Freshwater migration and feeding habits of juvenile temperate seabass <Emphasis Type="Italic">Lateolabrax japonicus</Emphasis> in the stratified Yura River estuary,the Sea of Japan

Juvenile temperate seabass Lateolabrax japonicus were sampled along the Yura River estuary from April to July 2008 to determine their distribution and feeding habits during migration within a microtidal estuary. Throughout the sampling period, juvenile seabass were distributed not only in the surf zone, but also in the freshwater zone, and they were particularly abundant in areas with aquatic vegetation in the freshwater zone. This distribution pattern suggests that the early life history of the temperate seabass depends more intensively on the river (freshwater) than previously considered. Small juveniles in the freshwater zone fed on copepods and chironomid larvae and upon reaching a standard length (SL) of approximately 20 mm did they fed on mysids. In contrast, juveniles (approx. 17–80 mm SL) in the surf zone fed mainly on mysids.     

Influence of swimbladder inflation failure on mortality,growth and lordotic deformity in Pacific bluefin tuna,Thunnus orientalis, (Temminck & Schlegel) postflexion larvae and juveniles

This study examined the influence of swimbladder inflation (SBI) failure on mortality (Experiment 1), lordotic deformity (Experiment 2) and growth (Experiment 1, 2) in Pacific bluefin tuna, Thunnus orientalis, postflexion larvae and juveniles by producing larvae lacking inflated swimbladders. Experiment 1 was conducted for postflexion larvae to juveniles (from 18 to 30 days‐post‐hatch; dph). Mortality was not significantly different between the fish with (WIS) and without (WOIS) inflated swimbladders. Standard length (SL) and body weight (BW) were significantly smaller in WOIS than WIS. Moreover, the SBI was found in WOIS after postflexion stage. In Experiment 2, two examination trials were conducted on SBI, vertebral deformity and growth for juvenile stage. Lordotic deformities were found neither in the WOIS nor in the WIS. Although SL and BW were significantly smaller in WOIS than WIS at 22 dph, after 37 dph no significant differences were found between them. The results of Experiment 1 and 2 indicate that SBI failure causes growth retardation until juveniles of 30 dph; however, it does not cause growth retardation in juveniles after 37 dph and mortality of postflexion larvae and juveniles. Moreover, SBI failure did not cause lordotic deformity in PBT differently from many other fish.     

Winter monsoon promotes the transport of Japanese temperate bass Lateolabrax japonicus eggs and larvae toward the innermost part of Tango Bay,the Sea of Japan

Northwesterly cold winds characteristic of the East Asian Winter Monsoon (EAWM) dictate winter climatic conditions over the Japanese Archipelago. Japanese temperate bass Lateolabrax japonicus is a commercially important coastal fish that spawns offshore in winter and uses shallow waters as nursery habitats. To investigate the effects of EAWM on the planktonic period of L. japonicus, eggs, larvae, and juveniles were quantitatively collected in Tango Bay on the Sea of Japan side in winter and spring from 2007 to 2017. Although eggs occurred close to the mouth of the bay, planktonic larvae occurred further inside as they developed. The horizontal distribution of planktonic larvae, combined with water velocity data obtained from mooring observations, indicated that planktonic larvae are transported south‐ to westward through Ekman current and an anticyclonic circulation, which are driven by northwesterly winds. To evaluate survival during the planktonic period in each year class, the abundance of benthic larvae/juveniles was divided by winter total landings of Lateolabrax spp. (proxy of the spawning stock size). This survival index exhibited a positive correlation with the northwesterly component of winter winds, and a negative correlation with winter air temperature (average from December to February, Spearman's correlation, p < .05). There was, however, no significant correlation with winter water temperature or winter freshwater discharge in the bay. We conclude that northwesterly cold winds of EAWM play a critical role in transporting L. japonicus eggs and larvae toward nursery habitats, specifically beaches and estuaries fringing the innermost part of Tango Bay.     

