Effect of dietary supplementation with apple cider vinegar and propionic acid on hemolymph chemistry, intestinal microbiota and histological structure of hepatopancreas in white shrimp, Litopenaeus vannamei.

This experiment was conducted to evaluate the effects of dietary supplementation of Apple cider vinegar (ACV) and propionic acid (PA) on biochemical parameters of hemolymph, intestinal microbiota and histology of hepatopancreas in white shrimp (Litopenaeus vannamei). Five experimental diets were evaluated in this study including diets supplemented with 1.0, 2.0 and 4.0% of ACV, 0.5% propionic acid, and a control diet with no supplements. Shrimps (initial weight of 10.2 ±â€¯0.04 g) in triplicate groups with the density of 25 shrimps per tank were fed the diets for 60 days. At the end of the feeding trial, shrimps fed with ACV and PA supplemented diets had significantly higher total protein level than those fed the control diet (P < 0.05). The number of Vibrio spp., R-cells (lipid storage cells) of hepatopancreas and cholesterol level in shrimps fed the diets containing ACV and PA were lower compared to the control group (P < 0.05). However, there was no remarkable variations in glucose concentration, B-cell number and tubule diameter among the experimental diets (P > 0.05). In addition, shrimps fed the ACV diets had significantly lower total heterotrophic marine bacteria compared to the control or PA groups, and the lowest bacterial number was observed in shrimp fed 4% ACV supplemented diet (P < 0.05). Supplementation of 2 and 4% ACV as well as 0.5% PA in the diet led to a significantly higher calcium concentration than the control treatment (P < 0.05). The lowest triglyceride concentration was observed in the shrimps fed diets containing 2.0 and 4.0% ACV, which resulted in 15 and 20% reduction, respectively (P < 0.05). Overall, the findings indicates that ACV and PA possess antimicrobial activity and demonstrate beneficial effects on health status, so they can be potentially used as feed additive in the feeding of L. vannamei.

Identification, tissue distribution and orexigenic activity of neuropeptide F (NPF) in penaeid shrimp.

The neuropeptide Fs (NPFs) are an invertebrate subgroup of the FMRFamide-like peptides, and are proposed by some to be the homologs of vertebrate neuropeptide Y. Although there is some information about the identity, tissue distribution and function of NPFs in insects, essentially nothing is known about them in crustaceans. We have identified and characterized NPF-encoding transcripts from the penaeid shrimp Litopenaeus vannamei and Melicertus marginatus. Two transcripts were identified from each species. For each shrimp species, the two transcripts differed from one another by the presence or absence of an insert in the portion of the open reading frame that encodes the NPF peptide. The two NPF isoforms are identical in L. vannamei and M. marginatus, with their predicted structures being KPDPSQLANMAEALKYLQELDKYYSQVSRPRFamide and KPDPSQLANMAEALKYLQELDKYYSQVSRPSPRSAPGPASQIQALENTLKFLQLQELGKLYSLRARPRFamide. RT-PCR tissue profiling showed both transcripts are broadly distributed within the nervous system of each species. The transcript encoding the shorter NPF was detected in some, but not all, midgut samples. The transcript encoding the longer NPF was absent in the midgut of both species, and neither transcript was detected in their skeletal muscle. Juvenile L. vannamei fed on a diet supplemented with the shorter NPF exhibited a marked increase in food intake relative to control individuals that did not receive the supplement; the NPF-fed shrimp also showed a significant increase in growth relative to the control group. Our data suggest that NPF is present in both the nervous system and midgut of penaeid shrimp, functioning, at least in part, as a powerful orexigenic agent.

A crustacean Ca2+-binding protein with a glutamate-rich sequence promotes CaCO3 crystallization.

The DD4 mRNA of the penaeid prawn Penaeus japonicus was shown previously to be expressed in the epidermis adjacent to the exoskeleton specifically during the post-moult period, when calcification of the exoskeleton took place. The encoded protein possessed a Ca2+-binding site, suggesting its involvement in the calcification of the exoskeleton. In the present study, an additional ORF (open reading frame) of 289 amino acids was identified at the 5' end of the previous ORF. The newly identified part of the encoded protein included a region of approx. 120 amino acids that was highly rich in glutamate residues, and contained one or more Ca2+-binding sites. In an immunohistochemical study, signals were detected within calcified regions in the endocuticular layer of the exoskeleton. Bacterially expressed partial segments of the protein induced CaCO3 crystallization in vitro. Finally, a reverse transcription-PCR study showed that the expression was limited to an early part of the post-moult period, preceding significant calcification of the exoskeleton. These observations argue for the possibility that the encoded protein, renamed crustocalcin (CCN), promotes formation of CaCO3 crystals in the exoskeleton by inducing nucleation.