ABSTRACT
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.
ABSTRACT
The Chinese mitten crab, Eriocheirsinensis, is an important farmed crustacean species in China, outranking other farmed crabs in yield and economic importance. An infection called "milky disease", caused by the yeast, Metschnikowiabicuspidata, has emerged in E. sinensis farms in northeast China and has caused progressive economic losses. The diseased crabs present with opaque, whitish muscles and milky hemolymph. Currently, there are no effective drugs to treat the infection. Clarifying the transmission route of M. bicuspidata would help to treat and prevent the disease. We investigated the effects of three different M. bicuspidata infection methods (feeding, immersion, and cohabitation) on E. sinensis. All three infection methods led to a high infection rate in healthy crabs. After 35 d, the infection rate was 76.7%, 66.7%, and 53.3% in the feeding, immersion, and cohabitation groups, respectively. Diseased crabs exhibited the typical symptom of hemolymph emulsification, with a high pathogen load of M. bicuspidata. The yeast was not detected in the oocytes of infected crabs. Fertilized embryos, zoea larvae, and megalopae of infected ovigerous crabs tested negative for yeast, indicating that direct transmission from mother to offspring does not occur. Our results highlight avenues for the prevention and control of this yeast.
ABSTRACT
The Chinese grass shrimp (Palaemonetes sinensis) was found with a white turbidity appearance in the Panjin area. After dissection, typical symptoms of milky disease with hemolymph emulsification and noncoagulation were observed; however, the pathogen was unknown. In this study, we aimed to isolate the pathogen of the diseased P. sinensis. We found that the pathogen could grow on the fungal medium Bengal red, and microscopic examination showed that it reproduced by budding. Molecular identification of the isolated and purified yeast strain LNMB2021 based on 26S rDNA sequence showed that the pathogenic pathogen was Metschnikowia bicuspidata (GenBank OK094821), with 98.74% homology with M. bicuspidata strain LNES0119 (GenBank OK073903) and 98.56% with M. bicuspidata strain Liao (GenBank MT856369). The results of an artificial infection test showed that M. bicuspidata caused the same clinical symptoms in P. sinensis, and the isolated pathogen was still the same, which proved that P. sinensis was a new host of M. bicuspidata. Histopathological analysis showed that there were obvious pathological changes in the hepatopancreas and muscle tissue of the diseased P. sinensis. Identification of the pathogen is essential for the prevention and control of the disease and the healthy culture of P. sinensis. Furthermore, considering the transmissibility and cross-host transmission of M. bicuspidata, its risk of infecting other aquatic animals deserves high attention.
