Analysis of the diet of the long‐snouted seahorse Hippocampus guttulatus by 18SrDNA amplification of prey in faeces

Breeding in captivity for research or exhibition (e.g. in aquaria) can replace the capture of wild specimens of endangered species and allow controlled reinforcement of wild populations. With this aim, diet analysis and establishing the adequate prey are determinant factors for breeding success. However, non‐invasive approaches such as faecal DNA analysis are advisable for analysing the diet of these species. Therefore, the aim of the present study was to demonstrate the usefulness of faecal DNA analysis by specific PCR amplification of prey DNA for assessing the diet of the seahorse Hippocampus guttulatus. In a comparison of the suitability of different genes (COI, 18SrRNA and 28SrRNA), 18SrRNA was found to be the most suitable for designing specific primers for the prey types fed to seahorses (Artemia, Palaemonetes and Mysidae). The technique was assessed in feeding experiments in which prey ingestion was recorded daily, and faeces were collected for DNA extraction and presence/absence PCR analysis. Amplification of the prey DNA in faeces was consistent with the sequence of prey supplied (prey eaten the day before was always detected). Differences in the time between feeding and detection in faeces suggested prey‐specific gut passage times, which were shorter for Palaemonetes than for Mysidae. This fact highlights the importance of feeding trials to avoid overestimating the consumption of prey with long gut retention when PCR‐based methods are used. This molecular technique is thus applicable for studying the feeding behaviour of captive seahorses and could be adapted for use in other marine species.     

Diseases of captive yellow seahorse Hippocampus kuda Bleeker,pot‐bellied seahorse Hippocampus abdominalis Lesson and weedy seadragon Phyllopteryx taeniolatus (Lacépède)

Seahorses, pipefish and seadragons are fish of the Family Syngnathidae. From 1998 to 2010, 172 syngnathid cases from the Toronto Zoo were submitted for post‐mortem diagnostics and retrospectively examined. Among the submitted species were yellow seahorses Hippocampus kuda Bleeker (n = 133), pot‐bellied seahorses Hippocampus abdominalis Lesson (n = 35) and weedy seadragons Phyllopteryx taeniolatus (Lacépède; n = 4). The three most common causes of morbidity and mortality in this population were bacterial dermatitis, bilaterally symmetrical myopathy and mycobacteriosis, accounting for 24%, 17% and 15% of cases, respectively. Inflammatory processes were the most common diagnoses, present in 117 cases. Seven neoplasms were diagnosed, environmental aetiologies were identified in 46 cases, and two congenital defects were identified.     

Effect of Different Live Prey on Spawning Quality of Short‐Snouted Seahorse,Hippocampus hippocampus (Linnaeus, 1758)

The importance of a suitable diet for reproduction has been recognized as one of the major factors in ornamental aquaculture. In seahorses, mysids have been described as preys in the wild. Also, Artemia has been usually employed for rearing fish, including syngnathids. Therefore, the aim of this work was to evaluate the effect of these live preys on the spawning quality of Hippocampus hippocampus. After 108 d, no differences were found in adults concerning all biological parameters evaluated, but broodstock fed on mysids showed better results than Artemia regarding number of spawning events (12 vs. 3), brood sizes (233.50 ± 59.04 vs. 68.00 ± 57.97 juveniles), and newborn seahorses standard length (10.61 ± 0.64 vs. 8.75 ± 1.32 mm). The better nutritional quality of mysids, overall in Docosahexanoic acid, could be one of the main responsible factors. However, mysids stock is conditioned by natural catches and rearing techniques are little known. Another alternative would be to combine them with Artemia in mixed diet. Further research must be done concerning mysids breeding techniques to delineate their employment as a sustainable prey for seahorse aquaculture. This trial showed for the first time the effect of mysids to enhance reproduction efficiency in H. hippocampus.     

Effects of the diet on seahorse (Hippocampus hippocampus) growth,body colour and biochemical composition

The short‐snouted seahorse (Hippocampus hippocampus) is a benthic fish that bases its survival on camouflage with the environment; therefore, both in wild and in captivity specimens, differences in coloration have been observed. This study was conducted to study the effect of five different diets, based on Artemia and frozen mysis, on fish survival, growth, skin colour parameters and biochemical composition. After 102 days of feeding different diets, no effect was observed on fish survival, growth, protein, ash or humidity content, except for colour changes on yellow and red fish. Differences on seahorse lipid and total carotenoids content were obtained in the experiment because of experimental diets. Fish eating Artemia showed the highest colour pattern response during the trial, with a maximum yellowness attained of 21. Additionally, a plateau of the skin colour expression for Artemia‐feeding fish was reached on day 180. A reversion of the yellowish skin colour was determined after a change of diet.     

