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.     

Effects of delayed first feeding on larval growth,survival and development of the sea cucumber Apostichopus japonicus (Holothuroidea)

The time of first feeding is an important factor for establishing successful exogenous feeding in aquaculture hatcheries as it is crucial for larval survival and growth. The effects of delayed first feeding by the sea cucumber Apostichopus japonicus larvae were investigated. Groups of larvae were fed immediately after completion of the digestive tract (fed control) or starved for 24, 48, 72, 108 and 120 h post fertilization. The growth rates of A. japonicus larvae were the same as those of the control group when first feeding was delayed for less than 72 h. When the period of starvation was beyond 72 h, growth rates were reduced. The fed control group had the highest percentage of late auricularia larvae (70.0%) on day 11, followed by the groups starved for 24, 48, and 72 h, respectively, which had 35.00%, 43.48% and 41.67% of late auricularia larvae. When larvae were starved for more than 72 h, the percentage of late auricularia larvae never exceeded 12.0%. Most larvae halted in development and were deformed before reaching metamorphosis when starved for 108 and 120 h. Prolonged exposure to starvation (more than 48 h) resulted in a significant decrease in percentage of larvae reaching settlement. The point‐of‐no‐return (PNR) of A. japonicus larvae was calculated 110.4 h. Ratios of stomach length to total length, stomach width to total width were more sensitive to starvation than other larval morphometrics. To avoid mortality and obtain adequate growth and development, A. japonicus larvae in commercial culture should be fed within 48 h of the completion of the digestive tract.     

5-Hydroxytryptamine enhances larval development of the giant freshwater prawn,Macrobrachium rosenbergii

The effect of 5-hydroxytryptamine (5-HT) on larval development of the giant freshwater prawn, Macrobrachium rosenbergii, was investigated. Four days after hatching, larvae were exposed to 5, 50 or 100 μg/ml 5-HT in 12 ppt commercial seawater for a period of 2 or more days. Continuous exposure to 50 μg/ml or 100 μg/ml 5-HT over 2 days reduced the survival significantly and all larvae died by 13 days post-hatch (after 9 days of exposure). On the contrary, there was no significant difference in the survival of larvae exposed to various concentration of 5-HT for 2 days compared to that of the control. Exposure to 5-HT for 2 days stimulated growth and developmental rate of the larvae. Furthermore, pigmentation of the exoskeleton was also induced within 1 h of exposure. The larvae exhibited bright-orange color on their eyestalks, cephalothoraxes and abdomens. Exposure to 100 μg/ml 5-HT for 2 days produced the highest growth, developmental rate and the strongest pigmentation. The average body lengths 25 days post-hatch of larvae exposed to 50 μg/ml and 100 μg/ml 5-HT for 2 days were 3.5 ± 0.6 mm and 4.3 ± 0.7 mm, respectively, compared to 2.8 ± 0.4 mm that of the control. Furthermore, the rate of transformation of larvae to postlarvae during 30–40 days post-hatch was significantly higher in those exposed to 50 μg/ml and 100 μg/ml 5-HT compared to that of the control. The present study indicates that brief exposure of early stage larvae to 50–100 μg/ml 5-HT enhances growth, developmental rate and pigmentation of the giant freshwater prawn larvae.     

Effect of nitrite on larval development of giant river prawn Macrobrachium rosenbergii

The effects of ambient nitrite concentrations on larval development of giant river prawn Macrobrachium rosenbergii were evaluated. The trials were conducted in two phases: phase 1, larvae from stages I through VIII and phase 2, larvae from stage VIII until post-larvae. In both phases larvae were kept in water with nitrite (NO₂-N) concentrations of 0, 2, 4, 8 and 16 mg/L. Oxygen consumption was analyzed for larvae in stage II at nitrite concentrations of 0, 4, and 8 mg/L. Survival, weight gain, larval stage index and metamorphosis rate decreased linearly with increasing ambient nitrite concentration. However, there was no significant difference between larvae subjected to 0 and 2 mg/L NO₂-N. In phase 1, there was total mortality at 16 mg/L NO₂-N, while in phase 2 larval development stopped at stage X in this treatment. The oxygen consumption in stage II increased significantly at NO₂-N concentration from 0 to 4 mg/L, but there was no difference between 4 and 8 mg/L NO₂-N. In conclusion, increasing ambient nitrite up to 16 mg/L NO₂-N delays larval development, reduces larval growth rate and causes mortality, whereas no significant effect occurs for levels below 2 mg/L NO₂-N. However, the establishment of a general safe level of nitrite to M. rosenbergii hatchery may be difficult due to the great variability in larvae individual sensitivity.     

