Microbial Diversity and Community Structure of Chinese Fresh Beef during Cold Storage and Their Correlations with Off-Flavors.

To investigate the diversity and dynamics of microorganisms in Chinese fresh beef (CFB) without acid discharge treatment during cold storage, high-throughput sequencing was employed to analyze the CFB refrigerated for 0, 3, 7, and 10 days. The results showed that the community richness of the fungi and bacteria decreased significantly. However, the diversity decreased in the early stage and increased in the later stage. At the phylum level, Ascomycota (74.1-94.1%) and Firmicutes (77.3-96.8%) were the absolutely dominant fungal and bacterial phyla. The relative abundance of both fungal and bacterial phyla displayed a trend of increasing and then decreasing. At the genus level, Candida (29.3-52.5%) and Lactococcus (19.8-59.3%) were, respectively, the dominant fungal and bacterial genera. The relative abundance of Candida showed a trend of increasing and then decreasing, while Lactococcus possessed the opposite trend. KEGG metabolic pathways analysis suggested that carbohydrate metabolism, membrane transport, and amino acid metabolism were the major metabolic pathways of bacteria. Bugbase prediction indicated the major microbial phenotype of bacteria in CFB during cold storage was Gram-positive (17.2-31.6%). Correlation analysis suggested that Lactococcus, Citrobacter, Proteus, and Rhodotorula might be the main microorganisms promoting the production of off-flavor substances in CFB. This study provides a theoretical basis for the preservation of Chinese fresh beef.

The Contribution of Microorganisms to the Quality and Flavor Formation of Chinese Traditional Fermented Meat and Fish Products.

Guizhou sour meat and sour fish, Chaoshan fish sauce, Sichuan sausage and bacon, Cantonese sausage, Jinhua ham, and Xinjiang air-dried beef are eight representatives of Chinese traditional fermented meat and fish products (FMFPs), which are favored by Chinese consumers due to their high nutritional value and quality. The quality of the spontaneously fermented Chinese traditional FMFP is closely correlated with microorganisms. Moreover, the dominant microorganisms are significantly different due to regional differences. The effects of microorganisms on the texture, color, flavor, nutrition, functional properties, and safety of Chinese traditional FMFPs have not been not fully described. Additionally, metabolic pathways for flavor formation of Chinese traditional FMFPs have not well been summarized. This article describes the seven characteristic Chinese traditional FMFPs and correlated dominant microorganisms in different regions of China. The effects of microorganisms on the texture, color, and flavor of Chinese traditional FMFPs are discussed. Furthermore, the metabolic pathways of microbial regulation of flavor formation in Chinese traditional FMFPs are proposed. This work provides a theoretical basis for improvement of Chinese traditional FMFPs by inoculating functional microorganisms isolated from Chinese traditional fermented foods.

The Volatile Flavor Substances, Microbial Diversity, and Their Potential Correlations of Inner and Surface Areas within Chinese Qingcheng Mountain Traditional Bacon.

The objective of this study was to explore the microbial diversity, volatile flavor substances, and their potential correlations in inner and surface Chinese Qingcheng Mountain traditional bacon (CQTB). The results showed that there were 39 volatile flavor substances in inner and surface CQTB detected by headspace solid-phase microextraction and gas chromatography-mass spectrometry (HS-SPME-GC-MS). Moreover, significant differences in volatile flavor substances between the inner and surface CQTB were observed. Sixteen key volatile flavor substances were screened (OAV > 1), including guaiacol, nonanal, ethyl isovalerate, and others. High-throughput sequencing (HTS) result indicated that Firmicutes, Proteobacteria, and Actinobacteria were the predominant bacterial phyla, and Ascomycota and Mucoromycota were the predominant fungal phyla. Staphylococcus, Psychrobacter, and Brochothrix were the predominant bacteria, and Debaryomyces, Penicillium, and Mucor were the predominant fungal genera. Spearman correlation coefficient analysis suggested that Apiotrichum and Lactobacillus were closely and positively correlated with the formation of key phenol compounds. The present work demonstrates the microbial diversity and related volatile flavor substances and their potential correlations in CQTB and provides a theoretical basis for the development of microbial starter culture and green processing of CQTB.

Potential Correlation between Microbial Diversity and Volatile Flavor Substances in a Novel Chinese-Style Sausage during Storage.

A novel Chinese-style sausage with Chinese traditional fermented condiments used as additional ingredients is produced in this study. The aim of this study was to investigate the microbial community's structure, the volatile flavor substances and their potential correlation in the novel Chinese sausage. High-throughput sequencing (HTS) and solid-phase microextraction-gas chromatography-mass spectrometry (GC-MS) were, respectively, used to analyze the microbial diversity and volatile flavor substances of the novel Chinese-style sausage during storage. The results showed that Firmicutes, Proteobacteria and Actinobacteria were the predominant bacterial genera, and Hyphopichia and Candida were the predominant fungal genera. A total of 88 volatile flavor substances were identified through GC-MS, among which 18 differential flavor compounds were screened (VIP > 1), which could be used as potential biomarkers to distinguish the novel sausages stored for different periods. Lactobacillus exhibited a significant negative correlation with 2,3-epoxy-4,4-dimethylpentane and acetoin and a significant positive correlation with 2-phenyl-2-butenal. Hyphopichia significantly positively correlated with ester. Leuconostoc significantly positively correlated with ethyl caprate, ethyl palmate, ethyl tetradecanoate and ethyl oleate while it negatively correlated with hexanal. This study provides a theoretical basis for revealing the flavor formation mechanisms and the screening of functional strains for improving the flavor quality of the novel Chinese-style sausage.

