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Compositions and Methods for Microbe Storage and Delivery

Inactive Publication Date: 2009-02-12
CONJUGON
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0028]While the invention is not limited to use with any particular medical device, in some preferred embodiments, the medical device comprises a urinary catheter. In some embodimen

Problems solved by technology

No mode of treatment is known to eliminate chronic, subclinical infections or to prevent intercurrent, clinically important infections.
Most measures that have been tested have not shown effectiveness in randomized clinical trials, however, and some are not applicable to patients with a neurogenic bladder.
However, these technologies have not been confirmed to be effective in randomized clinical trials (Maki and Tambyah, supra).
Additionally, antimicrobial treatment of asymptomatic CAUTI has been associated with the emergence of drug-resistant organisms, complicating management when a symptomatic infection does occur.
Given the difficulty of eradicating bacteriuria in a patient with long-term bladder catheterization, the problem of chronic bacteriuria and recurrent UTI in catheter-dependent persons is not likely to be resolved by the use of antimicrobial agents.
However, existing methods of pre-inoculation of the urinary tract are cumbersome.

Method used

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  • Compositions and Methods for Microbe Storage and Delivery
  • Compositions and Methods for Microbe Storage and Delivery

Examples

Experimental program
Comparison scheme
Effect test

example 1

Preparation of Freeze-Dried E. coli HU2117; Protocol 1

[0120]The goal of the investigation was to examine the effects of different excipients and conditions for lyophilization of HU2117, so as to maintain an effective cell concentration and level of viability of cells freeze-dried in a composition comprising a gelling agent.

Cell Preparation

[0121]Two flasks of cells (Flask A and Flask B) were grown, each from 1 ml of seed stock inoculated into 1 L modified EZ Rich Defined Glycerol medium, incubated at 37±1° C. for 8 hrs with constant shaking at 250 RPM. At the end of 8 hours, the OD600 of Flask A was 2.53 and the OD600 of Flask B was 1.11.

[0122]The cells were collected by centrifugation at 4° C., at 6000 RPM for 8 min. The pelleted cells were washed twice with 0.9% saline and once with 10 mM citrate buffer, pH 7.0. The cells pelleted from each liter of culture were resuspended into 2-3 ml of 10 mM citrate buffer, pH 7.0, for a final volume of approximately 5 ml.

[0123]The concentratio...

example 2

Preparation fo Freeze-Dried E. coli HU2117; Protocol 2

Cell Preparation

[0143]Two flasks of cells (Flask A and Flask B) were grown, each from 1 ml of seed stock inoculated into 1 L Modified EZ Rich Defined Glycerol medium, incubated at 37±1° C. for 8 hrs with constant shaking at 250 RPM. At the end of 8 hours, the OD600 of Flask A was 1.89 and the OD600 of Flask B was 1.53.

[0144]The cells were collected by centrifugation at 4° C., at 6000 RPM for 8 min. The pelleted cells were washed twice with 0.9% saline and washed once with 10 mM citrate buffer, pH 7.0.

[0145]The cells pelleted from each liter of culture were resuspended into 2-3 ml of 10 mM citrate buffer, pH 7.0, for a final volume approximately 5 ml.

[0146]The concentration of the resuspended cells was determined using plate counts. Resuspended cells from Flask A had a viable cell concentration of 1.2±0.3×1011 CFU / ml, and resuspended cells from Flask B had a viable cell concentration of 7.7±0.3×1010 CFU / ml. The cells from Flask A ...

example 3

Preparation of Freeze-Dried E. coli HU2117; Protocol 3

Cell Preparation

[0169]One 2 liter flask of cells was grown from 1 ml of seed stock inoculated into 1 L Modified EZ Rich Defined Glycerol medium, incubated at 37±1° C. for 8 hrs with constant shaking at 250 RPM. At the end of 8 hours, the OD600 was 2.2±0.03.

[0170]The cells were collected by centrifugation at 4° C., at 6000 RPM for 8 min. The pelleted cells were washed twice with 0.9% saline and washed once with 10 mM citrate buffer, pH 7.0.

[0171]The pelleted cells were resuspended into 2-3 ml of 10 mM citrate buffer, pH 7.0, for a final volume of approximately 10 ml.

[0172]The concentration of the resuspended cells was determined using plate counts. The resuspended cells had a viable cell concentration of 4.9×1010 CFU / ml.

Lyophilization

[0173]For each test, 0.5 ml of resuspended cells were mixed with 1.5 ml of an excipient selected from the list below and 10 ml of 2% autoclaved hydroxyethyl cellulose (HEC).[0174]Excipients (shown as ...

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Abstract

The present invention relates to the field of bacteriology. In particular, the invention relates to compositions of probiotic microbes and methods for making and using such compositions, e.g. in the treatment and prevention of catheter associated urinary tract infections.

Description

FIELD OF THE INVENTION[0001]The present invention relates to the field of bacteriology and probiotic therapies. In particular, the invention relates to novel compositions (e.g., probiotic microbe preparations) and methods of using the same (e.g., for coating surfaces, such as catheters). In some embodiments, the present invention comprises a freeze-dried composition comprising a microbe that can be reconstituted to form a gel, e.g., a medical lubricant, containing viable, colonogenic microbes.BACKGROUND OF THE INVENTION[0002]Bacteriuria and pyuria are uniformly present in patients who have indwelling urinary catheters. Antimicrobial therapy may transiently eradicate the bacteria, but bacteriuria promptly recurs, and the infecting bacteria become progressively resistant to antibiotics. No mode of treatment is known to eliminate chronic, subclinical infections or to prevent intercurrent, clinically important infections.[0003]Universal guidelines intended to prevent or delay catheter-a...

Claims

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Application Information

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IPC IPC(8): A61K35/00A61P13/00C12N1/20A61K35/74A61K35/741
CPCA61K9/19A61K35/741A61K35/74A61P1/00A61P13/00A61P31/04Y02A50/30A61K35/744A61L29/085A61L29/16A61L2420/06
Inventor SUZUKI, HIDEKIBRAICO, SALVATORE
Owner CONJUGON
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