Improvement of Matrinxã, Brycon amazonicus,Larviculture by Exposing Eggs to Triiodothyronine

This work evaluated the effect of triiodothyronine (T₃) on larviculture of matrinxã, Brycon amazonicus. Oocytes of three females were pooled, fertilized with pooled semen of two males and separated in four batches that were immersed in triiodothyronine solutions as follows: M₁ (control – water); M₂ (0.01 mg/L T₃); M₃ (0.05 mg/L T₃); and M₄ (0.1 mg/L T₃). Triiodothyronine did not affect fertilization rate and number of hatched larvae. Weight of hatched larvae was significantly higher in treatments M3 and M4, as well as among larvae sampled at Day 12 in all treatments. After 12 d of rearing, biomass gain was higher in the hormone treatments (M₁ 688 ± 569 mg; M₂ 2436 ± 562 mg; M₃ 3572 ± 569 mg; and M₄ 4129 ± 770 mg). In general, coefficients of variation of weight (CV_w) and length (CV_l) did not differ among treatments and cannibalism was registered between 36 and 72 hours post‐hatching (h.p.h.) without differences among treatments. Larval survival increased in the hormone treatments (M₁ 26.5%; M₂ 37.6%; M₃ 40.6%; and M₄ 40.8%). The results indicate that the immersion of matrinxã eggs in triiodothyronine can promote beneficial effects to its larviculture and indicate promising perspectives for culture of this tropical species.     

The influence of triiodothyronine (T<Subscript>3</Subscript>) on the early development of piracanjuba (<Emphasis Type="Italic">Brycon orbignyanus</Emphasis>)

This paper reports the triiodothyronine’s (T₃) effects on the early growth and survival of piracanjuba (Brycon orbignyanus) produced from fertilized eggs hormone exposed. The study was carried out in two phases. In the first phase, eggs divided in 6 batches were immersed in T₃ solutions: 0.01; 0.05; 0.1; 0.5 ppm; 1 ppm and control (no T₃). After a 15-min immersion, eggs were transferred to incubators where larvae were kept up to 72 h after hatching. Larval weight, length and yolk sac volume were determined every 12 h. Sixty and 72 h after hatching, larvae exposed to 0.5 ppm T₃ were significantly heavier than the others, and those exposed to 1 ppm T₃ showed the lowest weight. The yolk sac absorption was not affected. In the second experimental phase, the resulting fry from the first phase were stocked into 3 boxes per treatment (5 larvae L⁻¹) and fed with plankton, fish larvae and feed prepared in the hatchery (48% CP) in the first 3 days, plankton and feed from the 4th to the 10th day and only feed in the next (last) 5 days. Fry weight, length and specific growth rate were determined at 1, 5, 10 and 15 days. Survival was calculated in the last day. In the 15th day, fry length did not differ among treatments but the weight of the control group was higher. Higher survival in the T₃-treated groups suggested lower predation among fry. The results allowed us to conclude that there was no expressive effect of T₃ on the growth, but it improved the survival of the piracanjuba progeny.     

Stimulatory effect of thyroid hormones improves larval development and reproductive performance in alligator gar (Atractosteus spatula) and spotted gar (Lepisosteus oculatus)

Development of alligator gar (Atractosteus spatula) and spotted gar (Lepisosteus oculatus) larvae was evaluated after exposure of embryos to thyroid hormones (TH) by two different methods of administration. For the first method, alligator gar embryos were placed in a water bath with triiodothyronine (T₃; 1 ppm for 2 h), and control treatment embryos were placed in a water bath without T₃. For the second method, spotted gar adult males and females were injected with different TH during induced spawning: a group was injected with T₃ [20 mg kg^?1 body weight (BW)], a second group with thyroxine (T₄; 20 mg kg^?1 BW), a third group with thyroid‐stimulating hormone (TSH; 4 IU kg^?1BW) and a control group with dimethyl sulfoxide. Both methods resulted in increases in the concentrations of T₃ and T₄ in embryos of both species. An increase in the alkaline enzymatic activity of TH‐treated larvae was observed as well as an increase in the RNA:DNA, RNA:Dry weight and DNA:Dry weight ratios at hatching. Some positive effects of TH application were: a significant increase in the hatching rate of spotted gar larvae from TH‐injected parents and a higher percentage of successful spawns. Evenness of larval sizes and a higher survival rate were observed in alligator gar larvae. Snout development was accelerated by TH treatment in both species.     

