Effects of a Novel Effects of a Novel Compound on Fungal Compound on Fungal

ContaminationContaminationEstelle LevetinEstelle Levetin

Novel CompoundNovel Compound Composition not totally revealedComposition not totally revealed Described as Catalyst Altered WaterDescribed as Catalyst Altered Water Dilute solution of electrolytesDilute solution of electrolytes Working name “pH Balancer”Working name “pH Balancer” Reported to inhibit mold and bacteria Reported to inhibit mold and bacteria

and other beneficial effects and other beneficial effects pH subsequently measured at 12.63pH subsequently measured at 12.63

Other Compounds TestedOther Compounds Tested In addition to pH Balancer, we were In addition to pH Balancer, we were

asked to test various amendments:asked to test various amendments: 50/50 Solution50/50 Solution 50/50 Plus Solution50/50 Plus Solution pH Balancer Plus pH Balancer Plus

MethodsMethods Saturation experimentsSaturation experiments Spore germination experimentsSpore germination experiments pH experimentspH experiments Pretreatment experimentsPretreatment experiments Spray applicationSpray application Growth in liquid cultureGrowth in liquid culture

Ceiling Tile SaturationCeiling Tile Saturation Soaking ceiling tile squares (CTS) in Soaking ceiling tile squares (CTS) in

50% and 100% solutions of pH 50% and 100% solutions of pH Balancer inhibited fungal growthBalancer inhibited fungal growth

pH Balancer

50/50 Solution50/50 Plus Soln

Control0

2

4

6

8

Colo

ny d

iam

(cm

)

Growth of Stachybotrys chartarum on ceiling tiles with and without 50% test solutions

pH Balancer

50/50 Solution50/50 Plus Soln

Control0

1

2

3

4

Colo

ny d

iam

(cm

)Growth of Aspergillus versicolor on ceiling

tiles with and without 50% test solutions

pH Balancer

50/50 Solution50/50 Plus Soln

Control0

1

2

3

Colo

ny d

iam

(cm

)

Growth of Penicillium sp. on ceiling tiles with and without 50% test solutions

Alternaria alternata

Stachybotrys chartarum

Cladosporium cladosporoides

Aspergillus versicolor

Penicillium sp.

Solution Mean colony diameter (cm)*

50/50 Solution 0.23 2.33 1.10 0.90 0.23

50/50 Plus Solution 0.17 0.77 0 0.70 0

Control 6+ 6+ 4.40 4.38 3.42

*Although CTS are 6 cm in length and width, diagonal measurements are approximately 8.5 cm. Growth of the colony beyond 6 cm is indicated as 6+ in the table but plotted as 6.0 in the graphs.

Growth of fungi on Ceiling Tile Squares saturated with 100% solutions of test

compounds

5-Feb 12-Feb 18-Feb 26-Feb 4-Mar

Mean colony diameter (cm)

Alternaria-Control 0 5.28 6.00 6+ 6+

Alternaria-pH Bal 0 0 0 0 0

Alternaria-Bal PLUS 0 0 0 0 0

Stachybotrys-Control 0 2.87 4.77 5.72 6.00

Stachybotrys-pH Bal 0 0 0 0 0

Stachybotrys-Bal PLUS 0 0 0 0 0

Cladosporium-Control 0 1.57 2.23 3.13 3.90

Cladosporium-pH Bal 0 0 0 0 0

Cladosporium-Bal PLUS 0 0 0 0 0

Aspergillus-Control 0 1.33 2.13 2.90 3.60

Aspergillus-pH Bal 0 0 0 0 0

Aspergillus-Bal PLUS 0 0 0 0 0

Penicillium-Control 0 1.78 2.37 3.97 4.10

Penicillium-pH Bal 0 0 0 0 0

Penicillium-Bal PLUS 0 0 0 0 0

Growth of fungi on Ceiling Tile Squares saturated with 100% solutions of test compounds

Sheetrock SaturationSheetrock Saturation Comparable experiments on Comparable experiments on

sheetrock squares produced very sheetrock squares produced very different resultsdifferent results

7-Jan

14-Jan

21-Jan

28-Jan

Stachybotrys-Control

Stachybotrys-pH Bal-50%Alternaria-Control

Alternaria-pH Bal-50%0

2

4

6

8

Growth of Stachybotrys and Alternaria on Sheetrock with 50% Solution of pH Balancer

17-Ja

n

24-Ja

n

31-Ja

n

7-Feb

14-F

eb

Aspergillus-ControlAspergillus-pH Bal-50%

Cladosporium-ControlCladosporium-pH Bal-50%0

2

4

6

Growth of Aspergillus and Cladosporium on Sheetrock with 50% Solution of pH Balancer

21-Jan 28-Jan 4-Feb 11-Feb 18-Feb

Mean colony diameter (cm)

