HYGIENE SURVEY IN ROMANIAN BAKERIES ALINA DOBRE, IBA, ROMANIA & SATU SALO, VTT, FINLAND INTRODUCTION Hygiene survey was performed in six Romanian bakeries. The work was carried out at VTT Technical Research Centre of Finland (Espoo, Finland) and IBA (Institutul de Bioresurse Alimentare, Bucharest, Romania) from October 2008 to February 2009. The main aim of the work was to detect microbial contaminants from bakery environment and process line and evaluate hygiene state in Romanian bakeries. 1 (9)
SAMPLING IN BAKERIES Surface samples from process environment, equipment, packaging material and protective clothing of workers: the amount of aerobic bacteria, yeasts and moulds, Enterobacteriaceae, coliforms and Escherichia coli. Swab samples taken from process environment: the presence of Listeria spp., L. monocytogenes and Salmonella spp. Water samples: the amount of aerobic bacteria, yeasts and moulds, Enterobacteriaceae, coliforms, E. coli, Bacillus cereus and bacterial spores Air samples: the amount of aerobic bacteria, yeasts and moulds Raw material and product samples: the amount of aerobic bacteria, yeasts and moulds, Enterobacteriaceae, coliforms, E. coli, B. cereus and bacterial spores and presence of Listeria spp., Listeria monocytogenes and Salmonella spp. SAMPLING TOOLS The surface and workers samples were taken aseptically using four various Petrifilms total aerobic bacteria, yeast and moulds, Enterobacteriaceae, coliforms and Escherichia coli. Petrifilm 3M TM Enterobacteriaceae Count Plate Petrifilm 3M TM E.coli Count Plate Petrifilm 3M TM Yeast and Mould Count Plate Petrifilm 3M TM Aerobic Count Plate 37 0 for 48 h 25 0 for 120 h 25 0 for 72 h Method: The premoistened Petrifilms were pressed against surfaces at each sampling point. 2 (9)
SAMPLING TOOLS Air sampling was performed with MAS 100 air sampler. Air samples (100 l and 250 l) were collected directly on agar plates for aerobic bacteria and yeast and mould counts. The plates for aerobic bacteria were incubated at 25 C for 72 h and the plates for yeast and mould counts at 25 C for 120 h. MAS 100 air sampler SAMPLING TOOLS Water samples were taken from sources that are important for the production process but also for the hygiene program. Method of analysis: by filtering 100 ml of water samples through filters (450 m pore size). filters were placed aseptically on 4 various agar plates depending on the microbiological indicator tested: Aerobic bacteria plate count agar plates 25 0 C for 72 h Yeast & Mould potato dextrose agar 25 0 C for 120 h Bacterial spores plate count agar 30 0 C for 72 h Bacillus cereus chromogenic Bacillus cereus agar 30 0 C for 48 h In addition water samples were analysed on Petrifilms using 1 ml of water For bacterial spores the water samples were subjected to heat treatment at 80 0 C for 20 min; filtered and cultured on Plate Count Agar and Aerobic Count Petrifilm and incubated at 30 0 C for 72 h. 3 (9)
Properly cleaned surfaces should not contain coliforms and the amount of total aerobic bacteria should also be below 100 colony forming units (cfu) / 20 cm². The presence of coliforms indicate poor hygiene. The surfaces that are not in direct contact with food must also be cleaned in order to avoid cross contamination. Microbiological water quality is influenced by the hygienic status of hoses and taps, so is an important point of control for the hygienic program. For maintaining air quality is important to separate areas and distribution of air flow from high hygiene areas to lower hygiene areas. based on experience obtained in hygiene surveys performed at VTT and recommendations found in literature Surface samples Quality limits EC Ecoli E. coli 20 cm 2 ECcoliforms Coliforms 20 cm 2 EB Enterobacteriaceae 20 cm 2 AC Aerobic bacteria 20 cm 2 YM ** Yeasts 30 cm 2 YM** Moulds 30 cm 2 Good quality Coliforms & Enterobacteriaceae in raw cereal grains and flours do not imply mishandling < 1 < 1 < 1 20 1 1 Adequate / not recommended 20 > 100 1 >.. 30 1 > 30 Poor > 1 > 1 > 1 > 100 > 30 > 30 ** sum of yeasts and moulds together needs to fit the category 4 (9)
