Rise and metabolic roles of Vibrio during the fermentation of crab paste.

Microbial community may systematically promote the development of fermentation process of foods. Traditional fermentation is a spontaneous natural process that determines a unique nutritional characteristic of crab paste of Portunus trituberculatus, However, rare information is available regarding the development pattern and metabolic role of bacterial community during the fermentation of crab paste. Here, using a 16S rRNA gene amplicon sequencing technology, we investigated dynamics of bacterial community and its relationship with metabolites during the fermentation of crab paste. The results showed that bacterial community changed dynamically with the fermentation of crab paste which highlighted by consistently decreased α-diversity and overwhelming dominance of Vibrio at the later days of fermentation. Vibrio had a positive correlation with trimethylamine, hypoxanthine, formate, and alanine while a negative correlation with inosine and adenosine diphosphate. In contrast, most of other bacterial indicators had a reverse correlation with these metabolites. Moreover, Vibrio presented an improved function potential in the formation of the significantly increased metabolites. These findings demonstrate that the inexorable rise of Vibrio not only drives the indicator OTUs turnover in the bacterial community but also has incriminated the quality of crab paste from fresh to perished.

ATP catabolism and bacterial succession in postmortem tissues of mud crab (Scylla paramamosain) and their roles in freshness.

The seafood microbiome is highly diverse and plays an essential role in the spoilage of seafood. Nevertheless, how such a diverse microbiome influences freshness of mud crab (Scylla paramamosain) remains unclear. Here we investigated the postmortem ATP catabolism and succession of the bacterial community in the hepatopancreas and muscle of S. paramamosain using a high-performance liquid chromatography method and 16S rRNA gene amplicon sequencing. Our results showed a tissue-dependent change in ATP catabolism determinized the differences in the changes of nucleotide freshness indices of hepatopancreas and muscle over postmortem time of mud crab. The muscle K value could be used as an optimal nucleotide freshness indicator for the freshness of mud crab, with a proposed threshold of 20%. From a microbiota perspective, a more significant bacterial community change was observed in the muscle than in the hepatopancreas. These changes could result in a close relationship between ATP and its catabolites and microbial taxa in the muscle. Moreover, Photobacterium, Peptostreptococcaceae, average path distance, OTU richness, and Shannon index of muscle bacterial community markedly contributed to K value. These findings suggest that the mud crab of 4 h postmortem at room temperature is still edible. Notably, the importance of microbial community composition and interaction for the spoilage of mud crab should be carefully considered.

Tissue-specific metabolic responses of Cyprinus flammans to copper.

Copper (Cu) contamination is serious in China, with ≤2.76 mg/L in some waters. Exposure to Cu causes a high toxicity to the aquatic organisms and subsequent ecological risk. To understand fish responses to Cu exposure, we analyzed the metabonomic changes in multiple tissues (gill, liver, and muscle) of Cyprinus flammans using an nuclear magnetic resonance-based metabonomic technique. Our results showed that metabolic alterations are dose-dependent. No significant metabolic alterations in three tissues of fish are caused by 0.25 mg/L Cu. However, 1.53 mg/L Cu caused changes of energy-related metabolites and amino acids, which we suggest are due to enhanced metabolic acidosis in gill and muscle, decreased tricarboxylic acid cycle activity in muscle, increased gluconeogenesis from amino acids in liver, and improved glycogenesis in liver and muscle. The Cori cycle between liver and muscle is concurrently triggered. Furthermore, high concentration of Cu resulted in the alteration of choline metabolism such that we hypothesize that Cu induces membrane damage and detoxification of CuSO4 in gill as well as altered osmoregulation in all three tissues. Choline-O-sulfate in gill may be used as a biomarker to provide an early warning of Cu exposure in C. flammans. Moreover, Cu exposure caused alterations of nucleoside and nucleotide metabolism in both gill and muscle. These findings provide a new insight into the metabolic effects of Cu exposure on C. flammans and highlight the value of metabonomics in the study of metabolic metal disturbance in fish.

Isolation and characterization of phenanthrene-degrading Sphingomonas paucimobilis strain ZX4.

Phenanthrene-degrading bacterium strain ZX4 was isolated from an oil-contaminated soil, and identified as Sphingomonas paucimobilis based on 16S rDNA sequence, cellular fatty acid composition, mol% G + C and Biolog-GN tests. Besides phenanthrene, strain ZX4 could also utilize naphthalene, fluorene and other aromatic compounds. The growth on salicylic acid and catechol showed that the strain degraded phenanthrene via salicylate pathway, while the assay of catechol 2,3-dioxygenase revealed catechol could be metabolized through meta-cleavage pathway. Three genes, including two of meta-cleavage operon genes and one of GST encoding gene were obtained. The order of genes arrangement was similar to S-type metapathway operons. The phylogenetic trees based on 16S rDNA sequence and meta-pathway gene both revealed that strain ZX4 is clustered with strains from genus Sphingomonas.

