Reference

AlllDb E b 1 1 S

cations

NBSIR 81-2246

Biodeterioration of Standard

Reference Materials

G. J. Olson, W. P. Iverson, and F. E. Brinckman

Chemical Stability and Corrosion Division

Center for Materials Science

U.S. Department of Commerce

National Bureau of Standards

Washington, DC 20234

-QC

100

.1156

81-2246

1981

Prepared for Office of Standard Reference Materials

National Bureau of Standards

Washington, DC 20234

NATIONAL BUR^u

QV 9TANDAJU>8

not ace

OlLIOO

NBSIR 81-2246

•i* ' BIODETERIORATION OF STANDARD

REFERENCE MATERIALS

G. J. Olson, W. P. Iverson, and F. E. Brinckman

Chemical Stability and Corrosion Division

Center for Materials Science

U.S. Department of Commerce

National Bureau of Standards

Washington, DC 20234

April 1981

DISTRIBUTION OF THIS DOCUMENT IS UNLIMITED

Prepared for Office of Standard Reference Materials

National Bureau of Standards

Washington, DC 20234

U.S. DEPARTMENT OF COMMERCE, Malcolm Baldrige, Secretary

NATIONAL BUREAU OF STANDARDS, Ernest Ambler, Director
.

' 1

,

: '-v

"i '

.
CONTENTS

Page

List of Figures iv

List of Tables v

1. Introduction 1

2. Experimental Procedures 4

2.1 Enumeration of Microorganisms 4

2.2 Elemental Analysis 4

2.3 Exposure and Storage of SRMs 5

2.4 Mercury Binding 5

2.5 Chromium 5

3. Results and Discussion 5

4. Conclusions and Recommendations 9

5. References 11
LIST OF FIGURES

Page

1 Dual tube growth/absorbing system for measurement of

volatile microbial metabolites 4

2 Freeze dried urine rehydrated with sterile and non-sterile

distilled water 5

iv
LIST OF TABLES

Page

1. Elements, transformations, and SRMs 3

2. Numbers of microorganisms in urban particulate 5

3. Numbers of microorganisms in freeze dried urine 6

4. Binding of mercury to cells in urine 7

5. Growth of microorganisms on SRMs exposed to outdoor air

then stored at varying relative humidity 8

v
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'
Biodeteri oration of Standard Reference Materials

G. J. Olson 1

, W. P. Iverson, and F. E. Brinckman

Chemical Stability and Corrosion Division

Center for Materials Science

National Bureau of Standards

Washington, DC 20234

Several National Bureau of Standards Standard Reference

Materials have been examined for susceptibility to biodeterioration. Several of these materials were attacked by bacteria

and fungi when stored at elevated humidity after exposure to

outdoor air. Some SRMs underwent deterioration after following certificate instructions for handling. Suggestions for some certificate revisions are made.

Key words: bacteria; biodeterioration; biotransformations;

deterioration of materials; element volatilization; fungi;

metals; microorganisms; Standard Reference Materials.

1. Introduction

The validity and reliability of Standard Reference Materials certified for elemental composition in a given host matrix require that homogeneity of the SRM, and the concentrations of the certified elements,

will remain unaltered over a reasonable shelf life. This demands that

no chemical or biological process will occur that alters the homogeneity

or certified concentration of the specified elements or that alters the

matrix so as to reduce homogeneity or change concentrations of those

elements.

Many chemical or physical processes may occur during storage or

use of SRMs which can result in composition changes. These include

thermal cycling, photolysis, adsorption, volatilization, contamination,

atmospheric oxidation, hydrolysis, and the like. Such occurrences have

been studied in certain SRMs (R. Alvarez, personal communication). A more subtle form of deterioration, which has not received adequate

attention by analysts, may result from contamination by microbiota such

as bacteria or fungi. Such biodeterioration may be initiated by microbiota originally present in the particular matrix employed, as with

biological or environmental reference materials. Alternatively, contamination may be introduced subsequent to certification and sale to the

1 1979-81 NRC-NBS Postdoctoral Research Associate

1
user, who may introduce microbiological contamination in following

inadequate instructions for use or by using conventional handling procedures without adequate sterile controls.

