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Milky disease caused by Metschnikowia bicuspidata fungus has significantly harmed the Chinese mitten crab Eriocheir sinensis aquaculture industry. However, the effect of M. bicuspidata infection on the metabolism and intestinal flora of the crab remains unclear. In this study, we aimed to explore the changes in the metabolism and intestinal flora E. sinensis after 48 h of infection with M. bicuspidata, using metabolomic and metagenomic analyses. Metabolomic analysis results revealed 420 significantly different metabolites between the infected and control groups, and these metabolites were enriched in 58 metabolic pathways. M. bicuspidata infection decreased the levels of metabolites related to amino acid biosynthesis, the tricarboxylic acid cycle, as well as lysine, histidine, linolenic, arachidonic, and linoleic acid metabolism. These results indicated that M. bicuspidata infection significantly affected the energy metabolism, growth, and immunity of E. sinensis. The results of metagenomic analysis showed that the anaerobes and ascomycetes populations significantly increased and decreased, respectively, after M. bicuspidata infection. These changes in intestinal flora significantly upregulated metabolic and synthetic pathways while downregulating immunity-related pathways. The results of integrated metabolomic and metagenomic analyses showed that 55 differentially expressed genes and 28 operational taxonomic units were correlated with 420 differential metabolites. Thus, the intestinal flora changes caused by M. bicuspidata infection also affected the metabolites. This study provides novel insights into the metabolic-and intestinal microflora-based effects of M. bicuspidata infection in E. sinensis, as well as a theoretical basis for the interaction between fungi and crustaceans.

Metschnikowia bicuspidata is a globally distributed pathogenic yeast with a wide range of aquatic hosts. A new strain, M. bicuspidata LNES0119, isolated from the Chinese mitten crab Eriocheir sinensis, has caused a serious reduction in production and marked economic loss for the aquaculture industry in China. Therefore, the whole-genome sequence of M. bicuspidata LNES0119 was sequenced using Illumina and Oxford Nanopore technology; whole-genome annotation and comparative genomic analyses of this pathogen were performed as well. A high-quality genome of M. bicuspidata LNES0119 was 16.13 Mb in size, with six scaffolds and six contigs, and encoded 5,567 putative predicted genes. Of these, 1,467 genes shared substantial homology with genes in the pathogen–host interactions database. Comparative genomic analyses of three M. bicuspidata strains and one non-pathogenic yeast, M. aff. pulcherrima, showed 331 unique genes in M. bicuspidata LNES0119, 30 of which were putatively related to pathogenicity. Overall, we identified several meaningful characteristics related to pathogenicity and virulence that may play essential roles in the infection and pathogenicity of M. bicuspidata LNES0119. Our study will aid in identifying potential targets for further exploration of the molecular basis of the pathogenicity of M. bicuspidata as well as the therapeutic intervention of M. bicuspidata infection.

Background: Subgenotype C4 of enterovirus 71 (EV71) is the predominant agent of Hand Foot and Mouth disease (HFMD) circulating in the mainland of China. For the first time, a subgenotype C2 of EV71 named SY30-2 was isolated from a HFMD case in Beijing, China. Since it is uncertain whether antibodies raised against subgenotype C4 of EV71 can protect C2 EV71, it is important to monitor and check the presence of cross-reactive antibodies against new EV71 subgenotypes. To find out the causes for the different NtAb, this study is to investigate the relationships between amino acid residue variations and cross-reactive antibodies against EV71 subgenotypes C2 and C4.Methods: Nucleotide and amino acid sequences from full-length genome sequence of SY30-2 were compared to EV71 reference strains. A microneutralization test was used to detect neutralizing antibody (NTAb) in the sera of subgenotype C4 of EV71 infected cases against SY30-2 and FY17 (a C4 isolate). The 3D structure of the viral capsid protein of SY30-2 was constructed.Results: Genome sequence and similarity plot analyses showed that SY30-2 shared the highest identity with subgenotype C2 of EV71 strains in every fragment of the genome. While the microneutralization test result showed that children infected with subgenotype C4 of EV71 had higher NTAb titers against FY17 than SY30-2 (p < 0.001). The amino acid sequence comparison revealed that four amino acid residues VP1-22, VP1-31, VP1-249 and VP3-93 were highly conserved in subgenotype C4 of EV71 compared with the corresponding amino acid residues on subgenotype C2 of EV71 (p < 0.05). Furthermore, the 3D-structure of viral capsid protein showed that VP1-22, VP1-31 and VP3-93 were located on the surface of virion.Conclusion: This is the first report of an EV71 subgenotype C2 isolated from HFMD in Beijing, China. Only a few antigenic variations on subgenotype C2 of EV71 could have led to a great decrease in NTAb titer. Thus, imported new genotypes and subgenotypes of EV71 should be closely monitored. The efficacy of available vaccines against new viruses should be evaluated as well.

Bacterial pathogens have been confirmed as the major cause of acute diarrhea among outpatients in China. In this study, 370 stool samples from the patients aged from 15 to 87 years old with diarrhea were collected over 12 months (from May 2016 to April 2017) in two hospitals in Shunyi, Beijing. Bacterial isolation was performed for the common enteric pathogens: Campylobacter, Salmonella, Shigella, Diarrheagenic Escherichia coli, and Vibrio parahaemolyticus for 370 samples. The filtration method was used for the Campylobacter isolation in this study. The prevalence and molecular characterization of the Campylobacter were investigated. The isolation ratio for Campylobacter, Salmonella, Shigella, Diarrheagenic E. coli, and V. parahaemolyticus was 7.0% (26/370), 6.2% (23/370), 0.3% (1/370), 7.3% (27/370), and 10.3% (38/370), respectively. Based on the isolation result, Campylobacter positive cases presented in almost every month of the whole year and the isolation ratio was the highest among the tested pathogens during October to March. There was no significant difference between genders of Campylobacter positive cases. More Campylobacter positive cases presented dehydration compared with those who were positive for Salmonella. Twenty-six Campylobacter isolates were obtained in this study and 24 of these were Campylobacter jejuni. The antibiotic susceptibility tests indicated that 83.3% (20/24) of the isolates exhibited resistance to three or more types of antibiotic. Twenty STs were identified for the 26 Campylobacter isolates and four novel STs were identified in this study. No clonal cluster was found among these isolates. This is the first study for Campylobacter isolated using the filtration method in China which indicated the Campylobacter infection might be seriously under-ascertained in the diarrheal patients in China.