Periphyton characteristics influence the growth and survival of Holothuria scabra early juveniles in an ocean nursery system

Floating hapas (fine mesh net enclosures) are a cost‐effective ocean nursery system to culture post‐metamorphic Holothuria scabra to release size. The growth of periphyton biofilm on hapas is a natural food source for early juveniles. This study investigated the effects of periphyton quality (i.e. chlorophyll‐a, phaeopigment, total biomass, autotrophic index or AI), water quality (nutrients, chlorophyll‐a) and environmental parameters (temperature, rainfall) on the temporal variation in the growth and survival of early juvenile (~3 mm) H. scabra reared in floating hapas. Five trials where the juveniles were reared for 60 days each in the eutrophic coastal waters of Bolinao, the Philippines were conducted during different months over 2 years. Significant differences in the growth and survival of juveniles among trials were found. Absolute growth rates (AGR) ranged widely (0.01–0.09 g/day). Trials with high AGR of juveniles (0.07–0.09 g/day) during the first 30 days of rearing had significantly higher chlorophyll‐a (chl‐a) in biofilm (15.9–27.5 mg/m²) and lower AI. Conversely, during the subsequent 30 days, trials with high AGR of juveniles (0.06–0.11 g/day) had significantly lower chl‐a and higher AI. Multivariate analyses showed that chl‐a in biofilm, AI and nutrients in the water column are good indicators of periphyton quality and juvenile growth rates in floating hapas. Further, this study validates the expansion of the feeding mode of juveniles from primarily grazing on microalgae, to feeding on detritus and heterotrophs as they grow. These results are important in optimizing ocean nursery systems.     

Mouth Growth and Prey Selection in Juveniles of the European Long‐snouted Seahorse,Hippocampus guttulatus

Seahorses are ambush predators that swallow the prey through their tubular snout. In order to better understand mouth growth and its effect on prey preference by juveniles of the seahorse Hippocampus guttulatus, experimental assays were carried out by feeding juveniles (0–30 – d after male's pouch release [dar]) on a mixture of rotifers, Brachionus plicatilis; Artemia nauplii; and copepods; Acartia tonsa. Mouth development in juveniles was accomplished by growth of upper and lower jaws, which were linearly correlated with juvenile length, whereas mouth width (MW) and height (MH) grew exponentially with juvenile size. Additionally, MW and MH were exponentially correlated, so that the transversal section of the mouth increased linearly with juvenile age and length. Gut content and Ivlev' selectivity index demonstrated a permanent rejection of rotifers for the whole experimental period and a clear preference of juveniles for copepods from 0 to 15 dar and for Artemia nauplii afterwards. Results also suggest that juveniles are able to ingest larger prey than those provided as the limiting factor in prey ingestion was not MW, as for many marine fish larvae, but mouth area. A tentative feeding schedule to successfully feed H. guttulatus juveniles from 0 to 30 dar is proposed.     

Applying the ontogeny of digestive enzyme activity to guide early weaning of pigfish,Orthopristis chrysoptera (L.)

Weaning marine fish larvae from live prey to a dry microdiet is an important step towards optimizing the commercial production, but early weaning is constrained by the lack of sufficient digestive enzymes at first feeding. This study quantified the activity of five digestive enzymes throughout the larval period of pigfish (Orthopristis chrysoptera [L.]) to assess ontogenetic changes in digestive abilities, and then trials were conducted that determined the optimal time for weaning. The activity of all digestive enzymes was low or undetectable at first feeding (3 days post hatching, dph; 2.5 mm standard length, SL). A substantial increase in activity occurred at 5.7 mm SL (17 dph), 6.9 mm SL (21 dph), 7.7 mm SL (23 dph), 8.4 mm SL (25 dph) and 11.2 mm SL (30 dph) for bile salt‐dependent lipase, trypsin, chymotrypsin, amylase and acid protease respectively. During the weaning experiment, larvae were co‐fed live prey and microdiet beginning 15 dph (4.8 mm SL). Live prey was withdrawn from the diet at 24, 28, 32 or 36 dph, with the control receiving live prey and microdiet throughout (to 43 dph). There were no significant differences in mean final SL among treatments, but survival was significantly lower when larvae were weaned at 24 dph compared to 32–43 dph. Based on the digestive enzyme activity and survival, weaning larval pigfish at 32 dph (11.7 mm SL) when reared at 24°C is recommended.     

Effect of cultivated copepods (Acartia tonsa) in first‐feeding of Atlantic cod (Gadus morhua) and ballan wrasse (Labrus bergylta) larvae

The aim of this study was to compare the nutritional composition and effects of short periods with cultivated copepod nauplii versus rotifers in first‐feeding. Atlantic cod (Gadus morhua) and ballan wrasse (Labrus bergylta) larvae were given four different dietary regimes in the earliest start‐feeding period. One group was fed the copepod Acartia tonsa nauplii (Cop), a second fed enriched rotifers (RotMG), a third fed unenriched rotifers (RotChl) and a fourth copepods for the seven first days of feeding and enriched rotifers the rest of the period (Cop7). Cod larvae were fed Artemia sp. between 20 and 40 dph (days posthatching), and ballan wrasse between 36 and 40 dph, with weaning to a formulated diet thereafter. In addition to assessing growth and survival, response to handling stress was measured. This study showed that even short periods of feeding with cultivated copepod nauplii (7 days) had positive long‐term effects on the growth and viability of the fish larvae. At the end of both studies (60 days posthatching), fish larvae fed copepods showed higher survival, better growth and viability than larvae fed rotifers. This underlines the importance of early larval nutrition.     

