Initiation of carp spermatozoa motility and early ATP reduction after milt contamination by urine

Fish sperm collected by stripping males is frequently contaminated by urine. In this study, carp milt mixed with urine (0.5–7.5% of volume) was studied in order to evaluate the changes of some motility parameters (percentage of motile spermatozoa, velocity and beat frequency) and the ATP content of spermatozoa. In the absence of urine contamination, spermatozoa had an ATP content in the range of 8–9 nmol/10⁸ spermatozoa, an initial velocity of 100–160 μm s⁻¹ and a flagellar beat frequency around 30–50 Hz, 10 s after a 1/2000 dilution in an activating medium (45 mM NaCl, 5 mM KCl, 30 mM Tris–HCl, pH 8.0, osmolality <160 mosM kg⁻¹). In contrast, when milt was contaminated with 7.5% of urine for 1 h, the ATP content was 4–5 nmol/10⁸ spermatozoa and most spermatozoa had low initial velocity (30–100 μm s⁻¹) and flagellar beat frequency (10–30 Hz). It appears that the low osmolality of urine was responsible for the degradation in the quality of carp spermatozoa by an early activation in the collecting tube which induced an early reduction of the intracellular ATP store. From a practical point of view, milt contamination by urine during stripping can be avoided by first pressing the abdomen before sampling and then collecting the remaining urine by means of a catheter introduced into the urinary bladder.     

Age-Related Sperm Quality of Captive Striped Bass Morone saxatilis

Three groups of captive-reared striped bass Morone saxatilis ages 1, 3 and 12 yr, were examined for age-related changes of sperm characteristics including short-term storage. All groups had similar ranges of the following parameters (mean× SEM): expressible milt (5.6× 0.5 mI/kg body weight (BW) to 7.5× 2.1 mL/kg BW), percentage of motile sperm (55× 6% to 60× 2%), duration of sperm motility (69× 3 sec to 72× 5 sec) and percentage of viable sperm (91× 2% to 93× 2%). Compared to the 1 and 12-yr-old fish, the 3-yr-old fish produced the greatest number of spermatozoa (1,190× 370× 10⁹ spermatozoa/kg), sperm concentration (120× 8 × 10⁹ spermatozoa/mL) and spermatocrit (74× 4%). In addition, during short-term storage at 4 C, extender-preserved sperm samples of the 3-yr-old group showed a significantly higher ( P < 0.05) percentage of motile sperm and duration of sperm motility, compared to the other two groups. This suggests that short-term storage may be affected by the age of the male fish. Sperm longevity of the 3-yr-old group was successfully maintained for as long as 15 d, longer than that of the 1-yr-old group (9 d) and 12-yr-old group (7 d). Overall, the 3-yr-old fish appeared to have superior sperm quality than the 1 or 12-yr-old fish based on higher sperm production and increased sperm longevity.     

Short-term storage and cryopreservation of milt from Atlantic salmon and sea trout

Experiments on short-term preservation of sperm were performed with Atlantic salmon (Salmo salar). Fertility was maintained for up to 10 days when 2 mm thick samples were stored at 0° C under an oxygen atmosphere in the presence of antibiotics (125 IU penicillin and 125 μg streptomycin per ml sperm). Fertility was completely lost after 24 days. Sperm stored without antibiotics fertilized 100% of eggs after 6 days.

Cryopreservation was carried out with milt from Atlantic salmon and sea trout (Salmo trutta). Semen mixed with extender was frozen on dry ice (pellets) with subsequent storage in liquid nitrogen. Sperm pellets were thawed in a 0.12-M NaHCO₃-solution (10° C) before insemination. The suitability of an extender as previously described by Stoss and Holtz and of a 0.3-M glucose solution with the addition of 10% DMSO, was tested on two different batches of sperm and eggs in Atlantic salmon and sea trout. In addition, the extender earlier reported by Mounib and an aqueous solution of 10% DMSO were only used in Atlantic salmon with one batch of gametes.

