Whiteleg shrimp Penaeus vannamei farming in clear water recirculating aquaculture systems (RAS) is relatively recent, and consequently, knowledge on the shrimp dietary demands is still insufficient, particularly in the initial developmental stages. This study aimed at assessing the dietary protein requirement of whiteleg shrimp post-larvae (PL) in a clear-water recirculating aquaculture system (RAS). Six microdiets were formulated to contain 34%, 44%, 49%, 54%, 58%, and 63% crude protein (P34, P44, P49, P54, P58 and P63, respectively) and were evaluated in triplicates. Whiteleg shrimp PL (3.2 mg wet weight) were reared for 21 days in a clear-water RAS at Riasearch Lda. At the end of the feeding period, the optimal protein requirement was estimated at 47.1%, 46.4%, 47.2%, and 44.0% for weight gain, relative growth rate (RGR), feed conversion ratio (FCR), and survival, respectively. PL fed the P54, P58, and P63 diets achieved significantly higher final body weights than those fed P34. PL fed P34 showed significantly lower RGR and survival and significantly higher FCR values than those fed the remaining diets, suggesting that low protein diets may not be adequate to be used in this stage of shrimp development and/or for the clear-water RAS husbandry conditions. Moreover, diet P34 seemingly reduced the overall antioxidant status of the PL when compared to P44, P49, and P54. However, the P34 diet seems to have stimulated the PL immune mechanisms when compared to P44, P49, and P54, possibly due to increased levels of fish and algae oil. Similarly, despite the good growth performances, a diet containing 63% of protein also seemed to have compromised the overall shrimp PL antioxidant status and stimulate their immune system. Shrimp fed diet P54 showed an apparent overall superior antioxidant status when compared to the remaining diets, evidencing that using protein inclusion levels up to 54% in aquafeeds not only potentiates growth performances and survival but also can potentially be beneficial to the health status of P. vannamei PL grown in a clear-water RAS. Hence, results from this study suggest that a minimum of approximately 47% of protein should be considered when tailoring microdiets for whiteleg shrimp PL grown in a clear-water RAS, but inclusion levels up to 54% can be used with benefits to the PL antioxidant status.

Dietary additives have the potential to stimulate the whiteleg shrimp immune system, but information is scarce on their use in diets for larval/post-larval stages. The potential beneficial effects of vitamins C and E, β-glucans, taurine, and methionine were evaluated. Four experimental microdiets were tested: a positive control diet (PC); the PC with decreased levels of vitamin C and E as negative control (NC); the PC with increased taurine and methionine levels (T + M); and the PC supplemented with β-glucans (BG). No changes in growth performance and survival were observed. However, post-larvae shrimp fed the NC had lower relative expressions of pen-3 than those fed the PC, suggesting that lower levels of vitamins C and E may impact the shrimp immune status. Lipid peroxidation levels dropped significantly in the BG compared to the PC, indicating that β-glucans improved the post-larvae antioxidant mechanisms. Furthermore, when compared with the NC diet, PL fed with BG showed significant increases in tGSH levels and in the relative expression of crus and pen-3, suggesting a synergistic effect between vitamins C and E and β-glucans. Amongst the additives tested, β-glucans seems to be the most promising even when compared to a high-quality control diet.

The green tips of Salicornia ramosissima are used for human consumption, while, in a production scenario, the rest of the plant is considered a residue. This study evaluated the potential of incorporating salicornia by-products in diets for juvenile European seabass, partially replacing wheat meal, aspiring to contribute to their valorization. A standard diet and three experimental diets including salicornia in 2.5%, 5% and 10% inclusion levels were tested in triplicate. After 62 days of feeding, no significant differences between treatments were observed in fish growth performances, feeding efficiency and economic conversation ratio. Nutrient digestibility of the experimental diets was unaffected by the inclusion of salicornia when compared to a standard diet. Additionally, salicornia had significant modulatory effects on the fish muscle biochemical profiles, namely by significantly decreasing lactic acid and increasing succinic acid levels, which can potentially signal health-promoting effects for the fish. Increases in DHA levels in fish fed a diet containing 10% salicornia were also shown. Therefore, the results suggest that salicornia by-products are a viable alternative to partially replace wheat meal in diets for juvenile European seabass, contributing to the valorization of a residue and the implementation of a circular economy paradigm in halophyte farming and aquaculture.

The identification of novel feed materials as a source of functional ingredients is a topical priority in the finfish aquaculture sector. Due to the agrotechnical practices associated and phytochemical profiling, halophytes emerge as a new source of feedstuff for aquafeeds, with the potential to boost productivity and environmental sustainability. Therefore, the present study aimed to assess the potential of Salicornia ramosissima incorporation (2.5, 5, and 10%), for 2 months, in the diet of juvenile European seabass, seeking antioxidant (in the liver, gills, and blood) and genoprotective (DNA and chromosomal integrity in blood) benefits. Halophyte inclusion showed no impairments on growth performance. Moreover, a tissue-specific antioxidant improvement was apparent, namely through the GSH-related defense subsystem, but revealing multiple and complex mechanisms. A genotoxic trigger (regarded as a pro-genoprotective mechanism) was identified in the first month of supplementation. A clear protection of DNA integrity was detected in the second month, for all the supplementation levels (and the most prominent melioration at 10%). Overall, these results pointed out a functionality of S. ramosissima-supplemented diets and a promising way to improve aquaculture practices, also unraveling a complementary novel, low-value raw material, and a path to its valorization.

Whiteleg shrimp (Penaeus vannamei) is currently facing significant challenges related to severe disease outbreaks. As the importance of the host–microbiota relationship is being revealed, modulating this relationship has become a key strategy in disease management. Xylooligosaccharides (XOS)—short-chain sugar molecules—have been gaining attention for their potential health benefits in the prebiotics field. In this study, an XOS-rich extract from Salicornia ramosissima was incorporated into shrimp feeds to evaluate its impact on gut health, with the main focus on gut proteomics and microbiota. XOS were incorporated at 0.1% (XOS_0.1) and 1% (XOS_1) concentrations, and a 14-day feeding trial, followed by a bacterial challenge with Vibrio harveyi, was performed. The effects of XOS were evaluated by assessing zootechnical parameters, gene expression in the hepatopancreas, and gut microbiota and proteome. The results showed no significant differences in zootechnical parameters and gene expression after the 14-day trial between animals fed XOS diets and control. However, shrimp fed XOS_1 showed an increased diversity of the microbial communities in the gut when compared with those fed control. Also, known shrimp gut symbionts, such as Ruegeria, Leisingera, and Demequina, were significantly enriched in groups fed XOS after the feeding trial. XOS also modulated the regulation of proteins in the gut. Nevertheless, stressful conditions appear to alter the effects of XOS and the dynamics of gut bacteria. Further studies are warranted to understand the impacts of long-term inclusion of XOS extracts, especially on health-related parameters and disease resistance.