Evaluation of bacterial contamination and

control methods in soluble metal working

fluids.

Geneviève Marchand, Jacques Lavoie, Louise Racine, Nancy Lacombe, Yves

Éric Bélanger ASP métal électrique
Christian Lemelin GLD entreprises
Jean Desroches / Magnus

This is an electronic version of an article published in

Journal of Occupational and Environmental Hygiene,
Volume 7, Issue 6 June 2010 , pages 358 – 366 DOI:
10.1080/15459621003741631
Journal of Occupational and Environmental Hygieneis
available online at: http://oeh.informaworld.
com/soeh/content~db=all~content=a921030176~frm=titleli
nk
Evaluation of bacterial contamination and control methods in soluble metal working fluids

Geneviève Marchand, IRSST

Jacques Lavoie, IRSST
Louise Racine, IRSST
Nancy Lacombe IRSST
Yves Cloutier IRSST
Éric Bélanger ASP métal électrique
Christian Lemelin GLD entreprises
Jean Desroches / Magnus

Correspondence and reprint requests to

Geneviève Marchand, IRSST

Institut de recherche Robert-Sauvé en santé et en sécurité du travail (IRSST)
505, boul. De Maisonneuve Ouest
Montréal (Québec), H3A 3C2
Phone: (514) 288-1551 #316
Fax: (514) 288-6097
e-mail: marchand.genevieve@irsst.qc.ca

Key words: Metal working fluids, bacteria, contamination, control

ABSTRACT
Introduction: In the United States, 1.2 million workers are exposed to metalworking fluids.
During operations aerosols are produced, and airborne contaminants can be inhaled. Biocides are
used to control the bacterial content of metalworking fluids. They can create health-related
problems and their efficiency remains to be proven. Objectives: The objectives of this project
were to verify whether rigorous cleaning according to a standard protocol could reduce microbial
contamination and whether the use of biocides with different spectra could reduce the bacterial
population. Method: Four similar machine producing similar components were evaluated. A
specific treatment was applied to each machine. The machine used as a control was thoroughly
cleaned at the beginning, did not undergo any major cleaning afterward and was operated
without the use of any biocide. A major cleaning is a protocol described and recommended by
the fluid manufacturer. It was performed at the beginning on each of the three other machines.
Two of these machines were subsequently treated with biocides weekly. The fluid samples from
the four lathes were collected weekly during a six-month period. Total bacterial and cultivable
Gram-negative were analyzed for each sample. Results: Major cleaning of the machines (120-4)
did not significantly reduce the concentration of bacteria contained in the cutting fluids when
compared to the control machine (120-3) which did not undergo major cleaning. The
concentrations of total bacteria were in the 106 CFU /mL range for these two lathes. However, a
reduction in the total number of fluid changes was observed for this machine. Bacterial flora in
the cutting fluids was significantly controlled through the use of biocides. The concentrations of
bacteria were in the 103-105 CFU/mL range for the lathes with the use of biocides. Conclusion:
Since thorough cleaning is insufficient and biocides are recognized as being responsible for some
worker health problems, other avenues for controlling the bacterial flora in the cutting fluids
should be evaluated in order to reduce worker exposure to their bacterial contaminants.

