Novel use of culturomics to identify the microbiota in hospital sink drains with and without persistent VIM-positive Pseudomonas aeruginosa

Abstract In hospitals, Verona Integron-encoded Metallo-beta-lactamase (VIM)-positive Pseudomonas aeruginosa may colonize sink drains, and from there, be transmitted to patients. These hidden reservoirs are difficult to eradicate since P. aeruginosa forms biofilms that resist disinfection. However, little is known on the composition of these biofilms. Therefore, culturomics was used for the first time to investigate the viable microbiota in four hospital sink drain samples with longstanding VIM-positive P. aeruginosa drain reservoirs (inhabited by high-risk clone, sequence type ST111), and four drain samples where VIM-positive P. aeruginosa was not present. Microbial load and composition varied between samples, yielding between 471–18,904 distinct colonies and 8–20 genera. In two VIM-positive drain samples, P. aeruginosa was the most abundantly-isolated microorganism, and found in combination with other Gram-negative bacteria, Citrobacter, Enterobacter, or Stenotrophomonas. P. aeruginosa was in low abundance in the other two VIM-positive samples, and found with Gram-positive cocci (Enterococcus and Staphylococcus) or Sphingomonas. In VIM-negative drain samples, high abundances of Gram-negative non-fermenting bacteria, including Acinetobacter, non-aeruginosa Pseudomonas spp., Acidovorax, Chryseobacterium, Flavobacterium, and Sphingobium, as well as Candida, were cultured. Although additional experiments are needed to draw more firm conclusions on which microorganisms enable or inhibit VIM-positive P. aeruginosa persistence, our data provide unique insights into the microbial compositions of sink drain inlets.

Download Full-text

Treatment of Bloodstream Infections Due to Gram-Negative Bacteria with Difficult-to-Treat Resistance

Antibiotics ◽

10.3390/antibiotics9090632 ◽

2020 ◽

Vol 9 (9) ◽

pp. 632 ◽

Cited By ~ 2

Author(s):

Matteo Bassetti ◽

Antonio Vena ◽

Chiara Sepulcri ◽

Daniele Roberto Giacobbe ◽

Maddalena Peghin

Keyword(s):

Pseudomonas Aeruginosa ◽

Antibiotic Resistance ◽

Treatment Options ◽

Bloodstream Infections ◽

Gram Negative Bacteria ◽

Beta Lactamase ◽

Gram Negative ◽

Extended Spectrum Beta Lactamase ◽

Carbapenem Resistant ◽

Rising Incidence

The rising incidence of bloodstream infections (BSI) due to Gram-negative bacteria (GNB) with difficult-to-treat resistance (DTR) has been recognized as a global emergency. The aim of this review is to provide a comprehensive assessment of the mechanisms of antibiotic resistance, epidemiology and treatment options for BSI caused by GNB with DTR, namely extended-spectrum Beta-lactamase-producing Enterobacteriales; carbapenem-resistant Enterobacteriales; DTR Pseudomonas aeruginosa; and DTR Acinetobacter baumannii.

Download Full-text

A TAXONOMIC STUDY OF SOME GRAM-NEGATIVE, NON-FERMENTING BACTERIA

Canadian Journal of Microbiology ◽

10.1139/m63-064 ◽

1963 ◽

Vol 9 (4) ◽

pp. 499-507 ◽

Cited By ~ 3

Author(s):

Franklin P. Koontz ◽

John E. Faber

Keyword(s):

