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Amplification was performed in a MJ Mini Personal Thermal Cycler (Bio-Rad Laboratories Inc. USA). Fingerprinting, rRNA gene amplification, sequence analysis, treatment of amplification products, digital photo documentation and Computer-assisted analysis of the DNA patterns were performed as described previously (Olstorpe et?al. 2008). Moulds were identified based on morphological characteristics, following the procedures and taxonomic keys of Samson et?al. (2004). The data consisted of repeated measurements on the same units. Therefore, mixed linear models were used for analyses (Fitzmaurice et?al. 2004; Littell et?al. 2006). The dependence http://www.selleckchem.com/products/Fludarabine(Fludara).html between observations over time was modelled using an autoregressive covariance structure of the first order. For all response variables, the models included the effects of treatment, time and the interactions between these. The mixed procedure of the SAS (1999) package was used http://www.selleckchem.com/products/Nolvadex.html for analyses. Analyses were performed with the help of the Department of Economy, Applied Statistics and Mathematics, SLU, Uppsala. The number of LAB in the different diets was between 8��3 and 9��0?log units (CFU?g?1). This level was stable in the diets during the fermentation period and no significant difference was found between the treatments and the time points of measurements, indicating that the number of LAB was not influenced by diet preparation or start of backslopping (Table?1). The number of yeasts was not affected by the different treatments (P? https://en.wikipedia.org/wiki/Chlormezanone occasions with numbers close to the detection limit (102?CFU?g?1 grain). As their number did not increase during the undisturbed initial fermentation or after starting the backslopping, we assume that detected moulds arose from fungal spores present in the ingredients that survived the fermentation, but not from actively growing fungi. The amount of total aerobic bacteria was stable during the undisturbed fermentation period in all treatments. However, their number was higher in diet WWDGsc1 and diet WWDGsc5 than in diet WWDG. After backslopping, the number of total aerobic bacteria decreased in WWDGsc1 and increased in WWDGsc5. To survey the aerobic bacteria flora, we selected five colonies randomly from TGEA quantification plates grown from each triplicate of all three treatments at the last sampling occasion, and identified them by 16S rRNA gene sequencing. The test showed that 39 of 45 tested colonies belonged to LAB species, including the species Ent.? faecium (found nine times within the 45 isolates), Lactobacillus brevis (1), Lactobacillus fermentum (4), Lactobacillus mucosae (3), Lactobacillus panis (1), Lactobacillus paracasei (2), Lact.?plantarum (18) and Ped.?pentosaceus (1).