sink drains and patients

2020 ◽  
Author(s):  
B Constantinides ◽  
KK Chau ◽  
TP Quan ◽  
G Rodger ◽  
M Andersson ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Resistance ◽  
Resistance Genes ◽  
Bacterial Infections ◽  
Wide Spectrum ◽  
Klebsiella Oxytoca ◽  
Clinical Disease ◽  
E Coli ◽  
Antimicrobial Resistance Genes ◽  
Wide Range

ABSTRACTEscherichia coli and Klebsiella spp. are important human pathogens that cause a wide spectrum of clinical disease. In healthcare settings, sinks and other wastewater sites have been shown to be reservoirs of antimicrobial-resistant E. coli and Klebsiella spp., particularly in the context of outbreaks of resistant strains amongst patients. Without focusing exclusively on resistance markers or a clinical outbreak, we demonstrate that many hospital sink drains are abundantly and persistently colonised with diverse populations of E. coli, Klebsiella pneumoniae and Klebsiella oxytoca, including both antimicrobial-resistant and susceptible strains. Using whole genome sequencing (WGS) of 439 isolates, we show that environmental bacterial populations are largely structured by ward and sink, with only a handful of lineages, such as E. coli ST635, being widely distributed, suggesting different prevailing ecologies which may vary as a result of different inputs and selection pressures. WGS of 46 contemporaneous patient isolates identified one (2%; 95% CI 0.05-11%) E. coli urine infection-associated isolate with high similarity to a prior sink isolate, suggesting that sinks may contribute to up to 10% of infections caused by these organisms in patients on the ward over the same timeframe. Using metagenomics from 20 sink-timepoints, we show that sinks also harbour many clinically relevant antimicrobial resistance genes including blaCTX-M, blaSHV and mcr, and may act as niches for the exchange and amplification of these genes. Our study reinforces the potential role of sinks in contributing to Enterobacterales infection and antimicrobial resistance in hospital patients, something that could be amenable to intervention.IMPORTANCEEscherichia coli and Klebsiella spp. cause a wide range of bacterial infections, including bloodstream, urine and lung infections. Previous studies have shown that sink drains in hospitals may be part of transmission chains in outbreaks of antimicrobial-resistant E. coli and Klebsiella spp., leading to colonisation and clinical disease in patients. We show that even in non-outbreak settings, contamination of sink drains by these bacteria is common across hospital wards, and that many antimicrobial resistance genes can be found and potentially exchanged in these sink drain sites. Our findings demonstrate that the colonisation of handwashing sink drains by these bacteria in hospitals is likely contributing to some infections in patients, and that additional work is needed to further quantify this risk, and to consider appropriate mitigating interventions.

scholarly journals CAPSULE PRODUCTION PATTERN OF UROPATHOGENIC ESCHERICHIA COLI OF URINARY TRACT INFECTION PATIENTS IN KIRKUK HOSPITALS

2020 ◽  
Vol 17 (35) ◽  
pp. 621-627
Author(s):  
May Ali Hussien UMRAN ◽  
Sumaya Najim AL-KHATEEB
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract Infection ◽  
Urinary Tract ◽  
Tract Infection ◽  
Clinical Manifestations ◽  
Uropathogenic Escherichia Coli ◽  
E Coli ◽  
Intestinal Pathogens ◽  
Production Pattern ◽  
Living Organisms