Perhaps the most critical factor in the metabolism of the microbial

cells present in contaminated SRMs is water availability. The most useful measurement of water availability is "water activity" (aw

), defined

as the ratio of the equilibrium vapor pressure of water over the

material in question to that of pure water at the same temperature [1].

Growth of most microorganisms occurs only when aw

> 0.90, however,

xerophiles, (usually yeasts and molds) which grow under relatively dry

conditions (a = 0.85 to 0.60), exist [1]. Microorganisms do not grow

at aw

< 0.60. Most biologically susceptible SRMs are freeze dried, and

therefore, unless rehydration occurs, either accidentally or by design,

they should not experience biological deterioration. Rehydration is

possible, however, during customer use while dispensing and weighing or

as a result of failure to close a bottle cap tightly. Condensation of

moisture on a refrigerated SRM is very possible when it is moved to a warm laboratory.

Microbial or fungal attack on substrates containing trace (10 ^ to

-9 -1

10 g g ) quantities of a particular element in a natural or introduced (spiked) form may well result in measurable losses or translocations of the element over relatively short periods of time. Much of

the current environmental literature devotes considerable attention,

for example, to the biological transformations, transport, and reaccu- mulation in new reservoirs (often biological) of many trace toxic

elements, especially those frequently certified in needed SRMs [2, 3].

Fortunately, modern measurement techniques and methods are available

which are uniquely suited to assess the formation of certain molecular

species containing elements of concern even if this occurs slowly and

at very low concentrations. Such methods permit diagnostic use of such

element-containing metabolites as kinetic markers for use in establishing degradation rates and predicting form and trends of deterioration

of suspected SRMs. Table 1 lists some examples of microbially catalyzed

transformations of certain elements certified in some of the SRMs

potentially susceptible to biodeterioration. Volatile products are emphasized since these could lead to greatest loss of certified trace

elements from SRMs.

The present paper reports preliminary work designed to survey a group of currently produced NBS SRMs which we regard as susceptable to

biodeterioration, and for which certain elements, which we know to be

especially susceptible to such biotransformations [2, 3], have been

certi f ied.

2
Table 1. Examples of microbial transformations (especially volatile

products) of elements and some SRMs certified for these

elements potentially susceptible to such biodeterioration.

Volatile species formed may be lost to the atmosphere

resulting in alteration of certified element composition.

Element SRM Certified Microbial Transfor- Typical Environments

for Element mations of Element Where Transformation

Occurs

As 1571, 1648 (CH

3

) 3

As, AsH

3

, seawater [4],

(CH-^AsH as gases; sewage [5],

As0

2

in solution or sediments [6,7]

sol id

Hg 2672, 1645 Hg° , (CH ) Hg as tissue and urine [8],

f , gases; CH

3 Hg in sediments [9-12],

solution or solids feces [13],

water [14,15]

Cr 1569, 1570, +++ . Cr in solution waters and sedi1571 ments [16,17]

S 1621 H

? S, (CH^)? S, CH^SH, soils [18],

CS

?

, (CH 3

) ? S

? as manure [19],

gases waters and sediments

[20,21]

Se 1567 (CH^)

? Se, (CH^)? Se?

, soils and sewage [22]

(CH

3

) 2

Se

2

, Se° (all waters [23]

volatile)

Pb 1571 (CH

3

)^Pb (volatile), bogs [24],

organoleads sediments [25]

Sn

* 2672 (CH^SnH,, (CH,)

a

Sn, waters and sediments

other methyl tin com- [26,27,28]

pounds (most are volatile)

Certified for total Hg not Sn.

3
2. Experimental Procedures

2.1 Enumeration of Microorganisms

Microorganisms in SRMs were enumerated by surface or pour plating

[29] using trypticase soy agar or a modified iron-peptone agar [30]

with glucose (0.5 g L

^ ) substituted for gluconate. Incubation was at room temperature (about 22 °C) or 37 °C.

2.2 Elemental Analysis

Mercury in urine was quantified using a manual cold vapor procedure [31] employing a flameless atomic absorption spectrophotometer

(Mercometer, Anti-Pollution Control Technology Corporation). Volatile

mercury was detected and identified using a tandem gas chromatographatomic absorption system [32]. Chromium was determined using a PerkinElmer model 360 atomic absorption spectrophotometer equipped with a graphite furnace. Chromium volatilization experiments were performed

using a dual tube absorption system [33] shown in figure 1.