Pseudomonas Aeruginosa ◽

Morphological Characteristics ◽

Taxonomic Study ◽

Gram Negative Bacteria ◽

General Description ◽

Gram Negative ◽

Fermenting Bacteria ◽

Somatic Antigens ◽

Antigenic Heterogeneity ◽

Produce Acid

An investigation of 110 cultures of Gram-negative bacteria which failed to produce acid in conventional carbohydrate media was undertaken to determine their taxonomic position.Thirty-seven strains had peritrichous flagella, 30 were monotrichous, 8 were lophotrichous, 3 were either monotrichous or lophotrichous, and 32 strains were non-flagellated.Flagella stains and reaction in Hugh and Leifson"s O/F medium were used to divide the strains into general groups.Sixty-five strains were inert in O/F medium. Forty of these resembled the general description for the genus Alcaligenes. Seventeen strains resembled Alcaligenes biochemically, but were either monotrichously or lophotrichously flagellated. Several strains resembled Vibrio percolans.Nine strains resembled Achromobacter biochemically by oxidatively utilizing pentoses only.Twenty-three strains resembled Pseudomonas aeruginosa. All produced acid oxidatively in the O/F medium.Thirteen strains produced acid fermentatively in O/F medium.Serological studies indicated antigenic heterogeneity. Several strains shared flagellar and somatic antigens, while others were related by either somatic or flagellar components. Seventy-eight strains did not react with any somatic antisera and 54 strains did not react with any flagellar antisera. There was little correlation between antigenicity and biochemical or morphological characteristics.

Download Full-text

Opr86 Is Essential for Viability and Is a Potential Candidate for a Protective Antigen against Biofilm Formation by Pseudomonas aeruginosa

Journal of Bacteriology ◽

10.1128/jb.02004-07 ◽

2008 ◽

Vol 190 (11) ◽

pp. 3969-3978 ◽

Cited By ~ 45

Author(s):

Yosuke Tashiro ◽

Nobuhiko Nomura ◽

Ryoma Nakao ◽

Hidenobu Senpuku ◽

Reiko Kariyama ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Biofilm Formation ◽

Protective Antigen ◽

Morphological Changes ◽

Bacterial Pathogen ◽

Membrane Vesicles ◽

The Other ◽

Gram Negative Bacteria ◽

Potential Candidate ◽

Gram Negative

ABSTRACT Pseudomonas aeruginosa is an opportunistic bacterial pathogen that is one of the most refractory to therapy when it forms biofilms in the airways of cystic fibrosis patients. To date, studies regarding the production of an immunogenic and protective antigen to inhibit biofilm formation by P. aeruginosa have been superficial. The previously uncharacterized outer membrane protein (OMP) Opr86 (PA3648) of P. aeruginosa is a member of the Omp85 family, of which homologs have been found in all gram-negative bacteria. Here we verify the availability of Opr86 as a protective antigen to inhibit biofilm formation by P. aeruginosa PAO1 and several other isolates. A mutant was constructed in which Opr86 expression could be switched on or off through a tac promoter-controlled opr86 gene. The result, consistent with previous Omp85 studies, showed that Opr86 is essential for viability and plays a role in OMP assembly. Depletion of Opr86 resulted in streptococci-like morphological changes and liberation of excess membrane vesicles. A polyclonal antibody against Opr86 which showed reactivity to PAO1 cells was obtained. The antibody inhibited biofilm formation by PAO1 and the other clinical strains tested. Closer examination of early attachment revealed that cells treated with the antibody were unable to attach to the surface. Our data suggest that Opr86 is a critical OMP and a potential candidate as a protective antigen against biofilm formation by P. aeruginosa.

Download Full-text

MICROBIOLOGICAL SCREENING OF OTORRHOEA FROM PEOPLE COMING TO HOSPITAL IN MAHAJANGA

International Journal of Clinical and Biomedical Research ◽

10.31878/ijcbr.2019.61.07 ◽

2020 ◽

pp. 25-28

Author(s):

Rivo RAKOTOMALALA ◽

Patrick RANDRIANANDRAINA ◽

Tsiriniaina RAMAVOSON ◽

Fiacre RAMISARIMANANA ◽

Ainamalala Catherine RAZAFINDRAKOTO ◽

...

