Salinity-induced quiescence in eggs of the calanoid copepod Acartia tonsa (Dana): a simple method for egg storage

We report the effect of salinity and temperature on the viability of stored culture-based subitaneous eggs of the calanoid copepod Acartia tonsa for use of copepods in fish larvae culture. Quiescence induction was recorded at 17 and 25 °C, in salinities from 0 to 30. Quiescence was strongly induced at 0 salinity and partially at 5 in both temperatures. Eggs incubated at 0 salinity for up to 12 days at both temperatures showed a decline in the fraction able to be induced into quiescence by abrupt salinity changes. The hatching success of eggs that were able to enter quiescence stabilized after a 1-day incubation and remained ∼25% viable for 12 days in 17 °C. On the contrary, the 25 °C trial showed a gradual decline in viability until stabilizing ∼10% at day 7 and onwards. Longterm 17 °C incubation for 35 days showed that eggs remained quiescent with a viability of ∼14%. Hence, we recommend salinity storage of A. tonsa subitaneous eggs as a relevant shortterm technique, and a suitable alternative to the recently proposed cold storage of eggs when eggs are to be shipped from the copepod producer to a given fish larvae hatchery.     

Effects of adult stocking density on egg production and viability in cultures of the calanoid copepod Acartia tonsa (Dana)

The effect of stocking density of the calanoid copepod Acartia tonsa was evaluated in a 96 h rearing experiment. Possible density‐dependent egg production and egg viability were analysed at stocking densities of 100, 200, 300, 400 and 600 adults L⁻¹. Temperature, oxygen saturation and algal concentration were kept optimal. A non‐density‐dependent mortality rate of 15–19% day⁻¹ was documented. A non‐significant density‐dependent egg production was observed between 100 and 600 adults L⁻¹. The average egg production was 22.5±8.8 egg female⁻¹ day⁻¹ in all densities. The average egg hatching success was 84.7±4.8% and was never observed below 76.1%, with no significant differences across the stocking densities. Conclusively, as a practical recommendation for the aquaculture industry, copepod cultures with densities ranging from 100 to 600 adults L⁻¹ and presumably even more dense cultures are possible with the studied species obtaining a steady egg production and still high egg viability.     

Tolerance of un‐ionized ammonia in live feed cultures of the calanoid copepod Acartia tonsa Dana

Optimal water quality is considered as being a restriction for marine copepod cultures for live feed. There is a lack of knowledge on the water‐quality conditions in copepod cultures and the effect on copepods. Few studies have investigated the effect of ammonia on copepods, and fewer reports No Observed Effect Concentrations (NOEC) and Lowest Observed Effect Concentrations (LOEC), which provides safety levels before cultures are affected. This study investigates the tolerance of Acartia tonsa nauplii and adults to ammonia, using mortality as the endpoint after 24, 48 and 72 h of exposure. Nauplii were exposed to levels from 0 to 5127 μg NH₃ L^?1 and adults to levels from 0 to 8481 μg NH₃ L^?1. Nauplii NOEC was 30 μg NH₃ L^?1 and LOEC was 81 μg NH₃ L^?1. Adult NOEC was 477 μg NH₃ L^?1 and LOEC was 1789 μg NH₃ L^?1. 50% Lethal Concentrations (LC₅₀) for nauplii of 48 and 72 h was 1257 and 220 μg NH₃ L^?1. LC₅₀ for adults was 2370 (24 h), 972 (48 h) and 770 (72 h). Combining NOEC with excretion rates of NH₄/NH₃ a model was developed to calculate densities in batch cultures. We recommend that for batch cultures of A. tonsa, NH₃ is kept below NOEC for nauplii and that levels of NH₃ together with pH are monitored weekly.     

Influence of storage conditions on viability of quiescent copepod eggs (Acartia tonsa Dana): effects of temperature, salinity and anoxia

Copepods have proven to be an ideal source of live food for the production of marine fish larvae in aquaculture. Therefore, there is a need to develop new methods for production and storage of copepod eggs that can be hatched and used at fish farms. In the present study quiescent eggs of Acartia tonsa were stored for periods up to 35 weeks at different temperatures, salinities and oxygen conditions in a full factorial experiment. None of these storage conditions seemed to induce diapause in eggs even though this has been reported by other authors. The most promising storage conditions were those involving low temperature (<5°C), medium salinity (10–20 ppt) and anoxia. The practical aspects of these results for aquaculture are discussed.     