Influence of temperature on yolk resorption in common snook Centropomus undecimalis (Bloch, 1792) larvae

To determine the optimal rearing temperature for Centropomus undecimalis larvae during the yolk resorption period, changes in larval development were measured at four different temperatures (23, 25, 28 and 31°C). Yolk and oil‐globule volume was recorded for 25 larvae at four different times. This involved an initial measurement at hatch and at 24, 48 and 72 h posthatch (hph). Additional morphological measurements included standard length, body height and eye diameter. On average, at the end of the three trials, larvae reared at 25°C had a longer mean standard length than larvae reared at 23, 28 and 31°C. Larvae reared at 25°C also had more yolk and oil globule reserves than larvae raised at 28 and 31°C. The body height:length residuals were also the highest at 25°C (i.e. larvae had deeper or stockier bodies). The yolk sac was present up to 72 hph at 23 and 25°C, while it was entirely consumed after 48 hph in larvae held at 28 and 31°C. Larvae showed the fastest growth during the first 24 hph in all temperature treatments; this period corresponded to the highest energy consumption as determined by the decrease in yolk sac and oil‐globule volume. Eye diameter did not vary significantly with time during yolk‐resorption. We conclude that a temperature near 25°C is optimal for raising snook larvae during the yolk‐resorption period.     

Structural development of Pacific red snapper Lutjanus peru from hatching to the onset of first feeding

Successful rearing of larval fish requires culture conditions and feeding strategies matching the ontogenetic status of larvae. This study describes the external morphology and development of organs and structures involved in the feeding process (i.e. sensorial organs, mouthparts and digestive system) from hatching until first feeding in Pacific red snapper. Hatching occurred 26 h after fertilization at 26°C and total length (TL) was 2.45 ± 0.08 mm. The larvae showed an undifferentiated eye and digestive tract. At 48 hah, TL was 3.31 ± 0.12 mm. Yolk and oil globule were still present. The mouth was still closed, but the Meckel's, quadrate, hyoid and hyomandibular cartilages were present. The retina was formed by 5 layers, and a thin layer of pigment epithelium was observed in the outer nuclear layer (ONL). At 70 hah, TL was 3.44 ± 0.22 mm. A remnant of oil globule was still present. The mouth and anus were open. At 93 hah, the number of cones in the ONL of the retina have increased and there was more pigment in the pigment epithelial layer. A joint between Meckel's and the quadrate cartilage and also a joint between the hyomandibular cartilage and the skull were present. The presence of live feed was detected in the digestive tract of these larvae. Based on these observations, the Pacific red snapper larvae is functional to start ingesting live feed between the 3rd and 4th day after hatching.     

Embryonic development and morphology of eggs and newly hatched larvae of Pacific herring <Emphasis Type="Italic">Clupea pallasii</Emphasis>

The embryonic development and morphology of eggs and newly hatched larvae of the Pacific herring Clupea pallasii were described using laboratory-reared specimens originating from the Miyako Bay stock. The eggs were almost spherical in shape, 1.33–1.46 mm (mean: 1.38 mm) in diameter, and had a thick adherent chorion. They had a segmented pale yellow yolk, no oil globule, and a relatively wide perivitelline space. A subgerminal cavity was observed during the gastrula period, whereas the blastocoel did not appear. Mass hatching occurred by 271 h 45 min after fertilization, and the newly hatched larvae were 7.1–7.7 mm (mean: 7.5 mm) in total length with 53–56 myomeres at 9.6°C. The embryonic development of Pacific herring was substantially similar to that of zebrafish Danio rerio, American shad Alosa sapidissima, as well as Atlantic herring Clupea harengus, and generally followed the basic developmental pattern of teleosts. However, Pacific herring larvae hatched at a more developed stage than some other clupeoids, such as Japanese sardine Sardinops melanostictus, and the progressed developmental stage at hatching could be interpreted as an advanced adaptation.     