Retention mechanisms of white perch (Morone americana) and striped bass (Morone saxatilis) early-life stages in an estuarine turbidity maximum: an integrative fixed-location and mapping approach

The small‐scale distribution and retention mechanisms of white perch (Morone americana) and striped bass (M. saxatilis) early‐life stages were investigated in the upper Chesapeake Bay estuarine turbidity maximum (ETM). Physical measurements and biological collections were made at fixed‐location stations within the ETM during three research cruises in 1998 and two in 1999. Results were compared with mapping surveys of physical properties and organism distributions above, within, and below the ETM. Physical conditions at the fixed stations differed markedly among cruises and between years due to differences in freshwater flow and wind. In each year, striped bass and white perch larval concentrations were highest in waters of salinity 1–4. Larvae were more abundant in the ETM region in 1998, a high‐flow year, suggesting that the ETM provides favorable nursery habitat when low salinity waters and the ETM coincide in high freshwater‐flow conditions. In 1998, the earliest pelagic life stages of fish larvae (eggs, yolk‐sac larvae) and the copepod Eurytemora affinis, an important prey of feeding larvae, apparently were retained in deep, landward‐flowing water within the salt front and ETM region. Statistical analyses indicated that distributions of white perch and striped bass post‐yolk‐sac larvae were associated with E. affinis distributions and suggested that retention of larval fish could result from tracking prey. Comparing fixed‐station and mapping approaches demonstrates the importance of sampling at different spatial scales within the ETM region and suggests that larvae are faced with trade‐offs between selecting zones of high retention or high visual‐feeding success.     

The administration of Lactobacillus plantarum directly to food: An effective way to improve the survival and growth of juvenile lined seahorses (Hippocampus erectus)

Previously, we found Lactobacillus plantarum had probiotic effects on lined seahorse (Hippocampus erectus) juveniles by the addition Lactobacillus directly into rearing seawater containing juveniles and their food (i.e. copepods). However, how the juveniles consumed Lactobacillus, directly from water or from copepods in whose guts Lactobacillus may have accumulated by ingestion, is still unclear. To clarify this, three experiments were successively carried out in this study. In the first experiment, the manner by which juveniles consumed Lactobacillus, namely, directly from food, was discovered via a 2 × 2 factorial design with two factors of water and food, and two levels with and without L. plantarum. In the second experiment, a 5 × 4 factorial design with five levels of L. plantarum doses (D: 5, 10, 20, 40 and 80 × 10⁸ cfu/L) and four levels of enrichment time (T: 0.5, 1.0, 1.5 and 2.0 hr) was conducted to analyse lactic acid bacteria (LAB) contents in the enriched copepods under different treatments. In the third experiment, six kinds of Lactobacillus‐enriched copepods chosen from the second experiment that carried significantly different LAB contents were fed to juveniles, and it was found that the copepods containing LAB greater than 5 log₁₀colonies/g had the most improved effects for survival and growth. In conclusion, the effective manner of L. plantarum administration in seahorses is by the addition to their food. Additionally, D40 * T1.0, whose LAB content was more than 5 log₁₀colonies/g, is generally the most economic copepod treatment for improving survival and growth in H. erectus rearing.     

Feeding,egg production and laboratory culture of Schmackeria poplesia Shen (Copepoda: Calanoida)

Copepods are candidates with great potential as live prey for rearing fish larvae and juveniles in aquaculture; however, the techniques for a large‐scale culture of copepods are yet to be developed. In this study, we examined the effects of water temperature, salinity, prey concentration and algal species on the grazing and egg production rates of a calanoid copepod Schmackeria poplesia (Copepoda: Calanoida). The results showed that the grazing rate of S. poplesia was the highest when the copepods were cultured in seawater with temperature of 25 °C, salinity of 20 g L^?1, prey concentration at 10⁵ cells mL^?1 and supplied with Platymonas helgolandica as the prey. The egg production rates, however, was the highest when copepods were fed with a mixed prey of Isochrysis galbana and Phaeodactylum tricornutum (cell ratio 1:1, prey concentration 10⁵ cells mL^?1) at 25 °C, 20 g L^?1 of salinity. A 100 L cultural system was established to culture S. poplesia under the condition optimized for egg production. The total number of copepods increased 40–43‐fold with the production rates of 87–290 copepods L^?1 day^?1 in 14 days. This research was the first attempt for a large‐scale culture of S. poplesia and the technique established can be further applied in aquaculture.     

