Use of the freshwater rotifer <Emphasis Type="Italic">Brachionus angularis</Emphasis> as the first food for larvae of the Siamese fighting fish <Emphasis Type="Italic">Betta splendens</Emphasis>

Larvae of the Siamese fighting fish Betta splendens were reared on the mass-cultured small freshwater rotifer Brachionus angularis Laos strain (UTAC-Lao), Paramecia sp., and Artemia as live food sources. Larvae fed live food were found to have a significantly high survival rate (97.5–100%) 18 days after hatch (DAH) in comparison to the control unfed larvae, which died by 12 DAH. Rotifer-fed larvae were found to grow faster than paramecia-fed larvae. The fastest growth rate was observed in larvae fed a combination of rotifer and Artemia, with growth in these larvae increasing by 282% by 18 DAH [total length (TL) 11.3 ± 1.2 mm] relative to body measurements taken 3 DAH. The next fastest growth rate was observed in rotifer-fed larvae, with a 158% increase in growth observed by 18 DAH (TL 7.6 ± 0.5 mm). The paramecia-fed larvae were found to grow by only 54.3% (TL 4.6 ± 0.1 mm) during the same period.     

Early life history of yellow puffer,Chonerhinos naritus (Richardson, 1848) from Sarawak,Northwestern Borneo

The growth and morphological development including fins, spine distribution and pigmentation of larval and juvenile of hatchery‐reared yellow puffer, Chonerhinos naritus were described to provide essential information on the early life history of this species. The total length (TL) of newly hatched larvae was 3.42 ± 0.23 (mean ± SD) mm, reaching 5.66 ± 0.38 mm on 5 days after hatched (DAH), 7.80 ± 0.28 mm on 11 DAH, 9.88 ± 0.40 mm on 27 DAH and 10.92 ± 0.58 mm on 30 DAH. The yolk was completely absorbed in preflexion larvae at 4 DAH. The mouth opening started at 3 DAH of yolk sac larvae, while the teeth appeared starting from preflexion larvae at 7 DAH. Overall aggregate fin ray numbers including caudal fin attained full complement in postflexion larvae at 27 DAH. Several melanophores with appearance of small stellate were first appeared dorsally on the head of flexion larvae at 13 DAH, expanded at the dorsal region of the head, above the eye in juveniles at 30 DAH. The spines first appeared in preflexion larvae of C. naritus at 7 DAH, covering the ventral skin region below pectoral fin base and expanded to the ventral part of the body and nearly covered the whole abdomen region before the anus and below the eyes in juveniles. C. naritus remain as larvae for approximately 29 days, during which they metamorphose to the juvenile stage prior to sexual maturation. Observations in larvae development of C. naritus revealed similar characteristics with other Tetraodontidae species.     

Embryonic and larval development of black skirt tetra (Gymnocorymbus ternetzi,Boulenger, 1895) under laboratory conditions

The embryonic and larval development of black skirt tetra, Gymnocorymbus ternetzi, are described under controlled laboratory conditions. In addition, major histomorphological changes and the allometric growth patterns during larval development have been described. The laboratory‐reared broodstock, that is 1 year of age, were spawned. Hatching occurred 20–21 h after spawning at 24 ± 0.5°C. The cleavage was finished in 2 h and the early blastula stage occurred at 2:04 hours after spawning. The gastrulation started at 3:20 hours and 30% epiboly was observed at 3:34 hours after spawning. Eight‐somite stage was observed at 08:33 hours. And embryonic developmental stage was completed at 21 h after spawning. The newly hatched larvae were 1442 ± 14.3 μm in mean total length (TL). The mouth opened at 3 days after hatching (DAH). The yolk sac had been totally absorbed and the larvae started to swim actively within 3–4 days. Notochord flexion began at 11 DAH. The metamorphosis was completed and the larvae transformed into juveniles at 32 DAH. In this paper, the full developmental sequence from egg to juvenile of G. ternetzi is described for the first time.     