Comparative study of the effects of heavy metals on embryonic development of zebrafish

This study aimed to investigate the effects of heavy metals, such as copper, zinc, iron, cobalt, chromium, aluminium, manganese and molybdenum, on zebrafish egg hatching rate and the subsequent survival rate of the larvae. Each metal was used in the various concentrations of0.05 mg/L, 50 mg/L and 500 mg/L to detect the threshold limit of tolerance. The study's results showed significant differences in the hatching and survival rate of larvae between the control group and most of the experimental groups when adding heavy metals in various concentrations. The highest concentration (500 mg/L) of the induced heavy metals tested in this study resulted in total egg or larval mortality within 96 hr and significantly lower egg or larval life expectancy. Among the tested metals, zinc and molybdenum had the most significant impact on hatching time, in contrast to iron that did not differentiate the results from the control group. Copper, cobalt, chromium, aluminium and manganese did affect zebrafish eggs depending on their exposure concentrations, which differentiated the results from the control group. Also, the study's findings demonstrate a significant dependence of the hatched egg heart rate on the metal concentration, and in particular, higher concentrations of metals correspond to higher heart rate. Zinc, Molybdenum and copper was proved to be the most toxic heavy metals causing increased hazard rate to mortality up to 96 hr and shorter life expectancy. Finally, skeletal deformities that occasionally observed did not significantly affect the total life expectancy of the hatched eggs.     

EDCs‐induced glucocorticoid receptor related genes expression of the river pufferfish,Takifugu obscurus

Endocrine disrupting chemicals (EDCs) are of substantial concern when contaminating waters. We studied gene expression of larvae from river pufferfish, Takifugu obscurus exposed to EDCs. We cloned the full‐ or partial‐length cDNA sequence of genes related to the cortisol pathway, such as the glucocorticoid receptor (GR), mineralocorticoid receptor (MR), heat shock protein (HSP) 70 and 90, phosphoenolpyruvate carboxykinase (PEPCK) and the FK506‐binding protein 4 (FKBP52). The tissue and time‐dependent expression of mRNA was studied by real‐time PCR in T. obscurus exposed to two representative EDCs (bisphenol‐A and 4‐tert‐octylphenol). HSP70, HSP90, MR and FKBP52 mRNA expression level was much higher than GR and PEPCK expression. GR mRNA expression level was significantly upregulated after 48 h in both EDC‐treated fish groups compared with the control. Both groups also showed down‐regulation patterns after 48 h. The initial expression of PEPCK in both groups was down‐regulated after 24 h. In the bisphenol‐A treated group, the expression of FKBP52 showed substantially high up‐regulation from 6 to 48 h after exposure, and then strikingly reduced its expression level at 72 h. Our results provide insights into the EDCs metabolizing system of T. obscurus and offers baseline information for further research related to the possible use as a bio‐indicator of this commercially important aquaculture fish species.     

Acute toxicity of ammonia on juvenile cobia (Rachycentron canadum, Linnaeus, 1766) according to the salinity

Juvenile cobia (Rachycentron canadum) (total length 15.16 ± 0.92 cm and weight 19.26 ± 4.5 g) were exposed to different concentrations of ammonia–N (unionized plus ionized ammonia as nitrogen), using the static renewal method at different salinity levels of 5, 20, and 35‰ at pH 8.1 and 25°C. The 24, 48, 72, 96 h LC₅₀ values of ammonia–N for R. canadum juveniles were 60.28, 48.57, 37.42, 22.73 mg l⁻¹ at 35‰; 51.25, 43.63, 28.17, 19.05 mg l⁻¹ at 20‰; and 39.48, 25.31, 19.50, 8.13 mg l⁻¹ at 5‰, respectively. The 24, 48, 72, 96 h LC₅₀ values of NH₃–N (unionized ammonia as nitrogen) were 1.81, 1.46, 1.12, and 0.68 mg l⁻¹ at 35‰; 1.75, 1.49, 0.96, and 0.65 mg l⁻¹ at 20‰; and 1.52, 0.97, 0.71, and 0.31 mg l⁻¹ at 5‰, respectively. As the salinity decreased from 35 to 5‰, susceptibility of ammonia–N increased by 34.5, 47.88, 50.56, and 64.23% after 24, 48, 72, and 96 h exposure, respectively. Furthermore, we found that exposure of fish to ammonia–N caused an increase in oxygen consumption of 129.1, 157.5, and 192% and a decrease in the ammonia excretion level of 53.4, 38.2, and 23.3% with respect to the control.     

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.     

Effect of incubation temperature on the embryonic development and yolk‐sac larvae of the Pacific red snapper Lutjanus peru (Nichols & Murphy, 1922)

The effect of incubation temperature on embryonic development and yolk‐sac larva of the Pacific red snapper Lutjanus peru were evaluated by testing the effect of 26, 28 and 30°C, as this is the natural thermal interval reported during the spawning season of Pacific red snapper in the Gulf of California, Mexico. Sixteen developmental stages were observed. The incubation temperature affected the rate of development and time to hatching, being shorter at 30 than at 26°C, but no significant effect (P < 0.05) on larval length at hatching was registered. The depletion rate of yolk sac and oil globule was affected by incubation temperature particularly during the first 12 h post hatching (hph). At the end of the experiment (48 hph), significantly (P < 0.05) larger larvae were recorded at 26°C (TL = 3.22 ± 0.01 mm) than at 28° (TL = 3.01 ± 0.02 mm) and 30°C (TL = 2.97 ± 0.05 mm). Incubation of newly fertilized eggs at 26°C produces larger larvae, which may help to improve feeding efficiency and survival during first feeding.     