Rearing the spotted seahorse Hippocampus kuda by feeding live and frozen copepods collected from shrimp ponds

This study evaluated the use of live and frozen copepods collected from shrimp ponds for rearing juveniles of the spotted seahorse Hippocampus kuda. Protein and HUFA contents in frozen copepods were all higher than in Artemia nauplii, the conventional live food for seahorse juveniles. The results of this study showed that copepods can be used as feed for rearing seahorse fry and juvenile. The spotted seahorse showed obvious preference for live copepods and rarely fed on dead copepods on the tank bottom. Furthermore, the combination of frozen copepods and live Artemia nauplii resulted in highest growth and highest survival of the experimental seahorses. Further research on possible effects of DHA:EPA ratio on survival and growth of young H. kuda is recommended.     

Aspects of the feeding biology of the copepod Pseudodiaptomus hessei (Copepoda: Calanoida) under culture conditions

This study aimed to investigate the feeding behaviour (particularly gut fullness and evacuation, preying on rotifers and feeding preference in the water column) of the calanoid copepod, Pseudodiaptomus hessei, as a potential live feed species for aquaculture. Fed and starved, male and female P. hessei were fed rotifers (Brachionus plicatilis) in the presence and absence of microalgae for 24 hr. Starved copepods consumed more rotifers (11.31 ± 1.01, individual rotifers) than fed (8.06 ± 1.01, individual rotifers) while the number of rotifers consumed in the presence of microalgae was similar when fed or starved. Gut fullness and evacuation was determined by feeding copepods two different cell size microalgae species (Tetraselmis suecica and Isochrysis galbana). Gut fullness and evacuation percentage were observed under the dissecting microscope (as 0, 25, 50, 75 and 100%). Copepods fed on T. suecica (large cell) filled their guts more rapidly, while those fed on I. galbana (small cell) evacuated their guts faster. Feeding preference was determined using a multifactorial experiment where copepods were fed two microalgae species, T. suecica and I. galbana, each presented as benthic and planktonic food sources. P. hessei preferred to feed on planktonic microalgae first regardless of microalgae choice species for ±80 min, before shifting to benthic food source. This information can be used as baseline information for aquaculturists to rear the species as live feed for marine fish larvae.     

Survival,growth and biomass of mud polychaete Marphysa iloiloensis (Annelida: Eunicidae) under different culture techniques

Polychaetes are commercially exploited as fishing bait and supplemental diet to crustacean and finfish broodstock. However, scarcity of supply and biosecurity issues limits its use. Polychaete aquaculture has become the best alternative to wild resources. In this study, two different culture techniques (1‐sediment tank throughout the culture period and 2‐nursery tank with bioflocs for 30 days; thereafter, juveniles were transferred to sediment tank until harvest) using two diets [bioflocs (BF) and feed mill sweepings (FS)] were tested to determine the survival, growth and biomass of polychaete Marphysa iloiloensis (Glasby et al., Zootaxa, 2019, 4674) in the grow‐out. The 180‐day trial was conducted in 16 tanks (0.20 m²) stocked with 5,066 ± 575 M. iloiloensis trochophore larvae/tank which were assigned randomly into four culture treatments with four replicates each. The treatments were labelled as: (a) BFS – fed BF in sediment tank for 180 days; (b) BF + BFS – fed BF in nursery tank for 30 days, juveniles were transferred in sediment tank and fed BF for another 150 days; (c) FSS – fed FS in sediment tank for 180 days; and (d) BF + FSS – fed BF in nursery tank for 30 days, juveniles are transferred in sediment tank and fed FS for another 150 days. Results showed that polychaetes in BF + FSS had significantly higher survival than BFS. Polychaetes in both BF + FSS and FSS had significantly higher body weight than BF + BFS and BFS, while biomass in BF + FSS (175.73 ± 42.25 g/tank) was significantly higher compared to other treatments (p < .05). Polychaetes fed with FS had 61%–64% protein and 12%–13% fat, while those fed with BF exclusively had 66%–70% protein and 3.50%–4.50% fat. The findings indicate that FS whether or not BF is included in the diet can improve both survival and growth of polychaetes but significantly higher biomass can be achieved with the combination of BF and FS. Additionally, polychaetes in BF + FSS contained 64% protein and 13% fat. Therefore, BF can be used as an alternative diet to FS in the early life stages of M. iloiloensis and nursery rearing using BF is imperative to increase its biomass in the grow‐out production.     

