The association between the microbial contaminants in metalworking

fluids with the FeNO levels among machining industry workers

Nurul Maizura, Hashim1; *Zailina, Hashim1; Rukman Awang, Hamat2;
Hayati, Kadir3.
1

Department of Environmental and Occupational Health, Faculty of Medicine and

Health Sciences, Universiti Putra Malaysia,UPM, Serdang, Selangor, Malaysia.
2

Department of Medical Microbiology and Parasitology, Faculty of Medicine and

Health Sciences, Universiti Putra Malaysia,UPM, Serdang, Selangor, Malaysia
3

Department of Community Health, Faculty of Medicine and Health Sciences,

Universiti Putra Malaysia,UPM, Serdang, Selangor, Malaysia.
*Corresponding Author address and e-mail:
Department of Environmental and Occupational Health, Faculty of Medicine and Health
Sciences, Universiti Putra Malaysia,UPM, Serdang, Selangor, Malaysia,
zailina@upm.edu.my

The association between the microbial contaminants in metalworking

fluids with the FeNO levels among machining industry workers
Abstract
Introduction: Water based metalworking fluids (MWFs) are commonly used in
machining industries and are excellent media for microorganism growth. The aim
of this study was to determine the relationship between the airway inflammation
as indicated by the fractional exhaled nitric oxide (FeNO) with the microbial
contaminants in MWF aerosol and the workers reported health symptoms.
Methodology: This cross sectional study was carried out on 138 machining
workers exposed to MWF. The workers FeNO were measured using NIOXMINO instrumentation. The microbial assessments were carried out on the MWF
bulk samples and the aerosol in the work environment. DUO SAS SUPER
360TM sampler was used to sample the bacteria and fungus in the air. The data
were analysed using the SPSS Version 22.0.
Result and Discussion: Findings showed significantly different FeNO levels in
workers from different job sections (p=0.01). From the multivariate statistics,
there were also significant relationships between the high FeNO with their
closeness to the machines (p=0.03), more machines (p=0.02), high environmental
bacteria colonies (p=0.04), longer employment years (p=<0.001) and frequent
reporting of respiratory symptom such as cough (p=0.03).
Conclusion: Risk factors from the workplace such as the number of machines and
their closeness to them and high environmental bacteria colonies and longer
employment years had significant relationships with the airway inflammation
(FeNO). Exposure to MWF also resulted in significantly frequent cough.
Keywords: Metalworking fluids; airway inflammation and fractional exhaled
nitric oxide (FeNO)

1.0 Introduction
Metalworking fluids are used as lubricant, coolant, cutting fluids, machining
fluids or hydraulic oil (Ross, 2004). Metalworking fluids can helps in preventing metal
corrosion and act as a metal scrap remover while undergoes machining process
(NIOSH. 1998). It comprises of four types; straight oil, soluble oil, synthetic and semisynthetic metalworking fluids (NIOSH. 1998). The usage of metalworking fluids
potentially produced contaminants such as bacteria and fungi, and also soluble metal
ions such as chromium, nickel and cobalt in alloyed steel and hard metal (Suuronen et
al, 2007). These fluids are extensively used as industrial lubricants to facilitate the
lubricating and cooling of metalworking operations by reducing friction at tool-work
piece interfaces, thus reducing heat. MWF are also used to wash away waste metal
debris, subsequently improving the machining performance and as a result, prolonging
the life of the cutting tool.
The National Occupational Exposure Survey in 1981-1982 of National Institute
for Occupational Safety and Health (NIOSH) of United Stated reported that potentially
workers exposed to metalworking fluids was estimated around 1.2 million (Wenhai et.