Embryonic and larval development in the semelparous Nereid polychaete Hediste diversicolor (OF Müller, 1776) in Norway: Challenges and perspectives

Knowledge of broodstock manipulation, gametogenesis, artificial fertilization and larval nutrition is a prerequisite to reach a large‐scale production of the polychaete Hediste diversicolor. In this study, the characteristics of oogenesis and spermatogenesis and embryonic and larval development were observed. Moreover, the effect of temperature on embryonic development and effect of diets and sediment on earlier juvenile development were tested. Mature oocytes were characterized by a spherical shape with a diameter of about 200 μm. They showed radial symmetry with a large centric nucleus surrounded by four distinctive layers. Embryo cleavage started between 4 and 10 hr after fertilization, and developed to the swimming trochophore stage after 5–6 days after fertilization at 11°C. The rate of embryonic development increased with a temperature increase from 6.1 to 21.2°C. The trochophore larvae increased in length from day 9 and commenced differentiation into 3‐setiger stage larvae at day 12. The length growth of larvae until 7‐setiger stage was 0.43 mm/day when fed with fish feed, while those fed shellfish diet and smolt sludge both grew around 0.21–0.23 mm/day. The results suggest that fish feed is a superior diet compared to shellfish diets and smolt sludge for the early larval stage of H. diversicolor.     

Acute ammonia toxicity during early ontogeny of ide Leuciscus idus (Cyprinidae)

Acute ammonia toxicity was investigated in four developmental stages of the juvenile ide, Leuciscus idus: 1, 10, 20 and 30 days after the first feeding. Mean (±SD) total length of the larvae was 8.5 ± 0.3, 15.7 ± 0.7, 23.0 ± 2.0 and 29.7 ± 2.0 mm, and standard weight was 1.6 ± 0.3, 9.2 ± 5.5, 94.9 ± 31.0 and 196.0 ± 31.7 mg, respectively. The larvae used for toxicity tests were reared in the experimental, closed recirculating system. Groups of fishes (n from 7 to 10; in respect of fish size) were exposed to the ammonium chloride solutions in 1-L glass units. Water temperature was 25 ± 0.1 °C for both the rearing and the toxicity tests. pH was not adjusted and ranged between 8.4 and 8.7. The ammonium chloride solutions were renewed every 12 h. At the same time, dead larvae were counted and removed, and the pH and temperature measurements were taken. Each acute toxicity test duration was 96 h, and lethal concentration LC₁, LC₅₀ and LC₉₉ values were calculated for 24, 48, 72 and 96 h. The susceptibility of the ide larvae to ammonia decreased linearly with age up to 20th day and surprisingly increased during the next 10 days. The LC₅₀ (48 h) values ranged from 0.27 mg L^?1 of unionized ammonia nitrogen for 1 day after the first feeding (AFF) larvae to 1.42 mg L^?1 at day 20 after first feeding. The LC₅₀ (48 h) for 30 days AFF was as high as 0.67 mg L^?1. The critical level of the unionized ammonia nitrogen for ide larvae was suggested as 0.21 mg L^?1.     

Response of <Emphasis Type="Italic">Cyprinus carpio</Emphasis> to copper exposure: alterations in reduced glutathione,catalase and proteins electrophoretic patterns

This study was conducted to characterize the alterations in reduced glutathione (GSH) level, catalase (CAT) activity and proteins electrophoretic patterns in response to sublethal copper (Cu) exposure in Cyprinus carpio and to determine whether these responses are related to Cu accumulation in gills, chosen as target tissue. Fish were exposed to 0.1 and 1.0 mg/l Cu for 10 and 20 days. There were increasing level of Cu in the gill with increasing concentrations of metal in the exposure medium, and with increasing duration of exposure. GSH level and CAT activity increased in fish exposed to 1.0 mg/l Cu for both exposure periods, while no change was detected at the lower Cu concentration. Electrophoretic patterns of gill proteins by sodium dodecyl sulphate gel electrophoresis (SDS–PAGE) consist of 25-, 26-, 30-, 44- and 48-kDA medium molecular weight proteins (MMP) for five bands and 64-, 72-, 90- and 101-kDA high molecular weight proteins (HMP) for four bands in both control and treatment groups. The levels of 25-, 26- and 30-kDA MMP and 72- and 90-kDA HMP increased in response to Cu exposure. The present study demonstrated that Cu caused stress in fish gills and an acclimation with induction of GSH, CAT, MMP and HMP, which were important in the protection against metal damage, was observed.     

