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by claude@2026-07, 2026-07-03
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The paper develops and validates a high-throughput resazurin-based hemocyte viability assay to study bacterial pathogenicity in bivalves, aiming to overcome seasonal limitations in obtaining healthy oyster larvae for infection testing. Using eastern oyster hemocytes, the authors optimized the assay and determined LC50 values for Vibrio coralliilyticus RE22, Phaeobacter inhibens S4, and multiple hatchery-derived bacterial isolates, finding that the most virulent isolates had LC50 values from 53 to 195 MOI. Validation with oyster larvae showed that isolates with hemocyte LC50 <200 MOI caused larval mortality, whereas isolates with LC50 ≥690 MOI did not. The paper does not explicitly discuss endometriosis or adenomyosis; it was included in the corpus via a keyword match in the upstream search index.
Abstract
ABSTRACT Bacterial pathogens cause disease outbreaks in hatcheries, leading to larval mortality and significant economic losses. One of the major challenges in studying bacterial pathogenesis in larvae is the limited availability of healthy larvae throughout the year. This seasonal constraint delays testing and impedes progress in understanding bacterial infections, disease management, and the development of effective treatments for bivalve aquaculture. To address this challenge, a hemocyte viability assay was developed as an alternative for testing bacterial pathogenesis. A resazurin-based assay was optimized using eastern oyster hemocytes, and lethal concentration (LC) 50 values were determined for the bacterial pathogen Vibrio coralliilyticus RE22 (121 ± 2 multiplicity of infection, MOI), probiont Phaeobacter inhibens S4 (2,157 ± 56 MOI), and several bacterial isolates from shellfish hatcheries. The most virulent isolates had LC 50 values ranging from 53 to 195 MOI. To validate the assay, the isolates were tested on oyster larvae, confirming that bacterial isolates with LC 50 <200 MOI against hemocytes also caused larval mortality, while isolates exhibiting LC 50 values ≥690 MOI showed no adverse effects on oyster larvae. This high-throughput hemocyte viability assay provides a rapid and reliable alternative to larval assays, particularly during the off-season for hatcheries, facilitating year-round research on bacterial pathogenicity in bivalves.
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ABSTRACT
Bacterial pathogens cause disease outbreaks in hatcheries, leading to larval mortality and significant economic losses. One of the major challenges in studying bacterial pathogenesis in larvae is the limited availability of healthy larvae throughout the year. This seasonal constraint delays testing and impedes progress in understanding bacterial infections, disease management, and the development of effective treatments for bivalve aquaculture. To address this challenge, a hemocyte viability assay was developed as an alternative for testing bacterial pathogenesis. A resazurin-based assay was optimized using eastern oyster hemocytes, and lethal concentration (LC)50 values were determined for the bacterial pathogen Vibrio coralliilyticus RE22 (121 ± 2 multiplicity of infection, MOI), probiont Phaeobacter inhibens S4 (2,157 ± 56 MOI), and several bacterial isolates from shellfish hatcheries. The most virulent isolates had LC50 values ranging from 53 to 195 MOI. To validate the assay, the isolates were tested on oyster larvae, confirming that bacterial isolates with LC50 <200 MOI against hemocytes also caused larval mortality, while isolates exhibiting LC50 values ≥690 MOI showed no adverse effects on oyster larvae. This high-throughput hemocyte viability assay provides a rapid and reliable alternative to larval assays, particularly during the off-season for hatcheries, facilitating year-round research on bacterial pathogenicity in bivalves.
Competing Interest Statement
The authors have declared no competing interest.
Footnotes
Section on results and discussion were updated
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