Two in vitro methods for screening potential parasiticides against Ichthyophthirius multifiliis using Tetrahymena thermophila

Ichthyophthirius multifiliis (Ich) is a ciliate parasite that infects many species of freshwater fishes worldwide and causes heavy economic losses in aquaculture. Currently, parasiticides for controlling this parasite are limited, and few pond‐practical chemical therapies exist. Hence, the search for new parasiticides is urgently needed. One challenge confronting the screening of potential parasiticides is the difficulty in raising enough parasite for efficacy testing as Ich is an obligate parasite. This study used species of Tetrahymena, Ich‐related and cultivable ciliate protozoa, to evaluate two in vitro methods to test parasiticides. Plate counting and MTS assays (CellTiter 96^® AQ_ueous Non‐Radioactive Cell Proliferation Assay) were used to compare lethal concentrations or median lethal concentrations (LC₅₀) of copper sulphate, formalin and malachite green between T. thermophila and Ich theronts or between T. thermophila and Ich tomonts. The parasiticides that killed T. thermophila have been demonstrated to kill theronts or tomonts. These in vitro methods using T. thermophila can be used to screen novel parasiticides against Ich.     

Flavobacteria colonizing the early life stages of hatchery‐incubated Chinook salmon Oncorhynchus tshawytscha (Walbaum 1792) are markedly diverse

Flavobacterial diseases are significant impediments to hatchery‐based fishery conservation and aquaculture productivity worldwide. Recent studies revealed a multitude of novel flavobacteria within the reproductive fluids and unfertilized eggs of feral Chinook salmon Oncorhynchus tshawytscha broodstock, some of which were associated with systemic disease. Herein, embryonated eggs/fry from these broodstock were assayed for flavobacteria while in incubator stacks in three hatcheries over 2 years, as was the water entering hatchery incubators. Overall, >65% of sampled eggs and 38% of fry were colonized by flavobacteria. One hundred and ninety‐one egg and fry‐associated flavobacterial isolates were characterized phenotypically and via 16S rRNA gene sequencing and phylogenetic analyses, revealing that the majority fell into 22 clades (i.e., 15 Flavobacterium spp. groups and seven Chryseobacterium spp. groups) that varied in presence by facility. Although some matched previously described fish‐pathogenic species, the majority were distinct from all described flavobacteria and likely represent novel species. Of concern, iodophor disinfection at the commonly utilized dose/duration for egg‐surface disinfection did not eliminate flavobacteria. Results also implicated maternal routes of infection and source water for some flavobacteria. In total, study findings underscore the complexity of flavobacterial ecology within hatchery environments and highlight the need for improved hatchery biosecurity practices.     

Quantifying key parameters related to the life cycle of Caligus rogercresseyi

The salmon louse Caligus rogercresseyi (Boxshall and Bravo 2000) is a common ectoparasite of farmed salmonids in Chile. Sea lice can negatively impact the growth of hosts, adversely affecting aquaculture productivity. Unlike Lepeophtheirus salmonis (Krøyer, 1838), whose life cycle parameters have been well studied due to its importance in the Northern Hemisphere, for C. rogercresseyi no single source exists that quantifies the parameters required to model this ectoparasite's life cycle. Given that different species of sea lice have substantially different biological characteristics, it is important to parameterize the life cycle of C. rogercresseyi using appropriate observational data, rather than simply trying to adapt parameters developed for L. salmonis. Using data from existing literature, we quantified the development and survival rates for each stage in the C. rogercresseyi life cycle. We illustrate how development rates are affected by water temperature and explore the important impacts of salinity on rates of survival. We present equations that can be used to model development periods and survival proportions given certain water temperature and salinity profiles. While key parameters to quantitatively model the life cycle of C. rogercresseyi are presented, further research is required to adequately model the complete population dynamics of this ectoparasite on Chilean salmon farms and consequently to support decision-making to achieve effective control and mitigation.     