INTRODUCTION
Metalworking fluids (MWFs) have been used in the metal cutting industry for 200 years.1 In the
United States, 1.2 million workers are exposed to MWFs.2 Microbial contamination of these
fluids has been reported in several studies.3-10 This is a major concern for the manufacturing
industry because this contamination can lead to losses in the intrinsic quality of the cutting
fluids.2-11 Microbial growth can also result in or lead to the accumulation of toxins and allergens
in the fluids. 12 During machining, aerosols are generated and the contaminants contained in the
fluids can therefore be found in the air and inhaled by the workers. A study by Wang13
demonstrated that the cutting fluids used in these industries can be contaminated with very high
concentrations of fine particulates and contain microbial compounds and organisms. Many health
problems involving direct exposure to cutting fluids have been reported: contact dermatitis, skin
eruptions and sinusitis are some examples. 2, 14-19 An increase in respiratory health problems,
with extrinsic allergic alveolitis as an example, has also been observed for several years. 20-23
High bacterial concentrations usually found in MWFs create a health hazard for workers exposed
to aerosols originating from these fluids. Since health problems have been linked to the use of
cutting fluids, researchers have attempted to identify and quantify the microbial load in order to
establish dose and effect relationships for the health of exposed workers. 5,6,8,10 Cleaning and
fluid management through the use of biocides are two methods that could be used to control
bacterial concentration in MWFs, even if one study concluded that the cleaning protocols
currently used in industry are ineffective and that very high levels of bacterial contamination,
including some pathogens, have been found in the fluids, despite rigorous fluid management
protocols and systems. 5 NIOSH recommends the complete emptying of the machining tools or
lathes, and states that special attention should be paid to the biofilms that could be found on the
inner surfaces of pipes and at the bottom of fluid reservoirs. These biofilms are believed to
contribute to the rapid contamination of any new fluids if they are not destroyed during
cleaning. 5 One problem with the use of biocides is that most of them can release formaldehyde.
Formaldehyde is a well-known irritant for the respiratory tract and a known carcinogenic
agent. 2,24 This research project studied two methods of controlling bacterial proliferation in the
fluids, namely the use of thorough cleaning of the machinery and the systematic application of
biocides. Its prime objective was to compare the concentration of bacteria found in MWFs in
relation to the type of cleaning performed (basic or major). The second objective was to compare
the concentration of bacteria measured in the cutting fluids in relation to the use of a biocide
utilization protocol (alternate or single).

METHODOLOGY

Machinery
Four fundamentally similar lathes from one plant were selected. They were all manufactured by
Mori Seiki and were a mix of models SL250 and SL303 (Mori Seiki Ltd, Nagoya City, Japan).
Each lathe was equipped with a separate MWF reservoir with a maximum capacity of 125 liters.
No central tank system was used in this plant. All the lathes were used to manufacture similar
cast iron components with identical cutting tools. They were operated 24 hours a day and 7 days
a week. The cutting fluid used on all the machines was a soluble oil-based product purchased
from Magnus (Magnus, Boucherville, Canada).

Treatments
A different treatment was assigned to each lathe at the beginning of the study. The four different
treatments are presented in Table I. Basic cleaning consisted of removing the fluid, metal
deposits and other waste, and feeding the machine with new fluid. It was not a thorough cleaning
and consisted of the cleaning of the parts that were accessible. Major cleaning requires the
rigorous application of a protocol described by the fluid manufacturer, which requires
approximately ten hours of work for each lathe. This type of cleaning requires that all accessible
parts of the lathe, such as the reservoir, be meticulously cleaned first, and that a product called a
“machine cleaner” (Magnus, Boucherville, Canada) be circulated afterward through the lathe to
help loosen deposits and biofilms in the hard-to-reach parts. This operation must be repeated
until the cleaning fluid is free of metal debris and deposits. The machine cleaner is then removed
and replaced with new MWF. This cutting fluid is diluted to a concentration of 8% (8 parts oil in
92 parts water) according to the manufacturer's recommendations.
TABLE I. Summary of the treatments planned for each machine
Machine Type of cleaning Biocides
120-3 Basic cleaning Without biocide
120-4 Major cleaning Without biocide
125-3 Major cleaning Magnicide biocide
125-4 Major cleaning Grotan BK and Magnicide
biocides, alternately

Because of production constraints, unplanned cleaning maintenance was performed on the lathes
during the study. When routine cleaning was needed for a lathe, basic cleaning was performed
but the MWFs were replaced only when necessary for the machine's operation based on
empirical criteria. Table II provides a description of the unplanned maintenances that were
required for the entire duration of the project.
TABLE II. Summary of the unplanned but required treatments
Machine Cleaning Fluid replacement First maintenance
n n week of project
120-3 3 3 5
120-4 3 2 6
125-3 2 0 8
125-4 3 0 5
n: number