Organogenesis of exocrine pancreas in sharpsnout sea bream (<Emphasis Type="Italic">Diplodus puntazzo</Emphasis>) larvae: characterization of trypsin expression

The ontogeny and differentiation stages of digestive systems related with trypsin expression in larvae of sharpsnout sea bream, Diplodus puntazzo, were investigated from hatching to 40 DAH (days after hatching), and total lengths and weights of larvae were determined. Histologic and enzymatic techniques were used to explain the functional development of the pancreas including trypsin activity. The pancreas was identified as a compact structure located in the region slightly posterior to the liver. At 3 DAH, first anus and then mouth opened. Incipient pancreas secretion polyhedral cells could be first observed as zymogen granules. During larval metamorphosis, the pancreas became diffuse, spreading throughout the mesentery in proximity to the stomach, the anterior intestine and the pyloric caeca. The specific activity of trypsin (42.54 ± 6.8 mU/mg protein⁻¹) was found as early as after hatching at larvae size of 2.87 ± 0.34 mm at 0 DAH. Activity further increased until 10 DAH, especially after exogenous feeding. The highest trypsin activity was detected at 25 DAH as 119.26 ± 11.6 mU/mg protein⁻¹. It is concluded that exocrine pancreas organogenesis is the main critical step in the development of digestive system that results in zymogen granules accumulation and increased trypsin activity.     

Effects of the timing of initial feeding on growth and survival of loach (<Emphasis Type="Italic">Misgurnus anguillicaudatus</Emphasis>) larvae

The effects of delayed first feeding on growth and survival and starvation on the point-of-no-return of loach Misgurnus anguillicaudatus larvae were studied by evaluating morphometric characteristics under controlled conditions. Larvae began to feed exogenously at 3 days after hatching (DAH) and the PNR occurred between 9 and 10 DAH at 23 ± 1.0°C. The experimental design included a conventional feeding regime with initial feeding from 3 DAH as a control, delayed first feeding for 4, 5 and 6 DAH. Morphometric characteristics (head depth, body depth, eye diameter, mouth diameter, musculature height, total length and yolk sac volume) were evaluated under different initial feeding time (3, 4, 5 and 6 days after hatching). Loach larvae initiated first feeding at 4, 5 and 6 days after hatching achieved comparatively lesser growth performance in all morphometric characteristics than that of 3 days at the end of the experiment. By day 6, significant differences were observed between 3 and 6 days initial feeding larvae for all morphometric characteristics except eye diameter and mouth diameter. Similarly, significant differences were noticed between 3 and 5 days initial feeding. However, there were no significant differences in head depth, body depth, eye diameter, mouth diameter, and total length between 3 and 4 days initial feeding until 12 DAH. After 15 days rearing, significant differences in all morphometric characteristics appeared between 3 and 4 days initial feeding and followed to the end of the experiment. It was also observed that the yolk absorption in loach larvae was completed by 6 days irrespective of the differences in the initial feeding. The yolk volume of 4 and 5 DAH larvae initiated first feeding at 3 days (0.0125 ± 0.0015; 0.0077 ± 0.0009 mm³) had significant differences compared with yolk volume of larvae initiated first feeding at 4 days (0.0081 ± 0.0011; 0.0039 ± 0.0004 mm³), 5 days (0.0079 ± 0.0010; 0.0017 ± 0.0002 mm³) and 6 days (0.0082 ± 0.0011; 0.0016 ± 0.0001 mm³). Survival rates of four treatments were estimated daily for 30 days and significant differences were observed between the treatments at the end of the experiment. The final survival rate was higher when the loach larvae initiated feeding at 3 days (75.9%) when compared with 4 days (31.8%), 5 days (14.5%) and 6 days (6.4%). The present study suggests that the first feeding of loach larvae should be initiated at 3 DAH for achieving better growth and survival or else bad growth performance will engender if the first feeding is delayed.     