The impact of dietary supplementation with astaxanthin on egg quality and growth of long snout seahorse (Hippocampus guttulatus) juveniles

This study investigated the effect of dietary astaxanthin supplementation on egg quality and juvenile growth of long snout seahorse (Hippocampus guttulatus). Captive breed seahorse broodstock were fed four diets composed of frozen shrimp [Atlantic ditch shrimp, Palaemonetes varians) used as a vector to deliver artificial diets with increasing levels of astaxanthin (0, 75, 100 and 125 mg kg^?1 dry weight)]. The results indicated that the astaxanthin uptake into eggs from the enriched shrimp diets was highly efficient. Females fed unsupplemented astaxanthin diet produced similar‐sized eggs with lower concentration of astaxanthin than females fed diets with astaxanthin. The lower concentration of astaxanthin in the eggs was correlated with the production of smaller juveniles in comparison with the juveniles hatched from parents fed supplemented astaxanthin diets. Juvenile growth and survival was limited by their size on release from the male's pouch as at the end of 28‐day postparturition juveniles produced with the diet with no astaxanthin were still significantly smaller (P < 0.05) than those produced from parents fed astaxanthin‐supplemented diets. These results demonstrate a significant benefit of dietary astaxanthin supplementation in long snout seahorse diets in terms of improved egg quality and juvenile growth and survival.     

Feeding selectivity of the seahorse,Hippocampus kuda (Bleeker), juveniles under laboratory conditions

This study examined the feeding selectivity of Hippocampus kuda juveniles under captive conditions and evaluates different food organisms that could be used to improve hatchery‐rearing of this species. Newly born H. kuda were reared for 10 days in 60‐L capacity tanks and fed rotifers (Brachionus rotundiformis), zooplankton (mostly Pseudodiaptomus annandalei and Acartia tsuensis) alone or both food sources. The size and amount of food ingested increased as seahorses grew. Selective feeding of seahorses appeared to change as they develop, preferring copepod adults over nauplii and rotifers. A. tsuensis was highly selected by juveniles over P. annandalei. Specific growth rate in terms of body weight (SGR‐BW, 15% day^–1) was the highest and mortality rate (9% at day 10) the lowest in seahorses fed a mixed food sources. Slowest growth rate (0.3% day^–1) and highest mortality rate (60% at day 7) were observed in seahorses fed rotifers alone. These results indicate that copepods are suitable food for seahorse juveniles, but a mixture of food organisms in the rearing tank environment enhances survivorship and growth of H. kuda, thus potentially providing a source of cultured rather than wild specimens for characterizing the life history of this threatened species.     

Establishing larval feeding regimens for the Forktail Blenny Meiacanthus atrodorsalis (Günther, 1877): effects of Artemia strain,time of prey switch and co‐feeding period

This study aimed to establish feeding strategies covering the whole larval period of the forktail blenny, Meiacanthus atrodorsalis, based on the standard hatchery feeds of rotifers and Artemia. Three purposely designed experiments were conducted to determine the appropriate times and techniques to transition larvae from rotifers onto Artemia nauplii of a Great Salt Lake (GSL) strain, and a specialty AF strain, as well as subsequent transition onto enriched metanauplii of GSL Artemia. With a 3‐day co‐feeding period, larvae adapted well to a transition from rotifers to newly hatched GSL Artemia nauplii as early as 5 days posthatching (DPH), and as early as 3 DPH when fed the smaller AF Artemia nauplii. However, prolonging the rotifer‐feeding period up to 11 DPH did not negatively affect survival. Larvae fed Artemia nauplii of the AF strain showed 17–21% higher survival, 24–33% greater standard length and body depth, and 91–200% greater dry weight, after 20 days relative to those fed nauplii of the GSL strain. Meanwhile, enriched Artemia metanauplii of the GSL strain were shown to be an acceptable alternative to AF Artemia nauplii for later larvae, producing similar survival and growth when introduced from 8 DPH. Based on our findings, we recommend feeding M. atrodorsalis larvae rotifers as a first food between 0 and 2 DPH, introducing AF Artemia nauplii from 3 DPH, followed by enriched GSL Artemia metanauplii from 8 DPH onward, with a 3‐day co‐feeding period between each prey change.     

