Introduction

In the great majority of patients with sporadic amyotrophic lateral sclerosis (SALS) the cause of the disease remains unknown, and the proportion of environmental or genetic factors that underlie SALS continues to be debated [1]. Recent findings that the sporadic form of ALS may be even more common than was previously thought [2] have again focussed interest on the possibility that environmental agents play a part in the pathogenesis of the disease. Occupation has been widely used as a surrogate for environmental exposure to toxicants (for a review of occupational studies in ALS, see Sutedja et al. (2009) [3]), since few individuals are likely to be aware of particular toxicants they have been exposed to during their working lives.

A feature of some previous studies of occupations in SALS has been that only the main (typically the longest-held) occupation of the individual has been considered [4-6], though some have used all lifetime occupations [7-10]. It could be argued that all such studies should take lifetime occupations into account, since even time spent in a relatively short-lasting work environment that exposes an individual to a toxicant could be relevant to the later appearance of a disease. For example, a short exposure to asbestos fibres can lead to mesothelioma many years after asbestos exposure. Using all occupations may be especially applicable to SALS, since a short exposure to a neurotoxicant such as inorganic mercury results in the toxicant remaining within motor neurons indefinitely in both animals [11] and humans [12].

Occupations can be coded for epidemiological studies using either country-specific codes (such as the Australian and New Zealand Classification of Occupations, ANZSCO) or the International Standard Classification of Occupations (ISCO). ISCO is useful since it allows comparisons of studies performed in different countries, but this classification considers occupations in groups only and does not extend to individual occupations, as does a classification such as ANZSCO.

Although a large number of occupations have been reported to be associated with SALS [6], no toxicant exposure that is present in the majority of these occupations, and that has the potential to cause SALS, has been proposed. Our study, looking at lifetime occupations in people with SALS and controls, has found that truck drivers in particular have an increased risk of SALS. We suggest that a plausible link between a number of occupational findings in SALS is exposure to diesel exhaust.

Methods

Results

Risk of SALS in occupations

A. Occupations with 5 or more individuals.

Odds ratios, confidence intervals, and p-values for all occupational titles where 5 or more SALS and control individuals were present are listed for both ANZSCO and ISCO in Table S2. Those occupational titles with statistically-significant ORs for SALS are listed below.

1. Major Group

Males

In ANZSCO, Professionals (OR 0.59) had a decreased risk of SALS. Technicians and Trade Workers (OR 1.47), Machinery Operators and Drivers (OR 1.96), and Labourers (OR 1.72) had increased risks of SALS.

In ISCO, Managers (OR 0.67) and Professionals (OR 0.56) had decreased risks of SALS. Machinery Operators and Drivers (OR 1.59), Labourers (OR 1.68), and Elementary Occupations (OR 2.07) had increased risks of SALS.

The trend for ANZSCO and ISCO male Major Group occupations which require higher skills and tasks to have a reduced risk of SALS, and conversely those occupations requiring lower skills and tasks to have an increased risks of SALS, can be seen in Figure 1.

Download:

• PPT
  PowerPoint slide
• PNG
  larger image
• TIFF
  original image

Figure 1. The risk of SALS in male Major Occupational Groups.

In both ANZSCO and ISCO classifications, occupations with higher skills/tasks (on the left of the graphs) tended to have lower risks of SALS, and those with lower skills/tasks (on the right of the graphs) higher risks of SALS. 1 = Managers, 2 = Professionals, 3 = Technicians and Trade Workers, 4 = Community and Personal Service Workers, 5 = Clerical and Administrative Workers, 6 = Sales Workers, 7 = Machinery Operators and Drivers, 8 = Labourers, 9 = Elementary Occupations. Bars = 95% confidence internals. * = p <0.05, ** = p<0.01, *** = p<0.001.

https://doi.org/10.1371/journal.pone.0080993.g001

Females

In both ANZSCO and ISCO, no ORs for these occupational titles reached significance.

2. Submajor Group

Males

In ANZSCO, Chief Executives, General Managers and Legislators (OR 0.32), and Business, Human Resource and Marketing Professionals (OR 0.41) had decreased risks of SALS. Electrotechnology and Telecommunications Trades Workers (OR 1.95), Road and Rail Drivers (OR 2.05), Construction and Mining Labourers (OR 2.50), Farm, Forestry and Garden Workers (OR 1.98), and Other Labourers (OR 2.10) had increased risks of SALS.

In ISCO, Business and Administration Professionals (OR 0.46), and Information and Communications Technology Professionals (OR 0.28) had decreased risks of SALS. Personal Service Workers (OR 3.18), Building and Related Trades Workers, excluding Electricians (OR 1.79), Electrical and Electronic Trades Workers (OR 2.10), Stationary Plant and Machine Operators (OR 1.60), Drivers and Mobile Plant Operators (OR 1.78), Agricultural, Forestry and Fishery Labourers (OR 2.17) and Labourers in Mining, Construction, Manufacturing and Transport (OR 1.91) had increased risks of SALS.