Figure 1. Dual growth tube/absorbing system for measurement of microbial metabolites. One tube contains sterile, deionized water,

the other contains brewers yeast (SRM 1569) and contaminating

fungal spores. Volatile metal-containing metabolites can be

measured by atomic absorption after partitioning into the

steri 1 e water.

4
2.3 Exposure and Storage of SRMs

Some of the SRMs were exposed to outdoor air (NBS grounds) for

periods of 30 to 60 min on successive days (total exposure 3.5 hr),

then were stored loosely capped at relative humidities of 100, 90, 70,

and 49 percent. The higher humidities were maintained using dessicator

jars containing calcium chloride solutions [34], and the lowest humidity

represented the laboratory ambient relative humidity which averaged

49 percent and ranged from 33 to 73 percent (April to June, 1980).

2.4 Mercury Binding

Mercury absorbed to cells growing in urine certified for mercury

(low level, SRM 2672) was quantified by passing 5.0 mL of contaminated

urine through a 25 mm, 0.45 pm membrane filter (Millipore Corp.) to

trap cells. After a 5.0 mL deionized water rinse, filters were digested

in 1.5 mL plastic centrifuge tubes with 0.2 mL HNOg at 80 °C for 30 min

to solubilize Hg. The volume was made to 1.0 mL with deionized water

and an aliquot of the digest was analyzed for total mercury [31]. Dry

weights of cell material were determined by filtration through 0.45 pm

filters, rinsing as before, followed by drying to constant weight at

105 °C. Duplicate bottles were run in these experiments, and duplicate

mercury determinations from each bottle were performed. Duplicate

bottles rehydrated with sterile water and analyzed as above served as controls.

2.5 Chromium Studies . Possible chromium volatilization from brewers yeast and orchard

leaves (SRMs 1569 and 1571), both certified for chromium, was investigated using a dual tube apparatus (fig. 1). The two SRMs were placed

in a 100 percent relative humidity chamber for three days, then 3 g

portions were added to 12 x 100 mm glass screw cap tubes which were connected to an identical tube (containing 3.0 mL deionized water) via

a thin glass tube through the caps. The SRMs were inoculated with

fungal spores taken from contaminated brewers yeast and orchard leaves

stored at high humidity. Aliquots of the deionized water were analyzed

at intervals for total chromium using atomic absorption spectrophotometry.

3. Results and Discussion

All SRMs were found to be sterile as received except for the urban

4 particulate SRM 1648 which contained more than 10 viable microorganisms

per gram (table 2). As a comparison, total counts of bacteria in soil

5 8

range from 10 per gram in poor soil to 10 per gram in rich soil [35].

Freeze dried urine (SRMs 2671 and 2672) was sterile in the dehydrated

form. Rehydration, however, with non-sterile water (though it may meet

the purity requirement of < 0.1 pg L

^ Hg set forth in the certificate)

resulted in microbial contamination of the material (fig. 2). NBS

5
Table 2. Numbers of viable microorganisms in several as received,

urban particulate (SRM 1648) vials. Numbers indicate

per g.

vial 1 4.00 X 10

'

2 3.45 X 10

3 3.20 X 10

'

4 2.05 X 10

Figure 2. Freeze dried urine rehydrated with sterile (left) and non- sterile (right) distilled water.

6
house distilled water used in the reconstitution of the urine did not

contain detectable mercury (< 0.1 pg L by cold vapor atomic absorption, see reference 31), but contained more than 500 viable microorganisms per mL. A sample of distilled water from a microbiology laboratory

at the University of Maryland contained 1600 viable microorganisms per

mL. At 1 °C the cells do not grow in the urine, however if stored at room temperature these cells utilized nutrients in the urine and grew

O well, numbering greater than 10 cells per mL in a few days (table 3).

