Objective:To investigate the characterization of the microbe contamination of common animal decoction pieces and typical bacteria isolated from animal decoction pieces. Methods: 16 kinds including 46 batches of animal decoction pieces were collected. In light of the methods described as Chinese Pharmacopoeia (2020), the total aerobic microbial count (TAMC), total yeast count (TYMC), and thermophilic bacteria of samples were detected, the control bacteria were isolated from the samples. Multiple linear regression analysis was used to explore the factors related to microbe contamination load. Typical bacteria in the samples on the selective isolation mediums were isolated, and identified by the 16S rRNA amplicon sequencing analysis. Results: The TAMC of 37.0% samples exceeded 104 CFU·g-1, the TYMC of 32.6% samples were higher than 103 CFU·g-1, and the thermophilic bacteria were detected in 60.9% of the samples. The mean of lg TYMC of samples without guts was higher than those containing guts (P=0.01), the unqualified rate of TYMC of the samples without guts was higher than those containing guts (χ2=7.59, P=0.00). The mean value of lg TYMC of unheated samples was higher than those heated (P=0.02), the unqualified rate of TYMC of unheated samples was higher than heated samples (χ2=3.99, P=0.04). Bile-resistant gram-negative bacteria were detected in 13.0% (6/46) of the samples, the bile-resistant gram-negative bacteria load of 8.7% (4/46) samples exceeded 104 MPN·g-1. Escherichia coli was detected in 8.7% (4/46) of the samples, and Pseudomonas aeruginosa was detected in one batch of Cicadae Periostracum. The results of multivariate linear regression analysis indicated that the TAMC load was correlated with the load of thermophilic bacteria (P=0.00). The morphology and Gram’s staining properties of typical bacteria were observed under a microscope, the results of 16S rRNA sequence analysis indicated that a total of 55 typical bacteria strains were obtained including 47.3% of Bacillus, 23.6% of Enterobacter, and 16.4% of Enterococcus from the samples. Opportunistic pathogens such as Bacillus cereus, and the members of Enterobacter cloacae complex were detected in the samples. Conclusion: Animal decoction pieces containing tissue show a microbe contamination pattern as high microbe load, high level of thermophilic bacteria contamination, low level of bile-resistant gram-negative bacteria contamination. Bacillus is the most common typical isolates in the samples, more attention should be paid on the opportunistic pathogens contaminated in the samples.
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