Egg hatching rate and fatty acid composition of Acartia bilobata (Calanoida,Copepoda) across cold storage durations

To investigate egg storage capacity of the copepod Acartia bilobata for aquaculture interest, we tested hatching success rate (HSR) of inclusive eggs (mixture of all egg types) after 4°C storage. The HSR peaked after 14 days storage when incubating at 28°C for 48 hr (85.8 ± 1.6%) and 72 hr (87.6 ± 0.9%), then gradually declined until 1 year (48 hr: 7 ± 0.6%; 72 hr: 19.4 ± 3.9%). Reallocation of fatty acid profile suggests that docosahexaenoic acid (DHA) is correlated with the HSR of A. bilobata eggs. Additionally, we investigated the HSR of diapausing eggs (unhatched eggs after 72 hr incubation of the inclusive eggs) after 4°C storage. Their HSR peaked after 14 days storage (48 hr:75.3 ± 3.5%; 72 hr:78.2 ± 2.1%), then gradually declined until 60 days (48 hr HSR:42.1 ± 2.3%; 72 hr HSR:53.0 ± 3.2%). Overall, we illustrated the hatchability of diapausing and quiescent eggs of A. bilobata after 4°C storage. The cold storage capacities were low (<60% HSR after 60 days), and it could be limited by the egg DHA content. Our findings provide implications for future studies aiming to improve cold storage techniques of tropical copepod eggs for aquaculture applications.     

How to increase productivity of the copepod Acartia tonsa (Dana): effects of population density and food concentration

In this study, we analysed the effect of population density and food concentration on the fecundity of a Mediterranean strain of Acartia tonsa to maximize egg production. During 4‐day feeding experiments, egg hatching success and faecal pellet production were also followed. The algae Rhinomonas reticulata was supplied at different concentrations corresponding to 250, 500, 1000, 1500, 2000 and 3000 μg C L^?1 day^?1 at the following adult copepod density: 40, 80 and 160 ind. L^?1. Our results show a positive relationship between algal concentration and egg production under all experimental conditions confirming that the quantity of food strongly limits A. tonsa fecundity. Maximum egg production (57 eggs per female) was reached at the lowest density and at the maximum food concentration. Percentage of egg hatching success was not dependent on the quantity of food used. At the same food concentration, an increase in population density from 40 to 80 ind. L^?1 induced an increase in faecal pellet production per couple which did not correspond to an increase in egg production, suggesting that higher energetic costs were shifted to swimming activity. Productivity of the A. tonsa Mediterranean strain is mainly limited by the quantity of food rather than by crowding conditions.     

The effects of temperature and photoperiod on egg hatching success,egg production and population growth of the calanoid copepod,Acartia grani (Calanoida: Acartiidae)

Calanoid copepods, including species of the genus Acartia, are commonly used as larval diets for marine finfish. This study aimed to determine the separate effects of water temperature (18, 22, 24, 28° ± 0.5°C) and photoperiod (24L:0D; 18L:6D; 12L:12D; 8L:18D; 0L:24D) on Acartia grani egg production (EP), hatching rate (EHR) and population growth. Egg production rate was not affected by the two abiotic parameters. A. grani eggs incubated at T24°C and T28°C were the first to achieve 50% hatching rate (23–25 hr), with significant differences at the end of the experiment (48 hr) between T28°C treatment (EHR 88 ± 5%) and T18°C treatment (EHR 65 ± 2%). However, different temperature regimes did not affect final number of individuals in population growth experiment. Still, when eggs were excluded from data, population at lower temperatures (18°C) was mainly composed by the nauplii stage (72%), while at higher temperatures (24°C and 28°C) more than 60% of the population was composed by copepodites and adults. A. grani subjected to long‐day photoperiods had significantly lower EHR (16.7% at 24L:0D; 20.8% at 18L:6D) than at short‐day photoperiods (52.6% at 6L:18D; 50.0% at 0L:24D). In population growth experiment, eggs were the most common life stage after 12‐day culture. Lowest population number was found at constant light conditions (665.0 ± 197.1), suggesting higher metabolic rates and depletion of energy reserves in long‐day conditions. This study expanded knowledge on the biological response of A. grani to separate temperature and photoperiod regimes, and provided ground to improve the culture of this potential life feed species for hatcheries.     