Transpositional feeding rhythm of loach <Emphasis Type="Italic">Misgurnus anguillicaudatus</Emphasis> from larvae to juveniles and its ontogenesis under artificial rearing conditions

The diel feeding rhythm and ontogenesis during early life stage of loach Misgurnus anguillicaudatus were investigated under experimental conditions (light: L 06:00–18:00, D 18:00–06:00 h). Morphological and behavioral developments of loach from newly hatched to 40 days after hatching were observed. Larvae were able to prey on daphnia 3–4 days after hatching at 23 ± 0.5°C. As the larvae grew, they showed an increasing feeding capacity and a distinct feeding rhythm. Feeding intensity and incidence for day-4 larvae were highest at 10:00 and 16:00 h. The highest levels of feeding intensity for day-12 larvae occurred at 08:00, 12:00, and 18:00 h as did feeding incidence. By day 20, when the larvae metamorphosed, the highest levels of feeding intensity occurred at 06:00, 18:00, and 24:00 h and were concurrent with the highest feeding incidence. After metamorphosis, feeding capacity had again increased considerably and, in contrast to the earlier stages before day 20, feeding intensity for day-30 juveniles peaked at 05:00 and 20:00 h, about 1–2 h after the maximum feeding incidence. The feeding rhythm of loach juveniles at day 40 was almost the same as the day-30 juveniles. The estimated maximum daily feeding rates were 43.1%, 33.4%, 19.0%, 12.8%, and 5.8% of body weight on days 4, 12, 20, 30, and 40, respectively. Thus, loach was found to have different feeding rhythms in the pre- and post-metamorphosis stages, with the highest feeding activity in daytime during the larval planktonic stage before metamorphosis, and intensely nocturnal feeding behavior during the juvenile benthic stage after metamorphosis.     

Embryonic and larval development of serpae tetra Hyphessobrycon eques (Steindachner, 1882)

In this study, the embryonic and larval development stages of one of the most important ornamental fish serpae tetra (Hyphessobrycon eques) are described. The early life stage is documented from fertilization until the beginning of the juvenile period. The fertilized eggs (the average diameter = 938.55 ± 35.20 µm) were incubated at a water temperature of 26 ± 0.5°C. The cleavage finished in 1:10 hr (=h) and the early blastula stage occurred at 1:26 hr post fertilization (hpf). The gastrulation started at 3:05 hpf, and 50% epiboly was observed at 3:25 hpf. Segmentation stage was monitored at 7:26 hpf. Embryonic developmental stage was completed and hatching occurred 20–21 hpf. The total length (TL) of newly hatched larvae was 2.64 ± 0.21 mm. The larval development of serpae tetra was divided into four different periods: Yolk‐sac larva (1–4 DAH, TL = 2.77 ± 0.09 mm ‐ 3.85 ± 0.11 mm), preflexion larva (5–12 DAH), flexion larva (13–15 DAH, TL = 5.78 ± 0.46 mm on the 15th day) and post‐flexion larva (16–30 DAH, TL = 10.7 ± 0.27 mm on the 28th–30th days). The mouth and anus are closed at 1 DAH. The mouth and anus opened at 4 DAH. Exogenous feeding started on the 4th day. The first gulping of the swim bladder was on days 3. The larva begins to swim freely, and the yolk sac was completely consumed at 4 DAH. Histological structures of the eye and brain of new hatched larva were clearly identified at 1 day after hatching (DAH). According to histological findings, the digestive system (stomach, intestine) started to develop and the liver could be seen on the ventral side of the swim bladder at 5 DAH. No histological difference was observed between the anterior intestine and the posterior intestine at 15–16 DAH. The larval metamorphosis was completed, and the larvae transformed into juveniles at 28–30 DAH.     

Effect of triiodothyronine (T3) in the larval development of the barber goby Elacatinus figaro

The barber goby Elacatinus figaro is an endangered fish, endemic to Brazil and very important to the aquarium trade. One of the bottlenecks for its production in captivity is the larviculture, which is characterized by high mortality rates, especially in the first week after hatching, and prior to metamorphosis. The experiment evaluated the effect of triiodothyronine hormone (T₃) on survival, growth and metamorphosis of barber goby. Larvae of 14 days after hatching (DAH) were immersed in three concentrations of T₃, in triplicate: TC (control) – 0; T01 – 0.01; T025 – 0.025 and T05 – 0.05 mg/L. An additional replicate of each treatment was performed for sampling for the histological analysis of the thyroid follicles in 24 DAH larvae. The survival rates in TC, T01, T025 and T05 were 24%, 54%, 36% and 37% respectively, without significant differences between the treatments (p > 0.05). Regarding larval growth, the highest length values were obtained in TC. In T025 and T05, larval metamorphosis was anticipated in up to 11 days in relation to TC and T01. In T01 larvae, follicles were numerous with the presence of reabsorption vesicles in the colloid periphery, indicating increased production of thyroid hormones (THs), associated with the larval metamorphosis process. In T05, a follicle reduction in number and size was observed, given evidence of the end of metamorphosis. The use of 0.025 and 0.05 mg/L of T₃ anticipated metamorphosis in barber goby larvae, shortening the larviculture period for this species.     