Consequently, depending on the number and kinds of microorganisms in

the water used to reconstitute the urine, the SRM composition could be

significantly altered. For example, volatilization of mercury by the

cells (see reviews 2, 3) or binding of mercury to the cells [36] could

alter concentrations and homogeneity of distribution of mercury. To

date we have not found the microflora of the NBS house distilled water

capable of volatilizing elemental mercury (Hg°) from urine certified

for mercury. Microorganisms present in University of Maryland distilled water, however, volatilized Hg°. After three days 0.43 ng Hg° per

mL was found in the headspace of the container rehydrated with non- sterile water. A sterile control showed no volatilization of mercury.

No other volatile mercury containing metabolites (table 1) were

detected. The possibility exists that microorganisms in other

distilled water systems can also volatilize mercury.

Table 3. Numbers of viable microorganisms in freeze-dried urine

(SRMs 2671 and 2672) reconsitiuted with NBS distilled water.

Storage was at 1 °C or 22 °C. Numbers are organisms per mL.

Urine certified

Time of storage (1 °C)

1 day 6 days 12 days

Hg - elevated level 6.5 X 10

2

6.0 X 10

2

5.

X 10

2

00 X 10

2

- low level 5.8 X 10

2

6.9 X 10

2

3.7 X 10

2

3.6 X 10

2

F - elevated level 9.6 X

CM O 9.4 X 10

2

8.X 10

2

00 CM X 10

2

- low level 7.5 X 10

2

7.5 X 10

2

3.6 X 10

2

3.5 X 10

2

Time of storage (22 °C)

0 1 day 6 days

elevated level - 4.3 x 10

2 >10

low level - 6.5 x 10

2 >10

elevated level - 1.02 x IQ

2 >10

low level - 5.2 x 10

2 >10

7
Microorganisms which grew in the mercury-certified urine SRM bound

a significant percentage of the mercury. After six days 21.4 to 24.2

percent of the mercury was bound to cells (cells were removed from

solution by filtration) in the low level mercury in urine SRM, representating a concentration factor of 1514 to 3300 by the microorganisms,

which generally formed a sediment in the bottom of the container

(table 4). Cells occurring in other distilled water systems could have

a higher or lower binding affinity for mercury which could influence

how quickly significant alterations in SRM composition may occur.

Table 4. Binding of mercury to cells growing in urine certified for

mercury (low level )--SRM 2672. Values listed are means of

duplicate runs. Bottles 1 and 2 were rehydrated with NBS

distilled water, bottle 3 with University of Maryland

distilled water.

Bottle Hg bound

(nq/mg cells)

Hg bound

cS

Concentration

Factor

1 67.16 24.2 1774

2 59.39 21.4 1514

3 101.88 22.9 3300

River sediment (SRM 1645), urban particulate (SRM 1648), brewers

yeast (SRM 1569), orchard leaves (SRM 1571), and oyster tissue (SRM

1566) were examined over a range of humidities for susceptibility to

attack by airborne contaminants. Visual and microscopic examination

showed severe fungal deterioration of brewers yeast, orchard leaves,

and oyster tissue after exposure to outdoor air for short periods of

time on successive days (3.5 hr total) followed by incubation at room temperature and 90 or 100 percent relative humidity. Fungal growth on oyster tissue stored at 70 percent relative humidity was also noted.

Plate counts of these high-humidity exposed materials were not possible

due to tremendous fungal growth which literally consumed the SRMs.

Plate counts were possible only with urban particulate. Data (table 5)

indicate that storage at elevated relative humidity (90 to 100 percent)

induces growth of microorganisms in this SRM, however at lower humidities cell numbers are no greater than those observed for previously

unopened urban particulate (tables 2 and 5). Other SRMs exposed to

outdoor air then stored loosely capped at ambient laboratory relative

humidity (average 49 percent, range 33 to 73 percent) did not show

development of microbial populations visually, microscopically or by

plate counting after six months of storage.

8
Table 5. Growth of microorganisms in SRMs exposed to outdoor air

then stored at varying relative humidity for six months.

SRM Relative Humidity Stored

AMB

a 70 90 100

orchard leaves - - + +

oyster tissue - + + +

brewers yeast - + + +

river sediment - - -

urban particulate0

2 .64 x 10

4 4.9 x 10

3

1.05 x 10

5 1.62 x

(-) Values indicate no colonies on plates.

(+) Values indicate dense growth on plates (severe biodeterioration).

