an investigation of domestic laundry in europe - habits ... - SIFO

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78An investigation into the domestic laundry habits in Europe4.3.4 Laundry itemsThe laundry items used are listed in table 4-5. All items are naturally soiled bybeing used or worn by members of a private household. Each laundry item isused by the same (member of a) household during the experiment. In this waythe contamination of the laundry items during the experiment is as identical aspossible. To evaluate cross-contamination, sterile samples are used. The fabricis 100% cotton. The samples are autoclaved for 15 minutes at 121°C.Table 4-4 Laundry itemsLaundry item Textile material Fabric BrandDiaper 100% cotton woven HemaDishcloth 100% cotton non-woven Albert HeijnSocks 61% cotton, 35% polyester, 4% elastan knitted HemaHandkerchief 100% cotton woven Hema4.3.5 SamplingMicro-organisms are not evenly spread on the laundry items. In order to obtaina more homogenous sample, a paper shredder is used to cut the items intosmall pieces (see also photo 4.1). The paper shredder is disinfected with alcohol(70%) and the small pieces are collected in a sterile bag. After mixing, 30of these pieces are used as a sample for determining the amount of microorganismsbefore washing. Additionally, 30 pieces are put in a sterile washingbag (a bag used for washing tabs) and used for the assessment of the hygienicquality after washing. The samples are kept for a maximum of three days inthe refrigerator.Picture 4-1 Sampling
Hygiene effects of laundry processes in Europe 794.3.6 WashingThe washing machine is loaded with a gamma sterilised cotton base load. Thecomposition of the base load is in accordance with IEC 60456 [9]. The washingbags with laundry samples (a sample of each laundry item is put in a separatewashing bag) and the washing bag with the sterile samples are added tothe wash load. Direct contact between the washing bags is avoided. A perkilogram base load of 12.5 ml defibrinated sheep blood (Biotrading) is addedto increase the soil level. The load weight, washing programme, washing temperature,water hardness and detergent used depend on the test.4.3.7 Enumeration and confirmation of micro-organismsAll samples are directly removed from the machine and put in a sterile stomacherbag at the end of the washing process. 50 ml of a buffered peptone solution(Oxoid, CM509) with a temperature of 30°C is then added. After 5 minutes,the bag is stomachered for 30 seconds. The obtained solution is dilutedin PFZ (Biotrading, K110B009) and used for enumeration. The same procedureis used for the enumeration and confirmation of micro-organisms in thedirty laundry samples.Enumeration of Enterobacteriaceae1 ml of the serial dilution is transferred to a plate and mixed with ca. 10 mlViolet Red Bile Glucose Agar (VRBGA) (Lab M LAB88). After solidificationof the agar, a topcoat is added using the same medium. The plates areincubated for 20-24h at 37°C. Suspected Enterobacteriaceae colonies arecounted after the incubation period. Confirmation is done by testing the presenceof oxidase (Bactidrop, Oxidase test R21540). Enterobacteriaceae arepurple colonies surrounded by a zone of purple precipitation and are oxidasenegative.Enumeration of Staphylococcus aureus0.1ml of the serial dilution is spread on a Baird Parker (BP) plate (Biotrading,K002P090) and incubated for two days at 37°C. Black, shiny colonies with ahalo were counted.Enumeration of Bacilli0.1ml of the serial dilution is spread on a MYP/PREP plate (Biotrading,K104P090) and incubated for one day at 30°C.
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