The bacterium Escherichia coli is one of the best free-living organisms studied in depth. It is a surprisingly diverse species, since some strains of E. coli live in the intestine of animals as harmless commensals, while other distinct genotypes, such as an enteropathogenic or enterohemorrhagic E. coli, for example, cause morbidity and death marked as human intestinal pathogens. The purpose of this study was to develop and validate a PCR assay for a known and suspected uropathogenic E. coli virulence factor (kpsMT) gene region to determine the distribution of the gene and its role in the development of clinical diseases of the urinary system. A total of 25 urine samples were collected from patients with urinary tract infection (UTI) at Azadi and Kirkuk hospitals in the city of Kirkuk, Iraq. Samples of both genders and different ages were collected from patients with suspected urinary tract infection according to the clinical manifestations and symptoms diagnosed by the examining physician. The samples were cultured and positive samples were subjected to the IMViC test to identify E. coli bacteria and subsequently identified using the Vitek 2 compact system. Among 25 samples, 24 (96%) showed positive results for bacterial cultural growth. Of these, 17 (68%) were identified as Escherichia coli. Of the total of 17 isolates, 14 from patients with mild urinary tract infection, and 3 from patients with Urosepsis. The kpsMT gene was present in 14 isolates (82.3%), including 11 (78.5%) isolates from patients with mild urinary tract infection, and 3 (100%) isolates from patients with Urosepsis. It was concluded that Escherichia coli is the most prevalent in urine tract infection samples. Due to the abundance of the kpsMT gene in uropathogenic Escherichia coli (UPEC), this gene plays an important role in developing UTI if it is not treated correctly and quickly; mild cases of UTI can turn into Urosepsis.

scholarly journals A Rapid Immunochromatography Test Based on Hcp1 Is a Potential Point-of-Care Test for Serological Diagnosis of Melioidosis

2018 ◽  
Vol 56 (8) ◽  
Author(s):  
Phornpun Phokrai ◽  
Wisansanee Karoonboonyanan ◽  
Nida Thanapattarapairoj ◽  
Chidchanok Promkong ◽  
Adul Dulsuk ◽  
...  
Keyword(s):  
Point Of Care ◽  
Clinical Manifestations ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Target Antigen ◽  
Northeast Thailand ◽  
Serum Samples ◽  
Serological Method ◽  
Healthy Donors ◽  
Wide Range

ABSTRACTMelioidosis is a fatal infectious disease caused by the environmental bacteriumBurkholderia pseudomallei. It is highly endemic in Asia and northern Australia but neglected in many other tropical countries. Melioidosis patients have a wide range of clinical manifestations, and definitive diagnosis requires bacterial culture, which can be time-consuming. A reliable rapid serological tool is greatly needed for disease surveillance and diagnosis. We previously demonstrated by enzyme-linked immunosorbent assay (ELISA) that a hemolysin-coregulated protein (Hcp1) is a promising target for serodiagnosis of melioidosis. In this study, we developed a rapid immunochromatography test (ICT) using Hcp1 as the target antigen (Hcp1-ICT). We evaluated this test for specific antibody detection using serum samples obtained from 4 groups of human subjects, including the following: (i) 487 culture-confirmed melioidosis patients from four hospitals in northeast Thailand; (ii) 202 healthy donors from northeast Thailand; (iii) 90 U.S. healthy donors; and (iv) 207 patients infected with other organisms. Compared to culture results as a gold standard, the sensitivity of ICT for all hospitals was 88.3%. The specificities for Thai donors and U.S. donors were 86.1% and 100%, respectively, and the specificity for other infections was 91.8%. The results of the Hcp1-ICT demonstrated 92.4% agreement with the Hcp1-ELISA results with a kappa value of 0.829, indicating that the method is much improved compared with the current serological method, the indirect hemagglutination assay (IHA) (69.5% sensitivity and 67.6% specificity for Thais). The Hcp1-ICT represents a potential point-of-care (POC) test and may be used to replace the IHA for screening of melioidosis in hospitals as well as in resource-limited areas.

scholarly journals Bioinformatic and Molecular Analysis of Inverse Autotransporters from Escherichia coli

mSphere ◽  
2019 ◽  
Vol 4 (4) ◽  
Author(s):  
Kelvin G. K. Goh ◽  
Danilo G. Moriel ◽  
Steven J. Hancock ◽  
Minh-Duy Phan ◽  
Mark A. Schembri
Keyword(s):  
Escherichia Coli ◽  
Cell Surface ◽  
Biofilm Formation ◽  
Secretion System ◽  
Content Type ◽  
E Coli ◽  
Wide Range ◽  
Type V Secretion ◽  
K 12 ◽  
Type V