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.     

Hormone-induced ovulation, natural spawning and larviculture of Brazilian flounder Paralichthys orbignyanus (Valenciennes, 1839)

Mature Brazilian flounders Paralichthys orbignyanus were captured in coastal southern Brazil and their reproduction in captivity was studied. Brazilian flounder will spawn naturally in captivity when the water temperature is around 23 °C and 14 h of light is provided daily. Females were induced for ovulation and hand stripping using human chorionic gonadotropin, luteinizing hormone‐releasing hormone analogue or carp pituitary extract. There was no need to inject males, as running milt was observed during the spawning season. Fertilization and hatching rates were above 80% independent of the hormone used. Notochord length at hatching was 2.18±0.07 mm for larvae hatching from naturally spawned eggs. Larvae were reared in salt water (30–35 g L^?1) at 24 °C and under continuous illumination. Larviculture was with green water (Tetraselmis tetrathele 50 × 10⁴ cells mL^?1). Rotifers (10–20 ind mL^?1) were offered as first food 3 days after hatching and gradually replaced by Artemia nauplii (0.5–10 ind mL^?1). Larvae settled to the bottom 20 days after hatching and completed metamorphosis within a week after that. The total length for newly metamorphosed juveniles was 12.9±2.2 mm and the mean survival was 44.8%. The results demonstrate the feasibility of producing Brazilian flounder fingerlings for stock enhancement or grow‐out purposes.     

Light intensity effects on early life stages of black sea bass, Centropristis striata (Linnaeus 1758)

The effects of four light intensities on growth and survival of first‐feeding stage black sea bass larvae Centropristis striata were investigated in a controlled‐environment laboratory. Fertilized eggs, obtained from LHRHa‐induced spawning of captive broodstock, were stocked (72 eggs L^?1) into twenty 15 L black tanks under light intensities of 100, 500, 1000 and 1500 lx, with five replicate tanks per treatment. The photoperiod was 12L:12D, the temperature was 20°C and the salinity was 35 g L^?1. Larvae were fed rotifers Brachionus rotundiformis from day 2 post‐hatching (d 2ph) at 5–10 rotifers mL^?1. Microalgae Nannochloropis oculata and Isochrysis sp. were added (1:1) daily to maintain a density of 300 000 cells mL^?1. Hatching success and larval growth and survival from d 2ph through d 15ph were monitored. Hatching success was 28–38% under all light intensities, and notochord length at hatching ranged from 2.8 to 3.0 mm, with no significant differences among treatments. By d 15ph, growth (mg wet weight) was significantly higher in the 1000 lx (0.914) and 1500 lx treatments (0.892) than in 100 lx (0.483), and a highly significant trend (P<0.01) towards increased survival with increasing light intensities was observed, from 1.3% at 100 lx to 13.9% at 1500 lx. Higher light intensities within the range of 100–1500 lx improved growth and survival of early larval black sea bass, suggesting that even higher light intensities may improve culture performance. This is consistent with conditions in shallow, near‐shore locations where eggs and larvae are distributed in nature.     

Thyroid hormones in brown trout (Salmo trutta) reproduction and early development

