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Hemocyanin-modulated niacinamide shapes the hemolymph microbiota of penaeid shrimp

ABSTRACT: In aquatic invertebrates, the hemolymph functions as a conduit between the host and its environment, hence, maintaining hemolymph microbial homeostasis significantly influences disease pathogenesis. This study examined the critical role of the respiratory glycoprotein hemocyanin as a host factor in modulating hemolymph microbial composition in penaeid shrimp. Shrimp depleted of hemocyanin followed by Vibrio parahaemolyticus infection had a disruption in the niacinamide salvage pathway, which consequently increased total bacteria and relative abundance of the conditional pathogens Vibrio and Shewanella in the hemolymph. Moreover, in vivo knockdown of hemocyanin followed by niacinamide supplementation restored hemolymph microbial homeostasis, mitigating the competitive advantage of Vibrio. Overall, our data present novel insights into the pivotal role of niacinamide metabolism in modulating shrimp hemolymph microbial composition through hemocyanin.

INSTRUMENT(S): Liquid Chromatography MS - negative - reverse phase, Liquid Chromatography MS - positive - reverse phase

PROVIDER: MTBLS8272 | MetaboLights | 2024-08-12

REPOSITORIES: MetaboLights

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Project description:The Manila clam (Ruditapes philippinarum) is a cultured bivalve species with high worldwide commercial importance. Nevertheless, diseases can cause high economical losses. For this reason, the study of immune genes in bivalve mollusks has increased in the last years. The present work describes the construction of the first R. philippinarum microarray containing immune-related hemocyte sequences and its application for the study of the gene transcription profiles of hemocytes from clams challenged with Vibrio alginolyticus through a time course. A comparative analysis of gene expression was conducted between R. philippinarum infected and non-infected by V. alginolyticus clam hemocytes. Clams (n=100) were notched in the shell next to the adductor muscles and injected with 100 Âµl of Vibrio alginolyticus, strain TA15, (10^8 UFC/ml in PBS) to mimic an intramuscular infection. Controls (n=100) were injected with 100 Âµl of PBS. After stimulation, clams were returned to the tanks and maintained at 15ÂºC until sampling at 3, 8, 24, and 72 hours after challenge Hemolymph (1 ml) was withdrawn from the adductor muscle of the clams with a 0.5mm diameter (25G) disposable needle. Hemolymph from four individuals was pooled and biological replicates were taken at each sampling point. Hemolymph was centrifuged at 4Â°C at 3000 g for 10 minutes. The pellet was resuspended in 250 Âµl of Trizol (Invitrogen). Total RNA isolation was conducted following the manufacturer's specifications in combination with the RNeasy mini kit (Qiagen) for RNA purification after DNase I treatment. Gene expression profiling was performed using an R. philippinarum oligo-DNA microarray of 13,671 probes based on single-colour detection (Cyanine-3 only). Microarrays were scanned with Agilent scanner G2565BA at a resolution of 5 microns; all slides were scanned twice at two different sensitivity settings (XDRHi 100% and XDRLo 10%); the scanner software created a unique ID for each pair of XDR scans and saved it to both scan image files. Feature Extraction (FE) 9.5 used XDR ID to link the pairs of scans together automatically when extracting data. The signal left after all the FE processing steps have been completed is ProcessedSignal that contains the Multiplicatively Detrended, Background-Subtracted Signal.

			Action	DRS
	NEG_Fa18BSY_G_Na1.raw	Raw
	NEG_Fa18BSY_G_Na2.raw	Raw
	NEG_Fa18BSY_G_Na3.raw	Raw
	NEG_Fa18BSY_G_Na4.raw	Raw
	NEG_Fa18BSY_G_Na5.raw	Raw

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Hemocyanin-modulated niacinamide shapes the hemolymph microbiota of penaeid shrimp

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