Study on an integrated eco‐aquaculture system for rearing the yellow seahorse,Hippocampus kuda Bleeker

Seahorse aquaculture is challenged by insufficient feeding supply and lack of biological and nutritional knowledge, resulting in low survival rate and poor economical profitability. We report here an integrated eco‐aquaculture system in which the yellow seahorse and its natural prey were co‐cultured with fertilized water and seaweed in cement ponds. In the first stage, urea (10 g m^?3) and chicken manure (50 g m^?3) were used to fertilize the cultured water, 5–7 days later, rotifer and micro‐crustaceans were flourishing. Then, seahorse juveniles were stocked at 200 ind m^?3. After 2 weeks, seaweed Gracilaria lichevoides was transplanted into the ponds to regulate water quality, light and to provide holdfast attachment for seahorses. The optimal density of G. lichevoides was controlled between 0.5 and 2 kg m^?3. The introduction of seaweed provides the habitat for natural food of seahorse. Within the cluster of the seaweed, small crustacean density was over 450 individuals per 100 g of fresh weight. Initial size of seahorse juvenile was 1.03 ± 0.091 cm, After the 146‐day rearing period, seahorse survival rates were 70.8%, 57.7% and 42.5%, and body standard lengths were 11.33 cm, 10.84 cm and 10.04 cm in the integrated eco‐aquaculture system (GFA) and in monoculture systems of FA (fertilized + feeding) and BA (only feeding) respectively. This system incorporated traditional Chinese aquaculture technique in which feedings nature food organisms were cultivated by fertilization and ecological regulation. Results from these preliminary experiments suggest that the integrated system could be technically feasible, suitable and exemplary.     

Grow-out of juvenile seahorse Hippocampus kuda (Bleeker; Teleostei: Syngnathidae) in illuminated sea cages

This paper examines the feasibility of rearing 10–15-day- and 0.7–1.5-month-old seahorse Hippocampus kuda in illuminated sea cages to continue existing hatchery protocols to mass produce H. kuda for trade and enhance depleted wild stocks in their natural habitats. Thawed Acetes (a planktonic crustacean abundant in inshore seas) was fed to juvenile seahorses in lighted and unlighted sea cages while one group in lighted cages was not fed Acetes . After 10–12 weeks of rearing, both mean body weight and stretch height increased in all treatment groups, with lighted cage-reared seahorses fed Acetes being heavier (2 g) and longer (8 cm) than the other two treatment groups. Although instantaneous growth rates declined during the rearing period, these were generally higher among Acetes -fed seahorses in lighted cages (0.02–0.07) compared with those in the unlighted cages with Acetes and lighted cages without Acetes feeding. Mean survivorship in all groups ranged from 9% to 74% after the trials, but mean survivorship of juveniles in lighted cages with Acetes feeding (9–74%) was consistently lower than the two treatment groups as a likely result of crustacean and piscine predators being attracted by light and the odour of leftover Acetes in the lighted cages. These results demonstrate that light-attracted zooplankton prey supplemented by Acetes feeding may provide essential nutrients for the growth of H. kuda juveniles in illuminated sea cages. With further improvement in the grow-out protocol, it may provide a possible alternative livelihood to seahorse fishers and sufficient seed to re-populate depleted wild stocks of H. kuda .     

Effects of culture conditions on feeding response of larval Pacific red snapper ( Lutjanus peru, Nichols & Murphy) at first feeding

Feeding incidence or number of larvae with preys (FIC) and intensity or number of prey per larvae (FIT) at first feeding of Pacific red snapper ( Lutjanus peru) larvae was investigated under different conditions: prey type (rotifer and copepod nauplius) and density, nauplii size, light intensity, water temperature, salinity and microalgae concentration. Rotifers were not consumed at any prey density and FIC increased significantly when a high nauplii density (10 > 1, 0.1 mL^?1) and light intensity (2000 > 1000, 500, 0 lx) were supplied. In a multifactorial experiment where light intensity (2000, 2500, 3000 lx), tank colour (grey and black) and prey type (nauplii and a mixed diet: rotifers and nauplii) were tested, a significant difference was found only for light intensity and prey type with a significant interaction between these factors. FIC was significantly higher with nauplii stage I–III than IV–VI and also at 25 °C than at 28 °C. Green water (0, 0.3 × 10⁶ or 1 × 10⁶ cells mL^?1) and salinity (25, 30, 35 gL^?1) did not affect FIC. FIT was not affected by any variables tested except in the density experiment where it was significantly higher at 10 nauplii mL^?1.     