based on experience obtained in hygiene surveys performed at VTT and recommendations found in literature EC Ecoli EC coliforms EB AC YM yeast YM moulds Workers Quality limits E. coli Coliforms Enterobacteriaceae Aerobic bacteria Yeasts Moulds 20 cm 2 20 cm 2 20 cm 2 20 cm 2 30 cm 2 30 cm 2 Good quality < 1 < 1 < 1 20 1 1 Adequate / not recommended 20 > 100 1 > 30 1 > 30 Poor > 1 > 1 > 1 > 100 > 30 > 30 based on experience obtained in hygiene surveys performed at VTT and recommendations found in literature EC Ecoli ECcoliforms EB AC YM YM AC Liquid (1 ml on Petrifilm) E. coli Coliforms Enterobacteriac Aerobic Bacterial Yeasts Moulds Quality limits eae bacteria spores ml ml ml ml ml ml ml Good quality < 1 < 1 < 1 < 100 < 100 < 100 < 100 Adequate Poor 1 1 1 100 100 100 100 PCA CBcA PCA PDA PDA Liquid (100 ml on filter) Aerobic bacteria B. cereus Bacterial spores Yeasts Moulds Quality limits 100 ml 100 ml 100 ml 100 ml 100 ml Good quality < 10 000 < 10 000 < 10 000 < 10 000 < 10 000 Adequate Poor 10 000 10 000 10 000 10 000 10 000 5 (9)
based on experience obtained in hygiene surveys performed at VTT and recommendations found in literature AIR PCA PCA PDA PDA PDA PDA Aerobic bacteria Aerobic bacteria Yeasts Moulds Yeasts Moulds Quality limits 100 litre 250 litre 100 litre 100 litre 250 l 250 l Good quality < 10 < 25 < 3 < 2 < 6 < 5 Adequate / not recommended 10 30 25 75 3 2 8 6 5 20 Poor > 30 > 75 > 8 > 20 Swabs Listeria Salmonella Quality limits pos/neg pos/neg Good quality neg neg Poor pos pos SURFACE SAMPLES RESULTS IN PERCENTAGES 164 surfaces were studied in bakeries, including environmental, non and food contact equipment samples. From all sampling points the amounts of aerobic bacteria, yeasts and moulds, Enterobacteriaceae, coliforms and Escherichia coli were studied EC Ecoli EC coliforms* EB* AC YM yeast YM moulds E. coli Coliforms Enterobacteriaceae Aerobic bacteria Yeasts Moulds 20 cm 2 20 cm 2 20 cm 2 20 cm 2 30 cm 2 30 cm 2 Good quality 94 70 71 32 62 21 Adequate 27 14 46 Poor 6 30 29 41 24 33 * coliforms and Enterobacteriaceae in raw cereal grains and flours do not imply mishandling 6 (9)
CONCLUSIONS VISUAL OBSERVATIONS In general 5 bakery factories were visually in good condition and one in adequate condition (mainly because of improper condition of walls, floors and ceilings). In all bakeries no unnecessary utensils were in the process area and the personnel was wearing protective clothing according to instructions. Compared to Finnish bakeries there was a lot of personnel in the factories. Personnel is one of the main contamination sources. More attention should be paid to the working routines and hygiene of the protective clothing. CONCLUSIONS WATER In five bakeries the amount of microbes in water varied in different sampling places and in one bakery all the water samples analysed were of good quality. Damaged, dirty hoses and taps can contaminate the water taken from the main water pipeline. The water posts available in the factory must all be in regular use to avoid accumulation of biofilms in the pipelines and hoses. Damaged gaskets, hoses etc. parts that wear must be maintained. 7 (9)
CONCLUSIONS AIR The quality of air was in three bakeries at same level in all parts of factory and the microbe level was similar with outdoor air. In three bakeries the air quality was better in packaging area than in other parts. In one bakery the total bacterial count of air inside the factory was smaller than in the outdoor air indicating that the doors and windows were kept closed as they should. The other five bakeries were told that the airborne contamination risks can be reduced by keeping windows closed, by installing air filters, by separating areas and by arranging the air to flow from the high hygiene areas to the lower hygiene areas. CONCLUSIONS PATHOGENES In all of these six hygiene surveys neither Salmonella spp. nor Listeria monocytogenes were found. However Listeria innocua was found in one sample. Raw materials and product samples were generally of good quality 8 (9)
ACKNOWLEDGEMENT Special thanks to: Dr. Gun Wirtanen (VTT) Dr. Nastasia Belc (IBA, Institutul de Bioresurse Alimentare ) Dr. Gabriela Nedita (IBA) Dr. Valeria Gagiu (IBA) Cooperation bakeries in Romania SAFOODNET project (FP6 022808 2006) 9 (9)