Insemination with cryopreserved Atlantic salmon sperm resulted in 36 to 91.3% eyed eggs (control = 100). The differences were caused by the type of extender and the batch of gametes employed. The very simple extender consisting of 0.3 M glucose and 10% DMSO only was the most successful. Results with cryopreserved sea trout sperm ranged between 38.6–54.8% eyed eggs, showing no difference between treatments.     

Cryopreservation of YamúBrycon siebenthalae Milt

The yamú Brycon siebenthalae is an endemic fish of the Orinoco river basin, but wild stocks are decreasing because of the disruption of their habitat. We evaluated a protocol for the cryopreservation of yamú sperm to contribute to the preservation of this endangered genetic resource. Milt was mixed with a cryoprotectant medium (5.5% glucose, 12% egg yolk, and 5%, 10%, or 15% dimethyl sulfoxide - DMSO) in a ratio 1:4 (milt:medium), stored in 0.5-mL French straws, frozen in nitrogen liquid vapor (-76 C), then immersed and stored in liquid nitrogen for 10 d or 12 mo. Motility of thawed spermatozoa was higher ( P < 0.001) in 10% DMSO medium than 5% DMSO or 15% DMSO mediums; but lower than the control ( P < 0.001). With sperm cryopreserved, the highest level of fertilization was achieved with 10% DMSO ( P < 0.001) after 10 d or 12 mo of cryopreservation. Fertilization of eggs inseminated with 6.4 × 10⁹ spermatozoa per g of eggs was higher ( P <0.05) than with 1.6 × 10⁹ spermatozoa per g of eggs. There was no difference (P > 0.05) in fertilization between insemination doses of 3.2 × 10⁹ and 6.4 × 10⁹ spermatozoa per g of eggs. Cryopreservation of yamu milt can be performed successfully with a simple medium combined with 10% of DMSO as cryoprotectant. The highest level of fertility was achieved using between 3 × 10⁹ and 6 × 10⁹ spermatozoa per g of fresh eggs.     

Production of Florida red tilapia in seawater pools: Nursery rearing with chicken manure and growout with prepared feed

Florida red tilapia (Oreochromis sp.) were reared in 23 m³ seawater (37 ppt) pools. Monosex males (1.3 g mean weight) were stocked at a density of 25 fish/m³ and reared to fingerling size (>10 g) in pools receiving either chicken manure applied at a rate of 105 kg/ha day⁻¹ or pelletized feed (30% protein) administered ad libitum. Following the nursery period, fingerlings in fed pools were reared through adult, marketable sizes.

After 20 days of nursery rearing, mean fish weights (5.7–9.6 g) and survival (77.5–98.6%) in manured pools ranged from less than to greater than values in fed pools (7.9–9.4 g and 95.5–98.2%). By day 33, while mean weights (11.3±0.4 g) and survival (84.5±5.2%) in manured pools were significantly less than those in fed pools (18.0±0.6 g and 95.9±1.4%), fingerling-size fish were obtained from manured pools at an overall productivity of 55 kg/ha day⁻¹.

After 170 days in fed pools, mean fish weight was 467±9 g, survival was 89.7±0.9%, and food conversion was 1.6±0.2. Daily weight gain achieved a maximum of 4.4 g day before a rapid decline in water temperature from 28–29°C to 24–25°C caused a loss of fish appetite and evidence of disease.

The results suggest that while nursery rearing of Florida red tilapia in seawater pools fertilized with chicken manure is feasible, considerable variability in fish performance among pools can be expected, despite identical management methods. In pools receiving prepared feed, high growth rates and survival through adult, marketable sizes suggests a potential for commercial production of Florida red tilapia in seawater.     

