National and international prehospital staff have an interest in obtaining thorough knowledge about the contamination level of prehospital uniforms and, thus, about the risk of infection to which patients are exposed in the prehospital setting. Several international studies have examined various aspects related to health staff uniforms including, among others, the presence of methicillin-resistant Staphylococcus aureus (MRSA) [1,2], the importance of the washing procedure for microbiological reduction [3] and the hygiene culture among health professionals [4]. These studies, however, have all concerned hospital or home care staff.

Studies have demonstrated that uniforms in healthcare services become contaminated with microorganisms, among others through the staff’s contact with patients [4]. Thus, a clinical study demonstrated the spreading of infection from the uniform to the patient and the transmission of infection from patient to uniform and then to another patient [5]. Several other studies have also shown that there is a risk of transmitting bacteria by direct contact [6]. This indicates that uniforms in prehospital healthcare services may also constitute a source of infection and thus a health risk for patients.

In the Region of Southern Denmark, more than 100,000 patients are transported by ambulances annually (quote by: Niels Thorup, Regional Director, Falck Denmark A/S, Region of Southern Denmark, 2014). The staff in these ambulances carries standardized uniforms consisting of a jacket and a pair of trousers that are washed in accordance with instructions based on an accredited standard. Presumably, the washing procedure reduces the number and types of microorganisms on clothes after washing [3]. However, our literature search revealed no evidence of the exact significance of washing on daily practice in Danish prehospital settings.

Furthermore, several studies show that hospital uniforms can be contaminated with microorganisms [1,7,8]. The most commonly occurring microorganisms are S. aureus and Enterococcus, among several others [1,4]. Still, our literature search produced no studies examining which microorganisms are involved in the bacteriological contamination of staff uniforms in Danish ambulance services.

We therefore examined the extent to which the ambulance staff uniforms are contaminated with potentially pathogenic microorganisms before washing. Specifically, we examined the potentially pathogenic bacterial contamination and the prevalence rate for E. coli, B. cereus, Clostridium, S. aureus, Enterococcus and Pseudomonas. We also examined to which extent the bacterial contamination was reduced through use of a washing detergent with and without acetic peroxide. All uniforms were washed at 60 degrees Celsius and tumble-dried for 1 hour.

A cross-sectional study was carried out at a random number of selected ambulance stations in the Region of Southern and Central Denmark. In accordance with standard procedures, uniforms were washed locally in the stations where the staff is employed. Practical sampling was applied, and the collected uniforms had all been used in the ambulance service within the past 24 hours. The collection took place over a period of 7 days. The staff was not blinded as they are usually responsible for the washing of uniforms.

A total of 30 ambulance staff uniforms were selected randomly after they had been worn during a shift lasting a minimum of 8 hours. Before washing, all 30 uniforms were contact-printed with specific 25 cm² agar plates or inoculated using swab sticks for bacterial mapping.

Prints were obtained from the torso, the thighs and the wrist area which previous studies have shown are the most contaminated locations on healthcare uniforms [4,9].

Several healthcare related studies have found that healthcare-related uniforms were contaminated with various potentially pathogenic bacteria and thus associated with a risk of patient infection [5,10,11]. However, no Danish study has previously examined the bacterial contamination of prehospital uniforms which leaves us with no results for comparison. However, since our results indicate that an average print from a prehospital uniform has similar or fewer potentially pathogenic bacterial colonies than found on uniforms from in-hospital studies [12], we suggest that the contamination level of uniforms in prehospital settings is, to some extent, comparable to the contamination level of in- hospital uniforms.

Our findings of potentially pathogenic bacteria correspond to previous findings in hospital-based studies [1,12]. Our results indicate that S. aureus, Enterococcus and Clostridium are present on used prehospital uniforms with a presumed bacterial susceptibility comparable to that of patient isolates; however, we did not collect patients’ samples and therefore cannot compare the results to those of patient isolates. Nevertheless, the present bacteria are already known to cause healthcare associated infections (HAIs).

