A small amount of bacterial spores with strong tolerance still exist in raw milk after sterilization and can multiply under certain conditions, causing a serious impact on product quality and consumer health. In this study, raw milk samples containing Bacillus were collected to isolate aerobic Bacillus and thermophilic aerobic Bacillus. The isolates obtained were identified based on their morphological, physiological, biochemical and molecular biological characteristics, and the heat resistance of the isolated thermophilic aerobic Bacillus was studied. Results showed that 10 strains of Bacillus were isolated from the milk samples, of which three were thermophilic aerobic Bacillus and seven were aerobic Bacillus. Among them, Bacillus coagulans and Bacillus clausii could tolerate heat treatment at 115 ℃ for 15 min. All Bacillus in raw milk could be killed by heat treatment at 121 ℃ for 20 min which decreased the number of spores by four orders of magnitude. The dominant Bacillus species in raw milk were Bacillus subtilis, Bacillus cereus, Bacillus pumilus, Bacillus licheniformis, Bacillus circulans, Bacillus gelatini, Bacillus amyloliquefaciens, Bacillus coagulans, and Bacillus clausii. Bacillus coagulans and Bacillus clausii had stronger heat resistance and could tolerate heat treatment at 115 ℃ for 15 min.

In recent years, the addition of probiotics to milk powder has gradually become a research focus. In order to ensure the product quality and safety, a microbiological detection method for Lactobacillus fermentum CECT5716 in milk powder has been established. It was found that the population of Lactobacillus fermentum in milk powder could be accurately counted by spreading serial 10-fold dilutions in buffered peptone water (BPW) onto MRS plates and culturing them under anaerobic conditions at (36 ± 1) ℃ for (48 ± 2) h, with clear discrimination from inoculated bifidobacteria. This method was more suitable for counting Lactobacillus fermentum than the Chinese national standard method (GB 4789.35?2016), and had the advantages of high discrimination capacity, short culture time, simple operation, good repeatability, and high accuracy. This method also included 16S rDNA sequence analysis of the strain for its subsequent molecular identification. There was no significant difference in the accuracy of this method versus the national standard method in detecting 20 samples prepared in our laboratory (P = 0.127).

The residues of ochratoxin A, deoxynivalenol, aflatoxins B1 and zearalenone in dairy feeds were simultaneously detected using biochip array technology. The method accuracy, precision and reproducibility were evaluated. The results obtained that the correlation coefficient of the calibration curve for each analyte was greater than 0.99. The recoveries for quality control sample and negative sample at two spiked levels varied from 80% to 120%. The coefficient of variance (CV) was less than 15% for accuracy and reproducibility and was less than 10% for reproducibility. Compared with high performance liquid chromatography, this method gave consistent results with a simpler sample pretreatment procedure. In conclusion, the biochip array method was simple and accurate and it could provide an efficient and reliable tool for mass screening of samples.

Enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) allows gene amplification using primers designed based on ERIC, which are widely present in living organisms, yielding products that are useful to characterize the genomic structure. This method has been widely used in bacterial classification and research on the genetic relationship of strains because of its merits such as simplicity, ease of implementation, good repeatability and time saving. This paper reviews the background to bacterial classification research and the principle and characteristics of ERIC-PCR technique, and summarizes the advantages and disadvantages of ERIC-PCR technique in bacterial genotyping and recent progress in its application to genotype common food-borne pathogenic bacteria. At the same time, future prospects for the application of ERIC-PCR in traceability analysis of microbial contaminants in dairy enterprises.