Characterization of rainbow trout milt collected with a catheter: semen parameters and cryopreservation success

Collection of fish milt by stripping risks the danger of milt contamination by urine. This may seriously influence milt characteristics and quality, including usefulness for cryopreservation. Urine contamination of milt may be avoided by using a catheter for sperm collection. The objectives of this study were to provide basic characteristics of milt collected with a catheter, to test the usefulness of this milt for cryopreservation, and to correlate characteristics of fresh and cryopreserved semen with sperm fertility rates. Milt from 25 rainbow trout Oncorhynchus mykiss (Walbaum) males were used. All samples were cryopreserved using the pellet method within 1 h of collection, using 0.6 m sucrose and 10% dimethyl sulphoxide (DMSO) as an extender. Catheterization resulted in semen of very good motility (> 90% motile spermatozoa) and high fertilization rates after cryopreservation (mean fertilization rate 81.8 ± 13.3% of control, at a sperm/egg ratio of 2.4 ± 0.3 × 10⁶). Osmolality of seminal plasma and concentrations of sodium, potassium and magnesium ions had low variability, which suggests that they are important for creating a stable environment for sperm storage in the sperm duct. Higher variability of certain seminal plasma characteristics, such as protein concentration and antiproteinase activity, suggests that these characteristics are related to individual semen features of particular males. A strong correlation of seminal plasma zinc concentration with protein concentration may reflect an importance of zinc in semen biology. Cryopreservation caused a significant release of protein and acid phosphatase from spermatozoa. Our results did not reveal any single characteristic of semen collected by catheter that could be used as a powerful predictor of cryopreservation success, presumably because all samples were of high quality.     

Sperm Characteristics of Precocious 1-year-old Male Striped Bass Morone saxatilis

Abstract.— Captive-reared, 10-mo-old, male striped bass Morone saxatilis were sampled monthly for testicular development between February and June 1994. One of the five males sampled in February showed precocious testicular development and had a gonado-somatic index (GSI) of 1.26%. while the other four fish had immature testes with a mean GSI ± s^x, of 0.17 ± 0.03%. Spermiating individuals were present from April to June. In April the average body weight (BW) of spermiating males was 65 ± 4 g and their GSI reached a mean value of 4.75 ± 0.52%. In June, milt collected from ten precocious males contained motile spermatozoa with a mean of 31 ± 7% of the sperm showing forward movement. Mean milt volume and sperm concentration were 1.67 ± 0.41 mL/kg BW and 92.3 ± 1.8 ± 10⁹ spermatozoa/mL. respectively. These data show that male striped bass reared in captivity can reach sexual maturity during their first year. This is one year earlier than previously reported for striped bass in mid-Atlantic regions.     

Changes in Sperm Production,Sperm Motility,and Composition of Seminal Fluid in Caspian Brown Trout,Salmo trutta caspius,Over the Course of a Spawning Season

This study was carried out to evaluate milt quality in male Caspian brown trout (Salmo trutta caspius) over the course of the winter spawning season. Milt samples were collected biweekly during December and January. Chemical composition of seminal fluid, sperm production (milt volume, sperm density, spermatocrit,) and sperm motility characteristics (percentage and duration of motility) were measured. Milt volume, sperm density, osmolality, seminal minerals (Ca²⁺, Mg²⁺, K⁺, Na⁺, Cl^?), and total protein gradually decreased over the spawning season. Glucose and triglyceride content of milt did not show significant changes over the spawning season. Milt pH and the percentage and duration of motility were comparatively stable, declining only at the end of the season. Significant positive correlations were found between sperm density and seminal minerals, total protein and spermatocrit; percentage of motile spermatozoa and seminal minerals, total protein; and duration of motility and K⁺, Cl^?, total protein, and pH. Results show that season has a significant influence on milt quality in male Caspian brown trout, with the best milt being available at the beginning of spawning season.     