Histological development of digestive tract in discus, <Emphasis Type="Italic">Symphysodon</Emphasis> spp. larvae

The present study provides information on the histomorphological development of digestive system of discus, Symphyosodon spp., larvae during the first month of life. Discus larvae are altricial at hatching, with an undifferentiated digestive tract and a large yolksac, which is completely consumed within 7 days. The mouth opens 3 days after hatching (DAH) and the larvae starts feeding on AF Artemia at 4 DAH when offered. At 3 DAH the digestive tract is differentiated with distinct esophagus, stomach anlage, and mid- and hindguts. At 5 DAH, discus larvae is an active feeder, equipped with partly developed jaws and ossified gill arches and an inflated swim bladder. The liver and pancreas are present and supranuclear inclusion vacuoles (SIV) appear in the hindgut for the first time. Gastric glands in stomach were first observed 7 DAH and proliferated by 11–13 DAH. SIV were a common feature in the midgut and hindgut epithelium until 15–23 DAH. Therefore, exclusive use of artificial diets should be postponed until 2–3 weeks after hatching.     

Effects of antibiotic‐induced differences in bacterial load on growth and shape of early larval European seabass (Dicentrarchus labrax L.)

The aim of the present study was to quantify the effects of antibiotic‐induced differences in bacterial load on the size and shape of fish larvae, using Dicentrarchus labrax of day after hatching (DAH) 3 as a case study. They were split in two treatment groups and reared in 50 ml vials until DAH 14, with the control treatment (“NA”) including larvae reared in filtered autoclaved seawater without antibiotics, while the second (“A”) included larvae reared in filtered autoclaved seawater with rifampicin, ampicillin, kanamycin, trimethoprim and gentamicin, with a concentration of 10 mg/L each. They were sampled for bacterial presence on DAH 4, 7 and 14, and had their mortalities recorded, their total lengths, gut lengths, anal body depths, eye diameters, head depths, yolk sac lengths and yolk sac depths measured, and their outlines analysed on DAH 7 and 14. Treatment NA exhibited the highest mortalities on DAH 14. The antibiotics had a significant size effect, yielding larvae with larger total length on DAH 7 and 14, larger bodies on DAH 7, and on DAH 14 larger anal body depth and greater variance in body size. Their effect on outline shape was also significant in both age classes, with increasing differences from DAH 7 to DAH 14. On DAH 7, “A” specimens were more uniform in their dorso‐ventral development, and on DAH 14 “NA” had more slender shapes. The beneficial total length and size effects and the witnessed shape effects might be associated with the low bacterial presence.     

Weaning of the wedge sole <Emphasis Type="Italic">Dicologoglossa cuneata</Emphasis> (Moreau): influence of initial size on survival and growth

The weaning phase can be decisive in fish-culture viability. In this work, the relationship between the initial size and weaning success has been studied in wedge sole (Dicologoglossa cuneata). For each age (30, 50, and 70 days after hatching, DAH), two to three sizes were selected, and all were put on the same feeding schedule for 20 days. Each batch (three replicates) was sampled at 1, 10, and 20 days. Specific growth rate (SGR) and survival were compared at the end of the co-feeding period, after 10 days on dry feed only. The best results for survival and growth were found with the smallest larvae, and vice versa. The SGRs and survival rates recorded during the co-feeding period were higher (0.8–15.6 day⁻¹ and 68.3–97.8%) than those from the dry-food phase (0.9–4.7 day⁻¹ and 56.3–66.7%). Successful weaning (survival = 65% and SGR = 9.3 day⁻¹) is possible with 30 DAH larvae (7.6–8.1 mm and 3.9–4.6 mg). In conclusion, the most effective weaning would be possible at 30 DAH, implying significant Artemia savings (25–50%).     