Gravid brown trout (Salmo trutta) females were injected with various doses of a synthetic gonadotropin-releasing hormone analog (GnRHa), given with or without an injection of triiodothyronine (T₃), in order to investigate the potential of T₃ (a) to enhance the stimulatory effect of GnRHa on ovulation, and (b) to enhance the growth and survival of the produced progeny. From the time the hormonal treatments were initiated until ovulation was detected 5–38 days later, endogenous plasma T₃ levels increased from an average of 3.6 to 11.6 ng ml⁻¹. Injection with 20 mg T₃ kg⁻¹ body weight, further elevated plasma T₃ levels at ovulation (16.0 ng ml⁻¹. Mean time to ovulation was reduced significantly in fish injected with 10 μg kg⁻¹ of GnRHa, whereas treatment with lower doses was ineffective. Injection with T₃ did not enhance the ovulatory response of brown trout to GnRHa. Unfertilized eggs obtained from T₃-injected females had a higher T₃ content, suggesting a transfer of T₃ from the maternal circulation into the oocytes. Maternal T₃ injection had no effect on egg fertilization rates, embryo survival to eyeing and hatching, or the prevalence of abnormal larvae at the time of hatching. Length and weight gain of the progeny during yolk absorption was also not influenced by maternal T₃ treatment. At the completion of yolk-sac absorption, progeny from females injected with T₃ had a higher prevalence of skeletal abnormalities than controls. The results suggest that in teleosts like brown trout, which have high endogenous circulating T₃ levels, treatment of females with T₃ does not enhance responsiveness to GnRHa and it has the potential for deleterious effects on their offspring.     

Spawning behaviour,early development and first feeding of the bluestriped angelfish [Chaetodontoplus septentrionalis (Temminck & Schlegel, 1844)] in captivity

Successful natural spawning of Chaetodontoplus septentrionalis in captivity from 19 March to 11 May, 2008 is described for the first time. A single male dominates a harem of two females, spawning with each at dusk, from 10 min before to 20 min after sunset. Each female laid an average 119 × 10³ eggs during the spawning period. Fertilized eggs were spherical, buoyant and had a diameter of 0.83 ± 0.02 mm (mean ± SD). Embryonic development lasted 15–18 h at 28.1 °C. Newly hatched larvae were 1.60 ± 0.07 mm in total length (TL) with 27 myomeres. Larvae completed yolk absorption within 3 days post hatching (ph) at 3.01 ± 0.08 mm TL. Ten days ph, the larvae had attained 3.95 ± 0.12 mm TL. Larvae were fed either 100% s‐type rotifers (Brachionus rotundiformis), 100% copepods (Microsetella sp.), a combination of the two (50%:50%) or without live feed (starved control) to determine the effect of live feed on the survival rate. The survival was significantly (P<0.001) higher in larvae fed a combination of diet than the others. These results indicate that C. septentrionalis is a potential species for captive breeding programs and the use of a combination of diet (s‐type rotifers and copepods) may be a suitable first food for the larvae.     

Physiological responses associated with capture and crowding stress in matrinxãBrycon cephalus (Gunther, 1869)

The dynamics of plasma cortisol, blood glucose, plasma chloride and liver glycogen were investigated in matrinxã (Brycon cephalus) submitted to capture and various periods of crowding. A total of 400 fish (700±22 g weight) were distributed in four ponds divided into four 50‐m² squares (25 fish/square, 350 g L^?1), where they were acclimated for 30 days. On the sampling day, after 24 h without food, all fish from three squares were transferred to the fourth square. Six fish were sampled before the procedure (control group, zero time) and 1, 3, 6 and 24 h after the capture and crowding. Each sampling was performed in a different pond to prevent additional stress. Fish were anaesthetized and blood and liver collected for biochemical analysis. Water temperature, pH, dissolved oxygen, alkalinity, ammonia and nitrite levels were within acceptable levels for matrinxã rearing. Slight but not significant increases were verified in plasma cortisol and blood glucose levels, as were decreases in plasma chloride and liver glycogen levels. The results suggest that matrinxã is highly tolerant to the procedures of capture and short‐term crowding.     

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.     