ABSTRACT Proteins secreted by the type V secretion system possess multiple functions, including the capacity to mediate adhesion, aggregation, and biolfilm formation. The type V secretion system can be divided into five subclasses, one of which is the type Ve system. Proteins of the type Ve secretion system are also referred to as inverse autotransporters (IATs). In this study, we performed an in silico analysis of 126 completely sequenced Escherichia coli genomes available in the NCBI database and identified several distinct IAT-encoding gene families whose distribution varied throughout the E. coli phylogeny. The genes included three characterized IATs (intimin, fdeC, and yeeJ) and four uncharacterized IATs (here named iatA, iatB, iatC, and iatD). The four iat genes were cloned from the completely sequenced environmental E. coli strain SMS-3-5 and characterized. Three of these IAT proteins (IatB, IatC, and IatD) were expressed at the cell surface and possessed the capacity to mediate biofilm formation in a recombinant E. coli K-12 strain. Further analysis of the iatB gene, which showed a unique association with extraintestinal E. coli strains, suggested that its regulation is controlled by the LeuO global regulator. Overall, this study provides new data describing the prevalence, sequence variation, domain structure, function, and regulation of IATs found in E. coli. IMPORTANCE Escherichia coli is one of the most prevalent facultative anaerobes of the human gut. E. coli normally exists as a harmless commensal but can also cause disease following the acquisition of genes that enhance its pathogenicity. Adhesion is an important first step in colonization of the host and is mediated by an array of cell surface components. In E. coli, these include a family of adhesins secreted by the type V secretion system. Here, we identified and characterized new proteins from an emerging subclass of the type V secretion system known as the inverse autotransporters (IATs). We found that IAT-encoding genes are present in a wide range of strains and showed that three novel IATs were localized on the E. coli cell surface and mediated biofilm formation. Overall, this study provides new insight into the prevalence, function, and regulation of IATs in E. coli.

Escherichia coli O157 Diversity with Respect to Survival during Drying on Concrete

2003 ◽  
Vol 66 (5) ◽  
pp. 780-786 ◽  
Author(s):  
S. M. AVERY ◽  
S. BUNCIC
Keyword(s):  
Escherichia Coli ◽  
Survival Rate ◽  
Phage Type ◽  
Survival Rates ◽  
Pfge Type ◽  
Meat Production ◽  
Escherichia Coli O157 ◽  
Production Chain ◽  
E Coli ◽  
Wide Range

Shiga toxin (Stx)–producing Escherichia coli O157 isolates (n = 123) were divided into groups according to origin, genotype (pulsed-field gel electrophoresis [PFGE] type, or ribotype), type of Stx produced, or phage type (PT). The survival rate ([number of CFU after 24 h of drying/number of CFU before drying] × 100) for each isolate was determined in triplicate after drying on concrete for 24.0 h. The overall mean survival rate among the 123 E. coli O157 isolates studied was 22.9%, but there was a wide range of responses to drying on concrete, with a minimum of 1.2% and a maximum of 61.9% of the initial inocula being recovered after drying. Among the groups, those isolates that originated from cases of human disease were, on average, significantly more sensitive (P &lt; 0.001) to drying (with a mean survival rate of 15.3%) than isolates from the other three sources (with mean survival rates of 27.7, 26.0, and 22.9% for meats, bovine or ovine feces, and bovine hides, respectively). When the isolates were grouped by genotype, three of the PFGE types were, on average, significantly more resistant to drying than two other PFGE types were, and similarly, significant differences in average resistance to drying between groups of E. coli O157 with different ribotypes were seen. There were no differences between the abilities of isolates producing different Stxs (Stx 1 or Stx 1 and Stx 2) to survive drying. E. coli O157 isolates of PT4, PT21/28, and PT32 survived drying on concrete better than groups of other PTs did. Since the E. coli O157 isolates had various abilities to survive drying on concrete, drying could contribute to a kind of E. coli O157 natural selection along the meat chain. This possibility may have significant meat safety implications if a range of E. coli O157 isolates are simultaneously exposed to drying at any point along the meat production chain. Those E. coli O157 isolates that are more able to survive drying could be more likely to pass farther along the meat chain and ultimately reach consumers.

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