Females

In both ANZSCO and ISCO, no ORs for these occupational titles reached significance.

3. Minor Group

Males

In ANZSCO, Chief Executives, General Managers and Legislators (OR 0.32) had decreased risks of SALS. Fabrication Engineering Trades Workers (OR 2.92), Truck Drivers (OR 2.27), Construction and Mining Labourers (OR 2.50), Food Process Workers (OR 3.19), Farm, Forestry and Garden Workers (OR 1.98), and Miscellaneous Labourers (OR 2.32) had increased risks of SALS.

In ISCO, Finance Professionals (OR 0.43) had a decreased risk of SALS. Building Frame and Related Trades Workers (OR 2.11), Heavy Truck and Bus Drivers (OR 1.99), Agricultural, Forestry and Fishery Labourers (OR 2.18), and Mining and Construction Labourers (OR 2.47) had increased risks of SALS.

Females

Female Cooks (OR 3.65) in ISCO had increased risks of SALS. No statistically-significant ORs for these occupational titles were found in ANZSCO.

4. Unit Group

Males

In ANZSCO, Accountants (OR 0.32) had a decreased risk of SALS, while Truck Drivers (OR 2.26) had an increased risk of SALS.

In ISCO, Accountants (OR 0.31) had a decreased risk of SALS, while Heavy Truck and Lorry Drivers (OR 2.10), and Building Construction Labourers (OR 3.31) had increased risks of SALS.

Females

Cooks had an increased risk of SALS in both ANZSCO (OR 3.64) and ISCO (OR 3.65).

5. Occupation Group (ANZSCO only)

Males

In ANZSCO, Truck Drivers had an increased risk of SALS, with 30 SALS (8.1%) and 13 control (3.6%) individuals being truck drivers (OR 2.36, 95% CI 1.21 to 4.59, p = 0.012).

Females

No statistically-significant female ORs were found in these occupational titles.

B. Occupations with fewer than 5 individuals.

The relative frequency of occupational titles with fewer than 5 SALS or control individuals are available in Spreadsheet S1 for males, and Spreadsheet S2 for females. The percentage difference between SALS and controls for each ANZSCO occupation can be seen in ascending or descending order by sorting the Frequency Difference column on the Excel spreadsheets; negative values indicate decreased frequencies of SALS, and positive values increased frequencies of SALS.

In males, occupations that appeared to be particularly more frequent for SALS were Metal Fabricator, Cook, Builder's Labourer, Delivery Driver and Fencer; those particularly less frequent for SALS were Accountant, Chief Executive or General Manager, and Sales and Marketing Manager. In females, occupations that were particularly more frequent for SALS were Retail Manager, Cook, and Fruit or Nut Picker; those particularly less frequent for SALS were Bar Attendant, and Nanny.

Discussion

Two major findings have come from this study of lifetime occupations as potential risk factors for SALS. Firstly, occupations requiring higher skills and tasks tended to have decreased risks of SALS, whereas occupations requiring lower skills and tasks tended to have increased risks of the disease. Secondly, the majority of occupations with significantly increased risks of SALS, and in particular truck drivers, were likely to be exposed to diesel exhaust during their working hours. Of note, occupations with decreased risks of SALS, being mostly managerial and indoors, would be less likely to be exposed to diesel exhaust.

The stand-out occupation in this study as regards an association with SALS was truck driving. This was the only occupation to remain positively associated with SALS down to the smaller numbers of individuals who were present in the most detailed ANZSCO occupational group. Of interest, a 1980 study of 504 World War II veterans with ALS and 504 matched military controls also found that truck (and tractor) driving was the only individual occupation that had excess numbers of ALS patients [17]. Truck drivers are exposed to a number of potentially injurious environmental agents [18], but the most studied of these has been exposure to diesel exhaust [19-25]. The predominance of diesel use in trucks can be seen in figures from the Australian Bureau of Statistics Motor Vehicle Census for 2012 (www.abs.gov.au/ausstats/abs@.nsf/mf/9309.0), which shows that the percentage of passenger vehicles using diesel was 7%, light commercial vehicles 44%, campervans 62%, buses 76%, non-freight carrying trucks 83%, light rigid trucks 91%, and articulated trucks 98%.

Factors contributing to the level of diesel exhaust exposure of truck drivers are whether windows are open or closed (with higher exposure when windows are open), warmer compared to cooler weather (with warmer weather increasing the exposure) and exposure during daytime city driving (when exposure is higher than during long-haul driving in the evening on suburban or rural roads) [24]. The age of the truck also contributes to the exposure, since fuel exhaust can leak through the rubber seals of the driver compartment in older trucks [22].

