Use of prophage free host for achieving homogenous population of bacteriophages: New findings

Kumar, Gaurav; Sundarrajan, Sudarson; Paul, Vivek Daniel; Nandini, S.; Rajendran, Saravanan; Hariharan, Sukumar; Sriram, Bharathi; Padmanabhan, Sriram

doi:10.1016/j.virusres.2012.07.026

Cited by 7 publications

(10 citation statements)

References 38 publications

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“…The following day, after an overnight incubation at 37°C, plates displaying confluent lysis were selected and 3 ml of SM buffer and 2% (vol/vol) chloroform were added before incubation at 37°C for 4 h. High-titer phage solution was removed from the plates, centrifuged (8,000 ϫ g, 10 min) to remove cell debris, and then filter sterilized (pore size, 0.22 m) and stored at 4°C. Both phages were propagated in the prophage-free isolate S. aureus RN4220 (34) to avoid potential contamination with mobilized phages (35). Sensitivity assay.…”

Section: Methodsmentioning

confidence: 99%

Combined Use of Bacteriophage K and a Novel Bacteriophage To Reduce Staphylococcus aureus Biofilm Formation

Alves

Gaudion

Bean

et al. 2014

Appl Environ Microbiol

149

128

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e Biofilms are major causes of impairment of wound healing and patient morbidity. One of the most common and aggressive wound pathogens is Staphylococcus aureus, displaying a large repertoire of virulence factors and commonly reduced susceptibility to antibiotics, such as the spread of methicillin-resistant S. aureus (MRSA). Bacteriophages are obligate parasites of bacteria. They multiply intracellularly and lyse their bacterial host, releasing their progeny. We isolated a novel phage, DRA88, which has a broad host range among S. aureus bacteria. Morphologically, the phage belongs to the Myoviridae family and comprises a large double-stranded DNA (dsDNA) genome of 141,907 bp. DRA88 was mixed with phage K to produce a high-titer mixture that showed strong lytic activity against a wide range of S. aureus isolates, including representatives of the major international MRSA clones and coagulase-negative Staphylococcus. Its efficacy was assessed both in planktonic cultures and when treating established biofilms produced by three different biofilm-producing S. aureus isolates. A significant reduction of biofilm biomass over 48 h of treatment was recorded in all cases. The phage mixture may form the basis of an effective treatment for infections caused by S. aureus biofilms.

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Section: Methodsmentioning

confidence: 99%

Combined Use of Bacteriophage K and a Novel Bacteriophage To Reduce Staphylococcus aureus Biofilm Formation

Alves

Gaudion

Bean

et al. 2014

Appl Environ Microbiol

149

128

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“…Phages' resistance to adsorption could be due to a variety of factors including the loss of phage receptor molecules on hosts; physical barriers such as capsules masking the receptor molecules; and restriction-modification mechanisms such as phage-genome uptake blocks, super-infection immunity and restriction modification [14].…”

Section: Introductionmentioning

confidence: 99%

Determining the Minimum Inhibitory Concentration of Bacteriophages: Potential Advantages

Vipra¹,

Desai²,

Junjappa³

et al. 2013

AiM

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The minimum inhibitory concentration (MIC) is the concentration at which an antibacterial agent experiences the complete inhibition of organism growth. Bacteriophages represent a rich and unique resource of anti-infectives to counter the growing world-wide problem of antibiotic resistance. In this study, we compared the host range of lytic bacteriophages and temperate phagesbelonging to various genera, namely Staphylococcus, E. coli and Salmonella, with a range of clinical isolates using two methods: the classical agar overlay method and a newly developed MIC method. MIC was only observed with isolates that were susceptible to phage infection, which correlated with the agar overlay assay, whereas no MIC was detected with isolates that were resistant to phage infection. The simple MIC method was useful in determining phage adsorption and host range, and detecting possible prophage contamination in phage preparations. Interestingly, this method was also applicable to strain differentiation through phage susceptibility testing using a 96-well, high throughput format that proved to be easy, cost-effective, fast and reliable.

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“…Phages K and 44AHJD were amplified in S. aureus strains RN4220 and KB600, as described previously [27]. Briefly, the propagating hosts were grown at 37˚C in LB broth to an absorbance at 600 nm of ~0.8 and then infected with the respective phages at a MOI of 0.1 and further incubated for 4 h. Phage was harvested following centrifugation of the culture lysate at 3000 × g for 10 min to remove the cell debris.…”

Section: Bacteriophage Propagation Enumeration and Host Range Determentioning

confidence: 99%

“…While phage K was propagated using RN4220 as a propagating host, phage 44AHJD was propagated using KB600 host since we have earlier observed that 44AHJD requires endolysin supplementation for propagation in RN4220 [27]. Both the phages were amplified to a titer of −1 × 10 10 PFU/mL in bioreactors using the protocol described earlier [27] and the host range of both the phages were assessed on a panel of Staphylococcus aureus isolates that included both MSSA and MRSA.…”

Section: Bacteriophage Amplification Enumeration and Host Range Detementioning

confidence: 99%

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Therapeutic Potential of Staphylococcal Bacteriophages for Nasal Decolonization of <i>Staphylococcus aureus</i> in Mice

Narasimhaiah¹,

Asrani²,

Palaniswamy³

et al. 2013

AiM

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Bacteriophages represent a rich and unique resource of anti-infectives to counter the global problem of antibiotic resistance. In this work, we assessed the bactericidal activity of two virulent staphylococcal phages, K and 44AHJD, against S. aureus isolates of clinical significance, and tested their efficacy in vivo. The phage cocktail lysed >85% of the clinical isolates tested. Both the phages were purified by ion-exchange column chromatography following propagation in bioreactors. The purity profiles of the ion-exchange purified phages were compared with those of phages purified using cesium chloride density gradient ultracentrifugation, and infectiousness of the purified phages was confirmed by plaque forming assay. The in vivo efficacy of a phage cocktail was evaluated in an experimental murine nasal colonization model, which showed that the phage cocktail was efficacious. To our knowledge, this is the first report of phage use in an in vivo model of nasal carriage.

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Use of prophage free host for achieving homogenous population of bacteriophages: New findings

Cited by 7 publications

References 38 publications

Combined Use of Bacteriophage K and a Novel Bacteriophage To Reduce Staphylococcus aureus Biofilm Formation

Combined Use of Bacteriophage K and a Novel Bacteriophage To Reduce Staphylococcus aureus Biofilm Formation

Determining the Minimum Inhibitory Concentration of Bacteriophages: Potential Advantages

Therapeutic Potential of Staphylococcal Bacteriophages for Nasal Decolonization of <i>Staphylococcus aureus</i> in Mice

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Use of prophage free host for achieving homogenous population of bacteriophages: New findings

Cited by 7 publications

References 38 publications

Combined Use of Bacteriophage K and a Novel Bacteriophage To Reduce Staphylococcus aureus Biofilm Formation

Combined Use of Bacteriophage K and a Novel Bacteriophage To Reduce Staphylococcus aureus Biofilm Formation

Determining the Minimum Inhibitory Concentration of Bacteriophages: Potential Advantages

Therapeutic Potential of Staphylococcal Bacteriophages for Nasal Decolonization of &lt;i&gt;Staphylococcus aureus&lt;/i&gt; in Mice

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Therapeutic Potential of Staphylococcal Bacteriophages for Nasal Decolonization of <i>Staphylococcus aureus</i> in Mice