Effects of turbidity on survival of larval ayu and red sea bream exposed to predation by jack mackerel and moon jellyfish

We conducted laboratory experiments to examine the effects of turbidity on the survival of red sea bream Pagrus major and ayu Plecoglossus altivelis altivelis larvae when exposed to either visual (jack mackerel juveniles) or tactile (moon jellyfish) predators. The experiments were conducted in 30-l tanks with three different levels of turbidity obtained by dissolving 0, 50, or 300 ppm kaolin. Predators were introduced to the experimental tanks followed by larvae of either red sea bream (mean ± standard deviation 6.1 ± 0.3 to 11.4 ± 2.1 mm standard length) or ayu (6.6 ± 0.3 and 24.4 ± 1.8 mm). When exposed to jack mackerel, the mean survival rate of larvae was significantly higher in 300 ppm treatments compared with the other turbidity levels. When exposed to moon jellyfish, however, there was a less marked difference in the survival rates among different turbidity treatments. Survival rates of ayu larvae exposed to moon jellyfish were generally lower than those of red sea bream. Our study indicates that anthropogenic increases of turbidity may increase the relative impact of jellyfish predation on fish larvae.     

Use of common salt affects aggressiveness in matrinxã larvae (Brycon amazonicus)

Aggressiveness is potentiated during the production of fish of commercial interest due to the densification and confinement, and this may affect the welfare and health of these animals. We compared the effect of salinity on aggressiveness, locomotor activity and body cortisol level in matrinxã larvae (Brycon amazonicus) submitted to three experimental treatments: 0, 2 and 4 g of common salt/L. For each treatment, the larvae were collected at 24, 36, 48, 60 and 72 HAH (hours after hatching) and then transferred to 5‐L aquariums for 30 min to analyse behavioural categories. The use of common salt increased the frequency of aggressive interaction, cannibalism, mortality and locomotion in all observation periods. Body cortisol levels did not differ between treatments until 48 HAH, but showed a significant increase at 72 HAH in the 4 g salt/L. There was a positive correlation between aggressive interactions and locomotor activity. Negative correlation was observed between these parameters and cortisol up to 60 HAH; however, they were positively correlated at 72 HAH. We concluded that the use of common salt stimulates the display of aggressive behaviour and locomotor activity and that the effect of salt is dependent on the developmental phase of the larvae of B. amazonicus.     

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.     

Alterations in blood electrolytes of a freshwater catfish <Emphasis Type="Italic">Heteropneustes fossilis</Emphasis> in response to treatment with a botanical pesticide, <Emphasis Type="Italic">Nerium indicum</Emphasis> leaf extract

The present study aimed at investigating the effects of Nerium indicum leaf extract on the blood electrolytes of Heteropneustes fossilis for short- and long term. Fish were subjected to Nerium indicum leaf extract for short term (11.27 mg/L i.e. 0.8 of 96 h LC₅₀) and long term (2.81 mg/L i.e. 0.2 of 96 h LC₅₀). Fish were killed on each time intervals from control and experimental (Nerium indicum) groups after 24, 48, 72, and 96 h in short-term exposure and after 7, 14, 21, and 28 days in long-term experiment. Blood samples were analyzed for calcium and inorganic phosphate levels. Acute exposure of Nerium indicum leaf extract caused a progressive decrease in the serum calcium levels after 48 h in fish H. fossilis, which persists till the close of the experiment (96 h). The serum inorganic phosphate levels remain unaffected till 48 h in the Nerium indicum leaf extract–exposed fish. After 72 and 96 h, the levels exhibit a decrease. Chronic Nerium indicum leaf extract treatment provoked a decrease in serum calcium levels at day 14. This decrease continues till 28 days. The serum phosphate level of the Nerium indicum leaf extract–treated fish decreases on day 14 and 21. However, on day 28, the levels become close to the normal values. We conclude that Nerium indicum leaf extract exposure alters the blood electrolytes of the fish, thus causing physiological disturbances which might affect seriously the normal vital functions, growth rate, reproduction, and their survival in nature.