Diesel exhaust is composed of a complex mixture of gases and particulates [20,24]. A number of components of the exhaust, such as formaldehyde, benzene, hexane, and styrene may be involved in the pathogenesis of SALS [25]. Formaldehyde exposure has been associated with a risk of SALS in two studies [26,27]. Derivatives of both benzene and hexane cause neurofilamentous proximal axonal swellings in rodents [28], and proximal axonal swellings filled with neurofilaments have long been recognised as a pathological feature of ALS [29]. Some of these toxic agents may interact with genetic variants in people who are susceptible to SALS. For example, the cytotoxicity of the glutathione-depleting chemical styrene oxide varies with low or high superoxide dismutase 1 (SOD1) activity [30], and SOD1 mutations are one cause of familial ALS [31]. The potential neurotoxicity of a large number of components of hydrocarbon fuels has been reviewed by Ritchie et al. (2001) [20]

Experimental studies indicate that diesel exhaust can have deleterious effects on the CNS, including upregulation of the proinflammatory cytokine TNF-alpha [32] which can enhance motor neuron susceptibility to the excitotoxins [33] that are suspected to play a part in ALS [34], and upregulation of microglia [35] which also occurs in ALS [36]. Evidence concerning air pollution-induced neuroinflammation and alterations to the blood-brain barrier has been reviewed by Genc et al. (2012) [37].

Truck drivers were not the only workers in this study who had an increased SALS risk combined with a potential exposure to diesel exhaust. Eleven other occupations and occupational groups at risk of SALS took place in workplaces where diesel exhaust has been reported to be a potential hazard, and ten of these have previously been associated with SALS. Although Truck Drivers dominate the numbers in their respective ANZSCO Occupation, Unit, and Minor Groups, they are only a moderate part of The Submajor Group, and a small part of the Major Group. However, the p values for the odds ratio of the latter two occupations were smaller than the p values of the occupational titles that contain Truck Drivers alone (Table 2), suggesting that non-truck driver occupations within these titles also have an increased risk of SALS, possibility because related occupations in these groups are also likely to have exposure to diesel exhaust.

Diesel exhaust is not the only occupational hazard that truck drivers are exposed to. They are also prone to sleep apnoea [18], which can lead to intermittent hypoxia. Low ambient oxygen levels have been suggested to be a possible reason for an increased incidence of SALS in airline pilots [38], though exposure to jet fuel, or to diesel fuel used in aircraft refuelling trucks, could also be a reason for this increased risk [24]. However, in none of the other at-risk occupations in our study was there any indication that intermittent hypoxia is part of the workplace environment. It has been suggested that intermittent hypoxia could be a cause of the increased risk of SALS reported in fire fighters [39], but this increased risk could equally be due to diesel exhaust from fire trucks or inhalation of particulate matter from smoke exposure. Obesity is another disorder seen often in truck drivers [18], but in SALS an inverse relationship applies, with patients usually having a lower premorbid body mass index than controls [40].

Occupational exposure to diesel and other hydrocarbon fuel exhaust could explain the puzzling increase in SALS in military personnel. Three US studies have reported a doubling of SALS incidence after military service [5,41,42], and military personnel who saw active service had an increased risk of SALS compared to those who did not [42]. All service branches (army, navy and air force) had the increased SALS risk, despite being engaged in different duties, suggesting some common factor underlay the increased risk. Although two of the studies involved Persian Gulf war veterans [41,42], the third reported on servicemen before 1990 [5], which suggests that the SALS risk is not related to any particular war zone, but to the experience of war itself. One review of hydrocarbon fuel exposure, which paid particular attention to military uses, emphasised the widespread use of diesel and other hydrocarbon fuels in all service branches of the military during combat [20]. As well as the use of these fuels to power land, sea and air military vehicles, a number of unique applications lead to increased hydrocarbon fuel exposure. These include desert sand suppression with jet fuel during the Persian Gulf war (with rapid vaporisation of the fuel at the high local temperature and entry deep into the lungs when combined with fine desert sand), use of diesel fuel aerosols to visually obscure troops and equipment, diesel fuelling of unvented military tent heaters, dermal exposure to hydrocarbon fuels due to contamination of shower water, and the use of jet fuel as aircraft heat sinks [20]. Apart from accidental spills, possibly the greatest respiratory exposure to hydrocarbon fuel is suffered by military fuel workers operating without respiratory protection below the open access hatches of aircraft fuel tanks [20]. It is therefore of interest that the deployed:non-deployed risk ratio for SALS after the Persian Gulf war was higher in air force personnel than in other service branches [42]. Unfortunately, neither ANZSCO nor ISCO has adequate classifications for active military service, so this could not be assessed in the present study.