Range 33 to 73 percent, mean = 49 percent, NBS general purpose

laboratories

Urban particulate contains microorganisms initially, however,

counts after storage at ambient and 70 percent relative humidity

were either the same or lower than previously unopened bottles

(see table 2). Values listed are numbers of viable microorganisms per g.

Although exposure to outdoor air and incubation at elevated humidities probably represent "worst" case conditions in SRM handling it was necessary to quickly identify conditions leading to SRM biodeterioration in this preliminary survey. Microorganisms present in outdoor air

can be taken inside by ventilation systems or open windows. Elevated

laboratory humidities may occur during warm, humid weather (especially

with the advent of the energy crisis and resultant higher thermostat

settings) or as a result of steam or hot water use (autoclaving, glass- ware washing) even in laboratories which are served by controlled humidity ventilation systems. Future studies to measure SRM water activity

after different exposure times at various humidities will be instructive in determining biodeterioration potential.

No volatile chromium production has been detected from contaminated brewers yeast or orchard leaves. These studies are continuing and

will be expanded to cover additional elements and SRMs listed in table 1.

4. Conclusions and Recommendations

Improper storage of certain NBS Standard Reference Materials will

result in severe microbiological deterioration. The relative humidity

9
of storage and contamination by outside microorganisms are the two most

critical factors in deterioration of many of the SRMs examined. Urban

particulate is already laden with microorganisms which will grow at

elevated relative humidity. Rehydration of freeze dried urine will

result in contamination and microbial growth unless the urine is

rehydrated aseptically.

Under proper handling procedures set forth in most of the SRM

certificates, microbial deterioration of SRMs should not occur. Expo- sure to airborne microbial contaminants and elevated humidity either

deliberately (during weighing or removal from bottle) or accidentally

(not closing the lid) could result in SRM biodeterioration. Microbial

cultures, obtained by air sampling in several laboratories, have been

isolated and are being tested for ability to cause deterioration of

selected SRMs. How long an SRM must be exposed to a given relative

humidity for deterioration is also the subject of current investigations.

At present, however, we can recommend certain certificate modifications. For example, freeze dried urine (SRM 2671 and 2672) should be

rehydrated with sterile, mercury-free water. If sterile water is not

available, the rehydrated SRM must be refrigerated immediately and

should be used promptly. We recommend that rehydrated urine be discarded after 24 hr [37], to ensure microbial contaminants will not grow

to any extent in the SRM.

Studies designed to examine certified element composition changes

due to biodeterioration in other on the shelf and proposed SRMs

(table 1) are underway. These and other results will be the subject of

subsequent reports.

This work was sponsored by the Office of Standard Reference

Materials, National Bureau of Standards. We thank Michelle Leff for

technical assistance.

Certain commercial equipment, instruments, and materials are identified in this paper in order to adequately specify the experimental

procedure. Such identification does not imply recommendation or endorsement by the National Bureau of Standards, nor does it imply

that these items are necessarily the best available for the purpose.

10
5.

REFERENCES

1. Christian, J. H. B. , Reduced Water Activity (chapter 4), in Microbial Ecology of Foods, Volume 1, Factors Affecting Life and Death

of Microorganisms, Academic Press, New York (1980).

2. Iverson, W. P. , and Brinckman, F. E. , Microbial Metabolism of

Heavy Metals (chapter 8), in Mitchell, R. (ed.), Water Pollution

Microbiology, John Wiley and Sons, New York (1978).

3. Summers, A. 0., and Silver, S. , Microbial Transformations of

Metals, Ann Rev. Microbiol. 32, 637 (1978).

4. Johnson, D. L. , Bacterial Reduction of Arsenate in Sea Water,

Nature 240, 44 (1972).

5. Cox, D. P. , and Alexander, M. , Production of Trimethylarsine Gas

from Various Arsenic Compounds by Three Sewage Fungi, Bull.

Environ. Contam. Toxicol. 9, 84 (1973).

6. Holm, T. R. , Anderson, M. A., Stanforth, R. R. , and Iverson, D. G.

,

The Influence of Adsorption on the Rates of Microbial Degradation

of Arsenic Species in Sediments, Limnol. Oceanogr. 25, 23 (1980).