Effect of temperature on energetic demands during the last stages of embryonic development and early life of Octopus vulgaris (Cuvier, 1797) paralarvae

This study describes the effect of seasonal average temperatures (14 and 18°C) in the Ría of Vigo, on the utilization of external yolk over the last five Naef stages of development (XV–XX) for Octopus vulgaris embryos. Also, the transference of the outer yolk to the inner yolk sac, and its use during embryonic development and early life by O. vulgaris paralarvae. Temperature had a marked effect on embryonic development, except during stages XV–XIX (until the second inversion) where development time was the same (14 days), regardless of temperature. There were no significant differences in outer yolk decrease between consecutive Naef stages at 14°C and 18°C. Contrary, significant differences at all Naef stages from XV to XIX (both, with or without outer yolk) were observed for inner yolk between temperatures. A higher accumulation of inner yolk in embryos at 14°C was observed, due to lower yolk consumption. Paralarvae incubated at both temperatures were maintained independently at starvation during 4 days. At 18°C, a reduced accumulation of inner yolk, especially during Naef stage XIX, was observed. In 24 h old paralarvae, there was already significant higher inner yolk content at 14°C than at 18°C. Unfed paralarvae at 18°C lost weight faster than those at 14°C, due to higher energetic requirements. Finally, from these results, we propose a paralarvae rearing protocol during the first days after hatching and during the last five Naef stage (XV–XX) at lower temperatures, since the energy requirements are lower during the initial maturation stage.     

Effects of supplementary fennel (Foeniculum vulgare) essential oil in diet on growth and reproductive performance of the ornamental fish,Convict cichlid (Cichlasoma nigrofasciatum)

This study evaluated the effects of diet containing 0 (C: control), 75 (D₁), 100 (D₂), 125 (D₃) and 150 (D₄) mg kg^?1 F. vulgare essential oil on growth and reproductive performance of C. nigrofasciatum. A total of 225 Convict cichlids female with mean weight (1.65 ± 0.02 g) were distributed into 15 glass aquaria (15 fish tank^?1) in triplicate treatments. During 40 days of experimental period, fish fed at 3% of their body weight daily. At the end of experiment, growth indices, gonadosomatic index and reproductive indices were determined. The best feed conversion ratio was obtained in D₄ (1.19 ± 0.03, P < 0.05). The addition of F. vulgare essential oil did not have any statistical effects on other growth indices (specific growth rate, weight gain and condition factor). Total survival rates in all treatments and control were uniformly high, ranging from 93% to 97% (P > 0.05). F. vulgare essential oil has no negative effect on survival rate of C. nigrofasciatum. There were no significant differences among protein content of fish body (P > 0.05). The lowest value of fat content (26.5 ± 0.5%) was observed in D_2, and it had significant difference with control and other groups (P < 0.05). The ash percentage in treatments fed with F. vulgare essential oil were higher than control fish (P < 0.05). The highest moisture content was detected in D₄ (72.68 ± 0.19%) and D₂ (71.23 ± 2%) groups. Among the used dosage, D₄ was the most effective dosage that could significantly increase GSI (11.06 ± 1.55%), fecundity (340 ± 21) and hatching ratio (92.33 ± 1.63%). There were no significant differences in diameter of eggs among various treatments (P > 0.05).     

Breeding and life cycle of the ornamental freshwater shrimp Neocaridina davidi in a biofilm‐based culture system

Neocaridina davidi is a popular shrimp in the aquarium industry; however, information regarding its husbandry is scarce. In this study, we investigated the contribution of biofilm to its life cycle, comprising three successive phases: (1) the evaluation of biofilm growth on plastic nets (PN), plastic sheets (PS) and agrovelo (AV); (2) the reproduction of adult shrimp to get juveniles (JI); and (3) the effects of biofilm on the survival and growth performance of JI. Trials were performed in aquaria with zero water exchange and natural environmental conditions. Biofilm was composed mainly of microalgae, diatoms, cyanobacteria and ciliates and used as the sole diet. Survival, biomass and biochemical reserves of JI reared in this culture system were significantly higher in the presence of PN and AV substrates. The occurrence of ovarian maturation and egg incubation of female shrimp in these treatments indicate that biofilm supplied the energy required for somatic growth and fecundity. Harvested females also displayed the size and the red pigmentation associated with premium pricing. Based on these results, it is concluded that N. davidi can complete the life cycle and display characteristic life history traits in a low‐cost biofilm technology system without losing economic value as ornamental species.     