Keyword(s):

Escherichia Coli ◽

Pseudomonas Aeruginosa ◽

The Other ◽

Coagulase Negative Staphylococcus ◽

Gram Positive ◽

Gram Negative ◽

E Coli ◽

Ear Infections ◽

Corynebacterium Sp ◽

Gram Positive Cocci

In whole, 56 patients were included. Amidst identified microorganisms were fungus (4,7%) and bacteria (95,3%) to which Gram negative bacilli represented 72,1% (n=44), Gram positive cocci 6,4% (n=10), Gram positive bacilli 8,2% (n=5) and Gram negative cocci 3,3% (n=2). Among these bacterias, Pseudomonas aeruginosa and Proteus sp were predominant, with respectively 41% (n=25), 23% (n=14). However, three cases of S. aureus reported, six with negative coagulase Staphylococcus, one with Escherichia coli, one with Klebsiella sp, one with Haemophilus sp, two cases with Neisseria sp and four cases with Corynebacterium sp. Two types of cultures were noticed, one of them monomorphic (91,1%, n=51) and the other polymorphic (8,9%, n=5) to which 3 associations of P. aeruginosa-Proteus sp, 1 association of P. aeruginosa- coagulase negative Staphylococcus and 1 association of P. aeruginosa- E. coli. No resistance to ciprofloxacin was observed with Pseudomonas, Neisseria sp, Haemophilus, and enterobacteria except for E. coli. No resistance to rifampicin was observed with S. aureus. However, the sensitivity of S. aureus to ciprofloxacin decreased (one bacterium out of three). The use of rifampicin or fluoroquinolones should be based on the type of ear infections,

Download Full-text

A study for Bacterial Infection in Acute Diarrhea Patients

Baghdad Science Journal ◽

10.21123/bsj.11.2.757-760 ◽

2014 ◽

Vol 11 (2) ◽

pp. 757-760

Author(s):

Baghdad Science Journal

Keyword(s):

Bacterial Infection ◽

Acute Diarrhea ◽

Biochemical Test ◽

The Other ◽

Gram Negative Bacteria ◽

Gram Negative ◽

E Coli ◽

Different Ages ◽

Public Laboratory ◽

First Time

500 samples of diarrhea stool were collected from different ages(less than 1year –upto30years) and for both genders from some patients in (Alwiya hospital for children, Al-kendi, central health public laboratory and some gavernarated labs) period(1/11/2009—1/10/2010). Kinds of bacteria and parasites agents were isolated and identified from patients with diarrhea. Nine species of gram negative bacteria from enterobacteriaceae were isolated, E. coli isolated are the higher ratio 4.8% of all, then Salmonella typhi4.6% while the lowest ratios is Citrobacterfreundii 0.4%, while the other identified species were be among the previous rotios. also Plesomonasshigelloides was isolated which concedride one of the bacterial local studies.many methods were depend on in order to identifiy; growth on media like (Enrichment ,differentials and differetials) media. Biochemical test represents with API20E, also. Some confirmed tests represent with a modern and important identical tools was used for first time to identify the entericbacteria, by (Rapid ID 32) usually used with mini API system where the results record automatically.

Download Full-text

Disinfection of Fruits with Activated Water

JOURNAL OF ADVANCES IN AGRICULTURE ◽

10.24297/jaa.v10i0.8462 ◽

2019 ◽

Vol 10 ◽

pp. 1864-1872

Author(s):

Prof. Teodora P. Popova

Keyword(s):

Antimicrobial Activity ◽

Aqueous Solutions ◽

Antimicrobial Properties ◽

The Other ◽

Gram Negative Bacteria ◽

High Antimicrobial Activity ◽

Gram Negative ◽

E Coli ◽

Number Of Microorganisms ◽

Lower Effect

The effect of ionized aqueous solutions (anolytes and catholyte) in the processing of fruits (cherries, morellos, and strawberries) for decontamination has been tested. Freshly prepared analytes and catholyte without the addition of salts were used, as well as stored for 7 months anolytes, prepared with 0.5% NaCl and a combination of 0.5% NaCl and 0.5% Na2CO3. The anolyte prepared with a combination of 0.5% NaCl and 0.5% Na2CO3, as well as the anolyte obtained with 0.5% NaCl, exhibit high antimicrobial activity against the surface microflora of strawberries, cherries, and sour cherries. They inactivate E. coli for 15 minutes. The other species of the fam. Enterobacteriaceae were also affected to the maximum extent, as is the total number of microorganisms, especially in cherries and sour cherries. Even stored for 7 months, they largely retain their antimicrobial properties. Anolyte and catholyte, obtained without the addition of salts, showed a lower effect on the total number of microorganisms, but had a significant effect on Gram-negative bacteria, and especially with regard to the sanitary indicative E. coli.