Optimum time for weaning South African <Emphasis Type="Italic">Scylla serrata</Emphasis> (Forskål) larvae from rotifers to <Emphasis Type="Italic">Artemia</Emphasis>

To determine the optimum time at which to wean Scylla serrata larvae from rotifers onto Artemia two experiments were conducted, approximately 1 month apart, using larvae from two different female crabs. In the first experiment, the larvae in three treatment groups, with nine replicates each, were fed rotifers for the first 8 days after hatching. Artemia were introduced on days after hatch (DAH) 0 – during the first zoeal instar (treatment R + A); on DAH 4 – during the second zoeal instar (treatment R4A); on DAH 8 – during the third zoeal instar (treatment R8A). In a control (ROT) larvae were fed with rotifers exclusively for 18 days until the completion of metamorphosis to megalopa. In the second experiment, the same four feeding schedules as in experiment 1 were used with an additional group of larvae (treatment AC) that were fed only on Artemia throughout the rearing period. Similar results were recorded in the two experiments. Larvae in treatments R + A and R4A performed significantly better than those in treatments R8A, ROT and AC. This was particularly evident when examining the proportion of zoeae which successfully completed metamorphosis to megalopa. Poor performance of larvae in treatments AC and ROT implied that rotifers are needed as a first food, but that rotifers alone do not fill the nutritional requirements of S. serrata larvae. Poor performance of larvae in treatment R8A suggested that the diet should be supplemented with Artemia before the end of the zoea 3 stage.     

Screening of enzyme activity for assessing the condition of larvae in the seven-band grouper <Emphasis Type="Italic">Epinephelus septemfasciatus</Emphasis> and devil stinger <Emphasis Type="Italic">Inimicus japonicus</Emphasis>

We conducted screening tests to determine whether enzyme activity is a suitable biomarker for assessing the physiological condition of marine fish larvae. The rearing experiments consisted of three trials, of which two were conducted using the seven-band grouper Epinephelus septemfasciatus for a period of 5 days after hatching (DAH), and one was conducted using the devil stinger Inimicus japonicus for 10 DAH. The trials were conducted under three different rearing-tank environments (shallow tank, intermediate tank, deep tank) in a water volume of 100 l and an aeration rate of 50 ml/min. We determined survival, surface death, growth, and enzyme activities (trypsin, esterase, and alkaline phosphatase). The highest survival rates and lowest surface deaths in both species occurred among the larvae grown in the deep tank. There was a significant and negative correlation between survival at 5 DAH and alkaline phosphatase activity at 0 DAH in the seven-band grouper. The same correlation was found between survival at 10 DAH and trypsin and alkaline phosphatase activity at 1 DAH in the devil stinger. Based on these results, we conclude that the activity of a specific enzyme is a candidate for assessing the physiological condition of marine fish larvae.     

An intensive hatchery rearing protocol for larvae of the bullseye puffer,Sphoeroides annulatus (Jenyns)

In this study bullseye puffer, Sphoeroides annulatus larvae were reared from hatching through to 1 or 2 months after weaning on an experimental scale in three replicate 600 L tanks and on three occasions during the spawning season (nine tanks in total). The rearing protocol used was green water (Nannochloropsis oculata and Isochrysis sp.) 100 000 cells mL^?1 from 0 to 11 days after hatch (DAH), 5–10 rotifers, Brachionus rotundiformis mL^?1 from 2 to 26 DAH, 0.1–1 Artemia mL^?1 from 21 to 34 DAH and weaning from 29 to 34 DAH. Survival to a month after weaning was 1%, a total of 3153 juveniles were produced with an average wet weight of 0.29 ± 0.07 g and a length of 27.5 ± 0.82 mm.     