The Scale‐up of Spotted Rose Snapper,Lutjanus guttatus,Larval Rearing at Mazatlan,Mexico

The scale‐up of spotted rose snapper, Lutjanus guttatus, larval rearing is described. Fertilized eggs (480,000) were obtained from a 1‐d harvest of a natural spawning captive broodstock acclimatized for 1 yr and 6 mo in two fiberglass tanks (18 m³). Fourteen hours after spawning, 89.6% of the collected eggs were floating, of which 96.2% were transparent with live embryos. Incubation at 25–26 C lasted 21 h, with 90.2 ± 2.1% hatching percentage of normal larvae. The percentage of viable larvae at 48 h after hatching was 79.7 ± 1.9%. Initial stocking density was 10.4 ± 1.0 larvae/L 2 days after hatching (d.p.h.). A total of 22,600 juveniles (1256 ± 170 juveniles/m³) were harvested from six 3‐m³ cylindrical fiberglass tanks. Average survival was 12.1 ± 1.1%. Final mean length and weight were 5.5 ± 0.05 cm and 2.24 ± 0.04 g, respectively. Growth expressed in total length was TL = 2.1476e^0.0543t (R² = 0.9911). Final mean biomass and condition factor were 2.8 kg/m³, 12.3% and 1.346. General length‐weight ratio was W = 0.05460 LT^2.2306.     

Live transportation of Macrobrachium rosenbergii (De Man) in chilled sawdust

Three cooling rates of 1.26±0.09°C h^?1 within 8 h (slow, T₁), 2.52±0.18°C h^?1 within 4 h (moderate, T₂) and 5.04±0.36°C h^?1 within 2 h (fast, T₃) were tested to cold‐anaesthetize farm raised Macrobrachium rosenbergii (De Man) (45–52 g) in each case from 25°C down to 15±1°C in a refrigerated chilling tank, provided with aeration. The cold‐anaesthetized prawns 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 15±1°C, for set durations of 6, 9, 12, 15 and 18 h. Survival was determined by revitalizing the prawns in aerated water with an initial temperature of 20°C, which was raised to 29±1°C within 3 h. The experiment was repeated using berried females acclimated to brackishwater of 12 g L^?1 salinity and the percentage survival recorded after live storage for durations ranging from 6 to 24 h at intervals of 3 h. Statistically valid safe durations for obtaining 100% survival of the cold anaesthetized and live stored prawns were determined using probit analysis at the three chilling rates tested, and were found to be 7.39, 6.98 and 4.54 h in the case of adult prawns, and 7.87, 8.17 and 6.43 h for berried females for T₁, T₂ and T₃ respectively. For practical purposes, the durations that yielded 95% survival rates were computed to be 16.47, 12.14 and 8.35 h in the case of adult prawns and 18.49, 19.02 and 11.11 h for berried females for T₁, T₂, and T₃ respectively. The berried prawns revitalized after live storage were incubated in tanks and the zoea larvae reared up to postlarvae (PL‐5), and compared against a control. No significant difference was found in larval hatch fecundity, survival rate and the production of PL L^?1 between the treatment and control, indicating that the method of cold anaesthetization and live storage of berried prawns could be used for successful transportation of broodstock.     

Hatching rate and larval growth variations in Pseudoplatystoma punctifer : maternal and paternal effects

In Pseudoplatystoma punctifer (e.g. Pseudoplatystoma fasciatum) larvae, parental effects on hatching, growth of initial stages and dry feed adaptation were evaluated as they could influence fry heterogeneity, which is responsible for the enhancement of cannibalism, and which remains one of the main factors of mortality during larval stages. A full factorial experiment was carried out with 3 females × 3 males producing nine families of full siblings, raised separately in triplicates into 30 L tanks at 28±0.5 °C in a water recirculating system. Paternal and maternal effects were observed on hatching success, yolk utilization efficiency and growth until 26 days post fertilization. Hatching success was generally over 80% except for one male × female combination (25%). Total length (TL) at hatching and during the first 4 weeks of exogenous feeding on live Artemia nauplii and dry feeds was determined in each family using digital photographs of larvae and nih image j analysis freeware. Mean TL was calculated for each family at each sampling time and analysed using multifactorial analysis of variance tests. These results indicate not only dam but also sire effects at very early developmental stages as well as in subsequent stages of P. punctifer.     

