scholarly journals Isolation of Four Lytic Phages Infecting Klebsiella pneumoniae K22 Clinical Isolates from Spain

2020 ◽  
Vol 21 (2) ◽  
pp. 425 ◽  
Author(s):  
Pilar Domingo-Calap ◽  
Beatriz Beamud ◽  
Justine Vienne ◽  
Fernando González-Candelas ◽  
Rafael Sanjuán

The emergence of multi-drug-resistant bacteria represents a major public-health threat. Phages constitute a promising alternative to chemical antibiotics due to their high host specificity, abundance in nature, and evolvability. However, phage host specificity means that highly diverse bacterial species are particularly difficult to target for phage therapy. This is the case of Klebsiella pneumoniae, which presents a hypervariable extracellular matrix capsule exhibiting dozens of variants. Here, we report four novel phages infecting K. pneumoniae capsular type K22 which were isolated from environmental samples in Valencia, Spain. Full genome sequencing showed that these phages belong to the Podoviridae family and encode putative depolymerases that allow digestion of specific K22 K. pneumoniae capsules. Our results confirm the capsular type-specificity of K. pneumoniae phages, as indicated by their narrow infectivity in a panel of K. pneumoniae clinical isolates. Nonetheless, this work represents a step forward in the characterization of phage diversity, which may culminate in the future use of large panels of phages for typing and/or for combating multi-drug-resistant K. pneumoniae.

Pharmaceutics ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1141
Author(s):  
Noha M. Elhosseiny ◽  
Tamer M. Samir ◽  
Aliaa A. Ali ◽  
Amani A. El-Kholy ◽  
Ahmed S. Attia

Neonatal sepsis is a leading cause of death among newborns and infants, especially in the developing world. The problem is compounded by the delays in pinpointing the causative agent of the infection. This is reflected in increasing mortality associated with these cases and the spread of multi-drug-resistant bacteria. In this work, we deployed bioinformatics and proteomics analyses to determine a promising target that could be used for the identification of a major neonatal sepsis causative agent, Klebsiella pneumoniae. A 19 amino acid peptide from a hypothetical outer membrane was found to be very specific to the species, well conserved among its strains, surface exposed, and expressed in conditions simulating infection. Antibodies against the selected peptide were conjugated to gold nanoparticles and incorporated into an immunochromatographic strip. The developed strip was able to detect as low as 105 CFU/mL of K. pneumoniae. Regarding specificity, it showed negative results with both Escherichia coli and Enterobacter cloacae. More importantly, in a pilot study using neonatal sepsis cases blood specimens, the developed strip selectively gave positive results within 20 min with those infected with K. pneumoniae without prior sample processing. However, it gave negative results in cases infected with other bacterial species.


2019 ◽  
Vol 39 (1) ◽  
pp. 97-98 ◽  
Author(s):  
Davin Ryanputra ◽  
Dingding Wang ◽  
Martin B. Lee ◽  
Boon Wee Teo ◽  
Pei Loo Tok

We report a rare case of carbapenemase-producing enterobacte-riaceae peritonitis in a patient undergoing automated peritoneal dialysis (APD). The PD catheter had to be removed as the patient remained unwell despite antibiotics. Antimicrobial resistance in PD peritonitis is a concern in this era of multi-drug resistant bacteria.


2020 ◽  
Vol 2020 ◽  
pp. 1-10 ◽  
Author(s):  
James T. P. Matshwele ◽  
Florence Nareetsile ◽  
Daphne Mapolelo ◽  
Pearl Matshameko ◽  
Melvin Leteane ◽  
...  

The potential antimicrobial properties of a tridentate polypyridyl ligand 4-butoxy-N,N-bis(pyridin-2-ylmethyl)aniline (BUT) 1 and its corresponding mixed ligand ruthenium complexes were investigated on drug-resistant and non-drug-resistant bacterial species. The ligand and its complexes were synthesized and successfully characterized by 1H NMR, UV/Vis, and FTIR spectra; ESI-MS; and magnetic susceptibility. Electronic spectra and magnetic susceptibility of these Ru(II)/(III) complexes suggest that they are of a low spin crystal field split, where the Ru(III) is a d5 and Ru(II) d6 low spin. These compounds were tested for antibacterial activity on two bacterial species: Staphylococcus aureus (S. aureus) and Klebsiella pneumoniae (K. pneumoniae), as well as their drug-resistant strains methicillin-resistant Staphylococcus aureus (MRSA) and multidrug resistant Klebsiella pneumoniae (MDR K. pneumoniae). All the compounds inhibited growth of the two non-drug-resistant bacteria and only one drug-resistant strain MRSA. However, only the ligands BUT and 2,2-dipyridylamine showed activity against MRSA, while all complexes did not show any antibacterial activity on MRSA. We observed large zones of inhibition for the Gram-positive S. aureus and MRSA bacteria, compared to the Gram-negative K. pneumoniae bacteria. DNA cleavage studies with gel electrophoresis showed denatured bacterial DNA on the gel from all the complexes, with the exception of the ligand, suggesting DNA nuclease activity of the complexes in the bacterial DNA.