Biocides were used only on lathes 125-3 and 125-4. Magnicide (Magnus, Boucherville, Canada)
is a chlorine-based biocide (5-chloro-2-methyl-4-isothiazolin-3-one) (2.63% a.i.) + (2-methyl-4-
isothiazolin-3-one (0.18% a.i.), while Grotan BK is a formaldehyde releaser (1,3,5-Tris (2-
hexahyethyl)hexahydro-1,3,5-triazine (85% a.i.) (Magnus,Boucherville,Canada). Biocides were
added systematically every Wednesday, starting with the fourth week of the project. Magnicide
was used every week on lathe 125-3, but both biocides, Magnicide and Grotan BK, were used in
turn every week on lathe 125-4. This protocol was chosen by the operators as recommended by
the manufacturers.
The refractive index of the fluid for all the lathes was monitored daily, directly in the plant, as an
indicator of the state of the MWF and of the cleaning requirements. Cleaning frequency became
a function of this index and varied from one machine to another. This measurement was
performed with a model RHB-3 refractometer with a scale from 0 to 32 RI (refractive index)
(Westover Scientific Inc., WA, USA) (results not shown). Maintenance technicians used this
index to decide when and what type of maintenance was necessary for proper operation of their
machine. Odor, loss of viscosity, and visible deposits in the collection tank were other criteria
used by the technicians to decide whether maintenance was necessary.
They also had to take into account the quality of production and the impact of maintenance on
performance.

Collection of Samples
Fluid samples were collected from each machine every Monday for six months. They were
collected at the same location in the fluid collection tank under the machines using a 25-mL
sterile pipette (Fisherbrand, Ontario, Canada) and transferred to 50-mL Corning tubes (Corning,
NY, USA). All the samples were shipped on ice to the laboratory and analyzed within 24 hours
as recommended by the Standard methods for the examination of water and wastewater.25
Precautions were taken to prevent freezing of the samples during shipping.
Air sampling of endotoxins was done at the end of the project to evaluate the increase in
endotoxin concentrations in the ambient air after the addition of the biocide. Sampling was done
using a 37mm cassette with fiber glass membrane from Gelman Sciences (Pall, Québec,
Canada). The flow rate was 2 liters/minute for a period of 180 minutes.

Bacterial Analysis and Identification

All samples were processed according to IRSST methods 264-3 and 341-1. 26-27 They were
mixed using a multi-tube vortexer mixer (VWR Scientific Products, Ontario, Canada) for at least
30 minutes.
The culture media used were trypticase soya agar (TSA) and MacConkey agar (Quélab,
Montréal, Canada). The incubation temperature was 37oC for a period of 48 hours. Spreads were
performed in triplicate, with a dilution factor ranging from 10-1 to 10-6, depending on the
samples. All dilutions were done with peptone water (fluid A).
After the counting of each sample, for which the number of colonies per plate was between 30
and 300, the dilution factor was used to calculate the number of colony forming units per
milliliter of fluid (CFU/mL). The three replicates were then averaged to calculate the
concentration of cultivable bacteria in CFU/mL for each sample.
Depending on the samples, from 1 to 16 colonies with a different appearance were subcultured in
a pure culture to allow identification. Subcultures were done on trypticase soya agars with or
without 5% sheep blood (Quélab, Montréal Canada). The bacteria were identified either by
comparing their fatty acid profiles obtained by gas phase chromatography with the database of
the Sherlock identification system from MIDI company (Newark, USA) , or from their
biochemical characteristics obtained using the Microscan system of Siemens (Illinois, USA).