Download Full-text

Extended Spectrum Beta Lactamase (ESBL), bla TEM,bla SHVand bla CTX-M,resistance genes in Community and Healthcare Associated Gram Negative Bacteria from Osun State, Nigeria

Infectious Disorders - Drug Targets ◽

10.2174/1871526520999200729181559 ◽

2020 ◽

Vol 20 ◽

Author(s):

Ganiyat Shitta ◽

Olufunmilola Makanjuola ◽

Olusolabomi Adefioye ◽

Olugbenga Adekunle Olowe

Keyword(s):

Antibiotic Resistance ◽

Resistance Genes ◽

Gram Negative Bacteria ◽

Beta Lactamase ◽

Multiple Antibiotic Resistance ◽

Esbl Production ◽

Gram Negative ◽

Extended Spectrum Beta Lactamase ◽

Healthcare Associated ◽

Esbl Producers

Background: Extended Spectrum Beta Lactamase (ESBL) production in gram negative bacteria confers multiple antibiotic resistance, adversely affecting antimicrobial therapy in infected individuals. ESBLs result from mutations in β-lactamases encoded mainly by the bla TEM,bla SHVand bla CTX-Mgenes. The prevalence of ESBL producing bacteria has been on the increase globally especially its upsurge among isolates from community-acquired infections. Aim: To determine ESBL prevalence and identify ESBL genes among clinical isolates in Osun State, Nigeria. Material and Methods: A cross-sectional study was carried out from August 2016 –July 2017 in Osun State, Nigeria. Three hundred and sixty Gram negative bacteria recovered from clinical samples obtained from both community and healthcare associated infections were tested. They included147 Escherichia coli(40.8%), 116 Klebsiella spp(32.2%), 44 Pseudomo-nas aeruginosa(12.2%) and23 Proteus vulgaris (6.4%) isolates. Others were Acinetobacter baumannii, Serratia rubidae, Citrobacter spp, Enterobacter spp and Salmonella typhi. Disk diffusion antibiotic susceptibility testing was carried out, isolates were screened for ESBL production and confirmed using standard laboratory procedures. ESBLs resistance genes were identified by Polymerase Chain Reaction (PCR). Results: All isolates demonstrated multiple antibiotic resistance. Resistance to ampicillin, amoxicillin with clavulanate and erythromycin was 100%, whereas resistance to Imipenem was very low (5.0%). : Overall prevalence of ESBL producers was 41.4% with Klebsiellaspp as the highest ESBL producing Enterobacteriacaea. ESBL producers were more prevalent among the hospital pathogens than community pathogens, 58% vs 29.5% (p=0.003). ESBL genes were detected in all ESBL producers with the blaCTX-Mgene predominating (47.0%) followed by blaTEM(30.9%) and blaSHVgene was the least, 22.1%. The blaCTX-Mgene was also the most prevalent in the healthcare pathogens (62%) but it accounted for only 25% in those of community origin. Conclusion: A high prevalence of ESBL producing gram negative organisms occurs both in healthcare and in the community in our environment with the CTX-M variant predominating. Efforts to control spread of these pathogens should be addressed.

Download Full-text

Clonal Clusters, Molecular Resistance Mechanisms and Virulence Factors of Gram-Negative Bacteria Isolated from Chronic Wounds in Ghana

Antibiotics ◽

10.3390/antibiotics10030339 ◽

2021 ◽

Vol 10 (3) ◽

pp. 339

Author(s):

Denise Dekker ◽

Frederik Pankok ◽

Thorsten Thye ◽

Stefan Taudien ◽

Kwabena Oppong ◽

...

