Dung V. Le

Bio: Dung V. Le is an academic researcher from Auckland University of Technology. The author has contributed to research in topics: Panopea zelandica & Geoduck. The author has an hindex of 5, co-authored 10 publications receiving 88 citations. Previous affiliations of Dung V. Le include Vietnam National University, Ho Chi Minh City.

Differential expression of novel metabolic and immunological biomarkers in oysters challenged with a virulent strain of OsHV-1.

Abstract: Early lifestages of the Pacific oyster (Crassostrea gigas) are highly susceptible to infection by OsHV-1 μVar, but little information exists regarding metabolic or pathophysiological responses of larval hosts. Using a metabolomics approach, we identified a range of metabolic and immunological responses in oyster larvae exposed to OsHV-1 μVar; some of which have not previously been reported in molluscs. Multivariate analyses of entire metabolite profiles were able to separate infected from non-infected larvae. Correlation analysis revealed the presence of major perturbations in the underlying biochemical networks and secondary pathway analysis of functionally-related metabolites identified a number of prospective pathways differentially regulated in virus-exposed larvae. These results provide new insights into the pathogenic mechanisms of OsHV-1 infection in oyster larvae, which may be applied to develop disease mitigation strategies and/or as new phenotypic information for selective breeding programmes aiming to enhance viral resistance.

Practical Fertilization Procedure and Embryonic Development of the New Zealand Geoduck Clam (Panopea Zelandica)

Abstract: Cultivation of the geoduck Panopea zelandica (Quoy & Gaimard, 1835) requires knowledge on embryonic development to produce spat in hatcheries. This study investigated the development of P. zelandica embryos at 15°C and 35 ppt and the optimal sperm:egg ratios for fertilization under hatchery conditions. Panopea zelandica broodstock were induced to spawn by serotonin injection. Sperm and eggs were collected and fertilization was conducted at sperm:egg ratios of: 50:1, 100:1, 500:1, 1000:1 and 10,000:1 over 40 min. The optimal sperm:egg ratio was <500:1 and the normal embryo yield at 3 and 18 h post-fertilization (hpf) ranged from 83–96%. Panopea zelandica eggs (~80 μm diameter) developed the first and second polar bodies within 15–20 and 50–55 min post-fertilization, respectively. The blastula appeared at ~8 hpf, including the XR and XL cells and the presumptive shell field depression. Gastrulation occurred at 12–18 hpf with organic material apparent at the shell field depression. The mid-stage trochophore, which appeared at around 35 hpf had an apical plate with an apical tuft. The shell field spread to form the periostracum, which expanded and folded into right and left segments covering the late trochophore. The early D-stage veliger appeared at 45 hpf with the soft body being enclosed by two valves and the appearance of the velum. These observations will serve as the basis for future analyses of P. zelandica embryogenesis and for optimization of commercial production of D-veliger larvae.

The Effects of Aragonite Saturation State on Hatchery-Reared Larvae of the Greenshell Mussel Perna canaliculus

Abstract: The major cultured mussel species Perna canaliculus is now supported by hatchery production, providing the opportunity to explore and optimize environmental parameters to enhance production. Other cultured bivalve larvae have demonstrated performance that is directly correlated to the aragonite saturation state (Ωar) of their tank water, with low or undersaturated water being detrimental and artificially elevated Ωar enhancing productivity. Trials were, therefore, designed to specifically explore Ωar sensitivity in preveliger (0–2 days old, prodissoconch I = “PD1″) and veliger (2–21 days old, prodissoconch II = “PD2″) stages of P. canaliculus separately. For the PD1 experiment, commercial incubation tanks (control Ωar 1.9) were modified to target Ωar 0.5 or 0.8 by elevating pCO2, or 2.9, 4.5, and ∼7 by the addition of sodium carbonate. In the control environment, 72.8% ± 2.9% of fertilized eggs formed viable “D” veligers within two days; an increased yield of 82.6% ± 3.8% in Ωar 4.5 was found to be nonsignificant. In comparison, only 12.7% of the Ωar ∼7 and <1% of the Ωar 0.5 and 0.8 eggs attained the veliger stage, with the remaining underdeveloped or malformed. By 2 days postfertilization, reactive oxygen species were significantly elevated in the undersaturated treatments, whereas DNA damage, lipid hydroperoxides, and protein carbonyls were significantly higher in the Ωar 0.5 and ∼7 treatments. Antioxidant enzyme levels were significantly lower in these extreme treatments, whereas Ωar 4.5 larvae showed elevated superoxide dismutase, glutathione reductase, and peroxidase levels. Carry-over effects persisted when veligers were transferred to control conditions, with no net recruitment from undersaturated Ωar, 29.4% of eggs surviving to pediveliger under control conditions, compared with 33.2% following Ωar 4.5 exposure or 1.9% from Ωar ∼7. In the PD2 veliger trial, linear shell growth halved in undersaturated water, but was unaffected by elevation of Ωar. Mortality rate was consistent across all treatments, suggesting relative resilience to different Ωar. It is recommended that hatcheries trial Ωar 4–4.5 enrichment in preveliger incubation water to improve yield and minimize oxidative stress. Preveliger stages present a potential survival bottleneck, and focused research exploring sensitivity to near-future ocean acidification is, therefore, needed.