Endotoxin Analysis
At the end of the final week of the project, six endotoxin samples were collected before and after
the addition of biocide. The samples were collected close to the opening of the lathes during
normal use. All samples were kept in a desiccation jar until their analysis. The analysis of the
endotoxin samples was performed using IRSST method 332. 28

Statistical Analyses
Data analyses have shown that after a log transformation, the distribution was very close to a
normal distribution. Medians were reported since large variations were observed between weeks.
An ANOVA analysis with Dunnett multiple comparison tests was used for comparison to the
control. Lathe 120-3 was used as the control because it was subjected to the maintenance
treatment most commonly carried out in this plant. All statistical tests were performed with a p
value of 0.05. NCSS 2004 software 29 was used for these statistical analyses.

RESULTS

Maintenance
All the lathes were cleaned two or three times. Lathes 120-3 and 120-4 each needed more than 2
fluid replacements, and no fluid replacement was necessary for lathes 125-3 and 125-4.

Bacterial Count
Bacterial concentrations found in the MWFs were below 10 CFU/mL for the 4 lathes at the
beginning of the study and after the fluid change. Figures 1 to 4 show all the bacterial
concentrations in the 4 lathes for the 6 consecutive months of the project. The INRS-France
recommended limit of 106 CFU/mL has been drawn on the graphic as a reference. 30 That value
was exceeded many times during the study for the lathes with no added biocide, but less often for
the other ones with the addition of biocides. Tables III and IV show the overall cultivable
bacteria concentrations and their geometric means obtained from growth on soya trypticase agar
(TSA) and MacConkey agar for the 6-month period.
TABLE III. Geometric means and median of the concentrations of total cultivable bacteria on
TSA in the cutting fluids of the four lathes
Machine n Median GM GSD
CFU/mL CFU/mL
120-3 26 2.7E+06 1.9E +05 3.3E +2
120-4 26 1.0E+06 3.3E +05 1.4E +2
125-3 23 7.5E+03 3.1E +03 1.3E +3
125-4 22 6.6E+04 6.9E +03 5.5E +2
GM: geometric mean; GSD: geometric standard deviation; n: number

TABLE IV. Geometric means and median of the concentrations of cultivable Gram-negative
bacteria on MacConkey agar in the cutting fluids of the four lathes
Machine n Median GM GSD
CFU/mL CFU/mL
120-3 26 1.6E+06 1.2E +5 2.5E +2
120-4 26 5.8E+05 1.9E +5 1.1E +2
125-3 22 2.7E+03 1.9E +3 1.1E +2
125-4 20 3.4E+04 3.1E +3 4.8E +2
GM: geometric mean; GSD: geometric standard deviation; n: number
 TSA
Mac Conkey
CFU/ml
100000000
fluid
10000000 change

1000000

100000

10000

1000
fluid
fluid
100 change
change
10

1
0 2 4 6 8 10 12 14 16 18 20 22 24 26

weeks

FIGURE 1. Weekly concentrations of cultivable bacteria found in the metalworking fluid of lathe 120-3 (control
machine) when cultivated on TSA and MacConkey media. The straight line indicates the recommended
concentration of 106 CFU/mL.

TSA
CFU/ml MacConkey

100000000

10000000

1000000

100000

10000

1000
fluid
100 cleaining change fluid
change
10

1
0 2 4 6 8 10 12 14 16 18 20 22 24 26

weeks

FIGURE 2. Weekly concentrations of cultivable bacteria found in the metalworking fluid of lathe 120-4 (major
cleaning) when cultivated on TSA and MacConkey media. The straight line indicates the recommended
concentration of 106 CFU/mL.
 TSA
CFU/m L Mac Conkey

100000000

10000000

1000000

100000

10000

1000

100
cleaning
10 cleaning

1
0 2 4 6 8 10 12 14 16 18 20 22 24 26

weeks

FIGURE 3. Weekly concentrations of cultivable bacteria found in the metalworking fluid of lathe 125-3 (major
cleaning and Magnicide biocide) when cultivated on TSA and MacConkey media. The straight line indicates the
recommended concentration of 106 CFU/mL.