al, 2006), while the global demand of metalworking fluids for 2010 was 2.2 million tons
and there was increased consumption of metalworking fluids in Asia reported from
2007 until 2010 with 3.3% annual rate (Byers, 2011). Usually high capacity usage of
metalworking fluids was related to automotive, commercial aircraft and heavy
machinery industries (Cohen et.al, 2006). During the application of metalworking
fluids, aerosol of metalworking fluids consists of metal, chemical residues or physical
contaminants such as chips and fines will dispersed into the surrounding during the
operation (NIOSH, 1998).Water-based MWFs are most frequently used in the
machining industry, thus the primary concern is the presence of contaminants that
encourage the growth of bacteria and fungi.
Exposures to MWFs were associated to various health diseases such as cancer,
respiratory outcome and skin disease (Park et al., 2009). Dermatitis is the most common
complaint associated with MWFs. Respiratory effects such as upper respiratory
irritation, asthma and hypersensitive pneumonitis (HP) are caused by exposure to
diluted MWFs, microbial contaminants and chemical contaminants of the fluids (Park et
al., 2009). Based on a National Occupational Exposure Survey (NOES) by the National
Safety and Health, it is estimated that 1.2 million workers are potentially exposed to
MWFs (NIOSH, 1983). Workers were exposed to the MWFs by inhalation of the diluted
fluids mist (aerosol) or skin absorption of fluids through contact with the MWFs during
the handling with the workpieces coated with fluids (NIOSH, 1998).
Fractional exhaled nitric oxide (FeNO) is increasingly used as a non-invasive
measure of airway inflammation (Johansson et. al., 2013). The aim of this study was to
determine the relationship between the airway inflammation as indicated by the
fractional exhaled nitric oxide (FeNO) with the microbial contaminants in MWF aerosol
and the workers reported health symptoms.
2.0 Methodology
2.1 Study population
The cross-sectional study was conducted at one of the factory located in Negeri
Sembilan, Malaysia. The studied factory is a bearing manufacturing factory in which
MWF were extensively used during the metal working process. The inclusion criteria in
this study were workers ranging in age between 20 and 58 years old, healthy and had
been employed for more than 6 months. After screening for specific criteria, 138
workers who fulfilled the inclusion criteria were randomly selected to participate in this
study. Approval from the factory and written consents from all participants were also
obtained. The study was approved by the Medical Research Ethic Committee,
University Putra Malaysia prior to data collection (FPSK (FR14) P002).
2.2 Workplace and Metalworking Fluids
There were about 4 main sections in the factory based on the production of products
namely the self-aligning ball bearing (SABB), deep groove ball bearing (DGBB),
spherical roller bearing (SRB), large size roller (LSR) and 12 subsections which were
SABB 1, SABB 2, SABB 3, DGBB 1, DGBB 2, DGBB 3, SRB 1, SRB 2, SRB 3, SRB
4, SRB 5 and LSR. All of the sections except for SRB used almost similar types of
MWF. The distribution of MWF used in each section is shown in Table 1 below. (Table
1)

2.3 Environmental sampling

The microbial air samplings were conducted by using DUO SAS SUPER 360
in order to identify bacteria and fungus concentrations in the environment. There were
two different types of agar plates that were used in this sampling which were tryptic soy
agar (TSA) for bacteria and sabouraud dextrose agar (SDA) for fungus. The air flow
speed was selected at 200 L/min. Both of the agar plates were incubated at 25 C then
the colonies were counted to estimate the bacteria and fungus concentrations. The
microbiological air sampling results were recorded and interpreted.
2.4 Bulk sampling (MWF)
Bulk samples were collected in sterile containers from 12 MWF tanks in various
work sections. These were serially diluted to tenfold dilution in 0.85% NaCl and
inoculated into 2% sabouraud dextrose agar (SDA) for fungus and into tryptic soy agar
(TSA) for bacteria. All plates were incubated at 25C, for 2 to 7 days. The colonies were
counted to estimate the bacteria and fungi concentrations at the various work sections.
2.5 Measurement of Exhaled Nitric Oxide (FeNO)
Degree of airway inflammation were tested by using fractional exhaled NO
(FeNO). FeNO of the respondents were measured using portable electrochemical
detection device (NIOX-MINO; Aerocrine AB, Sweden), according to publish
procedure for FeNO measurement (American Thoracic Society, 2005). Respondents
who have a cold on that particular day were not being measured. Respondents were
advised to avoid nitrate rich food 3 hours before the measurement and avoid strenuous
exercise 1 hour before the measurement. Nose clips were not be used during FeNO
measurement because blocking of the nasal passages will cause exhaled nitric oxide
accumulate in the nose and potentially leak into the exhaled air stream through the
posterior nasopharynx (American Thoracic Society, 2005). During inhalation phase,
respondents were required to breathe in through a disposable nitric oxide scrubbing
filter and exhale out into the room air twice before the FeNO measurement. For the third
time of breathing, respondents were required to breathe through the disposable filter and
exhale into FeNO analyzer. The exhaled FeNO measurements at a flow of 50 mL/s for
10 seconds were guided by a light and sound signal in order to assure steady flow
(Montella et al., 2011).
2.6 Data Analysis
All the statistical analysis were analysed using the Statistical Package for Social
Sciences (SPSS) Version 22.0. Kolmogorov-Smirnov statistics was used to test the
normality of all continuous variables. Descriptive analysis was used to explore and
analyse socio-demographic data and other related variables in terms of mean value,
standard deviation and frequency. The correlation test (Pearson Correlation) was
performed to determine the correlation between the FeNO Level with environmental
and MWFs microbial contaminants. For multivariate analysis, a multiple linear
regression and multiple logistic regression were used to determine the relationship
between the selected variables with the dependent variables.
3.0 Result & Discussion
Data of 138 respondents were obtained through an interview and a selfconstructed questionnaire in which socio-demography information and working
information were collected. Information on respondents age, BMI, employement years
and smoking status were some of the data collected, as shown in Table 2 and Table 3.