Ovarian fluid pH enhances motility parameters of rainbow trout (Oncorhynchus mykiss) spermatozoa

All evidence to date suggest that sperm motility is the primary determinant of fertilization success in externally fertilizing fish species. Ovarian fluid, which comprises 10–30% of the total egg volume in salmonids, enhances sperm motility with respect to swimming speed, trajectory and the duration of movement. It was recently demonstrated that there is individual variability in sperm motility enhancing potential of ovarian fluid of particular females. In the present study we examined the effect of particular ovarian fluids collected from 31 females on the sperm motility parameters of one male of rainbow trout (Oncorhynchus mykiss) using computer-assisted sperm analysis (CASA). During our experiment we also monitored the pH of ovarian fluid. We found that particular fluids differed in the ability to activate spermatozoa; sperm remained immotile in four fluids and exhibited 50–100% motility in 27 samples. The percentage of motile sperm, velocity and duration of movement positively correlated with ovarian fluid pH (r² = 0.34–0.62). These data strongly suggest that the pH of the ovarian fluid is the primary determinant of sperm motility in rainbow trout under natural conditions of fertilization.     

Motility Parameters of Rainbow Trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) Spermatozoa in Relation to Sequential Collection of Milt,Time of <Emphasis Type="Italic">Post-mortem</Emphasis> Storage and Anesthesia

The present study investigated the effects of sequential collection of milt, time of post-mortem storage and anesthesia on rainbow trout (Oncorhynchus mykiss) sperm motility parameters (using computer-assisted sperm analysis – CASA) as well as seminal plasma osmolality and sperm concentration. The post-mortem storage and time of anesthesia altered motility characteristics of rainbow trout sperm to different extents. The moderate impact of time of anesthesia was manifested in a shortened duration of sperm motility after 10 min exposure of fish to anesthetic. The prolonged post-mortem storage (≥40–60 min), in addition to lowering sperm motility duration, also significantly influenced sperm motility parameters, such as sperm velocities, percentage of motile sperm and sperm trajectory parameters. These results clearly demonstrate that when milt from sacrificed fish is used for sperm motility studies, the time of post-mortem storage significantly alters sperm motility characteristics. Since sperm motility rate and swimming velocity could predict fertilizing ability, detrimental effects of prolonged post-mortem storage may lead to reduced fertilization success. Sperm concentration and seminal plasma osmolality were lower in the first fractions and increased with successive collections of milt. It suggests the presence of urine contamination of the first milt fractions which were collected by stripping. Therefore, testing of sperm concentration and/or seminal plasma osmolality should be mandatory while handling stored milt.     

Effects of slow release gonadotropin releasing hormone analog on milt characteristics and plasma levels of gonadal steroids in greenback flounder, Rhombosolea tapirina

Spermiated male greenback flounder (Rhombosolea tapirina) were implanted with gonadotropin releasing hormone analog (GnRHa) pellets at different dosages to examine their effects on milt characteristics and plasma levels of testosterone (T), 11-ketotestosterone (11KT) and 17,20β-dihydroxy-4-pregnen-3-one (17,20βP). Milt and blood samples were collected for up to 28 or 42 days post-implantation (p.i.) in two separate experiments using fish that were slightly or moderately spermiated, respectively. In both experiments, fish treated with GnRHa pellets showed a consistent and significant increase in milt volume relative to controls, and the increase of milt volume was more rapid than the increase in numbers of spermatozoa. Spermatocrit (Sct) was significantly lower in GnRHa-treated fish than in controls. Plasma levels of T and 11KT were elevated at 7 and 14 days p.i. in slightly spermiated fish treated with GnRHa and elevated plasma T and 11KT levels were accompanied by increased milt volume early in the spermiation period. In contrast, there was no significant difference in plasma T levels between GnRHa-treated and control fish in fish that were moderately spermiated at the time of implant. Treatment with GnRHa tended to result in lower plasma levels of 11KT and higher levels of 17,20βP relative to controls, and there was a positive relationship between the elevation of plasma 17,20βP and the increase in milt volume in response to implantation of GnRHa pellet. Slow release GnRHa administration had no effect on sperm activity or pH and osmolality of seminal plasma in either experiment.     