CFU/mL TSA
Mac Conkey
100000000

10000000

1000000

100000

10000

1000

100 cleaning cleaning

cleaning
10

1
0 2 4 6 8 10 12 14 16 18 20 22 24 26

weeks

FIGURE 4. Weekly concentrations of cultivable bacteria found in the metalworking fluid of lathe 125-4 (major
cleaning and alternate Magnicide and Grotan BK biocide) when cultivated on TSA and MacConkey media. The
straight line indicates the recommended concentration of 106 CFU/mL.
The largest average concentrations were both observed for control lathe 120-3: 2,700,000
CFU/mL for total cultivable bacteria and 1,600,000 CFU/mL for Gram-negative bacteria. The
lowest average concentrations were measured on lathe 125-3, namely 7,500 CFU/mL for total
cultivable bacteria and 2,700 CFU/mL for Gram-negative bacterial flora.
Concentrations for each lathe were distributed normally once log transformed. A two-factor
ANOVA with no interaction between the factor and Dunnett multiple comparison statistical tests
were applied to the data. A p value of 0.05 was used for the tests. The results showed that both
the total cultivable bacteria and Gram-negative bacteria concentrations were significantly
different between the lathes and between weeks for each lathe. Table V gives the results for the
ANOVA test. This test allows the conclusion that even with the variation observed from week to
week, there was a difference between the treatments. The Dunnett test was applied to identify the
differences between the control lathe (120-3) and the other ones. This test demonstrated that
concentrations between lathe 125-3 and the control were significantly different. No significant
differences were observed between concentrations of cultivable total bacteria and Gram-negative
bacteria for both lathes using biocide bacterial identification.
TABLE V. ANOVA results
a) Total cultivable bacteria

Source Degree Sum of Mean Prob Power

Freedom squares square F-Ratio Level (Alpha=0.05)

A: treatment 3 49.82 16.61 4.68 0.004947* 0.877785

B: week 25 286.36 11.45 3.23 0.000067* 0.999768
S 68 241.17 3.547
Total (Adjusted) 96 602.39
Total 97

* Term significant at alpha = 0.05

b) Gram-negative cultivable bacteria

Source Degree Sum of Mean Prob Power

Freedom squares square F-Ratio Level (Alpha=0.05)

A: treatment 3 50.92 16.97 5.53 0.001891* 0.928302

B: week 25 276.03 11.04 3.60 0.000017* 0.999944
S 66 202.48 3.068
Total (Adjusted) 94 551.02
Total 95

* Term significant at alpha = 0.05

Figure 5 presents the different genera of bacteria identified in the fluids of all the lathes during
the 6-month period. The types of bacteria encountered were mainly saprophytic organisms found
in the environment. Both Gram-negative and Gram-positive bacteria were found in our samples.
Pseudomonas was most often observed in all the lathes. In fact, 73 of the 79 bacterial growth
samples contained bacteria of this genus (92% of the samples). Despite this widespread
occurrence, very diverse bacterial flora were observed in all the fluids. Genus Citrobacter,
Staphylococcus, Alcaligenes, Acinetobacter, Bacillus, Shewenella and Brevundimonas were
identified in more than one machine. The control lathe contained the largest diversity of bacterial
species (Table VI). Lathe 125-4, where the two biocides were added in turn, showed the least
diversity in bacterial flora.
week

Lathes Genus 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26
Acinetobacter
Alcaligènes
Bacillus
Brevundimonas
120-3 Citrobacter
Corynebacterium
Pseudomonas
Shewanella
Staphyloccus
Acinetobacter
Alcaligènes
Bacillus
Brevundimonas
120-4 Citrobacter
Corynebacterium
Pseudomonas
Shewanella
Staphyloccus
Acinetobacter
Alcaligènes
Bacillus
Brevundimonas
125-3 Citrobacter
Corynebacterium
Pseudomonas
Shewanella
Staphyloccus
Acinetobacter
Alcaligènes
Bacillus
Brevundimonas
125-4 Citrobacter
Corynebacterium
Pseudomonas
Shewanella
Staphyloccus