A sensitive period during first feeding for the determination of pigmentation pattern in Atlantic halibut, Hippoglossus hippoglossus L., juveniles: the role of diet

Six groups of Atlantic halibut, Hippoglossus hippoglossus L., larvae were offered calanoid cope-pods at different periods from days 11 to 25 after first feeding (1.13-3.20 mm myotome height) in order to establish at which stage normal pigmentation was determined. Artemia nauplii enriched with an oil emulsion were used prior to and after the copepod period. Control groups were fed on copepods or Artemia only. The Artemia diet initiated an earlier intake of food and higher initial growth compared to the copepod diet. After 50 days of feeding, the average dry weights of the fish fed on Artemia and copepods were quite similar to the copepod-fed fish, while the Artemia-fed fish were the smallest in size. The lowest frequency of normally pigmented juveniles was found in the Artemia-fed group (66.4%), while the copepod group showed almost 100% normal pigmentation. A significantly higher frequency of pigmentation was found in juveniles given a copepod diet close to the initiation of metamorphosis than those provided with an earlier copepod period of equal duration. A high degree of eye migration was found in all groups, but was lowest in the Artemia-fed group. The initial stage of eye migration was found to occur at a larger body size in fish given Artemia and copepods, or a copepod diet than in fish fed on Artemia alone. There was no significant correlation between eye migration and growth rates prior to metamorphosis, although the largest individuals exhibited the most complete eye migration. High pigmentation frequencies were obtained in fish with a low 22:6n-3:20:.5n-3 (DHA:EPA) ratio (< 1.0).     

Growth,survival rate and fatty acid composition of sterlet (Acipenser ruthenus) larvae fed fatty acid‐enriched Artemia nauplii

We evaluated the growth and survival rate of sterlet (Acipenser ruthenus) larvae fed Artemia nauplii enriched with Olioω3 or Red Pepper commercial emulsions (BernAqua NV, Belgium). Sterlet larvae, 0.022 ± 0.002 g body weight, were randomly assigned to one of three feeding regimes with two different feeding durations. After administering live feed for 7 or 14 days, larvae were weaned onto commercial food and reared to 36 days posthatching (28 days of feeding). There were no significant differences in body weight among groups at the end of the trial. A significantly higher survival rate (p < 0.05) was observed in larvae fed Artemia enriched with Red Pepper for 14 days compared to other feeding regimes. Based on the analysis of growth parameters, we can conclude that 7 days of live feeding to be sufficient for efficient rearing of sterlet larvae. And longer duration of live feeding with use of special enrichment can be recommended for a higher survival rate.     

Habitat utilization,feeding, and growth of wild spotted halibut <Emphasis Type="Italic">Verasper variegatus</Emphasis> in a shallow brackish lagoon: Matsukawa-ura,northeastern Japan

Habitat utilization, feeding, and growth of a rare pleuronectid flatfish, spotted halibut Verasper variegatus, were examined in a brackish lagoon in northeastern Japan: Matsukawa-ura. The distribution and date–length data of spotted halibut collected mainly from beam-trawl samplings during 1983–2008 indicated that age-0 juveniles [n = 25, 6.0–18.0 cm total length (TL)] and older spotted halibut (n = 71, 13.8–43.0 cm TL) inhabited almost the entire northern part of Matsukawa-ura. Comparative distribution surveys of spotted halibut, stone flounder, and marbled flounder during 1985–1989 revealed similar distribution patterns of these flatfishes; the highest densities were found around the central part of Matsukawa-ura, although significantly lower abundance was detected for spotted halibut. Linear growth equations of age-0 juveniles demonstrated that spotted halibut were able to achieve high growth after June, probably because abundant prey (e.g., mysids and gammarids) and suitable physical conditions (warmer water temperature and lower salinity) were present. The main prey items shifted from various crustaceans including mysids, caridean shrimps, and anomurans to brachyura Hemigrapsus spp. (≥20 cm TL). Our study shows that spotted halibut use the shallow brackish lagoon as an important nursery for juveniles, and also as a feeding ground for young and adults.     

