False kelpfish (Sebastiscus marmoratus) is one of the target species in artificial breeding in China, and is susceptible to infection by Cryptocaryon irritans, which is an obligate parasitic ciliate that lives in the epithelium of the fish gills, skin and fins. Here, we sought to understand the mechanisms of molecular immunity of S. marmoratus against C. irritans infection. We carried out an extensive analysis of the transcriptome of S. marmoratus immune-related tissues. A paired-end library was constructed from the cDNA synthesized using a Genomic Sample Prep Kit. Five normalized cDNA libraries were constructed using RNA from the control group and the four groups of C. irritans-infected fish. The libraries were sequenced on an Illumina Mi-Seq platform, and functional annotation of the transcriptome was performed using bioinformatics software. The data produced a total of 149,983,397 clean reads from five cDNA libraries constructed from S. marmoratus immune-related tissues. A total of 33,291 unigenes were assembled with an average length of 1768 bp. In eggNOG (Evolutionary Genealogy of Genes: non-supervised orthologous groups) categories, 333 unigenes (0.94%) were assigned to defense mechanisms. In the immune system process sub-categories of gene ontology (GO) enrichment analysis, with the passage of time post-infection, the number of differentially expressed genes (DEGs) was reduced from 24 h to 48 h but then increased from 72 h to 96 h. Specifically, the immune-related differentially expressed genes (IRDEGs), which belong to the KEGG (Kyoto encyclopedia of genes and genomes) pathways, such as the complement and coagulation cascades, chemokine signalling pathways and toll-like receptor signalling pathways were mainly observed at 24 h post-infection. Infection with C. irritans resulted in a large number of DEGs in the immune-related tissues of S. marmoratus. The rapid and significant response of the S. marmoratus immune signalling pathways following C. irritans infection may be associated with their involvement in the immune process.
In 2019, a disease outbreak struck a hybrid sturgeon farm (Huso dauricus ♀ × A. schrenckii ♂) in Tiantai, Zhejiang province, leading to the deaths of 8000 sturgeons. The sturgeons exhibited reduced appetite, lethargic and uncoordinated swimming, and physical signs such as reddish petechiae and ulcers on the body and fins. Hemorrhagic spots were observed on the kidneys, spleen, and gonads, alongside reddish intestines with hemorrhagic ascites in the abdominal cavity. ST-1902 was isolated and identified as Aeromonas hydrophila through physiological and biochemical characterization and 16S rDNA sequence analysis. The pathogenicity of ST-1902 was confirmed through a challenge test, with a median lethal dosage (LD50) of 7.9 × 106 CFU/IND. Histopathological examination showed hyperplasia and neoplasm-like changes in the epicedial mesothelial tissues, enlarged and necrosis renal tissue, and serious hemosiderosis in spleen and gills. Virulent genes (Aer, Epa, Alt, Hly, and Act) were detected in ST-1902, corresponding to typical β-hemolysis, extracellular protease, and enterotoxin. Moreover, antimicrobial experiment detection indicated ST-1902 is sensitive to quinolones and phenicols but resistant to sulfamethoxazole, aminoglycoside antibiotics with Sul1, and Intl and Ant (3”)-I. These results suggest that A. hydrophila was the causative agent of the sturgeon disease and highlight the emerging threat it poses to the sturgeon industry.
Kudoa (Myxosporea: Multivalvulida) parasites are critical pathogens in marine and freshwater fish associated with significant economic losses and reduced market prices caused by post-mortem myoliquefaction or numerous cysts on muscles. In the present study, large yellow croakers infected by Kudoa were found during fish disease surveillance in China in November 2020 and used for morphological observation and characterization using light DIC microscopy and electron microscopy. Numerous creamy-white oval plasmodia were observed in muscles and on the surface of brain cartilage, gill arches, and serosal surfaces. The spores were considerably longer and thicker than previously reported Kudoa, with protruding polar filaments (PFs) in the mature spores, fingertip-shaped apical projections (APs), and polar capsules. Phylogenetic analyses with SSU rDNA, LSU rDNA, and mitochondrial DNA showed that the Kudoa-infected sample (LcK-2020) had the highest similarity to Kudoa iwatai reported in Japan. Based on the morphological characterization and phylogenetic analysis, it could be concluded that the sample LcK-2020 was infected by Kudoa iwatai, which would be the first report of Kudoa iwatai infection in large yellow croaker in China.
Enterocytozoon hepatopenaei (EHP) is an important factor limiting the growth of Penaeus vannamei. Its simple transmission route and lack of effective control drugs often lead to great losses for the shrimp aquaculture industry. In this study, some basic information of EHP was provided, including EHP spore purification, germination promoting treatment, quantification determination of the key genes expression, and the activities determination of key enzymes in glycolysis pathway at the stage of spore germination. The result showed that light microscopic and scanning electron microscopic analyses confirmed KOH promoted germination. The expression of the glyceraldehyde 3-phosphate dehydrogenase gene was stable, with high-level expression in treatment groups. Hexokinase gene expression and enzyme activity increased after promoting germination treatment. These findings indicated that the glycolytic pathway is enhanced during EHP germination. The results suggest that we can prevent EHP from invading host cells by attempting to block the energy supply.