Total egg harvest by the calanoid copepod Acartia tonsa (Dana) in intensive culture – effects of high stocking densities on daily egg harvest and egg quality

Understanding the factors limiting copepod productivity in dense cultures is a prerequisite for the partial or entire replacement of Artemia and rotifers as live feed for finfish larvae. In dense cultures, high encounter rates between individuals may increase stress, cannibalism incidents and potentially trigger resting egg production. We conducted an experiment to evaluate the potential egg production and egg quality of Acartia tonsa stocked at densities ranging from 10 to >5000 ind. L^?1. Egg Production (EP), Delayed Hatching Eggs production (DHE), hatching success (HS), egg mortality and water quality were used as end points. In the present system, A. tonsa was raised at >5000 ind. L^?1 without affecting the mortality, confirming that attaining this high density in culture is possible. However, egg harvest reached an optimum of 12 000 egg L^?1 day^?1 at ~2500 ind. L^?1 indicating that increasing stocking density above this level is not of practical interest. Calculations showed that the loss in egg harvest at stocking densities <2500 ind. L^?1 is of 1.3% for every additional 100 adult copepods L^?1. The increasing adult density did not affected the proportion of DHE produced (~10% of harvest) but decreased significantly the HS, though not to a point that would be problematic in a commercial production. Understanding the biology of copepods when stocked at high density is important to improve copepod culture systems and increase egg harvest yields. Technical solutions such as the continuous separation of eggs from adults in the water column, recirculation and the continuous provision of food are seen as potential solutions.     

A method for collecting eggs of Pseudocapillaria tomentosa (Nematoda: Capillariidae) from zebrafish Danio rerio and efficacy of heat and chlorine for killing the nematode's eggs

Pseudocapillaria tomentosa is a common pathogen of zebrafish (Danio rerio) in research facilities. We developed a method to collect and concentrate the nematode eggs using a modified sugar centrifugation method and documented their normal development. Embryonating stages with blastomere formation followed by elongation of the embryo prior to larva formation cumulated in developed larvae inside the eggs and hatching after 5–10 day. We then evaluated the efficacy of heat and chlorine to kill them based on a larva development assay. Eggs were exposed to 40, 50, 60 °C for 30 min and 1 h. Chlorine treatment was performed at 100, 250, 500, 1000, 3000 and 6000 ppm for 10 min. Samples exposed to 40 °C for 30 min or 1 h showed incidences of larvated eggs similar to controls. In contrast, no larvation occurred with eggs exposed to either 50 or 60 °C for 30 min or 1 h. Remarkably, in repeated assays, samples exposed to low doses of chlorine (100, 250, 500 and 1000 ppm for 10 min) showed significantly higher incidence of larvation than controls. Eggs treated with 3000 ppm for 10 min did not develop larvae, and no eggs were found after 6000 ppm treatment.     

Effects of storage and incubation temperature on the viability of eggs, embryos and larvae in two strains of an African catfish, Heterobranchus longifilis (Siluriformes, Clariidae)

Two experiments, dealing with short‐term storage of ova and thermal conditions to optimize gamete and eggs management in hatcheries of the African catfish, Heterobranchus longifilis, were carried out. In the first experiment, ova collected by stripping from two strains of H. longifilis were stored for intervals up to 8 h at two temperature regimes: in a domestic refrigerator (3–5°C) and at ambient room temperature (20.5–22°C). In the second experiment, eggs were incubated from fertilization to hatching at different experimental temperatures (21, 25, 29, 32 and 35°C) to determine the effects of temperature on the kinetics of white egg appearance, hatching times and hatching quality. Gamete storage at warmer temperatures significantly prolonged viability irrespective of the strain used. In fact, the hatching rate for ova stored at 20.5–22 and 3–5°C for 5 h ranged between 75.2–79.3% and 6.5–9.4% respectively. Loss of viability was most noticeable after 6 h storage at ambient room temperature. Post‐storage viability significantly declined after 2 h exposure to the domestic refrigerator temperature. No hatching of normal larvae took place after 8 h post‐storage time. Results from the second experiment showed that time to maximum whitening of eggs was both strain‐ and temperature‐dependent. The time to maximum mortality of eggs was shorter in the Layo strain (LS) than in the Noun strain (NS), regardless of incubation temperature. The appearance of white eggs was shorter with increasing incubation temperatures. Hatching times decreased with increasing temperature, regardless of strain. Hatching took place from 21 to 27 h and 19 to 24 h after fertilization at temperature of 29°C, respectively, for NS and LS. The length of the hatching period was remarkably shorter for LS than NS at any tested incubation temperature, except 35°C. No hatching took place at 21°C. The highest proportion of normal larvae occurred at 25 and 29°C, respectively, for NS and LS. Hatching rate was highest at 25 and 29°C, respectively, for NS and LS. There was a significantly higher proportion of deformed larvae at 35°C regardless of the strain.     