1999 ◽  
Vol 556 ◽  
Author(s):  
C.-I Chen ◽  
A. Meike ◽  
Y.-J. Chuu ◽  
A. Sawvel ◽  
W. Lin

AbstractTransport of bacteria is investigated as part of the Large-Block Test (LBT), a thermally perturbed block of Topopah Spring tuff. Two bacterial species,Bacillus subtilisandArthrobacter oxydans, were isolated from the Yucca Mountain tuff. Natural mutants that can grow under the simultaneous presence of the two antibiotics, streptomycin and rifampicin, were selected from these species by laboratory procedures, cultured, and injected into the five heater boreholes of the large block hours before heating was initiated. The temperature, as measured 5 cm above one of the heater boreholes, rose slowly over a matter of months to a maximum of 142°C and to 60°C at the top and the bottom of the block. Samples were collected from boreholes located approximately 5 ft below the injection points. Double-drug-resistant bacteria were found in the collection boreholes nine months after injection. Biochemical tests support the identification ofB. subtilisin the collection boreholes. Double-drug-resistant microbes also appeared in the heater boreholes where the temperature had been sustainably high throughout the test. The number of double-drugresistant bacteria that were identified in the collection boreholes increased with time until the heater was deactivated. Negative indications in the collection holes after the heater was deactivated support the supposition that these bacteria were the species that were injected. An apparent homogeneous distribution among the collection boreholes suggests no pattern to the migration of bacteria through the block. The relationship between bacterial migration and the movement of water is not yet understood. These observations indicate the possibility of rapid bacterial transport in a thermally perturbed geologic setting. The implications for colloid transport need to be reviewed.


Author(s):  
Leoney Andonissamy ◽  
Suma Karthigeyan ◽  
Seyed Asharaf Ali

Introduction: The bacteria colonising the oral cavity and the dentures acquire drug resistance due to frequent usage of antibiotics systemically and application of mouth rinses and denture disinfectants locally. These multidrug resistant bacteria pose potential threat to the health of the patient as infections caused by them do not respond to conventional antibiotics. Aim: The present study aims at detecting the drug resistant bacteria in patients who wear complete dentures. Materials and Methods: The study is a descriptive study and follows laboratory invitro study design involving 30 complete denture patients. Swabs were collected from their oral cavity as well as complete denture surfaces. Antibiotic sensitivity tests were performed for the following bacteriae namely Viridans streptococci species, Staphylococcus aureus, Klebsiella pneumoniae and E.coli. Isolation of the bacteria were done by means of selective media and subjected to biochemical tests. The 16S rRNA sequencing was done to ascertain the microorganisms by which 20 isolates of each of the selective bacteria were obtained. The bacteria were classified as sensitive, intermediate sensitive and resistant based on antibiotic sensitivity tests. Those isolates which exhibited Multi-Drug Resistance (MDR) were visualised using SEM. Results:Viridans streptococci spp. (40%) and Staphylococcus aureus (25%) isolates were resistant to Amoxiclavulinic acid and Methicilin, whereas Klebsiella pneumoniae (30%) and (30%) E.coli isolates were most resistant to Cefotaxime and Doxicilin. Conclusion: Drug resistant bacteria have been identified from complete dentures and oral cavity in the present study. Antibiotic sensitivity tests, 16S rRNA sequencing and SEM are vital investigative tools to detect and to visualise drug resistant bacteria. Cell density, Extracellular Polymeric Substances (EPS) and capsule could be important factors for providing drug resistance.


2017 ◽  
Author(s):  
Andre Mu ◽  
Jason C. Kwong ◽  
Nicole S. Isles ◽  
Anders Gonçalves da Silva ◽  
Mark B. Schultz ◽  
...  

AbstractWhole-genome sequencing of microbial pathogens is revolutionising modern approaches to outbreaks of infectious diseases and is reliant upon organism culture. Culture-independent methods have shown promise in identifying pathogens, but high level reconstruction of microbial genomes from microbiologically complex samples for more in-depth analyses remains a challenge. Here, using metagenomic sequencing of a human faecal sample and analysis by tetranucleotide frequency profiling projected onto emergent self-organising maps, we were able to reconstruct the underlying populations of two extensively-drug resistant pathogens, Klebsiella pneumoniae carbapenemase (KPC)-producing Klebsiella pneumoniae and vancomycin-resistant Enterococcus faecium. From these genomes, we were able to ascertain molecular typing results, such as MLST, and identify highly discriminatory mutations in the metagenome to distinguish closely related strains. These proof-of-principle results demonstrate the utility of clinical sample metagenomics to recover sequences of important drug-resistant bacteria and application of the approach in outbreak investigations, independent of the need to culture the organisms.