Effects of controlled air exposure on the survival,growth, condition,pathogen loads and refrigerated shelf life of eastern oysters

The benefits of exposing eastern oysters to air during commercial culture have not been well‐characterized. An adjustable longline system (ALS) designed in Australia and recently adopted by the nascent aquaculture industry in the northern Gulf of Mexico, allows growing oysters at any position in the water column and is perfectly suited to study the benefits of air exposure. Four‐month old diploid oysters were deployed in an ALS and divided into three groups: 1) oysters exposed to air daily for 8–12 hr during low tide, 2) oysters exposed to air weekly (~24 hr once a week), and 3) oysters kept subtidally. Oyster mortality and growth rates, Perkinsus marinus load and condition index were then determined every 3 months over 2 years, while refrigerated shelf life and Vibrio vulnificus load were determined in summer and early fall of the second year. Summer mortalities were delayed, P. marinus infection intensities tended to be lower and condition index was significantly higher in oysters exposed to air daily compared with oysters exposed to air weekly or held subtidally. Shell heights of oysters exposed to air daily were lower for most of the study due to a lower growth rate during the initial sampling interval following deployment. No consistent differences were found in V. vulnificus loads or refrigerated shelf lives between the groups. It is recommended that ALS be set so that oysters are kept subtidally fall through spring to promote growth, and exposed to air daily during summer to delay P. marinus proliferation.     

Efficacy of single and multiple doses of fenbendazole against gill parasites (Dactylogyrus sp.) of Labeo rohita (Hamilton, 1822) and its physio‐metabolic effects on the fish

The study was conducted to investigate the efficacy and physio‐metabolic responses of single and multiple doses of fenbendazole (FBZ) in Labeo rohita against gill parasite (Dacylogyrus sp.). Two Isoenergetic (395 Kcal DE/100 g) and isonitrogenous (35% CP) purified diets viz. control (without FBZ) and treatment (0.2% FBZ) were prepared. The efficacy of FBZ was measured after administration of single dose (20 mg/kg b. wt.) on 0th day and multiple doses of FBZ, that is, 20 mg/kg b. wt. on 0th, 3rd and 7th day through diet. Although oral administration of FBZ at single dose could significantly reduce the parasite in fish in comparison with the control group, 100% efficacy of the drug was not achieved up to 15th day of treatment. On the other hand, multiple doses of FBZ could show almost 100% efficacy on 15th day in terms of elimination of parasites from gill. The drug induced oxidative stress in examined fish. There was no significant change in activities of hepatic and muscle aspartate aminotransferase and alanine aminotransferase of L. rohita up to 15th day after oral administration of FBZ at single dose. But both single and multiple doses of FBZ could significantly enhance the hepatic superoxide dismutase, catalase and glutathione‐S‐transferase activities in fish up to 7th day of drug administration and significantly decreased thereafter up to 15th and/or 30th day. It is concluded that oral administration of FBZ at multiple doses (20 mg/kg b. wt.) on 1st, 3rd and 7th day could be effective against Dactylogyrus sp. infection in L. rohita.     

