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The Impact of an Environmental Probiotic of the type BioZone onFungal Contaminated Air Filters.

Author: Herman Sabath, PhD, MPH, MSc, CMC

Assistant: Yuli Horesh

Introduction

1) General (Historical Background)

Since the beginning of the as" century, several serious epidemiological events have occurred caused by microbialgrowth in Built environments.

Among them are; Hospital Acquired infections (nosocomial) that afflicted Health care institutions.

Attempts by celebrated hygiene pioneers such as Dr. Semmelweiss, unfortunately, did not bare fruits.

However, major outbreaks of nosocomial events triggered an increased attention paid by managements, a

phenomenon that continues to this day.

In Israel, awareness on this topic increased following reported cases of nosocomial incidents (Between 5000 -

6000 Deaths) most recently reported by media and 40,000 hospitalized individuals have experienced microbialinfections.

As per WHO, hospital- acquired infections are also becoming increasingly challenging to treat as antibiotic resistant

bacterial strains emerge, particularly in ICU. WHO estimated that drug resistant microbes cause up to 60% of

nosocomial infections, and they could outpace the development of new drugs that can effectively treat microbial

infection.

According to the most recent figures from the CDC, an estimated 1.7 - 1.8 million nosocomial infections occurred

in the USA during 2002, resulting in 99.000 - 110.000 deaths, and adding billions in potentially avoidable

expenditure (59.4 Billion Dollars in 2011, excluding indirect cost such as loss of productivity and diminished quality

of life).

2) Purpose

It was this situation that brought a well-known Medical Center in the metropolitan area of Tel Aviv to initiate an

environmental study with the double purpose of evaluating the Indoor microbial quality ie Bacterial & Fungal

contaminants, with Qualitative and Quantitative determinations, and evaluating a unique, Non toxic mixture of

Probiotic bacteria as a potential preventive method for Indoor bacterial and fungal infections.

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3) Scope of Work

We performed a limited scope of work in order to determine the efficacy of a Probiotic treatment over fungal

contamination of two filtration units used in HVAC systems designed for Hospitals.

4) Methods

In the first stage of our survey, each one of the units was previously sampled and analyzed for existence of fungal

contaminants (QT and QL), by Hayes Microbial LAB, an accredited and certified environmental microbiology

laboratory In Virginia, USA.

In the second stage, following Probiotic treatment via a unique aerosolization method which distributes a

measured amount of the BioZone Probiotic mixture on infected areas at a predetermined schedule according to

needs. Fungal samples were collected and sent to an analytical, mycology LAB in the USA.

a) Creating an "lnvitro" environment in accordance with ISO 13697;2001 criteria.

b) Collection Media: agar samples- OGYE+ oxytetracycline Hydrochloride (@) 10 cm2

c) A Patented probiotic Mixture of Bacillus spp. (BioZone) which includes: Bacillus coagulans,

Bacillus lentus, Bacillus lichenijormis, and Bacillus pumilus. All Grade One safety level according tothe FDA.

d) Mikropor filter F8, Dimension: 592/592/292 mm

e) Intervals of Sampling/elapsed time:

• Control (0)

• 24 Hours

• 48 Hours

• 72 Hoursf) The collected samples were packed and shipped according to the environmental laboratory

requirements.

5) Results

We performed collection of samples from the main filters of a Hospital AHU (Air Handling Unit) and analyzed the

results at a certified accredited and registered environmental microbiology LAB (USA), and obtained the following

results as tabulated in table 1 & 2, representing the level of fungal contamination prior to "Biozone" application.

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Table 1 (Before)

Concentration of Fungal counts in units of CT/cm2* in filter nQ1 ofthe AHU

# Sample Location Cladosporium Penicillium

1 Center of Filter 84,000 38,000

2 Top Left of Filter 6 4,200

3 Bottom Right of Filter 2 2,400

• Penicillium

Filter no 1 - Before BioZone Awlication90,000

80,000

70,000

60,000

50,000

40,000

30,000

20,000

10,000

o

• Cladosporium

Center of Filter Top Left of Filter Bottom Right ofFilter

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Table 1 (After)

Concentration of Fungal counts in unites of count/ern" in filter nQ1 of the AHU

# Sample Location Cladosporium

1 Center of Filter NFD**

2 Top Left of Filter NFD**

3 Bottom Right of Filter 100

120 ~------------------------------------------Filter no 1 - After BioZone Application100 +-----------------~~--------