PLoS ONE ◽  
2021 ◽  
Vol 16 (1) ◽  
pp. e0244673
Author(s):  
Julalak C. Ontong ◽  
Nwabor F. Ozioma ◽  
Supayang P. Voravuthikunchai ◽  
Sarunyou Chusri

Multidrug resistant Enterobacterales have become a serious global health problem, with extended hospital stay and increased mortality. Antibiotic monotherapy has been reported ineffective against most drug resistant bacteria including Klebsiella pneumoniae, thus encouraging the use of multidrug therapies as an alternative antibacterial strategy. The present works assessed the antibacterial activity of colistin against K. pneumoniae isolates. Resistant isolates were tested against 16 conventional antibiotics alone and in combination with colistin. The results revealed that all colistin resistant isolates demonstrated multidrug resistance against the tested antibiotics except amikacin. At sub-inhibitory concentrations, combinations of colistin with amikacin, or fosfomycin showed synergism against 72.72% (8 of 11 isolates). Colistin with either of gentamicin, meropenem, cefoperazone, cefotaxime, ceftazidime, moxifloxacin, minocycline, or piperacillin exhibited synergism against 81.82% (9 of 11 isolates). Combinations of colistin with either of tobramycin or ciprofloxacin showed synergism against 45.45% (5 in 11 isolates), while combinations of colistin with imipenem or ceftolozane and tazobactam displayed 36.36% (4 of 11 isolates) and 63.64% (7 of 11 isolates) synergism. In addition, combinations of colistin with levofloxacin was synergistic against 90.91% (10 of 11 isolates). The results revealed that combinations of colistin with other antibiotics could effectively inhibit colistin resistant isolates of K. pneumoniae, and thus could be further explore for the treatment of multidrug resistant pathogens.


2019 ◽  
Vol 12 (2) ◽  
pp. 91
Author(s):  
Laumaillé ◽  
Dassonville-Klimpt ◽  
Peltier ◽  
Mullié ◽  
Andréjak ◽  
...  

The lack of antibiotics with a novel mode of action associated with the spread of drug resistant bacteria make the fight against infectious diseases particularly challenging. A quinoline core is found in several anti-infectious drugs, such as mefloquine and bedaquiline. Two main objectives were set in this work. Firstly, we evaluated the anti-mycobacterial properties of the previous quinolines 3, which have been identified as good candidates against ESKAPEE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter spp. and Escherichia coli) bacteria. Secondly, a new series 4 was designed and assessed against the same bacteria strains, taking the pair of enantiomers 3m/3n as the lead. More than twenty compounds 4 were prepared through a five-step asymmetric synthesis with good enantiomeric excesses (>90%). Interestingly, all compounds of series 3 were efficient on M. avium with MIC = 2–16 µg/mL, while series 4 was less active. Both series 3 and 4 were generally more active than mefloquine against the ESKAPEE bacteria. The quinolines 4 were either active against Gram-positive bacteria (MIC ≤ 4 µg/mL for 4c–4h and 4k/4l) or E. coli (MIC = 32–64 µg/mL for 4q–4v) according to the global lipophilicity of these compounds.


F1000Research ◽  
2021 ◽  
Vol 10 ◽  
pp. 444
Author(s):  
Muzaheed Muzaheed ◽  
Naveed Sattar Shaikh ◽  
Saeed Sattar Shaikh ◽  
Sadananda Acharya ◽  
Shajiya Sarwar Moosa ◽  
...  

Background  The presence of Extended-spectrum β-lactamase (ESBL) positive bacteria in hospital setting is an aggravating influential factor for hospitalized patients, and its consequences may be hazardous. Therefore, there is a need for rapid detection methods for newly emerging drug-resistant bacteria. This study was aimed at the molecular characterization of ESBL-positive Klebsiella pneumoniae isolates recovered from clinical samples.   Methods  A total of 513 K. pneumoniae isolates were obtained from various clinical samples during June 2019 to May 2020. The collected isolates were investigated for antimicrobial susceptibility (antibiogram), and PCR and DNA sequencing were performed to analyse the ESBL genes.   Results  Among the 513 isolates, as many as 359 (69.9%) were ESBL producers and 87.5% were multi-drug resistant, while none had resistance to imipenem. PCR scored 3% blaTEM, 3% blaSHV, and 60% blaCTX-M-15 genes for the tested isolates.   Conclusion  The study showed that CTX-M-15 was the major prevalent ESBL type among the isolates. Additionally, all the isolates were susceptible to carbapenems. Screening and detection of ESBL tests are necessary among all isolates from the enterobacteriaceae family in routine microbiology laboratory to prevent associated nosocomial infections. A larger study is essential to understand molecular epidemiology of ESBL producing organisms to minimize morbidities due to these multidrug resistant organisms.


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