Effect of larval and prey density, prey dose and light conditions on first feeding common dentex (Dentex dentex L.) larvae

Initial larval stocking density, prey density, daily prey ration and light conditions (light intensity and photoperiod) were tested for common dentex larval rearing under experimental conditions. Experiments continued until the first peak of larval mortality. The best results in larval survival were obtained with an initial stocking density of between 10 and 40 larvae L^?1, fed with at least 10 rotifers mL^?1, maintaining ratios of 500–1000 rotifers larva^?1, with one or two adjustments of prey density per day. The use of more than 2000 rotifers larva^?1 or three daily adjustments of live prey density had negative effects on larval survival. The best light conditions for common dentex larval rearing were found using a photoperiod of 24 h L:0 h D and an intensity of at least 3.4 μmol m^?2 s^?1.     

The first‐feeding response of larval southern bluefin tuna,Thunnus maccoyii (Castelnau, 1872), and yellowtail kingfish,Seriola lalandi (Valenciennes, 1833), to prey density,prey size and larval density

We investigated the first‐feeding success of two species: southern bluefin tuna (Thunnus maccoyii) and yellowtail kingfish (Seriola lalandi) to determine if similar culture parameters can be used for both, especially when S. lalandi are held in the same tanks as prey for T. maccoyii. The feeding performance (proportion and intensity) was examined in three short‐duration (4 h) experiments: prey density, prey size and larval density. Increasing prey density from 0.5 to 25 rotifers mL^?1 increased the proportion of T. maccoyii and S. lalandi larvae feeding. Prey size alone did not affect feeding in either species. Seriola lalandi had a decreased proportion of larvae feeding when larval density reached 50 larvae L^?1 concurrent with a gradual increase in feeding intensity between 2 and 50 larvae L^?1. In T. maccoyii, there was no pattern to the effect of larval density on the proportion of larvae feeding. The overall feeding performance of larvae was higher in T. maccoyii than S. lalandi. Increased prey density improved the first‐feeding ability of T. maccoyii and S. lalandi larvae. The effect of larval density on S. lalandi feeding requires further investigation, to ensure that they remain feeding when provided as prey in T. maccoyii culture. The identification of factors in this study, which increase first‐feeding success, will improve the culture of both species.     

Effects of photoperiod,light intensity,turbidity and prey density on feed incidence and survival in first feeding yellow tang (Zebrasoma flavescens)(Bennett)

Zebrasoma flavescens (Bennett) aquaculture is limited by high mortality during first feeding. Photoperiod, light intensity, turbidity and prey density are culture parameters that have been shown to affect feed incidence and survival in some food fish species, offering a logical starting point to improve first feeding in Z. flavescens. This study aimed to determine the effect of photoperiod, light intensity, turbidity and prey density on feed incidence and survival in larval yellow tang age 3 DPH to 5 DPH (days post hatch). Larvae were reared in four photoperiods (24L:0D, 16L:8D, 12L:12D, 0L:24D), four light intensities (1,500, 3,000, 4,500 and 6,500 lx), three turbidity ranges (0 cells/ml, 100,000–200,000 cells/ml and 400,000–600,000 cells/ml) and four prey densities (1, 3, 6 and 9 per mL). Photoperiod at 16L:8D and 12L:12D significantly increased feed incidence; 16L:8D significantly increased survival. Light intensities at 3,000 and 4,500 lx significantly increased feed incidence. Larvae reared in 400,000–600,000 cells/ml fed and survived significantly better than those in clear water. Larvae in 1 per mL fed and survived significantly less than those fed at 6 per mL.     