Effect of different chilling rates for cold anaesthetization of Penaeus monodon (Fabricius) on the survival, duration and sensory quality under live storage in chilled sawdust

The present work was carried out with a view of determining the chilling rates for cold anaesthetization and live storage of Penaeus monodon (Fabricius) in chilled sawdust, for live transportation. Three chilling rates of 1.38 ± 0.16 °C h^?1 within 8 h (slow), 2.76 ± 0.32 °C h^?1 within 4 h (moderate) and 5.52 ± 0.64 °C h^?1 within 2 h (fast) were tested to cold anaesthetize 144 farm‐raised P. monodon (22–25 g) in each chilling rate at 15 g L^?1 salinity, from 25 °C down to 14 ± 1 °C in plastic net boxes kept in a refrigerated chilling tank provided with aeration. Eight cold‐anaesthetized shrimps subjected to each chilling rate were packed in an insulated cardboard box (triplicate) between two layers of moist and chilled (2–3 °C) sawdust, and kept inside a chilled storage cabinet at 14 ± 1 °C for durations of 16, 20, 24, 28, 32 and 36 h. Survival was determined by revitalizing the shrimps in aerated water with an initial temperature of 20 °C, which was raised to 28 ± 1 °C at 2.7 °C h^?1 within 3 h. Statistically valid, safe durations for obtaining 100% survival were 22.9 ± 1.1, 19.1 ± 0.4 and 14.6 ± 1.1 h, using probit analysis at the slow, moderate and fast chilling rates respectively. The weight loss (1.5–8.8%) due to cold anaesthetization and subsequent revitalization of the packed shrimp was insignificant. Sensory evaluation showed significant improvement in general appearance, and also in colour as well as in flavour of the meat when compared with that of the freshly harvested dead shrimps. The texture as well as odour of the raw and cooked meat between treated and untreated shrimps was unaffected. The effects of chilling rates and shipping durations on sensory quality were insignificant. One hundred per cent survival was obtained for 24, 20 and 16 h for the slow, moderate and fast chilling rates respectively. Percentage survival of the shrimps at different durations was significantly different among the chilling rates, but pair‐wise comparison revealed that the slow and moderate chilling rates were identical. Hence, the moderate chilling rate, which took only 4 h, can be considered the optimum, though the choice of different chilling rates depends on the duration of live storage desired.     

The influence of temperature on advective loss of Atlantic cod (Gadus morhua) eggs from the inshore environment

We use a simple model of temperature-dependent egg development and mortality to develop several hypotheses concerning the effect of temperature on the occurrence of eggs of Atlantic cod, Gadus morhua , in Placentia Bay, Newfoundland. Predictions from this exploratory model were tested through a series of synoptic ichthyoplankton surveys throughout spawning and postspawning periods during 1997 and 1998. Although several egg mortality relationships were explored through the simulation, a constant mortality rate best represented the pattern observed in the two years of data. Peaks in late stage egg densities occurred in August of both 1997 and 1998 and were apparently decoupled from egg production peaks in April. We observed a decrease in mortality and the distance dispersed during egg development with increases in water temperature. We suggest that the effects of predation are small relative to the advective effects within this system, and that the interaction between advection and temperature-dependent vital rates of eggs may have dramatic consequences for coastal retention of propagules produced by inshore spawning events.     

Changes in lipid content, fatty acid composition and lipid class composition of eggs and developing larvae (0–40 days old) of cultured common dentex (Dentex dentex Linnaeus 1758)

Total lipid content, fatty acid (FA) composition and lipid class composition of common dentex eggs spawned at different times and larvae reared under different culture conditions until 40 days posthatch (dph) were analysed to get a general pattern of lipid composition during larval development. Two groups of larvae were kept under starvation to compare their FA composition with that obtained from normally fed larvae. To compare FA use or accumulation during larval development, results were grouped according to the developmental stage of the larvae instead of age in days posthatch. Saturated and monounsaturated FAs decreased along larval development, while polyunsaturated fatty acid (PUFA) content increased. The ratio of docosahexaenoic acid (DHA)/eicosapentaenoic acid shifted from 4 to 5 in early developmental stages to lower than 1 after metamorphosis. Arachidonic acid levels remained constant along larval development. Larvae kept 6 days under starvation consumed most of their n-3 PUFA while conserving the DHA to values at day 0. The results presented here are useful for the design of nutritional experiments, because there were differences detected in terms of lipid and FA composition between developmental stages with higher differences mainly found in first-feeding larvae and early developmental stages.