Immunity in Molluscs: Recognition and Effector Mechanisms, with a Focus on Bivalvia

Abstract: The study of molluscan immune systems, in particular those of bivalve molluscs (e.g., clams, oysters, scallops, mussels), has experienced great growth in recent decades, mainly due to the needs of a rapidly growing aquaculture industry to manage the impacts of disease and the wider application of -omic tools to this diverse group of invertebrate organisms. Several unique aspects of molluscan immune systems highlighted in this chapter include the importance of feeding behavior and mucosal immunity, the discovery of unique levels of diversity in immune genes, and experimental indication of transgenerational immune priming. The development of comparative functional studies using natural and selectively bred disease-resistant strains, together with the potential but yet to be fully developed application of gene-editing technologies, should provide exciting insights into the functional relevance of immune gene family expansion and molecular diversification in bivalves. Other areas of bivalve immunity that deserve further study include elucidation of the process of hematopoiesis, the full characterization of hemocyte subpopulations, and the genetic and molecular mechanisms underlying immune priming. While the most important aspects of the immune system of the largest group of molluscs, gastropods (e.g., snails and slugs), are discussed in detail in Chap. 12, we also briefly outline the most distinctive features of the immune system of another fascinating group of marine molluscs, cephalopods, which include invertebrate animals with extraordinary morphological and behavioral complexity.

Metabolomics Study of Immune Responses of New Zealand Greenshell™ Mussels (Perna canaliculus) Infected with Pathogenic Vibrio sp.

Abstract: Vibrio coralliilyticus is a bacterial pathogen which can affect a range of marine organisms, such as corals, fish and shellfish, with sometimes devastating consequences. However, little is known about the mechanisms involved in the host-pathogen interaction, especially within molluscan models. We applied gas chromatography-mass spectrometry (GC-MS)-based metabolomics to characterize the physiological responses in haemolymph of New Zealand Greenshell™ mussels (Perna canaliculus) injected with Vibrio sp. DO1 (V. coralliilyticus/neptunius-like isolate). Univariate data analyses of metabolite profiles in Vibrio-exposed mussels revealed significant changes in 22 metabolites at 6 h post-infection, compared to non-exposed mussels. Among them, 10 metabolites were up-regulated, while 12 metabolites were down-regulated in infected mussels. Multivariate analyses showed a clear distinction between infected and non-infected mussels. In addition, secondary pathway analyses indicated perturbations of the host innate immune system following infection, including oxidative stress, inflammation and disruption of the TCA cycle, change in amino acid metabolism and protein synthesis. These findings provide new insights into the pathogenic mechanisms of Vibrio infection of mussels and demonstrate our ability to detect detailed and rapid host responses from haemolymph samples using a metabolomics approach.

Copper-induced immunomodulation in mussel (Perna canaliculus) haemocytes.

Abstract: Copper is a common contaminant in aquatic environments, which may cause physiological dysfunction in marine organisms. However, the toxicity mechanisms of copper in marine bivalves is not fully understood. In this study, we applied an integrated approach that combines flow cytometry and Gas Chromatography-Mass Spectrometry (GC-MS)-based metabolomics to characterize cellular and molecular mechanisms of copper immunotoxicity in New Zealand Greenshell™ mussel (Perna canaliculus) haemolymph. Flow cytometric results showed significant increases in haemocyte mortality, production of reactive oxygen species and apoptosis (via alteration of caspase 3/7 and mitochondrial membrane potential) of haemocytes exposed to increasing total concentrations of Cu2+ (62.5, 125.0 and 187.5 μM) compared to a low Cu2+ concentration (25.0 μM) and control (0.0 μM). In addition to flow cytometric data, our metabolomics results showed alterations of 25 metabolites within the metabolite profile of Cu2+-exposed haemolymph (125 μM) compared to those of control samples. Changes in levels of these metabolites may be considered important signatures of oxidative stress (e.g., glutathione) and apoptosis processes (e.g., alanine, glutamic acid). This study provides insights into the cellular and molecular mechanisms of oxidative stress and apoptosis in marine bivalves and highlights the applicability and reliability of metabolomic techniques for immunotoxicological studies in marine organisms.