Impact of sublethal di‐n‐butyl phthalate on the aquaculture fish species Nile tilapia (Oreochromis niloticus): histopathology and oxidative stress assessment

Phthalates such as di‐n‐butyl phthalate (DBP) and their esters are widely used plasticizers, their ubiquitous presence in daily life, inevitably leads to their restricted use due to important environmental pollution and health impacts and endocrine disruption potential. The aim of this study was to examine the effects of a sublethal concentration of 10 mg L^?1 DBP on haematocrit (HCT) values, gills and liver histology, malondialdehyde (MDA, 2‐thiobarbituric acid‐TBA reactivity) and reduced glutathione (GSH) levels in gills and liver tissue as oxidative stress biomarkers in the aquaculture fish species Nile tilapia (Oreochromis niloticus) after 24 (DBP‐24) and 96 (DBP‐96) h exposure. No differences were found between per cent HCT values in the 24 h exposure groups (P > 0.05). Response of antioxidant defence systems in liver and gill tissues of the fish were dependent on exposure duration and changed to a higher extent during 96 h. MDA levels in liver tissue increased in DBP treated fish in comparison to the control fish. However, the differences between the exposure and control groups were not significant (P > 0.05). A statistically significant decrease (P > 0.05) was recorded in gill MDA levels in the DBP‐96 group when compared to the control and DBP‐24 groups. The liver GSH levels were unchanged in the DBP treated fish. However, GSH levels were significantly lower in the gill tissue of the DBP‐96 group. Exposure to DBP caused several degenerative changes in the histology of gill and liver tissue. Gills displayed hyperaemia, epithelial lifting, oedema, talengiectasia, epithelial hyperplasia and fusion of secondary lamellae, whereas in liver several circulatory anomalies (hyperaemia, blood congestion and sinosoid dilatation) and vacuolization of hepatocytes were observed. Histopathological results demonstrated that the gills were more affected than the liver perhaps due to their direct contact with DBP.     

The effect of flow on the competition between the alien racer goby and native European bullhead

The racer goby is an invasive Ponto‐Caspian fish spreading throughout Europe. They threaten a native species of similar biology, the European bullhead, by displacing them from shelters. These shelters are necessary for reproduction as well as for protection against predators and hydrodynamic forces. However, abiotic conditions may strongly modify the outcome of an interspecific competition in the wild. Nevertheless, little is known of the effect of flow velocity on the competition between these rheophilic species, although this factor is crucial for their distribution in the field. We video‐recorded fish behaviour for 2 h in single‐species and mixed‐species pairs in the presence of single shelters at three flow velocities: 0, 10 (a velocity preferred by the racer goby) and 30 cm·s^?1 (a velocity greater than preferred by the racer goby) to determine whether the invader can deprive the native species of its shelter. At the flow of 0 and 10 cm·s^?1, the racer goby exhibited aggressive behaviour towards bullhead, and this restricted the time spent by the bullhead in the shelter. Moreover, although the flow of 30 cm·s^?1 inhibited racer goby aggression, the time spent by the bullhead in the shelter in interspecific competition was still reduced when compared to intraspecific controls. Our results suggest that under natural conditions, the racer goby displace bullheads from their shelters even at flow velocities greater than optimal for the racer goby.     

Effects of fish farming on microbial planktonic communities in the middle Adriatic sea

The effects of farming on planktonic microbial communities were investigated at the coastal sea bass/sea bream farm in the oligotrophic middle Adriatic Sea. Analyses of nutrients, chlorophyll a, cyanobacteria (Prochlorococcus), heterotrophic bacteria and nanoflagellates (both pigmented and heterotrophic) were carried out on samples taken during six seasonal cruises at different water depths. Farming activity increased the natural concentrations of some nutrients (ammonium, nitrite, phosphate, dissolved inorganic nitrogen) and changed their seasonal pattern. The values still remained typical for oligotrophic environ‐ments demonstrating no risk for eutrophication. Enhanced nutrient supply provoked an immediate increase in abundances of both autotrophic and heterotrophic microbial groups (except pigmented nanoflagellates) and chlorophyll a. The effect of farming was more visible for the heterotrophic microbial component. Results from this study reveal a potential common pattern of microbial response to farming in the oligotrophic environments. It also suggests the importance of heterotrophic microbial web in transferring the matter and energy released from the fish farms in nutrient poor environments.     