7. McBride, B. C. , and Wolfe, R. S. , Biosynthesis of Dimethyl arsine

by Methanobacterium. Biochemistry 1_0, 4312 (1971).

8. Magos, L. , Tuffery, A. A., and Clarkson, T. W. , Volatilization of

Mercury by Bacteria, Brit. J. Industr. Med. 22, 294 (1964).

9. Stary, J. , Havlik, B. , Kratzev, K. , Prasilova, J. , and Hanusova, J.

,

Mercury Circulation in Aquatic Environment. Part 3: Mercury

Transformations in Sediments, Acta Hydrochim. Hydrobiol. 8, 29

(1969).

10. Jensen, S. , and Jernelov, A., Biological Methylation of Mercury

in Aquatic Organisms, Nature 223 , 753 (1969).

11. Berdicevsky, I., Shoyerman, H. , and Yannai , S. , Formation of

Methyl mercury in the Marine Sediment under in vitro Conditions,

Environ. Res. 20, 325 (1979).

12. Olson, B. H. , and Cooper, R. C. , In situ Methylation of Mercury in

Estuarine Sediment, Nature 252 , 682 (1974).

13.

Edwards, T. , and McBride, B. C. , Biosynthesis and Degradation of

Methylmercury in Human Faeces, Nature 253 , 462 (1975).

14.

Havlik, B. , Stary, J. , Prasilova, J. , Kratzer, K. , and Hanusova, J.

,

Mercury Circulation in Aquatic Environment. Part 2: Metabolism

of Methyl and Phenyl Mercury in Phytoplankton, Acta. Hydrochim.

Hydrobiol. 7, 401 (1979).

11
15. Akagi , H. , Mortimer, D. C. , and Miller, D. R. , Mercury Methylation

and Partition in Aquatic Systems, Bull. Environ. Contam. Toxicol.

23, 372 (1979).

16. Lebedeva, E. V. , and Lyalikova, N. N. , Reduction of Crocoite by

Pseudomonas chromatophi la sp. nov. , Microbiology 48, 405 (1979).

17. Bopp, L. H. , Ehrlich, H. L. , Friello, D. A., and Chakrabarty, A. M.

,

Plasmid Coded Chromate Resistance in a Hudson River Isolate,

Abstracts of the Annual Meeting of the Am. Soc. Microbiol., p. 138

(1979).

18. Banwart, W. L. , and Bremner, J. M. , Formation of Volatile Sulfur

Compounds by Microbial Decomposition of Sulfur-Containing Amino

Acids in Soils, Soil Biol. Biochem. 7, 359 (1975).

19. Banwart, W. L. , and Bremner, J. M. , Identification of Sulfur Gases

Evolved from Animal Manures, J. Environ. Qual. 4, 363 (1975).

20. Aneja, V. P. , Overton, J. H. , Cupitt, L. T. , Durham, J. L. , and

Wilson, W. E. , Carbon Disulfide and Carbonyl Sulfide from Biogenic

Sources and their Contributions to the Global Sulfur Cycle, Nature

282 , 493 (1979).

21. Zinder, S. H. , and Brock, T. D. , Microbial Transformations of

Sulfur in the Environment (chapter 10), in Nriagu, J. 0. ed.

,

Sulfur in the Environment. Part II: Ecological Impacts,

John Wiley and Sons, New York (1978).

22. Reamer, D. C. , and Zoller, W. H. , Selenium Biomethylation Products

from Soil and Sewage Sludge, Science 208 , 500 (1980).

23. Chau, Y. K. , Wong, P. T. S. , Silverberg, B. A., Luxon, P. L. , and

Bengert, G. A. , Methylation of Selenium in the Aquatic Environment,

Science JJ32, 1130 (1976).

24. Harrison, R. M. , and Laxen, D. P. H. , Natural Source of Tetraalkyllead in Air, Nature 275 , 738 (1978).

25. Wong, P. T. S. , Chau, Y. K. , and Luxon, P. L. , Methylation of Lead

in the Environment, Nature 253 , 263 (1975).

26. Jackson, J. A., Blair, W. R. , Brinckman, F. E. , and Iverson, W. P.

,

Gas Chromatographic Speciation of Methyl stannanes in the Chesapeake

Bay Using Purge and Trap Sampling with a Tin-Selective Detector,

submitted for publication.