Experimental culture of larvae,post‐larvae and juveniles of the West Indian top shell,Cittarium pica (Linnaeus 1758)

The West Indian top shell, Cittarium pica, is an endangered vetigastropod of ecological and commercial value from the Caribbean. In order to assess the use of aquaculture as a tool for its sustainable production and conservation, embryos were produced in hatchery and experimentally cultured until juveniles under different conditions. Embryos were incubated under two temperatures (25 and 29°C) and six densities (0.3, 0.6, 1.0, 4.0, 9.0 and 37.0% of bottom‐coverage). Larvae were reared under different temperatures (25 and 27°C), densities (0.5, 1, 1.5 and 10 larvae/ml) and culture systems (static and down‐welling). Post‐larvae were obtained in three different settling conditions, and the early juveniles were cultured supplying fresh seaweed (Laurencia obtusa and Padina gymnospora), natural multi‐specific biofilm and Cylindroteca sp. biofilm. The growth and survival of embryos and larvae were not affected by temperature, but they were affected by density. Higher values were obtained at low densities (<1% of embryos bottom‐coverage and <1 larvae/ml), except for the growth of larvae, which was similar among treatments. Higher larval survival was recorded using the static culture system, and higher percent of post‐larvae recovered was associated with lower density of crawling veliger (0.2 and 0.1/ml), while its higher growth rate was related to the use of biofilms with conspecific mucus and low water‐flow (50%/h). The type of food tested did not affect the juvenile growth, but higher survival was obtained in those fed with seaweed than with multi‐specific biofilm.     

Influence of feeding regime and temperature on development and settlement of oyster Ostrea edulis (Linnaeus, 1758) larvae

Under controlled conditions of food density and temperature, larval performances (ingestion, growth, survival and settlement success) of the flat oyster, Ostrea edulis, were investigated using a flow‐through rearing system. In the first experiment, oyster larvae were reared at five different phytoplankton densities (70, 500, 1500, 2500 and 3500 μm³ μL^?1: ≈1, 8, 25, 42 and 58 cells μL^?1 equivalent TCg), and in the second, larvae were grown at four different temperatures (15, 20, 25 and 30°C). Overall, larvae survived a wide range of food density and temperature, with high survival recorded at the end of the experiments. Microalgae concentration and temperature both impacted significantly larval development and settlement success. A mixed diet of Chaetoceros neogracile and Tisochrysis lutea (1:1 cell volume) maintained throughout the whole larval life at a concentration of 1500 μm³ μL^?1 allowed the best larval development of O. edulis at 25°C with high survival (98%), good growth (16 μm day^?1) and high settlement success (68%). In addition, optimum larval development (survival ≥97%; growth ≥17 μm day^?1) and settlement (≥78%) were achieved at 25 and 30°C, at microalgae concentrations of 1500 μm³ μL^?1. In contrast, temperature of 20°C led to lower development (≤10 μm day^?1) and weaker settlement (≤27%), whereas at 15°C, no settlement occurred. The design experiments allowed the estimation of the maximum surface‐area‐specific ingestion rate  = 120 ± 4 μm³ day^?1 μm^?2, the half saturation coefficient {X_K} = 537 ± 142 μm³ μL^?1 and the Arrhenius temperature T_A = 8355 K. This contribution put a tangible basis for a future O. edulis Dynamic Energy Budget (DEB) larval growth model.     

Effects of feeding live or frozen prey on growth, survival and the life cycle of the cuttlefish, Sepia officinalis (Linnaeus, 1758)

The effects of feeding live or frozen grass shrimp (Palaemonetes varians) to the cuttlefish, Sepia officinalis, were determined in two experiments. During Experiment I, two populations of 30 cuttlefish (aged 90 days old) were fed either live or frozen grass shrimp. Cuttlefish fed live shrimp grew larger, matured earlier, had a shorter life cycle (255 days) than the ones fed frozen shrimp (282 days), and had lower mortality. Females from the group fed frozen shrimp matured a month later but were significantly larger, 130.9 ± 38.5 g, compared to 74.2 ± 16.0 g, laid larger eggs, 0.47 ± 0.11 g, compared to 0.28 ± 0.10 g, and had higher individual fecundity (411 eggs female⁻¹, compared to 150 eggs female⁻¹). Newly born hatchlings from both groups had similar weights. During Experiment II, six replicates of 15 cuttlefish (50 days old) were used, three for each of the two diets tested. The exact same amount of live or frozen shrimp was provided to both populations twice a day. No differences in growth and feeding rates or food conversions were found at the end of the experiment. During the first week, cuttlefish fed frozen shrimp grew larger, and had higher conversion rates, compared to the ones fed live shrimp. Mortality was higher for the group fed live shrimp (36.6%) in Experiment II, mainly occurring during the last week. Mortality for cuttlefish fed frozen shrimp in Experiment II was 2.2%. Results obtained here indicate that freezing the grass shrimp only had a negative effect on the survival of S. officinalis in Experiment I. This revised version was published online in August 2006 with corrections to the Cover Date.