The effect of β‐1,3‐glucan derived from Euglena gracilis (Algamune™) on the innate immunological responses of Nile tilapia (Oreochromis niloticus L.)

Algamune^? is a commercial additive produced from Euglena gracilis, providing a rich source of the β‐1,3‐glucan paramylon. Isolated kidney phagocytes of Nile tilapia were incubated with graded doses (0, 8, 16, 32, 64 and 128 μg/ml) of Algamune^? and purified paramylon to gauge their ability to elicit the production of reactive oxygen species. A linear response was observed for extracellular superoxide anion for both sources but only Algamune^? for intracellular superoxide anion. After corroborating the immunostimulant properties ex vivo, a feeding trial was conducted to evaluate the dietary supplementation of Algamune^? (0, 100, 200, 400 and 800 mg/kg of diet) for Nile tilapia. Fish were fed for 3 weeks, after which, fish were sampled for blood and head kidney phagocytes. The remaining fish were challenged with Streptococcus iniae. Macrophage extracellular superoxide anion production was significantly elevated in fish fed diets with 200 mg of Algamune^? kg^?1 when compared to fish fed the basal diet. Even though the disease challenge did not show statistical differences, it is worth mentioning that fish fed intermediate doses of Algamune^? had lowest numerical mortality values. Therefore, Algamune^? was demonstrated to enhance some immunological responses of tilapia both in ex vivo and in vivo.     

Reproductive cycle of the rock oyster,Striostrea prismatica (Gray, 1825) from two locations on the southern coast of Ecuador

The reproductive cycle of the rock oyster Striostrea prismatica was determined at two fishing areas, General Villamil and Ayangue (located on the southern coast of Ecuador), between May 2012 and April 2013. Monthly sampling campaigns were performed at both locations. The tissues were histologically examined to determine gonadal index (GI), oocyte development, follicular area coverage and sex ratio. Surface seawater temperature, salinity and chlorophyll a concentration were measured during samplings. Our results show a similar annual reproductive pattern at both locations. The GI reached maximum values during the summer. Oysters reached highest ripeness in January and February, while spawning occurred in February–March. Gametogenesis was linked to a consistently increasing follicular area (from 1.3 ± 0.7% to 85.8 ± 7.8%) and associated to surface seawater temperature. Spawning coincided with warm water temperature fluctuations and a seawater salinity decrease. No correlation was found with Chlorophyll a concentration. The sex ratio of sampled populations was 1:1, suggesting that oysters sizing more than 10 cm in shell length present a stable sex proportion in the population. The diameter of mature oocytes was significantly reduced (32.7%) during histologically preparations in comparison to fresh oocytes. Our study provides useful information of environmental factors that may control the observed gametogenesis and spawning activity of S. prismatica.     

Effect of environmental conditions on the seasonal and inter‐annual variability of small pelagic fish abundance off North‐West Africa: The case of both Senegalese sardinella

The objective of this study was to assess the effect of environmental variations on the abundance of Sardinella aurita and Sardinella maderensis in Senegalese waters in the upwelling system. Monthly data indicating the abundance of sardinella were first estimated from commercial statistics, using Generalized Linear Model from 1966 to 2011. Abundance indices (AIs) were then compared with environmental indices, at the local scale, a Coastal Upwelling Index (CUI) and a coastal Sea Surface Temperature (SST) index, and on a large scale, the North Atlantic Oscillation (NAO), the Atlantic Multidecadal Oscillation (AMO) and the Multivariate El Niño Southern Oscillation Index (MEI), using correlations and times series analyses. The results showed that the abundance of sardinella is determined by a strong seasonal pattern and inter‐annual fluctuations. The abundance of S. aurita peaked in spring and in autumn, whereas that of S. maderensis peaked in the warm season (July–September). The trend of the sardinella abundance was significantly correlated with the CUI, especially in autumn and spring. Interannual fluctuations of S. maderensis and S. aurita abundance are, respectively, driven by the precocity and the duration of the upwelling season that is attributed to distinct migration patterns. Both sardinella species also respond with a delay of around 4 years to the winter NAO index and the autumn CUI, and the AMO index, respectively, both related to migration patterns. The wide variations in sardinella biomass are caused by variations in environmental conditions, which should be considered in the implementation of an ecosystem‐based approach in sardinella stocks management.     

