DAVIS JOANNE T (US)
HINZE GILBERT THEO (ZA)
| US5674537A | 1997-10-07 | |||
| US3616355A | 1971-10-26 | |||
| US4724059A | 1988-02-09 |
| 1. | l. |
| 2. | A composition for treating Legionella Pneumophila comprising an electrochemically activated anioncontained aqueous solution. |
| 3. | A composition as claimed in claim 1, wherein the anioncontaining solution is obtained from the electrolysis of an aqueous solution of a salt. |
| 4. | A composition as claimed in claim 1, wherein the anioncontaining solution is produced by an electrochemical reactor, the electro chemical reactor including a through flow, electrochemical cell having two caxial cylindrical electrodes with a caxial diaphragm between them so as to separate an annular inter electrode space into a catalytic and an analytic chamber. |
| 5. | A composition as claimed in claim 1, wherein the solution has a redox potential of above + 600 mV and a pH of about 6,57,5. |
| 6. | A method for treating Legionella Pneumophila comprising the steps of atomising a suitable dosage of an electrochemically activated, anioncontaining aqueous solution; and dispensing the atomise dosage of aqueous anioncontaining solution into an atmosphere to be treated. |
| 7. | The use of an electrochemically activated anioncontaining aqueous solution in the preparation of a medicament for use in the treatment of Legionella Pneumophila in humans. |
Background. o the Invention : So-called Legionnaire's disease became known in 1970'after an outbreak of a serious respiratory disease, diagnosed as mavir. Q ceen caused bv Legioneiia oneumoohila. Current treatment in humans includeã Erythromycin with the addition of Rifampicin in non-responding cases. haveawidenaturaldistributioninwaterandtheirgrowthlegionellab acteria is othermicro-organisms,includingPseudomonasspecies,whichby provide nutriments and protect them from adverse conditions, including the effet of biocidal treatment of water.
The Legionella bacteria can infect humans by means cf an aerosol, moving into the breathing zone of persons and deposition of the aerosol into the lungs.
Other sources of infection include recreational waters, residential and industrial waters, air-conditioning systems, humidifiers, respiratory therapy apparats, dental water supply lines ana resuscitation systems.
Current conr,-ol mesures against infectior. include super heating, hyper- chlorination and chlorine aasification. Howwer, no operatina, maintenance, cleaning and decontamination procedures presently exist that are generally regarded as safe work practices.
Obiect of the Invention: It is accordingly the object of this invention to provide a composition for treating Legionella as well as an associated method for treating same.
Brief Summarv of the Invention According to a first aspect of the invention there is provided a composition for Pneumophilacomprisinganelectro-chemicallyactivatedtreatingLe gionella anion-containing aqueous solution.
The anion-containing solution, or so-called anolyte, may be obtained from the electrolysis of an aqueous solution of a salt. The salt may be sodium chloride.
In particular it may be non-iodated sodium chloride or potassium chloride.
The anion-containing solution and the associated cation-containing solution may be produced by an electro-chemical reactor or so-called electrolysis device.
The electro-chemical reactor may include a through flow, electro-chemical cell having two c-axial cylindrical electrodes with a c-axial diaphragm between them so as to separate an annular inter electrode space into a catalytic and an analytic chamber.
The anolyte may have a redox potential of above + 600 mV and preferably about +750 mV and may have a pH of about 6,5-7,5. The anolyte may include any one of more or radical anion species from the group consisting of ClO; C10- ; HC10; OH; HO, - ; H, O,; ;03 S, 082-and C12062-.
According to a second aspect of the invention there is provided a method for treating Legionella Pneumophila comprising the steps of atomising a suitable dosage of an electro-chemically activated, anion-containing aqueous solution; and dispensing the atomise dosage of aqueous anion-containing solution into an atmosphere to be treated, the aqueous solution being substantially as herein defined.
According to a third aspect of the invention there is provided the use of an electro-chemically activated anion-containing aqueous solution in the preparation of a medicament for use in the treatment of Legionella Pneumophila in humans.
Detailed Description of Preferred Embodiment: A preferred embodiment of the invention will now be described by means of three non-limiting examples only.
An electro-chemical reactor, including a through flow, electro-chemical cell having two co-axial cylindrical electrodes with a c-axial diaphragm between them so as to separate an annular inter-electrode space into a catalytic and an analytic chamber, was used to produce anolyte and catholyte solutions.
Example 1 Anolyte solution with varying characteristics was used as shown in the respective examples.
A series of trials have been conducted whereby various dilutions of aqueous anion-containing solutions have been seeded with Legionella pneumophila (Serotype 1) organisms and the microcidal effects of treatment with anolyte have been observe after incubation for a period of 4 days (96 hours) at a temperature of C. The efficacy of the treatment with anolyte at the various dilutions and times of exposure was established by the presence or absence of Legionella cultures on the infecte BCYE culture medium.
Three replicates of each of the dilutions and of the control groups were seeded with a pure culture of Legionella Pneumophila (Serotype 1), resulting in counts of above 7 million parts per millilitre (TNTC).
Samples were collecte at pre-determined time intervals and transferred onto the growth medium before being incubated for 4 days (96 hours) at i37°C.
As can be seen from the Table for Example 1, anolyte was microcidal at levels between +998 mV and +407 mV (i. o. w. at a dilution rate of more than 1-10).
Example 2 Further tests were then conducted to narrow down the ranges of efficacy using a reducing-oxidation potential (ORP) as the monitoring (measuring) and on a similar basis as set out in Example 1.