Keyword(s):

Molecular Epidemiology ◽

Virulence Factors ◽

Iron Uptake ◽

Chronic Wounds ◽

Sub Saharan Africa ◽

Gram Negative Bacteria ◽

Beta Lactamase ◽

Secretion Systems ◽

Gram Negative ◽

E Coli

Wound infections are common medical problems in sub-Saharan Africa but data on the molecular epidemiology are rare. Within this study we assessed the clonal lineages, resistance genes and virulence factors of Gram-negative bacteria isolated from Ghanaian patients with chronic wounds. From a previous study, 49 Pseudomonas aeruginosa, 21 Klebsiellapneumoniae complex members and 12 Escherichia coli were subjected to whole genome sequencing. Sequence analysis indicated high clonal diversity with only nine P. aeruginosa clusters comprising two strains each and one E. coli cluster comprising three strains with high phylogenetic relationship suggesting nosocomial transmission. Acquired beta-lactamase genes were observed in some isolates next to a broad spectrum of additional genetic resistance determinants. Phenotypical expression of extended-spectrum beta-lactamase activity in the Enterobacterales was associated with blaCTX-M-15 genes, which are frequent in Ghana. Frequently recorded virulence genes comprised genes related to invasion and iron-uptake in E. coli, genes related to adherence, iron-uptake, secretion systems and antiphagocytosis in P. aeruginosa and genes related to adherence, biofilm formation, immune evasion, iron-uptake and secretion systems in K. pneumonia complex. In summary, the study provides a piece in the puzzle of the molecular epidemiology of Gram-negative bacteria in chronic wounds in rural Ghana.

Download Full-text

The antimicrobial potential of cannabidiol

Communications Biology ◽

10.1038/s42003-020-01530-y ◽

2021 ◽

Vol 4 (1) ◽

Author(s):

Mark A. T. Blaskovich ◽

Angela M. Kavanagh ◽

Alysha G. Elliott ◽

Bing Zhang ◽

Soumya Ramu ◽

...

Keyword(s):

Bacterial Infections ◽

Modern Medicine ◽

Gram Negative Bacteria ◽

Gram Negative ◽

New Class ◽

Clostridioides Difficile ◽

Excellent Activity ◽

Induce Resistance ◽

First Time

AbstractAntimicrobial resistance threatens the viability of modern medicine, which is largely dependent on the successful prevention and treatment of bacterial infections. Unfortunately, there are few new therapeutics in the clinical pipeline, particularly for Gram-negative bacteria. We now present a detailed evaluation of the antimicrobial activity of cannabidiol, the main non-psychoactive component of cannabis. We confirm previous reports of Gram-positive activity and expand the breadth of pathogens tested, including highly resistant Staphylococcus aureus, Streptococcus pneumoniae, and Clostridioides difficile. Our results demonstrate that cannabidiol has excellent activity against biofilms, little propensity to induce resistance, and topical in vivo efficacy. Multiple mode-of-action studies point to membrane disruption as cannabidiol’s primary mechanism. More importantly, we now report for the first time that cannabidiol can selectively kill a subset of Gram-negative bacteria that includes the ‘urgent threat’ pathogen Neisseria gonorrhoeae. Structure-activity relationship studies demonstrate the potential to advance cannabidiol analogs as a much-needed new class of antibiotics.

Download Full-text

Ultrastructural studies of Chlamydia psittaci 6BC variant strains. I. Ultrastructure of the surface layers of egg-passaged 6BC strain

Canadian Journal of Microbiology ◽

10.1139/m73-142 ◽

1973 ◽

Vol 19 (8) ◽

pp. 887-894

Author(s):

Linda Poffenroth ◽

J. W. Costerton ◽

Nonna Kordová ◽

John C. Wilt

Keyword(s):

Chick Embryo ◽

Microscopic Examination ◽

Electron Microscopic Examination ◽

Chlamydia Psittaci ◽

Gram Negative Bacteria ◽

Surface Layers ◽

Electron Microscopic ◽

Gram Negative ◽

Ultrastructural Studies ◽

First Time

Electron microscopic examination of a semipurified Chlamydia psittaci 6BC strain attenuated in chick embryo yolk sac revealed for the first time two morphologically distinct small elementary bodies which differ both in the ultrastructure of their surface layers and in their buoyant densities in sucrose gradients. Also, the morphology of the surface layers of the larger reticulate forms in cell-free systems is described for the first time. Many points of difference between the surface envelopes and internal structure of chlamydial particles and those of Gram-negative bacteria are discussed.

Download Full-text