Biochemical indexes and gut microbiota testing as diagnostic methods for Penaeus monodon health and physiological changes during AHPND infection with food safety concerns

Abstract

Severe shrimp disease outbreaks have a destructive impact on shrimp aquaculture and its associated downstream food processing industries. Thus, it is essential to develop proper methods for shrimp disease control, which emphasizes the importance of food safety. In this study, we performed biochemical tests and gut microbiome analysis using uninfected control and Vp AHPND ‐infected Penaeus monodon samples. Biochemical tests were performed to assess the phenoloxidase (PO) activity, respiratory Burst (RB) activity, nitrite concentration, superoxide dismutase (SOD) activity, total hemocyte count (THC), and total protein concentrations. Overall, upregulations were detected in these biochemical tests, which showed the activation of the immune response in P. monodon during acute hepatopancreatic necrosis disease (AHPND) infection, especially at 6 hpi and 12 hpi. Besides that, shrimp gut samples were collected and pooled ( n = 3), followed by DNA extraction, PCR amplification targeting the V3/V4 16S ribosomal RNA (rRNA) region, next‐generation sequencing (NGS), and bioinformatics analysis. Proteobacteria was the most abundant phylum in both samples. The Rhodobacteraceae family and Maritimibacter genus were proposed to be vital forshrimp health maintenance. Vp AHPND bacterial colonization and secondary Vibrio infections were postulated to have occurred based on the higher abundances of Vibrionaceae family and Vibrio genus in the Vp AHPND ‐infected sample. Firmicutes phylum together with Photobacterium and Aliiroseovarius genera were inferred to be pathogenic or related factors of AHPND infections. In conclusion, physiology (immune response activation) and gut microbiome changes of disease tolerant P. monodon during AHPND infection were identified. Both biochemical tests and 16S rRNA analysis are proposed as a combined strategy for shrimp health diagnosis for ensuring shrimp health maintenance, disease control, and food safety.

Gut microbiota of Pacific white shrimp ( Litopenaeus vannamei ) exhibits distinct responses to pathogenic and non-pathogenic Vibrio parahaemolyticus

“…Vibrionaceae has served as the signature for AHPND diagnosis ( 50 ). The higher abundances of the Vibrionaceae family and Vibrio genus were postulated to be caused by the colonization of the AHPND-causing Vp and related to secondary Vibrio infections ( 51 ). Apart from some highly virulent strains causing the primary disease, Vibrio spp.…”

Enterocytozoon hepatopenaei Infection in Shrimp: Diagnosis, Interventions, and Food Safety Guidelines

“…This is due to the fact that infected shrimp exhibit no clear clinical symptoms over a short period of time, and healthy shrimp may become infected with EHP by cohabiting with diseased shrimp. In addition, biochemical assays and 16S rRNA analyses are proposed as appropriate diagnostic methods to determine shrimp gut microbiome changes and health status in order to ensure food safety [ 36 ]. As a result, it is critical to develop an effective approach to detect EHP infection in shrimp, particularly in the early stages of infection.…”

Perspective on intestinal microbiota temporal changes of herbal additives treated shrimp in a natural aquaculture setting

“…Research on aquatic organisms, especially economically important aquaculture animals, has focused on disease-related topics, including pathogens such as Vibrio species (de Souza Valente et al, 2020;Gainza and Romero, 2020;Soo and Bhassu, 2022). Many non-pathogenic strains within the Vibrio genus are important for crustaceans due to their ability to produce chitinases (Sugita and Ito, 2006), which may support healthy gut function.…”