Effects of replacing dietary fish oil and squid liver oil with vegetable oils on the growth,tissue fatty acid profile and total carotenoids of the giant freshwater prawn, Macrobrachium rosenbergii

A feeding trial was conducted to investigate the complete substitution of either fish oil (FO) or squid liver oil (SLO) with crude palm oil (CPO), canola oil (CO) sunflower oil (SFO) or linseed oil (LO), as the sole added lipid source in diets fed to triplicate groups of giant freshwater prawn, Macrobrachium rosenbergii (initial weight = 0.42 ± 0.01 g) for 6 weeks. Prawns fed the CO or SLO diets showed significantly higher (P < 0.05) specific growth rate than those fed the FO or CPO diets. The feed conversion ratio of the prawns was significantly better when fed the CO diet, compared with the FO, CPO, SFO and LO diets. The muscle eicosapentaenoic acid content of prawns fed the vegetable oil (VO) diets were not significantly different (P > 0.05) from those fed the FO diet, although all VO‐based diets led to a significantly lower docosahexaenoic acid content compared with prawns fed the FO or SLO diet. The whole‐body total carotenoid content was significantly lower for prawns fed the SLO diet compared with prawns on the CO or CPO diets. The successful use of VO instead of marine‐based oils in prawn diets will likely reduce feeding costs associated with M. rosenbergii aquaculture.     

The investigation of the effects of different feeding strategies on the growth of dolphin cichlid (Cyrtocara moori Boulenger, 1902) fish

This study was designed to investigate the effects of restrictive and cyclical feeding on the growth parameters of dolphin cichlid (Cyrtocara moorii Boulenger, 1902). In the study, 300 dolphin cichlid with an initial average weight of 1.12 ± 0.23 g were placed in 15 aquariums (each of them were 60 L). The five experimental groups, were designed as; the control group (A), feeding until satiation twice a day for 90 days, starvation for 1 week (B), starvation for 2 weeks (C), starvation for 1 day/satiation for 3 days (D), starvation for 1 day/satiation for 1 day (E); and the study was carried out in three repetitions. At the end of the study, groups B, C and D showed partial compensatory growth. The highest weight gains and specific growth rates were observed in control group (A), followed by groups B, C and D (p > 0.05). The lowest weight gain was observed in group E (p < 0.05). The feed conversion ratio did not show any significant difference between the groups (p > 0.05). In terms of feed consumption ratio, Control, C and D groups consumed similar amounts of feed (p > 0.05), while B and E groups consumed less feed (p < 0.05) than the other groups.     

The effect of replacement of fish meal by yeast extract on the digestibility,growth and muscle composition of the shrimp Litopenaeus vannamei

Feeding trials were conducted to determine the dietary level of yeast extract (YE) for replacing dietary fish meal for evaluating whether yeast extract was superior to intact yeast as an alternative protein source for shrimp Litopenaeus vannamei. The basal diet (control, D0, containing 25% fish meal), was compared with five isonitrogenous and isoenergetic experimental diets [replacing 15% (D15), 30% (D30), 45% (D45), 60% (D60) or 100% (D100) of the fish meal in the basal diet with IYE]. The digestibility, growth and muscle composition of the shrimp were measured. The results showed that all replacement treatments displayed higher apparent digestibility of crude protein than did the control. The trypsinase activity in shrimp hepatopancreas increased significantly, whereas lipase activity decreased as the amount of dietary YE increased. The shrimp treated with D30 diet displayed the highest amylase activity in hepatopancreas. There was no significant difference in the weight gain (WG) and survival of shrimp between the control and the YE replacement treatments. Feed conversion ratio (FCR) increased as the dietary YE increased, and the FCRs of the D60 and the D100 treatments were significantly higher than that of the control (P < 0.05). The growth performance among the treatments was closely related to the similarity of the essential amino acids in the diets. There was no significant difference in muscle composition of the shrimp between control and other treatments. In conclusion, up to approximately 45% of the fish meal in shrimp diet can be replaced by yeast extract in the presence of supplemental fish oil, phosphorus and calcium.     