Lack of essential fatty acids in live feed during larval and post-larval rearing: effect on the performance of juvenile <Emphasis Type="Italic">Solea senegalensis</Emphasis>

Despite the large progress obtained in recent years, Senegalese sole (Solea senegalensis) production of high quality juveniles is still a bottleneck. This paper examines the effect of larval and post-larval lipid nutrition on juvenile performance and quality. Four dietary treatments were tested: A—enriched Artemia spp. (EA); B—non-enriched Artemia spp. (NEA); C—EA during the pelagic larval period and NEA after larval settlement; D—50% EA and 50% NEA. Juvenile fatty acid profile at 60 days after hatching (DAH) clearly reflected the larval and post-larval diet composition. Feeding sole larvae on NEA (poor in lipids and essential fatty acids-EFA) had a negative effect, reducing growth (total length and dry weight) after 30 DAH and decreasing digestive enzyme activity at the end of the rearing period (60 DAH). However, relatively good performance compared to the EFA-richest treatment (A) was obtained when larvae were fed 50% EA and 50% NEA (D) or even EA only during the pelagic larval period followed by NEA after larval settlement (C). Malpigmentation was not affected by the dietary regimes and its incidence was very low. However, skeletal deformities were prevalent, particularly in the caudal complex, independently of diet. The results confirm that Senegalese sole appear to have lower larval EFA requirements than most cultured marine species and potentially even lower requirements during the post-larval stage. The importance of studying the impact of early nutrition on later juvenile stages was clearly highlighted in this study.     

Change in nutrient composition of Artemia grown for 3–4 days and effects of feeding on‐grown Artemia on performance of Atlantic halibut (Hippoglossus hippoglossus,L.) larvae

Major challenges in culture of Atlantic halibut larvae have been slow growth during the late larval stages and inferior juvenile quality due to pigmentation errors and incomplete eye migration during metamorphosis. The hypothesis of this study was that feeding on‐grown Artemia would alleviate these problems. Artemia were grown for 3–4 days on Origreen or Origo. The growth and nutrient composition of Artemia nauplii and on‐grown Artemia were analysed, and both Artemia types were fed to Atlantic halibut larvae, on‐grown Artemia from 15 days post‐first feeding (dpff). The body length of Artemia increased with 20%–70% in response to on‐growing. In all experiments, protein, free amino acids and the ratio of phospholipid to total lipid increased, while lipid and glycogen decreased. The fatty acid composition improved in some cases and not in others. The micronutrient profiles were not negatively affected in on‐grown Artemia. All these changes are thought to be beneficial for marine fish larvae. The final weight of Atlantic halibut postlarvae was similar, and 90% of the juveniles had complete eye migration in both groups. It is concluded that the present version of Artemia nauplii probably covers the nutrient requirements of Atlantic halibut larvae.     

Feeding rhythm and growth of the tongue sole, Cynoglossus semilaevis Günther, during its early life stages

The feeding rhythm and growth characteristics of the early life stages of the tongue sole, Cynoglossus semilaevis Günther, were studied. Larvae began to prey on rotifers about 2–3 days after hatching at 23°C. About 12 days after hatching, larvae had grown to 8–9 mm in total length and were able to prey on Artemia larvae. As the larvae grew, they showed an increasing feeding capacity and a distinct feeding rhythm. Feeding intensity for day‐6 larvae was highest at 12:00 and 18:00 hours, about 2–3 h after the maximum feeding incidence. The highest levels of feeding intensity for day‐16 larvae occurred between 09:00 and 18:00 hours. By day 26, when the larvae had metamorphosed, feeding capacity had again increased considerably and, in contrast to the earlier stages, feeding intensity peaked at 18:00 and 24:00 hours concurrently with feeding incidence. Thus, tongue sole were found to have different feeding rhythms in the pre‐ and post‐metamorphosis stages, with the highest feeding activity in the daytime during the larval planktonic stage, and at night during the juvenile benthic stage. The estimated maximum daily feeding rates were 65%, 40% and 11% of body weight on days 6, 16 (larvae) and 26 (juveniles) respectively. Size variation increased markedly with development.     