Most of the respondents were from the age range between 20 years old to 29
years old which comprised of 40.6% from the total respondents and most of the selected
respondents were overweight (42.0%). About 43.5% of the total respondents were
smoker and 5.8% of them takes alcohol in their daily life (Table 2). Beside that, the
working informations of the respondents were also gathered as shown in Table 3. As for
current working information, majority of the respondents have been working in the
factory for 6 months to 5 years (47.1%) (Table 3).
Based on Table 4, there were about 12 work sections in which the metalworking
fluids were used in the machining section. Majority of the selected respondents worked
at DGBB 1 (13%). The distributions of respondents work station, number of number of
machine and distance between each MWFs machine are shown in the Table 4. Reported
health symptoms of the respondents were also collected during the interview (Table 5).
The levels of microbial contaminants were measured in different job sections.
The microbial levels were obtained by measuring the bacterial and fungal concentration
in the environment and from the MWFs machines. Table 6 showed the level of
microbial contaminants in different job sections. Based on 6, Section SABB has highest
bacterial and fungal level in the environment. After conducted the descriptive analysis,
the total mean value for the environmental bacteria and fungus in all job section were
285.83 cfu/m3 and 231.2 cfu/m3 respectively. Whereas, the total means value for
microbial contamination in MWF were 37916.7 cfu/ml (bacteria) and 38833.3 cfu/ml
(fungus) (Table 6).
The high level of microorganism in the environment and MWFs machine in
SABB job section might be due to many factors, such as the distance between the
metalworking fluids machine in the section. The distance between metalworking fluids
machine at SABB section were close to each other which were within 3 meter and less.
The bacteria and fungus levels in the air decreased when the distance between the
metalworking fluids machine were increased. Findings showed that the total number of
bacteria collected with a Reuter centrifugal sampler in the air surrounding machines was
inversely related to the distances from the machines (Laitinen et al., 1999).
Besides the distance of MWFs machine, the numbers of MWFs machine in the
job section were also contributed to the high level of microorganism in the environment
and MWFs machine. Job Section SABB 2 had the highest number of water-based
metalworking fluids machines in the factory. The higher the number of metalworking
fluids machines in a section, the higher would be the environmental contaminants. This
is due to the accumulation of microorganism concentration from the MWFs machine in
the job section.
Table 7 shows the distribution of FeNO level of respondents at different job
section. Result shows that SRB job section had the highest mean value of FeNO level
which was 34.46 ppb. Workers in SRB 1 had the highest mean value of FeNO level
which was 43.00 ppb. As shown in Table 7, a comparison test was conducted to
determine the significant difference between the FeNO levels between each sub-section.
One way ANOVA test shows there were significant difference between job sections for
FeNO level (p=0.01).The high mean values of FeNO level in SRB job section were due
to the high total number of sub-section and also respondents in the section. Besides that,
SRD job section had the highest number of machine which contributed to the high level

microbial contaminants. Studies shows, worker exposure to aerosols of fluids such as