: This genus was present in the cutting fluid for that week, without consideration of the concentrations.
FIGURE 5. Variation in bacterial diversity in the cutting fluid from week to week
Table VI: Bacterial diversity in the fluid samples obtained by growth on TSA
120-3 120-4 125-3 125-4
Acinetobacter lwofii Acinetobacter sp Alcaligenes sp Brevundimonas sp
Alcaligenes faecalis Alcaligenes spp Bacillus cereus Micrococcus luteus
Bacillus megaterium Bacillus licheniform Brevundimonas sp Pseudomonas
Bacillus sp Citrobacter freundii Citrobacter sp pseudoalcaligenes
Citrobacter freundii Escherichia coli Pseudomonas balearia Pseudomonas spp
Flavobacterium breve Providencia sp Pseudomonas spp Staphylococcus sp
Gordona sp Pseudomonas spp Pseudomonas stutzeri
Klebsiella oxytoca Pseudomonas stutzeri Staphylococcus sp
Micrococcus sp Shewenella putrefaciens Yersinia sp
Pseudomonas Staphylococcus xylosus
pseudoalcaligenes
Pseudomonas spp
Corynebacterium sp
Pseudomonas stutzeri
Shewenella putrefaciens
Staphylococcus sp
Staphylococcus warneri

Endotoxin Concentrations in the Air

The endotoxin concentrations measured in the ambient air before and after the addition of
biocides were 1.2±0.1 EU/m3 of air and 9.6±11.9 EU/m3 respectively.