Exposure to diesel exhaust could explain the observation that SALS is more common in higher, compared to lower, population densities [43], because diesel fume exposure is higher in inner city areas compared to suburban or rural areas [21]. On the other hand, the incidence of SALS is almost identical in men and women, with only a slight male predominance in most studies [44], which argues against a link to diesel exhaust, since most of the occupations exposed to diesel exhaust are undertaken by men. Another contrary finding is that the use of diesel engines has been increasing over time in many countries. For example, heavy trucks in the US switched to diesel engines in the 1950s [24], and in Australia the number of registered diesel motor vehicles has been increasing, with the latest Australian Bureau of Statistics figures showing an increase of 61% from 2007 to 2012. Most investigators, however, now agree that the incidence of SALS remains stable over time [44], which would not be expected if an increasingly common environmental toxicant was playing a part in SALS. This is a complex issue, however, since although diesel fuel use has been increasing, the types of diesel fuels have changed over time [20], and more efficient filters are now being used in diesel-powered vehicles.

As regards methodology, advantages of this study were that: (1) this is the largest case-control study of occupation in SALS to date; (2) controls were closely matched, limiting the likelihood of type I statistical errors; (3) two occupational classifications were used, which allowed classification of individual occupations, as well as allowing comparisons with other international studies; (4) an accurate diagnosis of SALS could be made from detailed neurological reports; and (5) having access to lifetime occupations allowed consideration of all (not only ‘usual’) occupations as potential risk factors.

Limitations of the study were that: (1) no corrections were made for multiple testing. We acknowledge that with this large number of occupations some will reach statistical significance by chance only. Rather than risk discarding potentially important results by correcting for multiple testing, however, we chose to examine all occupations that had significant associations with SALS for plausible biological reasons that could underlie these associations [45]; (2) females had fewer types of occupation, so numbers of individuals were limited in some female groups; (3) domestic duties were listed as an occupation by many (mostly female) individuals, and we had to assume that certain duties were undertaken as part of these duties to fit the classification codes (see Methods); (4) a number of individuals listed occupations that could not be classified into either of the two systems. The percentage of these was low and similar in cases and controls, and this factor was therefore unlikely to affect the results; (5) neither ANZSCO or ISCO could assess military duties in detail. For example, neither active military service, nor whether service was in the army, navy or air force, could be determined; (6) a difference in levels of education between patients and controls could lead to differences in the way the questionnaire was completed. However, as has been noted before in relation to this cohort [46], in Australia the level of literacy is high (http://www.abs.gov.au/ausstats), with free compulsory education until the age of 17 years. Close matching of cases and controls (e.g., partners) would add to the likelihood of similar education and social levels. Furthermore, a subset of ALS patients and controls from this cohort has previously been shown to have similar levels of education [46]. Finally, occupation itself is a major indicator of socioeconomic status [47] so when one is comparing disease rates between occupations they are likely to be from similar socioeconomic levels; (7) Voluntary participation in filling in the questionnaire could cause selection bias by favouring patients who were keen to find the underlying cause of their disease, and thus might be more prone to recall bias. No correction could be made for this, since no information was available on the size of the potential patient population for this study (no Australia-wide study of ALS incidence and prevalence has been published) [46].

Many people are exposed to environmental toxicants such as diesel exhaust, yet few get SALS, so simple exposure alone is unlikely to lead to the disease. Individual genetic susceptibility to one or more of the many toxicants in hydrocarbon fuels has been postulated as a reason why only some people exposed to these fuels suffer neurotoxic effects [20]. It has recently been suggested that stress increases the uptake of toxicants into human corticomotor neurons, and that this might lead to SALS [48]. Many of the occupations in this study that had an increased risk of SALS, as well as exposure to diesel exhaust, have been reported to be stressful, including truck driving [49]. A study examining stressors that occur at the same time as exposure to diesel exhaust could be one future avenue to explore in the search for underlying causes of SALS. Another possible study would be to assess early-life exposure to diesel exhaust as a possible risk factor for the later onset of SALS. This could be done by looking at the occupations of parents whose offspring later get SALS. Of interest, an association between parental occupational exposure to diesel exhaust and the onset of childhood brain tumours has recently been described [50].

In conclusion, two of the largest case-control studies of SALS have now found that truck drivers have a uniquely increased risk of the disease. Exposure to diesel exhaust is common in truck drivers, and exposure to this toxicant, as well as other hydrocarbon fuels, may underlie some of the epidemiological findings related to SALS.

Supporting Information

Acknowledgments

We thank participants for donating questionnaire data, neurologists for providing clinical details on their patients, Grant Tranter for assistance with statistical analyses, and Motor Neuron Disease Associations in each Australian state who have aided patient recruitment for this research.