Compensatory response of Hong Kong oysters to co-occurring stressors: Zinc oxide nanoparticles and low salinity exposure.

Zinc Oxide nanoparticles (ZnO NPs), due to their ubiquitous use in industrial and consumer applications, present potential risks to marine ecosystems and biota, especially oysters. The physiological and immunological health of marine species is highly dependent on salinity levels. However, the combined impact of lowered salinity and exposure to ZnO NPs, particularly on key marine species like oysters, is an area that requires more research. Our study aimed to examine these concurrent stressors' impacts on phenotypic markers, gill and hepatopancreas physiological indices, and hemocyte immune parameters of Crassostrea hongkongensis. We subjected six oyster cohorts to varied ZnO NPs concentrations and salinity levels over 21 days. Our findings reveal that individual exposure to ZnO NPs or diminished salinity disrupts oyster physiology, impacting metabolism, antioxidant capacity, immune response, and energy distribution through distinct mechanisms. Remarkably, low salinity constituted a more significant threat than isolated ZnO NPs. However, when confronted with combined stressors, oysters exhibited a compensatory response, attenuating individual stressors' detrimental effects. This adaptation was characterised by reduced apoptosis rates, increased calcium ion concentration in mature hemocytes, and a restoration of conditioned indices, hepatopancreas alkaline phosphatase, and gill catalase activity to baseline levels. Principal Component Analysis and Integrated Biomarker Responses validated this compensatory phenomenon. Partial Least Squares Pathway Model analysis underscored these stressors' profound implications on oyster health, primarily driven by stressor exposure rather than mere zinc concentrations, despite acknowledging zinc's immunosuppressive impact on oyster immunity. Our research emphasises the importance of assessing multiple stressors' cumulative effects on aquatic species' ecological resilience, accentuating the need for comprehensive analyses incorporating functional specificity among diverse organs and immune components, including gill, hepatopancreas, and the critical hemocytes.

Wild oyster population resistance to ocean acidification adversely affected by bacterial infection.

The carbon dioxide induced ocean acidification (OA) process is well known to have profound effects on physiology, survival and immune responses in marine organisms, and particularly calcifiers including edible oysters. At the same time, some wild populations could develop a complex and sophisticated immune system to cope with multiple biotic and abiotic stresses, such as bacterial infections and OA, over the long period of coevolution with the environment. However, it is unclear how immunological responses and the underlying mechanisms are altered under the combined effect of OA and bacterial infection, especially in the ecologically and economically important edible oysters. Here, we collected the wild population of oyster species Crassostrea hongkongensis (the Hong Kong oyster) from their native estuarine area and carried out a bacterial challenge with the worldwide pervasive pathogen of human foodborne disease, Vibrio parahaemolyticus, to investigate the host immune responses and molecular mechanisms under the high-CO2 and low pH-driven OA conditions. The wild population had a high immune resistance to OA, but the resistance is compromised under the combined effect of OA and bacterial infection both in vivo or in vitro. We classified all transcriptomic genes based on expression profiles and functional pathways and identified the specifically switched on and off genes and pathways under combined effect. These genes and pathways were mainly involved in multiple immunological processes including pathogen recognition, immune signal transduction and effectors. This work would help understand how the immunological function and mechanism response to bacterial infection in wild populations and predict the dynamic distribution of human health-related pathogens to reduce the risk of foodborne disease under the future climate change scenario.

Autophagy Dually Induced by AMP Surplus and Oxidative Stress Enhances Hemocyte Survival and Bactericidal Capacity via AMPK Pathway in Crassostrea hongkongensis.

Crassostrea hongkongensis (Hong Kong oyster) is an ecologically and economically valuable shellfish endemic to South/Southeast Asia. Due to ocean acidification and warming waters, they have become increasingly vulnerable to invading microbes including Vibrio parahaemolyticus, a significant foodborne human pathogen. In recent years, outbreaks of V. parahaemolyticus have emerged as a perennial phenomenon in parts of the world, necessitating to better understand the biology of host-pathogen interactions in this under-examined marine invertebrate. Although an immunologically relevant autophagy apparatus has been identified in Crassostrea gigas, an evolutionarily close mollusk cousin, the precise mechanistic details of C. hongkongensis autophagy during V. parahaemolyticus infection are still wanting. Here, we compellingly demonstrated that in vivo V. parahaemolyticus challenge robustly triggered autophagic signaling in C. hongkongensis hemocytes peaking at 6 h post-infection, which subsequently promoted bacterial clearance and dampened premature apoptosis. Simultaneously, a large surplus of adenosine monophosphate (AMP) and elevations in reactive oxygen species (ROS, specifically mitochondrial O2 - and cellular H2O2) formation were observed post-infection. Extrinsically applied AMP and ROS could synergistically induce AMP-activated protein kinase (AMPK) phosphorylation to stimulate downstream autophagic events. V. parahaemolyticus infection-induced autophagy was pharmacologically shown to be AMPK-dependent in vivo. Overall, our results establish autophagy as a crucial arm of host defense against Vibrio infections in mollusks, and provide new insights into the underappreciated roles of ROS and AMP as co-regulators of autophagy.