The fact that we found a lower prevalence of Enterococcus than in a previous study focusing on nurse uniforms [4] is probably explained by the fact that nurses’ tasks are not identical to those performed by staff in the prehospital setting. For example, prehospital staff may perform much fewer tasks related to personal care, e.g. hygiene following a restroom visit, etc., which are known to be associated with infection. Although we did not find E. coli or Pseudomonas, we cannot dismiss that prehospital uniforms could be contaminated with either of these microorganisms. Nevertheless, we assume that the contamination level and hence the risk of infecting patients is, indeed, limited.

Washing and tumble-drying is known to aid bacterial reduction, especially as prehospital uniforms dictate a maximum washing temperature of 60C. However, our results show that washing and tumble-drying in domestic machines for 1 hour, regardless of the choice of washing powder, did not eliminate all microorganisms. The washing machine, the tumble-dryer and the environment could all have caused this contamination, and it would have been beneficial if we had collected environmental samples which would have provides us with knowledge of a possible correlation between contamination of uniforms and contamination of machines and/or the environment. In addition, we could also have tested the temperature in the tumble dryer and the washing machine.

Unfortunately, our methodology has a number of limitations that must be addressed. Firstly, the results of the washing process are compromised given that the two groups compared did not have the same degree of contamination at baseline. However, we were unable to ensure this as this was an in situ study. Secondly, due to practicalities such as timeframe and financial limitations, we did not perform a sample size calculation, which could have ensured a sample size sufficiently large to obtain a significant difference. Therefore, we acknowledge that this study should be considered a pilot study, and that a larger future study requires further consideration.

There is also a risk of bias associated with the position of the prints before and after washing. Although we tried to ensure that prints were taken from the same location before and after washing, there is a margin of error associated with the outcome as we cannot guarantee that the prints were placed in the same location before and after washing. Another important issue that must be emphasized is the variation in our results caused by the use of domestic washing machines; however, we did intend to illustrate the result during the practical laundering and handling process in the prehospital setting. We used three uniform sampling locations which have previously been shown to be the most contaminated locations for assessing the mean degree of contamination of the uniforms, and we assessed the mean degree of contamination. The sampling technique itself was well known by the environmental laboratory performing the analysis, and its feasibility for that purpose is well-established in current literature. It was therefore decided not to further test its reproducibility.

As mentioned, the number of international studies on the effect of different washing methods is very limited [7,10], and no previous studies in the Danish ambulance services can substantiate our findings on bacterial contamination or on the quality of the used washing methods. At the same time, evidence of the risk associated with the transmission of microorganisms between staff and patients is very limited. Furthermore, no Danish correlation studies between bacterial CFU and incidence of infection in the prehospital setting have been performed. Nor have any studies sought to correlate different methods of washing with the incidence of infections. In Denmark, no comprehensive registration of HAIs is presently in place and therefore such analysis is not possible. We recommend that such registration be introduced to enable studies on the association between contamination degree and risk of infection.

Overall, further studies are needed on prehospital bacterial contamination and hygiene interventions. Increased evidence of contamination degrees, their underlying causes and the effect of hygiene interventions and procedures will, all things being equal, strengthen infection control in the prehospital setting.

The results of this study indicate the presence of a contamination level which - in several respects - underpins the need to comply with existing hygiene instructions in the prehospital setting. Daily change of uniform, washing at a minimum of 60 degrees Celsius and use of a disinfectant detergent (acetic peroxide) reduce the degree of bacterial contamination and thus reduce the risk that patients become infected. The results concerning a potentially pathogenic contamination level and the prevalence of specific microorganisms, including B. cereus, Clostridium, S. aureus and Enterococcus, stress the need for daily change of uniform and a high hygiene standard in daily prehospital work with patients to avoid increasing the risk that microorganisms are transmitted from ambulance uniforms to patients.