The effects of salinity on reproductive development and egg and larvae survival in the spotted scat Scatophagus argus under controlled conditions

In the present study, we report the first successful instance of controlled reproduction in Scatophagus argus, which has recently emerged as a new aquaculture resource. The controlled reproduction process for S. argus was optimized with regard to salinity acclimation. Gonadal maturation was affected by salinity in both sexes. Levels of plasma 17β‐estradiol (E₂) and 11‐ketotestosterone (11‐kT) were salinity dependent and increased significantly with the duration of acclimation. Plasma levels of gonadal steroids were higher in fish held at 25‰ salinity. The highest gonadosomatic indices (GSI), 15.1 ± 1.6 in the female and 6.4 ± 1.2 in the male, were also observed at 25‰ salinity. Nevertheless, the optimal salinity for S. argus embryonic development and larval culture was 15‰. Thus, the salinity requirement for gonadal maturation and early development are quite different. The use of advanced reproductive technologies combining salinity acclimation and stimulation of luteinizing hormone‐releasing hormone analog (LHRH‐A₂) resulted in a fertilization rate of 83.2%–91.3% and embryonic survival rates of over 90%. Embryos of S. argus at the 2‐cell, blastula, gastrula and pharyngula stages were observed. Most embryos hatched after 21.0 hr of incubation at 28.0 ± 1.0°C. The development of larvae into juveniles was completed at 40–45 days posthatch (dph). In this study, we provide information about the controlled reproduction of S. argus and identify the optimal environmental parameters for S. argus embryonic and larval culture, with the aim of developing reliable reproductive techniques for its mass production.     

The use of preserved copepods in sea bream small‐scale culture: biometric,biochemical and molecular implications

Considering the well‐known problems arising from the use of rotifers and Artemia as live prey in larval rearing in terms of fatty acid deficiencies, the aim of this study was to evaluate a partial or complete replacement of traditional live prey with preserved copepods during the larviculture of gilthead sea bream (Sparus aurata). Sea bream larvae were randomly divided into 4 experimental groups in triplicates: group A larvae (control) fed rotifers followed by Artemia nauplii; group B fed a combined diet (50%) of rotifers–Artemia and preserved copepods; group C fed rotifers followed by preserved copepods; and group D fed preserved copepods solely. Survival and biometric data were analysed together with major molecular biomarkers involved in growth, lipid metabolism and appetite. Moreover, fatty acid content of prey and larvae was also analysed. At the end of 40 days treatment, a stress test, on the remaining larvae, was performed to evaluate the effects of different diets on stress response. Data obtained evidenced a positive effect of cofeeding preserved copepods during sea bream larviculture. Higher survival and growth were achieved in group B (fed combined diet) larvae respect to control. In addition, preserved copepods cofeeding was able to positively modulate genes involved in fish growth, lipid metabolism, stress response and appetite regulation.     

Can we adjust a marine cyclopoid copepod to freshwater?—First step towards a ‘universal’ live feed product for fish and shrimp larvae

The copepod Apocyclops royi tolerates very low salinity while being highly capable of synthesizing n-3 fatty acids based on PUFA-poor diets. The feasibility of culturing A. royi in freshwater fed with baker's yeast was evaluated. The copepods were adjusted for 8 days by decreasing stepwise from salinity 20 to freshwater diluting with deionized water. Initially, the cultures exhibited adequate survival and reproduction. However, after 10 days in freshwater, they stopped reproducing presumably due to the lack of vital ions in the medium. Therefore, before continuing the freshwater cultures, we tested the survival of a batch of salinity 1 pre-acclimated adults and copepodites in various types of freshwater: deionized, bottled ‘mineral’ and tap water for 48 h. All copepods died in deionized water, but a significantly higher survival, 29% ± 11%, was found in mineral water versus 11% ± 8% in tap water. Hence, we continued culturing the copepods in freshwater using ‘Maglekilde spring’ water as diluter. The copepods regained and kept reproducing until day 71 where the experiment was terminated. After a 48 h freshwater versus salinity 20 exposure, nauplii survival was significantly less in freshwater 25 ± 12 versus 79 ± 20 (±STD) at salinity 20 but not different in mean body length. For the first time, we conducted NMR metabolomics revealing a large decrease in glycine betaine and a large increase in lactate in the copepods when challenged by freshwater versus salinity 20. There is obviously a ‘price to pay’ when culturing not only the copepods in freshwater, but also perspectives in developing a ‘universal’ A. royi live-feed product.