Enterocytozoon hepatopenaei (EHP) is a parasite in shrimp farming. EHP mainly parasitizes the hepatopancreas of shrimp, causing slow growth, which severely restricts the economic income of shrimp farmers. To explore the pathogenic mechanism of EHP, the host subcellular construction, molecular biological characteristics, and mitochondrial condition of Litopenaeus vannamei were identified using transmission electron microscopy (TEM), real-time qPCR, an enzyme assay, and flow cytometry. The results showed that EHP spores, approximately 1 μm in size, were located on the cytoplasm of the hepatopancreas. The number of mitochondria increased significantly, and mitochondria morphology showed a condensed state in the high-concentration EHP-infected shrimp by TEM observation. In addition, there were some changes in mitochondrial potential, but apoptosis was not significantly different in the infected shrimp. The qPCR results showed that the gene expression levels of hexokinase and pyruvate kinase related to energy metabolism were both upregulated in the diseased L. vannamei. Enzymatic activity showed hexokinase and lactate dehydrogenase were significantly increased in the shrimp infected with EHP, indicating EHP infection can increase the glycolysis process and decrease the oxidative phosphorylation process of L. vannamei. Previous transcriptomic data analysis results also support this conclusion.
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.
The large yellow croaker (Larimichthys crocea), one of the most economically valuable marine fish species in China, suffers significant economic losses in aquaculture due to infectious diseases caused by marine pathogens, such as Nocardia seriolae. The pathogenic mechanisms underlying N. seriolae infection in L. crocea and the host immune responses remain inadequately characterized. To investigate the molecular mechanisms of this infection, we conducted transcriptome sequencing on the head kidney tissues of L. crocea at 1, 3, 7, and 14 days post-infection with N. seriolae. In total, 421, 1052, 3215, and 2459 upregulated genes, along with 1853, 1777, 3718, and 3134 downregulated genes were identified, respectively. KEGG enrichment analysis revealed that differentially expressed genes were predominantly associated with immune and metabolic pathways. Notably, pathways involved in Toll-like receptor signaling, ECM–receptor interaction, cytokine–cytokine receptor interaction, and focal adhesion were significantly enriched, highlighting an immune response to N. seriolae infection in L. crocea. In addition, significant enrichment of the citrate cycle (TCA cycle) and oxidative phosphorylation pathways in metabolic processes suggests an upregulated ATP synthesis to meet the heightened energy demand associated with the immune response to infection. These findings contribute to a deeper understanding of the immune defense mechanisms in the head kidney of L. crocea against N. seriolae infection and elucidate aspects of N. seriolae pathogenicity.
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.