The effect of photoperiod and feeding frequency on larval of the Amazonian ornamental fish Pyrrhulina brevis (Steindachner, 1876)

This study aimed to evaluate the performance of Pyrrhulina larvae, Pyrrhulina brevis, under different photoperiods and feeding frequencies. A completely randomized design was used, with four replicates in a 5 × 2 factorial, with five photoperiods (6L:18D, 12L:12D, 16L:8D, 20L:4D, 24L:0D) and two feeding frequencies (two or four times a day). Four hundred larvae with an initial length of 4.6 ± 0.09 mm were randomly distributed in 40, 1‐L containers at a density of 10 larvae/L. Feeding was performed with Artemia nauplii in the ratio of 150 Artemia nauplii/larva/feeding. After 15 days, all larvae were euthanized for measurements of length (mm), final weight (mg), survival rate (%) and larvae uniformity in weight an length (%). The data were subjected to analysis of variance and subsequently, in case of significance, a Tukey test was performed to compare means. There was no interaction between photoperiod and feeding frequency on studied variables (P > 0.05). Likewise, there was no influence of feed rate on the same variables (P > 0.05). There was only influence of photoperiod, where the larvae subjected to longest photoperiods (24L:0D and 20L:4D) showed improved weight and length (P < 0.05). Therefore, a photoperiod of 20L:4E, with feeding twice a day is recommended.     

Antioxidant activity and lipid peroxidation in Artemia nauplii enriched with DHA‐rich oil emulsion and the effect of adding an external antioxidant based on hydroxytyrosol

Artemia nauplii catabolize polyunsaturated fatty acids (PUFA); in particular, they retroconvert docosahexaenoic acid (DHA, 22:6n‐3), so enrichment is a continuous quest towards increasing PUFA through the use of PUFA‐rich enrichment products. However, optimal conditions during enrichment (aeration, illumination and temperatures around 28°C) tend to accelerate autoxidation of PUFA and the formation of potentially toxic oxidation products. Water‐soluble antioxidants like the polyphenolic compound hydroxytyrosol (3,4‐dihydroxyphenylethanol), a polar molecule found in the water fraction resulting after the milling process of olives, arise as promising compounds to prevent oxidation during Artemia enrichments. We investigated the antioxidant activity and lipid peroxidation in Artemia nauplii during enrichment and the effect of adding an external antioxidant based on hydroxytyrosol during the enrichment with a PUFA‐rich emulsion (M70). For this purpose, the activity of antioxidant enzymes (catalase, superoxide dismutase, glutathione‐S‐transferase, glutathione peroxidase), as well as lipid peroxidation, was determined in enriched and unenriched Artemia nauplii. To validate antioxidant activity and lipid peroxidation, in a first experiment, nauplii were enriched with microalgae (Tetraselmis suecica), yeast (Saccharomyces cerevisiae) and M70 emulsion. In a second experiment, enrichment with a commercial emulsion (DC Super Selco), M70, and a combination of M70 and hydroxytyrosol (Hytolive, HYT) added as an external antioxidant were performed. The combination of M70 with HYT produced the best results, in terms of activity of antioxidant enzymes. The analysis of the fatty acids from total lipids showed that the addition of hydroxytyrosol preserved the DHA percentage of enriched nauplii.