## 2 Materials and Methods
### Analytical Methods  
The procedure followed in the current study required the following materials: a delimiting frame; sterile swabs on a wooden rod; and six EOs with 100% purity from the Young Living Essential Oils, used for their antifungal properties [71, 72, 73, 74, 75, 76, 77, 78, 79]. The types of EOs employed were: _Mentha piperita, Lavandula angustifolia, Citrus limon, Melaleuca alternifolia, Marjoram_ and _Origanum vulgare_. The products used consisted of: _Mentha piperita_, batch 103519, expiry date 7 August 2021; _Lavandula angustifolia_, batch 87491, expiry date 7 August 2021; _Citrus limon_, batch 103517, expiry date 7 August 2021; _Tea Tree_, batch 89075, expiry date October 2021; _Marjoram_, batch 86942, expiry date July 2022; and _Oregano_, batch 89094, expiry date September 2022. The material resources used consisted of: Sabouraud sterile culture medium for the yeast and mould isolation; an incubator ICT 18/FALC with a temperature range between 5 and 80 \({}^{\circ}\)C; a microbiological hood; glass slides and slides for the microscopic identification; microbiological handles; an optical microscope Micros Austria with binocular head series BIM-105B; an API yeast identification kit (r) 20C AUX; and a densitometer. Six different areas on the face of the traditional coat were examined, as shown in Figure 2. The working areas were delimited by means of a metal dial with a size of 25 cm\({}^{2}\). Small fragments (millimetres) of fur and sheepskin were removed with tweezers previously sterilized by the incandescent method for testing of the fungi concerned; so as to observe the effects that they might have had on the individual strands of material. The tests were conducted both before and after applying the EOs to each delimited area, although the primary samples were considered as blank references. After delimiting the surfaces on which the in-situ procedures were to be performed, the samples of the surface were taken. The samples were extracted using sterile swabs from all six areas of the sheepskin coat, both before and after the application of the Eos (see Table 1). Ten drops (the equivalent to 300 uL) of the corresponding EO were applied directly to the tested material surface in the centre of each delimited area [75]. After the application of the EOs, three samples were taken for each of the six different work areas delimited on the area of sheepskin to which the EOs were applied. The three samples were taken at pre-set intervals, with the first sample being taken 30 min after the administration of the EOs, the second test being administered after 24 h and the third after 48 h. Table 1 also shows the type of material examined and the type of EO applied to each of the delimited areas. The EO used for each type of material concerned was selected in accordance with the recommendations made in the literature, especially that of [PERSON] et al. [56], which state that certain EOs tend to be more effective than others when applied to certain types of material, such as those which constitute heritage objects. It must be noted that the air in the room in which the sheepskin coat was stored and examined was not circulated, and neither was the room ventilated for 48 h during the entire examination, so as to avoid contamination with other types of spores from the atmosphere that could, otherwise, have been introduced by currents of air moving through the room. The temperature was maintained between 22 \({}^{\circ}\)C and 24 \({}^{\circ}\)C, while the humidity was kept between 52% and 55%. With the EOs having been applied directly to the examined surfaces, all the samples were seeded on sterile Sabouraud culture media, with the plates used being incubated for 30 days at a temperature of 28 \({}^{\circ}\)C. The third stage of the procedure involved isolating and identifying the fungi developed on culture media.