Alternaria-Control 0 2.60 3.82 5.20 5.93

Alternaria-pH Bal 0 2.17 4.23 4.70 4.92

Alternaria-Bal PLUS 0 1.92 4.00 4.70 5.17

Stachybotrys-Control 0 3.00 4.52 4.85 5.13

Stachybotrys-pH Bal 0 0.85 2.30 3.92 4.30

Stachybotrys-Bal PLUS 0 1.00 2.80 3.87 4.47

Cladosporium-Control 0 3.33 4.17 5.12 5.50

Cladosporium-pH Bal 0 1.07 1.30 1.87 2.13

Cladosporium-Bal PLUS 0 1.38 2.10 2.55 2.83

Aspergillus-Control 0 3.50 3.80 4.50 4.60

Aspergillus-pH Bal 0 1.83 2.13 2.78 3.07

Aspergillus-Bal PLUS 0 2.27 2.67 3.37 3.48

Growth of fungi on Sheetrock Squares saturated with 100% solutions of test compounds

No inhibition on SheetrockNo inhibition on Sheetrock Could gypsum interfere with pH Could gypsum interfere with pH

Balancer?Balancer? Gypsum (calcium sulfate) used as a soil Gypsum (calcium sulfate) used as a soil

amendment for alkaline soilsamendment for alkaline soils Gypsum has a neutral pH but Gypsum has a neutral pH but

significant buffering capacity and will significant buffering capacity and will lower soil pHlower soil pH

pH Balancer has a pH of 12.63pH Balancer has a pH of 12.63

Solution pH of Solution

Alternaria alternata

Cladosporium cladosporoides

Stachybotrys chartarum

Aspergillus versicolor

Control – 1 100% MEB 5.19 90+ 90 90+ 90+

Control – 2 50% water 50% MEB

5.18 90+ 90 90+ 90+

Control – 3 75% water25% MEB

5.13 90+ 90 90+ 90+

50% pH Balancer50% MEB

11.82 0 0 50 46.7

75% pH Balancer25% MEB

12.28 0 0 0 0

50% Balancer Plus 50% MEB 11.80 0 0 32.3 5.7

75% Balancer Plus 25% MEB 12.22 0 0 0 0

Effect of pH Balancer on Spore Germination

pH Changes of Solutions Surrounding Sheetrock Squares

456789

1011121314

11-Feb 17-Feb 23-Feb 1-Mar

pH V

alue

WaterpH BalBal Plus

pH Changes of Solutions Surrounding Ceiling Tile Squares

456789

1011121314

11-Feb 17-Feb 23-Feb 1-Mar

pH V

alue

WaterpH BalancerBalancer Plus

Is it only a pH effect?Is it only a pH effect?3-

Mar

10-M

ar

17-M

ar

24-M

ar

31-M

ar

Alternaria pH Balancer

Alternaria 0.025M NaOH

Alternaria control01

23

4

5

6

Growth of Alternaria alternata on ceiling tile squares soaked with different solutions

pH Changes of Solutions with Ceiling Tile Squares

4

6

8

10

12

14

0 7 14 21

Days

pH V

alue

pH Bal

NaOH

pH Changes of Solution with Sheetrock Squares

4

6

8

10

12

14

0 7 14 21

Days

pH V

alue

pH Bal

NaOH

Pre-treatmentPre-treatment Ceiling Tile Squares were soaked in pH Ceiling Tile Squares were soaked in pH

balancer and allowed to dry to simulate its balancer and allowed to dry to simulate its use in manufacturing use in manufacturing

Tiles were stored for varying lengths of Tiles were stored for varying lengths of timetime

Tiles were later soaked with sterile distilled Tiles were later soaked with sterile distilled water and inoculated with fungiwater and inoculated with fungi

Pretreated tiles completely inhibited fungal Pretreated tiles completely inhibited fungal growthgrowth

Other experimentsOther experiments Spray applications of pH Balancer Spray applications of pH Balancer

failed to inhibit actively growing fungi failed to inhibit actively growing fungi on both substrateson both substrates

Growth of various fungi (including Growth of various fungi (including several plant pathogens) was several plant pathogens) was completely inhibited in liquid medium completely inhibited in liquid medium (50% malt extract broth and 50% pH (50% malt extract broth and 50% pH Balancer)Balancer)

ConclusionConclusion pH Balancer inhibited fungal growth pH Balancer inhibited fungal growth

on ceiling tileson ceiling tiles May be a useful addition to May be a useful addition to

manufacturing processmanufacturing process Other applications for pH Balancer Other applications for pH Balancer

require further testingrequire further testing

AcknowledgementsAcknowledgements Assistance of Claudia Owens, Laura Assistance of Claudia Owens, Laura

Wacker, and Christin Townsend is Wacker, and Christin Townsend is greatly appreciated.greatly appreciated.