microbe has been shown to be associated with specific occupational asthma, increased
respiratory symptoms, hypersensitivity Pneumonitis (Kennedy et. al., 1999). Most of the
respondents in the SRB 2 had higher employment years which contributed to the high
FeNO level of the respondent in the section due to prolong exposure (Table 7).
Correlation between the FeNO level with environmental and MWFs microbial
contaminants were showed in Table 8. Result shows that there is a significant
correlation between the FeNO level with environmental bacteria (r=0.17, p=
0.05).Based on previous study, worker exposure to aerosols of fluids such as microbe
has been shown to be associated with specific occupational asthma, increased
respiratory symptoms, hypersensitivity Pneumonitis (Kennedy et. al., 1999). According
to a study by Fornander et. al., in vitro test showed that a sample of used MWF taken
directly from a cutting machine at the factory had considerably higher inflammatory
promoting effect than unused MWF. Whether this is caused by higher amounts of
antibacterial formaldehyde, microbial growth and generation of pro inflammatory
products such as endotoxin or enrichment of particulate matter is not known. However,
it does illustrate that the MWF used in the production, over time, probably becomes
more likely to cause airway inflammation (Fornander et. al., 2013). Even though in our
study, there is no correlation between MWFs contaminants and FeNO level, but there is
correlation between environmental and MWFs contaminants. Thus, most of the
microorganism presences in that environment were emitted from MWFs machine in a
form of aerosol in which more likely to cause airway inflammation in the respondents.
It has previously been suggested that endotoxin contamination of MWF is the causative
agent responsible for inducing an inflammatory response in airways [(Gordon, 2004),
(DeLorme et. al., 2003)] (Table 8).
Table 9 shows the relationship between socio-demographic, lifestyle and
workplace information with the FeNO levels. Referring to the table, there were
significant relationships between employments years ( =8.93, 95% CI= 7.33 10.54,
p= <0.001), distance of machine ( = 0.79, 95%CI= 0.64 - 0.96, p= 0.03), numbers of
machines ( = 2.55, 95% CI= 1.11-5.82, p= 0.02), job section ( = 3.18, 95% CI= 1.456.98, p= 0.01), and environmental bacteria ( = 12.60, 95% CI=1.65 24.56,p= 0.04)
with the FeNO levels (Table 9).
Based on a study, the increase in bronchial responsiveness was associated with
duration of exposure to water-based MWFs (Kennedy et al., 1999). This is accordance
to our study, whereby high employments years (higher duration of exposure) were
related to high FeNO level (airway inflammation). Our finding also were consistent
with other study that the distance between MWFs machine were inversely related to the
FeNO level which is due to high microbial contaminants level. Finding showed that the
total number of bacteria collected with a Reuter centrifugal sampler in the air
surrounding machines was inversely related to the distances from the machines
(Laitinen et al., 1999). According to a study by Fornander et. al., bacterial
contamination of the MWF may have contributed to the airway inflammation.
In this study, job section of the respondents was also one of the risk factor for
high FeNO level. High FeNO level were indicated in respondents who were work in a
job section that had high environmental bacteria level, number of machine and closer
distance of MWFs machine. Besides that, the effect of smoking on FeNO levels in this

study should be discussed. In previous study,it has been shown that FeNO of steroid
smoker asthmatics is lower than non-smoker asthmatics and that generally smoking
reduces the levels of FeNO (Sato et. al.,2008). However, our result showed that
smoking does not affect the FeNO level.
Based on Table 10, there was significant relationship between FeNO levels with
cough. Airway symptoms were reported in 39% of the workers exposed to MWF
although the measured levels of MWF substances in the work place air were low.
Besides that, workers that operated machines using MWFs had significantly higher
prevalence of work-related nasal irritation and coughing compared to unexposed
workers at the same factory (Fornander et. al., 2013).According to Kennedy et. al.,
incidence rate for a clinically relevant increase in airway inflammation accompanied by
asthma like symptoms such as cough and chest tightness was over twice as high among
the machinists compared with other apprentices (Kennedy et al., 1999). Based on a
study, elevation of FeNO level helps to make a final diagnosis of chronic cough
(Gordan,2008). Another study by Sato et. al., also concluded that FeNO measurements
could be used as a diagnostic marker of prolonged cough, especially for the differential
diagnosis bronchial asthma and cough variant asthma from eosinophilic bronchitis
without asthma and others (Table 10).
4.0 Conclusion
In conclusion, this study shows that airway symptoms among machining
workers exposed to MWF is a common problem. Risk factors from the workplace such
as the number of machines and their closeness to them and high environmental bacteria
colonies and longer employment years had significant relationships with the airway
inflammation (FeNO). Exposure to MWF also resulted in significantly frequent cough.
Thus, FeNO test is useful for making the diagnosis and differential diagnosis of chronic
cough in clinic practices.
5.0 Acknowledgement
This research was funded by the E-Sciences grant scheme (MOSTI) with Project
code: 06-01-04-SF1203. The author would like to express her gratitude to all the
researchers who have contributed to this review paper
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