Characterization and phylogenetic analysis of a phenanthrene-degrading strain isolated from oil-contaminated soil.

Bacterium strain EVA17 was isolated from an oil-contaminated soil, and identified as Sphingomonas sp. based on analysis of 16S rDNA sequence, cellular fatty acid composition and physiological-chemical tests. The salicylate hydroxylase and catechol 2, 3-dioxygenase (C230) were detected in cell-free lysates, suggesting a pathway for phenanthrene catabolism via salicylate and catechol. Alignment showed that both of the C230 and GST genes of the strain EVA17 had high similarity with homologues of strains from genus Sphingomonas. The phylogenetic analysis based on 16S rDNA and C230 gene sequence indicated that EVA17 should be classified into genus Sphingomonas, although the two phylogenetic trees were slightly different from each other. The results of coamplification and sequence determination indicated that GST gene should be located upstream of the C230 gene.

Characterization of a strain of Sphingobacterium sp. and its degradation to herbicide mefenacet.

A bacterium (designated strain Y1) degrading acetanilide herbicide mefenacet was isolated from aerobic sludge. Based on the analyses of partial 16S rRNA gene, cellular fatty acid and BIOLOG-GN, and general physiological and biochemical characteristics, strain Y1 was identified as Sphingobacterium multivolum. Strain Y1 was able to degrade mefenacet used as sources of carbon and energy. Degradation of mefenacet was accompanied by producing the metabolites N-methylaniline and an unidentified compound with molecular weight 205, indicating a metabolic pathway of mefenacet initiated by hydrolysis of amido bond.

Influences of quinclorac on culturable microorganisms and soil respiration in flooded paddy soil.

OBJECTIVE: To investigate the potential effects of herbicide quinclorac (3,7-dichloro-8-quinoline-carboxylic) on the culturable microorganisms in flooded paddy soil. METHODS: Total soil aerobic bacteria, actinomycetes and fungi were counted by a 10-fold serial dilution plate technique. Numbers of anaerobic fermentative bacteria (AFB), denitrifying bacteria (DNB) and hydrogen-producing acetogenic bacteria (HPAB) were numerated by three-tube anaerobic most-probable-number (MPN) methods with anaerobic liquid enrichment media. The number of methanogenic bacteria (MB) and nitrogen-fixing bacteria (NFB) was determined by the rolling tube method in triplicate. Soil respiration was monitored by a 102G-type gas chromatography with a stainless steel column filled with GDX-104 and a thermal conductivity detector. RESULTS: Quinclorac concentration was an important factor affecting the populations of various culturable microorganisms. There were some significant differences in the aerobic heterotrophic bacteria. AFB and DNB between soils were supplemented with quinclorac and non-quinclorac at the early stage of incubation, but none of them was persistent. The number of fungi and DNB was increased in soil samples treated by lower than 1.33 micro x g(-1) dried soil, while the CFU of fungi and HPAB was inhibited in soil samples treated by higher than 1.33 microg x g(-1) dried soil. The population of actinomycete declined in negative proportion to the concentrations of quinclorac applied after 4 days. However, application of quinclorac greatly stimulated the growth of AFB and NFB. MB was more sensitive to quinclorac than the others, and the three soil samples with concentrations higher than 1 microg x g(-1) dried soil declined significantly to less than 40% of that in the control, but the number of samples with lower concentrations of quinclorac was nearly equal to that in the control at the end of experiments. CONCLUSION: Quinclorac is safe to the soil microorganisms when applied at normal concentrations (0.67 microg x g(-1)).

Effects of butachlor on microbial enzyme activities in paddy soil.

This paper reports the influences of the herbicide butachlor (n-butoxymethl-chloro-2', 6'-diethylacetnilide) on microbial respiration, nitrogen fixation and nitrification, and on the activities of dehydrogenase and hydrogen peroxidase in paddy soil. The results showed that after application of butachlor with concentrations of 5.5 micrograms/g dried soil, 11.0 micrograms/g dried soil and 22.0 micrograms/g dried soil, the application of butachlor enhanced the activity of dehydrogenase at increasing concentrations. The soil dehydrogenase showed the highest activity on the 16th day after application of 22.0 micrograms/g dried soil of butachlor. The hydrogen peroxidase could be stimulated by butachlor. The soil respiration was depressed within a period from several days to more than 20 days, depending on concentrations of butachlor applied. Both the nitrogen fixation and nitrification were stimulated in the beginning but reduced greatly afterwards in paddy soil.