Light intensity affects the survival and growth of matrinxã larvae,Brycon amazonicus (Spix & Agassiz, 1829)

We evaluated the effect of light intensity on survival rate and zootechnical performance in matrinxã larvae (Brycon amazonicus) that is an important species for fish farming in the Amazon region. For this, the larvae were submitted to three experimental stages : Stage I—10–72 hr after hatching (HAH), Stage II—72–168 HAH and Stage III—168–288 HAH. The animals were submitted to three treatments of different light intensities: low (±20 lx), intermediate (±200 lx) and high (±2,000 lx). The low light intensity increased the survival rate in 24, 48, 72, 120, 168 and 288 HAH. The size of the animals was homogeneous in 72 and 168 HAH in the low and intermediate treatments. The high light intensity increased the zootechnical parameters in 288 HAH. We suggest the use of low light intensity in the initial periods of observation (24 and 48 HAH) to reduce aggressiveness and increase the survival and homogeneity of the animals. On the other hand, it is possible to suggest an increase in the light intensity in the final period of observation (288 HAH), as the increase in intensity stimulated growth and affected the survival rate of the matrinxã larvae.     

On the breeding behaviour and reproduction of the freshwater prawn,Macrobrachium rosenbergii (de Man 1879) (Decapoda‐Crustacea) under laboratory conditions

The giant freshwater prawn, Macrobrachium rosenbergii is one of the most common decapod species, and now getting more attention from the aquaculturists world wide due to its high market demand. It is commercially important because of its size as well as its eating flesh qualities. The breeding behaviour, reproduction and hatching of this species were observed for about 8 months during 2009. Juveniles (0.55 ± 0.177 g total weight and 2.7 ± 0.12 cm, total length) were reared in rounded fibre glass tanks (1.3 m, diameter). Pre‐mating moult occurs in prawns once the ovaries ripen in their carapace cavity so as to transform the prawn into berried stage. Recorded incubation period ranged from 18 to 24 days. The number of eggs ranged from 2050 to 150 500 and the fecundity ranged from 435.2 to 3849.1 eggs. Number of hatched larvae ranged from 1825 to 123 410 larvae for females of 4.71 to 39.1 g respectively. The number of eggs carried by female prawn was directly proportional to its body weight (no. of eggs = 3441.3 wt. of female – 32 292, r² = 0.819). (fecundity = 82.066 wt. of female ? 235.04, r² = 0.7779; fecundity = 317.86 length of female ? 2651, r² = 0.833). Hatching rate ranged from 65% to 91%, and there was a correlation between number of hatched larvae and size of female parent (no. of larvae = 10 369 length of female – 102 965, r² = 0.8159; no. of larvae = 2792.9 wt. of female – 26 268, r² = 0.829). This study can greatly help in the management strategies of prawn hatcheries and improve its hatching technology.     

Is batch variability in hatching time related to size heterogeneity and cannibalism in pikeperch (Sander lucioperca)?

Larval size heterogeneity is known to induce cannibalism, and procedures to avoid larval size differences are consequently implemented already during egg incubation and hatching. We investigated the relation between larval development variability, size heterogeneity and cannibalism in pikeperch. Larvae were sorted into five groups according to the time of hatching during a 65‐h period. The larvae with different times of hatch were then reared separately or together during an 18‐day period. Late hatched larvae were longer (P=0.003) and had less yolk remaining (P<0.001) than early hatched individuals at the time of hatching. However, on 11 days post fertilization, the late‐hatching larvae tended to have larger yolk reserves than earlier hatched individuals (P=0.07). Furthermore, the next day, a lower proportion in the late fraction had switched to exogenous feeding (P=0.024). That larvae with a late hatching time developed slower suggests a positive relationship between the hatching time and the embryonic developmental rate. However, differences in the length and available yolk reserves at hatching between larval fractions with different hatching times suggest that hatching is not strictly associated with a specific developmental stage, and that factors other than the development rate of the embryo are involved in the timing of hatching.     