As is illustrated in the Table for Example 2, it is deduced that: 1. A contact time of about 5 minutes at about 750 mV and a contact time of about 30 minutes at about +607mV is completely microcidal against Legionella Pneumophila (Serotype 1); and 2. The microcidal effect of anolyte is directly proportional to the ORP of the dilution.
Example 3 P. aeruginosa and S. aureus (Methicillin resistant) strains were cultured overnight on blood agar plates. Both of these strains were obtained from clinical specimens obtained during routine laboratory investigations at the General Hospital in Johannesburg, South Africa.
The L. pneumophila strain was cultured for 3 days on BCYE agar as it is a slow-growing organism. This isolate was obtained from the ATCC (American type culture collection) reference stock cultures, designated ATCC 33155.
These plate cultures were used for preparation of the liquid suspensions in Ringer's solution.
A suitable inoculum of each of the 3 test strains was removed from the agar plates with a nichrome loop and emulsified in 1/40 strength Ringer's buffer.
These were then homogenised in a vortex mixer (the 1/40 Ringer's buffer is suitable for diluting the fastidious Legionella as well as the S. aureus and P. aeruginosa). Using a 0.5 McFarland's standard opacity tube, which is the equivalent to 150 million organisms/ml, the capacity of the three cultures in suspension was adjusted to an opacity to give a final count (i. e. after adding to the Ringer's solution or Ringer's-anolyte solution) of approximately 1 million colony forming units per ml (1 x 106 cfus per ml - called the "high count challenge). A second set was prepared with a 1/10 dilution (1 x 105 cfus -called the"low count challenge").
Anolyte solutions were prepared as follows : 1: 1 - 1 part Ringer's 1/40 + 1 part anolyte (2.0 mi + 2.0 ml) 1: 50-49 parts Ringer's 1/40 + 1 part anolyte. (4.9 + 2.0) 1: 100-1 part 1: 5 anolyte + 1 part Ringer's (2.0 ml + 2.0 ml) 1: 150-1 part anolyte + 2 parts Ringer's (1.0 ml + 2.0 ml).
These dilutions were distributed in 100u 1 quantities in 5 ml disposable plastic test tubes in triplicate for each set of organisms.
A thiosulphate neutraliser was made up by adding 2 crystals per 10 ml (which
is also the amount used in the British Public Health Service Laboratories or PHLS including that of John Lee's Legionella Unit) and distributed in IOAI quantities in plastic disposable test tubes.
All cultures were pre-tested microbiologically to determine whether any effect such as a decrease in the number of viable organisms recovered would occur, using any of the reagents such as thiosulphate neutraliser or a 30 minute exposure to Ringer's buffer.
This test was done in triplicate as follows : Test tube dilutions: Ringer's 1: 1 1: 5 1: 100 1: 150 100µl only anolyte anolyte anolyte anolyte To each set of test tubes containing either the anolyte dilutions or plain Ringer's (i. e. the control), 1 drop (10gel) of culture was added. As the same conditions were being applied to both the test and the control samples, no special calculation was required for volume adjustment from IOAI to IlOgI when the culture was added.
From the Ringer's only control tube, a further 1/100 dilution in Ringer's was made at the appropriate time interval (see below) to facilitate counting, should the original plate count be too high to observe individual cfus.
At the appropriate time in intervals, (5 mins and 30 mins post-exposure) lOtL 1 of organism in anolyte dilution/Ringer's only (control) was removed and mixed with the 10 jul of thiosulphate neutraliser. This"mix"was seeded onto a petri dish (blood agar for the S. aureus and P. aeruginosa and BCYE for the Legionella). the plates were spread over the entire surface with a sterile nichrome spreader.
The blood agar plates were incubated for 48 hours at 37°C aerobically and the BCYE plates at the same temperature for 5 days aerobically in a sealed jar with a very moist atmosphere.
Colonies were counted using a colony counter with a magnifying lens and a grid.
No significant difference in the number of cfus/ml of the untreated (control) organisms were obtained after (a) being left in Ringer's solution for a 30 minute period and (b) treatment with sodium thiosulphate when compare with counts taken immediately after preparation of the suspensions. Thus any drop in cfus was purely due to the effect of the anolyte.
The"high count challenge"dose gave the following numbers of cfus/ml : S. aureus 3.4 1 o6 cfus/ml P. aeruginosa 1.2 x 106 cfus/ml L. pneumophila 2.7 x 106 cfus/ml The"low count challenge"dose gave the following numbers of cfus/ml : <BR> <BR> <BR> <BR> <BR> S. aureus 2.9 x 105 cfus/ml<BR> <BR> <BR> <BR> <BR> <BR> <BR> P. aeruginosa 2.2 x 105 cfuml L. pneumophila 4.9 x 105 cfu/ml <BR> <BR> <BR> <BR> <BR> All cultures with a concentration of 105 cfus/ml showed no growth (became non-viable) after being exposed to any of the dilutions of anolyte (1: 1,1: 50, 1: 100,1: 150) for both the 5 and 30 minute periods.
The results of these cultures containing 106 cfus/ml treated in the same manner with anolyte dilutions were as set in the Table for Example 3.
It is envisage that the following methods of treatment could be used :
It is envisaged that the following methods of treatment could be used: 1. By dosing anolyte onto elements such as a condenser used in air- conditioning systems; 2. By fogging anolyte into air-conditioning ducts or into the atmosphere eg. in an operation theatre, etc.; and 3. By patients inhaling fogged anolyte, thereby exposing the Legionella organism to the anolyte in the alveoli of the lungs.
It will be appreciated that many variations in detail are possible without departing from the scope and/or spirit of the invention as claimed in the claims hereinafter.