DISCUSSION
The results show that no fluid replacement was done on lathe 125-3 and lathe 125-4 where the
major cleaning protocol was applied and biocides were used, even if the frequency of unplanned
maintenance operations varied from one machine to the next, because of production constraints.
Less fluid replacement was done when a major cleaning protocol was used on the lathe even
when no biocides were used.
The first cleaning operations took place after five weeks for all the machines. They were all
cleaned three times, except for lathe 125-3, which was cleaned only twice. Lathe 120-4, to which
a major cleaning protocol was applied, had its fluid changed at a later time than lathe 120-3,
which underwent a basic cleaning. More research is needed in a more controlled environment to
conclude that this is a more efficient way of cleaning.
The use of TSA and MacConkey agars as growing media does not allow complete identification
of the entire bacterial flora present in MWF. Nevertheless, it allows a major part of the bacterial
flora to be compared, and leads to sufficient confidence about the conclusions of this study. The
bacterial concentrations found were of the same order of magnitude as the ones reported by
Simpson 31 and Veillette et al. 5 The concentrations of cultivable bacteria on TSA and
MacConkey were directly affected by the use of biocides. Our study clearly demonstrated a
reduction in the average concentrations between both lathes 125-3 and 125-4 and the control
lathe when biocides were used. This is contrary to the conclusions of Dilger et al.3 who reported
that biocides do not reduce the capacity of the bacteria to multiply in the fluids. When a Dunnett
multiple comparison statistical test was performed to confirm this observation, a significant
reduction was demonstrated for lathe 125-3. Conclusions similar to ours were arrived at by
Linnainmaa et al. 7 for the use of triazine as biocide.
This study shows that the major cleaning protocol did not significantly decrease the average
concentrations of cultivable bacteria. Similar conclusions were stated by Veillette et al. 5
Comparison between concentrations of cultivable bacteria for the two lathes to which biocide
was added did not show a significant difference between the use of one biocide or two biocides
alternately. The alternate addition of the two biocides, namely Magnicide and Grotan BK, did
not yield average concentrations of bacteria that were below those obtained when a single
biocide was used. In fact, even if no statistical difference could be observed between the two
lathes using biocide, the concentrations found when two biocides were used appears to be
slightly higher than concentrations when only one biocide was used. The lathes where the two
biocides were used showed a smaller number of genera but higher concentrations of bacteria.
This could be explained by a least effective control of Grotan BK compared to Magnicide for the
genus Pseudomonas, which is the dominant genus in the lathes.
For all the lathes, important reductions in bacterial contents were observed after cleaning the
machine or changing the fluid. However, a rapid re-colonization was observed. This was also
reported by Veillette et al. 5 who noted that a re-colonization occurred after only 12 hours. When
no biocides were used, the concentrations more often exceeded the recommended 106 CFU/mL.
In the lathes where biocides were used, a reduction in the concentrations could also be observed
for each intervention, such as the addition of biocide or cleaning. In those lathes, concentrations
above 106 CFU/mL were less often observed, and they were most often observed towards the end
of the project. This could be an indication that even with the use of biocide, the life of the
MWFs is not infinite. Nevertheless, the use of biocide showed a clear improvement in MWF
lifetime compared to the 6 weeks observed for the control (120-3) and the 16 weeks observed for
the lathe with the extensive cleaning at week one (120-4).
Bacterial diversity was not uniform and varied with time and with the machine. This diversity
can be affected by the maintenance treatments and can also depend on competition for the
substrate found in such an environment. Figure 5 shows that bacterial diversity was greater in the
fluid with no biocide added. The lowest bacterial diversity was observed for lathe 125-4, when
two biocides were used alternately even if the measured concentrations were not significantly
lower than the control (lathe 125-3). The alternate use of biocides seems to have limited the
growth of some of the bacterial flora that were allowed to grow on the lathe using only one
biocide. This data suggests that some types of bacteria have more difficulty growing in an
alternating regimen of biocides. Other combinations of biocides should be investigated for
comparison. The susceptibility of the microbial groups should also be investigated in order to
establish the best combination of biocides. An alternate use of these two biocides may control
bacterial diversity better but not necessarily reduce the average concentration. As in other
studies, 3, 32-33 the genus Pseudomonas was observed as dominant in most of our fluid samples no
matter what treatment was used. Similar conclusions were reported by Rossmoore et al. 33 and
Dilger et al. 3 in cultivable analysis where neither rigorous cleaning nor the use of biocides seems
to have had an impact on the presence of this genus of bacterium in cutting fluids, other than to
reduce its concentrations when biocides are used.
There are no occupational exposure limits for endotoxins in the ambient air. A slight increase in
endotoxin concentration was observed after the addition of biocides. This slight increase in
endotoxin concentration could be the worst-case scenario. It might be caused by the cellular lysis
of the bacteria present in the cutting fluid, which liberated the endotoxins. The endotoxin
concentrations in the ambient air remained relatively low compared to other occupational
settings. The results have shown that the level of endotoxins measured in this study were all
below the average of 26 EU/m³ observed for 50 outdoor air samples, and well below the levels
as high as 5000 EU/m3 measured in a project on cotton mills. 34

CONCLUSION
Major cleaning of the machines did not significantly reduce the overall average concentration of
bacteria contained in the cutting fluids over a six-month period. MWF replacement was reduced
for the machines that underwent a major cleaning protocol and the use of biocides. The greatest
reductions in bacterial concentration were also observed for these machines. Cultivable bacterial
flora evaluated on TSA and MacConkey were shown to be controlled through the use of
biocides.
Biocide use clearly extends the lifetime of MWFs. This might lead to increased use by the
industry. However, because of health problems related to biocide use, other avenues for
controlling the bacterial flora present in cutting fluids, such as filtration or ozone treatment,
should be investigated. If biocides are used for microbiological control, proper fluid management
practices should be implemented with minimal utilization of these products.

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