GC-TOF-MS-Based Non-Targeted Metabolomic Analysis of Differential Metabolites in Chinese Ultra-Long-Term Industrially Fermented Kohlrabi and Their Associated Metabolic Pathways.

Fermented kohlrabi is a very popular side dish in China. Chinese kohlrabies industrially fermented for 0 years (0Y), 5 years (5Y), and 10 years (10Y) were employed and analyzed by non-targeted metabolomics based on GC-TOF-MS, and the differential metabolites were screened using multivariate statistical analysis techniques, including principal component analysis (PCA) and orthogonal partial least squares discrimination analysis (OPLS-DA). The results showed that 47, 38, and 33 differential metabolites were identified in the three treatment groups of 0Y and 5Y (A1), 0Y and 10Y (A2), and 5Y and 10Y (A3), respectively (VIP > 1, p < 0.05). The metabolites were mainly carbohydrates, amino acids, and organic acids. Furthermore, 13 differential metabolites were screened from the three groups, including L-glutamic acid, L-aspartic acid, γ-aminobutyric acid, and other compounds. Four metabolic pathways termed alanine, aspartate, and glutamate metabolism, arginine biosynthesis, arginine and proline metabolism, and glycolysis/gluconeogenesis were the most significant pathways correlated with the differential metabolites, as analyzed according to the Kyoto Encyclopedia of Genes and Genomes (KEGG). The odors for the three ultra-long-term industrially fermented kohlrabies were significantly different, as detected by E-nose. The present work describes the changes in metabolites between different ultra-long-term industrially fermented kohlrabies and the associated metabolic pathways, providing a theoretical basis for the targeted regulation of characteristic metabolite biosynthesis in Chinese fermented kohlrabi.

Coexistence of Bacillus thuringiensis (Bt)-transgenic and conventional rice affects insect abundance and plant fitness in fields.

BACKGROUND: As genetically modified (GM) crops are cultivated worldwide, concerns are emerging about the ecological consequences of the coexistence of transgenic and non-transgenic crops in fields. We first conducted field experiments using insect-resistant transgenic rice expressing Bacillus thuringiensis (Bt-transgenic rice) and its counterpart conventional rice (Oryza sativa L.) with or without insecticide spraying in 2013 and 2014. In 2015 and 2016, Bt-transgenic and conventional rice plants were employed in pure and mixed cages, with an infestation of the target insect (Chilo suppressalis) and with insecticide spraying as the control treatment to prevent target insect infestation. RESULTS: The presence of Bt-transgenic rice decreased the abundance of target insects but did not affect non-target insects and predators in fields. Compared with conventional rice, Bt-transgenic rice showed more empty seeds but comparable seed production in cages. The infestation of target insects significantly decreased the plant fitness of conventional rice in pure cages, but did not affect its fitness when conventional rice coexisted with Bt-transgenic rice. In mixed cages, the presence of Bt-transgenic rice decreased the abundance of target insects and the percentage of dead sheaths in conventional rice. CONCLUSION: The presence of Bt-transgenic rice benefits the growth and reproduction of non-transgenic rice in fields because of a decreased abundance of target insects. © 2018 Society of Chemical Industry.

Cut site selection by the two nuclease domains of the Cas9 RNA-guided endonuclease.

Cas9, the RNA-guided DNA endonuclease from the CRISPR-Cas (clustered regularly interspaced short palindromic repeat-CRISPR-associated) system, has been adapted for genome editing and gene regulation in multiple model organisms. Here we characterize a Cas9 ortholog from Streptococcus thermophilus LMG18311 (LMG18311 Cas9). In vitro reconstitution of this system confirms that LMG18311 Cas9 together with a trans-activating RNA (tracrRNA) and a CRISPR RNA (crRNA) cleaves double-stranded DNA with a specificity dictated by the sequence of the crRNA. Cleavage requires not only complementarity between crRNA and target but also the presence of a short motif called the PAM. Here we determine the sequence requirements of the PAM for LMG18311 Cas9. We also show that both the efficiency of DNA target cleavage and the location of the cleavage sites vary based on the position of the PAM sequence.

[Control efficacy of Trichogramma japonicum against Chilo suppressalis and Chilaraea auricilia].

In order to understand the relationships of the population increase capacities between egg parasitoid Trichogramma japonicum Ashmead (Hymenoptera: Trichogrammatidae) and two rice borers Chilo suppressalis Walker and Chilaraea auricilia Dudgeon (Lepidoptera: Pyralidae), and to evaluate the control efficacy of T. japonicum against C. suppressalis and C. auricilia, life table method was adopted to establish the experimental population life tables of the two rice borers and the population fecundity tables of T. japonicum on the two hosts, and to estimate the fecundity and parasitism capacity of T. japonicum on the two hosts. The generation period (T) of C. suppressalis and C. auricilia was 56.40 d and 47.80 d, and their intrinsic rate of natural increase (r(m)) was 0.0489 and 0.072; while the T of T. japonicum on C. suppressalis and C. auricilia was 9.75 d and 9.78 d, and the r(m) was 0.3154 and 0.3161, respectively. These parameters indicated that T. japonicum could control the two rice borers effectively.