80 +--------------------------------60 +-----------------------------40 +-------------------------------20 +--------------------------------

o +--------------r------------~---

• Cladosporium

Center of Filter Top Left of Filter Bottom Right of Filter

INTERNAnONAL SOCteT'I' OfINDOOR AIR OU.A1..ITV

ANOCL ••••ATE

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Table 2 (Before)

Concentration of Fungal counts in units of CT/cm2* in filter nQ2 ofthe AHU

# Sample location Alternaria Aspergilus Cladosporium

1 Center of Filter 2,000 78,000 40,000

2 Top Left of Filter. N/A N/A N/A

3 Bottom Right of Filter N/A N/A N/A

90,00080,00070,00060,00050,00040,00030,00020,00010,000

o

• Cladosporium

Aspergi Ius

• Alternaria

Center of Filter Top Left of Filter Bottom Right ofFilter

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Table 2 (After)

Concentration of Fungal counts in unites of count/ern" in filter nQ2 of the AHU

# Sample location Results

1 Center of Filter NFD**

2 Top Left of Filter NFD**

3 Bottom Right of Filter NFD**

1 ,-------------------------------------

1

1 +-------------------------------------1 +-------------------------------------1 +-------------------------------------1 +-------------------------------------o +-------------------------------------o +-------------------------------------o +-------------------------------------o +-------------------------------------o +-----------,------------r-----------,

Filter no 2 - After BioZone Aqglication BioZone

• Cladosporium

Aspergilus

Alternaria

Centerof Filter Top Leftof Filter Bottom RightofFilter

Observation: During the experiment, fluctuation in level of certain Fungi occurred in filter nQ1of theAHU i.e:

• After 24h of application, 300 - 1600 Penicillium• After 48h of application, NFD• After 72h of application, NFD

* CT/cm'- Counts per Square Meter

*. NFD - No Fungi Detected

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6) Summary

1) Our experiment is a pilot study, a preliminary step in searching for a solution that would involve

application of Nontoxic disinfectants mainly in leu and other sensitive locations in Hospitals.

2) The results we obtained by using SioZone Probiotics/EBA on two main filters of AHU were excellent and

encouraging. (See chapter results, before and after application of BioZone Probiotics).

3) Our hypothesis for the use of environmental Probiotics for an efficient control of microbial

(Bacterial/Fungal) growth, could be re-tested using our experimental model on a variety of objects such as

fomites, textiles, construction materials and a variety of porous and semi-porous materials, and similar to

those in hospitals, under similar environmental conditions such as; air temperature and relative humidity

(40% - 50% relative humidity and 1Soc- 22°c Air temperature)

4) It is in our intentions to go ahead with more experiments, using additional approaches/parameters such

as: decreased concentrations of the Probiotic mixture used in the current experiment, elapsed time

looking into the effect of shorter than 72 Hours (as it was also used by other researchers) and even more

important; to look into the more longer lasting effect of BioZone Probiotics application that has significant

positive economic implications.

5) All surface experiments executed using the Probiotic concept around the world, were done with Physical

Manual cleaning applications on surfaces. This is a first effectively proven experiment indicating that

BioZone type Probiotic can control and reduce surface Fungal contamination on Air Filters.

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Definitions

Nosocomial: Hospital acquired infections as any localized or systemic patient condition that; (a) resulted from the

presence of an infectious agent (Bacteria/Fungi) or its toxin(s) as determined by Laboratory testing; and (b) thecondition was not found to be present or incubating at the time of admission, UNLESSTHE INFECTION WAS

RELATEDTO A PREVIOUSCONDITION. (Source; the Public Health Law, Hospital Acquired Infection Reporting 2819)

Biosafe level 1: Non Pathogenic

QL: Qualitative

QT: Quantitative

CDC: Centers for disease Control and Prevention (USA)

WHO: World Health Organization (UN)

ICU: Intensive Care Unit

Environmental Probiotics: (Our Probiotic Mixture)

EBA - Ecological Balancing Agent

Probiotics: Non- pathogenic beneficial bacteria, members of the Bacillus genus(Bio-safety level one)

AHU: Air Handling Units of ventilation systems, equipped with HEPA and main filters.

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