Skeletal development and abnormalities of the vertebral column and fins in larval stage of hatchery‐reared American shad,Alosa sapidissima

The present study describes the skeletal development and the occurrence of deformities in American shad, Alosa sapidissima, larvae from hatching to 45 days after hatching (DAH). The ontogeny of the vertebral column started at 16 DAH (days after hatching), with the formation of the posterior neural and haemal arches, and was complete at 28 DAH. In comparison, the vertebral centra started to form at 16 DAH, with ossification being visible in all centra at 38 DAH. The pectorals were the only fins that formed before the onset of feeding (at 2 DAH), with ossification being complete at 45 DAH. The caudal fin formed at 5 DAH, with ossification being complete at 40 DAH. Dorsal and pelvic fin development began at 6 DAH and 20 DAH respectively. The ossification of both the dorsal and pelvic fins was visible at 45 DAH. The anal fin began forming at 14 DAH, and was complete at 30 DAH. The ossification of the anal fin was complete at 45 DAH. Overall, 22 types of skeletal deformities were detected in about 41% of individuals. Most anomalies were detected in the haemal region, while the fewest anomalies were detected in the anal fin. In addition, the frequency of deformities gradually increased with fish age at the different developmental stages. Our results are expected to contribute baseline information on how rearing conditions impact skeletal development, in addition to identifying potential causative factors of skeletal deformities.     

Otolith microstructure of brown sole <Emphasis Type="Italic">Pseudopleuronectes herzensteini</Emphasis>: validation of daily ring formation and the occurrence of microstructure denoting metamorphosis

Brown sole Pseudopleuronectes herzensteini larvae and juveniles were reared to validate daily otolith ring formation. At 15°C, a check (a distinct ring) formed on the sagittae and lapilli at 6 days after hatching, and clear increments regularly formed outside the check. For both otoliths, the relationship between the number of days after hatching and number of increments was linear, and the slope of the line was approximately 1; therefore, daily formation was validated. At 12°C, the check formed on the lapillus 8 days after hatching. Accessory primordia (AP) began forming on the sagittae of metamorphosing larvae, and the shape of the sagittae became complicated. AP were not formed on the lapillus; concentric rings were formed throughout larval and juvenile stages. Wide and obscure increments formed on the lapilli during metamorphosis (metamorphosing zone, MZ). Based on MZ, concentric rings that have formed on the lapilli of juveniles can be separated into larval and juvenile rings. The morphs of large juveniles’ lapilli were bilaterally asymmetric, and the blind-side lapilli were most suitable for otolith microstructure analysis. This study provides fundamental information for otolith microstructure analysis in wild brown sole.     

Influence of light intensity on feeding, growth, and early survival of leopard coral grouper (Plectropomus leopardus) larvae under mass-scale rearing conditions

This study investigated the effect of different light intensities on feeding, growth and survival of early stage leopard coral grouper Plectropomus leopardus larvae. Four different light intensities (0, 500, 1000 and 3000 lx) were used and larvae were kept under constant light conditions from 0 day after hatching (DAH) to 5 DAH. The larvae were fed a small S-type of Thai strain rotifers at a density of 20 individuals/mL from 2 DAH. The number of rotifers in larval digestive organ and total length of larvae were examined at 3 h intervals between 04:00 and 22:00 h on 3 DAH, and thereafter at 6 h intervals until the end of the experiment (5 DAH). Four experimental trials of the larval rearing were repeated using by 60 kL mass-scale rearing tanks. The results indicate that coral grouper larvae are visual feeders and their food intake increases with increasing light intensity. Food intake of larvae reared at 3000 lx was significantly higher than those reared at 0–1000 lx on 3 DAH despite being the first-feeding day (P < 0.01). On 4 DAH, total length of larvae reared at 3000 lx was significantly larger than those reared at the lower light intensities (0, 500 and 1000 lx), and thereafter light intensity significantly influenced larval feeding and growth until the end of the experiment. Survival on 5 DAH did not show a significant difference between light intensities, but survival rate at 3000 lx and 1000 lx had a tendency to be higher than those reared at the lower light intensities (0 and 500 lx). In contrast, larvae reared at 0 lx exhibited stagnant and/or negative growth. These results indicate that light intensity is significantly the factor affecting larval feeding, growth, and survival in coral grouper larvae under the rearing conditions.