27. Braman, R. S. and Tompkins, M. A., Separation and Determination

of Nanogram Amounts of Inorganic Tin and Methyl tin Compounds in

the Environment, Anal. Chem. 51, 12 (1979).

12
28. Hodge, V. F. , Seidel, S. L. , and Goldberg, E. D. , Determination of

Tin(IV) and Organotin Compounds in Natural Waters, Coastal Sediments, and Macro Algae by Atomic Absorption Spectrophotometry,

Anal. Chem. 51, 1256 (1979).

29. Rodina, A. G. , Methods in Aquatic Microbiology, University Park

Press, Baltimore, MD (1972).

30. Ferrer, E. B. , Stapert, E. M. , and Sokolski, W. T. , A Medium for

Improved Recovery of Bacteria from Water, Can. J. Microbiol. 9,

420 (1963).

31. U. S. Environmental Protection Agency, Methods for Chemical Analysis of Water and Wastes, EPA-625-/6-74-003 , Washington, DC (1974).

32. Blair, W. , Iverson, W. P. , and Brinckman, F. E. , Application of a Gas Chromatograph--Atomic Absorption Detection System to a Survey

of Mercury Transformations by Chesapeake Bay Microorganisms,

Chemosphere. 4, 167 (1974).

33. Brinckman, F. E. , Iverson, W. P. , and Blair, W. , Approaches to the

Study of Microbial Transformations of Metals, in Sharply, J. M.

,

and Kaplan, A. M. (eds.), Proc. 3rd Internat. Biodegradation Symp.

Applied Science Publishers, London (1976).

34. Dean, J. A., Lange's Handbook of Chemistry, 12th ed. , McGraw-Hill,

New York (1979).

35. Ehrlich, H. L. , Geomicrobiology, Marcel Dekker, New York (1981).

36. Nichols, D. H. , Mercury Binding to a Yeast, J. Gen. Microbiol. 30,

429 (1973).

37. Britt, E. M. , Needham, G. M. , Balows, A., and Truant, J. P. , Processing of Specimens and the Initiation of Primary Cultures and

Smears (chapter 4), in Blair, J. E. , Lennette, E. H. , and

Truant, J. P. (eds.), Manual of Clinical Microbiology, American

Society for Microbiology, Bethesda, MD (1970).

13
NBS-1 14A (rev. 2 -eci U.S. DEPT. OF COMM. BIBLIOGRAPHIC DATA

1. PUBLICATION OR

REPORT NO.

2. Performing Organ. Report No. 3. Publication Date

SHEET (See instructions) NBSIR 81-2246 April 1981

4.

TITLE AND SUBTITLE

BIODETERIORATION OF STANDARD REFERENCE MATERIALS

5. AUTHOR(S)

G. J. Olson, W. P. Iverson, F. E. Brinckman

6. PERFORMING ORGANIZATION (If joint or other than NBS. see instructions) J. Contract/ Grant No. national bureau of standards

DEPARTMENT 0 F COMMERCE • . Type of Report & Period Covered

WASHINGTON, D.C. 20234

9.

SPONSORING ORGANIZATION NAME AND COMPLETE ADDRESS (Street. City. State. ZIP)

Office of Standard Reference Materials

National Bureau of Standards

Department of Commerce

Washington, D. C. 20234

10.

SUPPLEMENTARY NOTES

Q

-

] Document describes a computer program; SF-185, FIPS Software Summary, is attached.

11.

ABSTRACT (A 200-word or less factual summary of most significant information. If document includes a significant

bi bliography or literature survey, mention it here) Several National Bureau of Standards Standard Reference Materials have been

examined for susceptibility to biodeterioration. Several of these materials were

attacked by bacteria and fungi when stored at elevated humidity after exposure to

outdoor air. Some SRMs underwent deterioration after following certificate instructions for handling. Suggestions for some certificate revisions are made.

12. KEY WORDS (Six to twelve entries; alphabetical order; capitalize only proper names; and separate key words by semicolons)

bacteria; biodeterioration; biotransformations; deterioration of materials;

element volatilization; fungi; metals; microorganisms; Standard Reference Materials.

13. AVAILABILITY 14. NO. OF

PRINTED PAGES

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