Larval development in European hake (Merluccius merluccius L.) reared in a semi-intensive culture system

Eggs of European hake (Merluccius merluccius L.) were stripped from fish caught at sea. Larvae were kept under semi‐intensive conditions at around 12°C. In addition, eggs were incubated in single wells at 9.2, 12.7 and 14.5°C, where hatching, development and survival were closely examined. During the larval stage, a total of 299 larvae were sampled to follow development and growth. In addition a small number of juveniles were sampled. Larvae hatched approximately 4 days after fertilization, and were 2.9 mm in total length (TL). At 6‐day post hatching (dph), the larvae were 4.1 mm (TL), the jaw apparatus was developed, and the larvae had started to feed. Most of the growth during the early larval period is restricted to the head, and there is almost no increase in length for the first 3–4 weeks post hatching. Teeth and pelvic fins appear at 25 dph. Development of unpaired fins at approximately 30 dph marks the start of the larval–juvenile transition. Weaning to formulated feed was accomplished 50 dph, when external morphology was similar to that of adult hake.     

Effect of dietary vitamin A on Senegalese sole (Solea senegalensis) skeletogenesis and larval quality

The effects of different levels of vitamin A (VA) in Senegalese sole larval performance and development were evaluated by means of a dietary dose–response experiment using enriched Artemia metanauplii as a carrier of this micronutrient. Larvae were fed from 6 to 27 days post hatch (dph) with enriched Artemia containing graded levels of total VA (1.3, 2.1, 4.5 and 12.9 µg VA mg^− 1 DW). The content of VA in live prey directly affected its accumulation in larvae and early juveniles. Retinyl palmitate accumulated during larval ontogeny, whereas retinol showed the opposite trend, decreasing from hatching until 41 dph and then remaining constant until the end of the study.In metamorphic larvae (10 and 15 dph), VA did not affect the number of thyroid follicles or the intensity of the immunoreactive staining of T₃ and T₄. However, at older stages of development (post-metamorphic larvae: 20, 30, 41 and 48 dph), VA decreased the number of thyroid follicles but increased their mean size and enhanced T₃ and T₄ immunoreactive staining. A dietary excess of VA did not affect either larval performance in terms of growth and survival or the maturation of the digestive system. However, the most remarkable impact of this morphogenetic nutrient was detected during skeletal morphogenesis. Dietary VA accelerated the intramembranous ossification of vertebral centrums, which led to the formation of a supranumerary haemal vertebra and a high incidence of fused and compressed vertebrae in fish fed 2.1, 4.5 and 12.9 mg VA mg^− 1 DW. In addition, VA also affected those structures from vertebrae and caudal fin formed by chondral ossification, leading to defects in their shape and fusions with adjacent skeletal elements. In particular, the caudal fin was the region most affected by the dietary treatments. In order of importance, the bones with more developmental anomalies were the modified neural and haemal spines, epural, hypurals and parahypural. The impact of systemic factors such as thyroidal hormones in skeletogenesis should not be neglected since present results revealed that an excess of dietary VA affected the levels of T₃ and T₄, which might have affected bone formation and remodelling, leading to skeletal deformities.     

Role of thyroid hormones in tilapia larvae (Oreochromis mossambicus): I. Effects of the hormones and an antithyroid drug on yolk absorption,growth and development

Treatment of one-day old yolksac larvae of tilapia (Oreochromis mossambicus) by immersion in 0.05 ppm T₄ or 0.01 ppm T₃ significantly accelerated the differentiation and growth of all the fins, particularly pectoral and tail fins. Both the treatments also significantly accelerated yolk absorption and transition to free-swimming activity in the larvae. The treatments also significantly accelerated the growth of the larvae, with T₃ at 0.01 ppm having a greater effect than T₄ at 0.05 ppm. The yolk conversion efficiency was found not to be significantly affected by the hormone treatments but the treated larvae exhibited an increased heart beat, suggesting metabolic stimulation by the hormones.On the other hand, yolk absorption and free-swimming activity were significantly delayed in tilapia larvae immersed in 25 ppm solution of an antithyroid drug, phenylthiocarbamide (PTC). PTC also retarded the growth of the larvae. T₄ (0.05 and 0.10 ppm) or T₃ (0.01 and 0.02 ppm) therapy removed the PTC-inhibition,albeit not completely, suggesting that thyroid hormones are involved in the larval growth and development of tilapia.