A laboratory scale recycling water unit for tilapia breeding

The technical features of a laboratory scale water recycling unit for experimental small scale tilapia breeding are described. Two units (1 and 2) were operated during a 6 month period, carrying a similar fish load (7·5 kg) and feeding rate (2% fish body weight/day). Unit 1 received natural illumination, while unit 2 was artificially illuminated (14/10 - light/dark cycle). Both units were equipped with a biological filter bed (substrate surface area, 3500 cm²). In unit 1, total ammonium and nitrite concentrations ranged from 0·05 to 0·5 mg liter⁻¹, while nitrate varied between 10–40 mg liter⁻¹. In unit 2 corresponding values were 0·15-3 mg liter⁻¹, 0·05–0·8 mg liter⁻¹ and 10–40 mg liter⁻¹. Temperatures ranged between 20–29°C and pH values between 7·5–6·9 in both units. Dissolved oxygen concentrations decreased gradually from 5·6 to 3·4 mg liter⁻¹ in unit 1 and from 5·6 to 2·6 mg liter⁻¹ in unit 2. Twenty-six spawnings occurred in unit 1 in March and April, while only eight spawnings occurred in unit 2, possibly because of the absence of sunlight. The significance of these results are discussed.     

Fertility of rainbow trout (Salmo gairdneri) gametes: Gamete viability in artificial media

A series of experiments was performed to test the effect of potassium ions (K⁺) on the storage of rainbow trout (Salmo gairdneri) sperm cells and eggs. Storage of eggs for up to 10 min in saline solutions, extender with K⁺ concentrations of ≤3.4 mM and coelomic fluid did not affect fertility. Fish Extender #6 had a detrimental effect on eggs within 15 s of application and reduced fertility to near zero within 10 min. Variation in the K⁺ concentration indicated that high (≥6.8 mM) concentrations decreased egg fertility while low (≤3.4 mM) concentrations had no effect on fertility. Insemination of eggs before the addition of Fish Extender #6 allowed storage for 14 h. Semen diluted (1:10) in coelomic fluid maintained fertility for 45 s, whereas semen diluted (1:1) in coelomic fluid maintained fertility for 60 min. Fish Extender #6 at K⁺ concentrations between 0 mM and 68 mM had no effect on fertility of semen diluted 1:2.5 after 1 h of storage.     

A diluent for sperm cryopreservation of gilthead seabream, Sparus aurata

Sperm of gilthead seabream, Sparus aurata, was diluted with solutions of different osmolarities and pH. The effect of the different diluents on sperm motility (intensity and percentage of motile sperm) was studied. Motility was induced as early as 10 s after mixing the sperm with diluents having an osmotic pressure higher than 500 mOsm/l. The intensity of motility decreased when the osmotic pressure was reduced, and was zero or significantly inhibited when the osmotic pressure of the diluent (300–380 mOsm/l) was close to that of the fish's seminal plasma (364.6±3.03 mOsm/l). The pH of the diluent did not have any effect on sperm motility (in a range of 6.8 to 8.9). A diluent which prevents spermatozoa motility (osmotic pressure 375 mOsm/l and pH 7.35) was successfully used to cryopreserve S. aurata sperm at −196°C. This diluent is considered promising for the long-term preservation of gilthead seabream sperm.     

Manipulation of ploidy in yellow perch (Perca flavescens) by heat shock, hydrostatic pressure shock, and spermatozoa inactivation

Heat shocks, hydrostatic pressure shocks, and ultraviolet radiation were evaluated for their efficacy as methods of manipulating ploidy in yellow perch (Perca flavescens). The most effective methods of inducing triploidy were heat shocks of 28–30°C applied at a time of initiation (TI) of 5 min postfertilization for durations of 10 or 25 min, and hydrostatic pressure shocks of 9000 or 11 000 psi applied at a TI of 5 min for a duration of 12 min. These treatments resulted in triploidy induction rates that ranged from 54–100%, and embryonic survival rates of 16–80%. Cold shocks of 0°C had no effect on the ploidy or survival of embryos. For perch, hydrostatic pressure shock offered several advantages over heat shock as a method of manipulating ploidy. The most effective methods of inducing tetraploidy were hydrostatic pressure shocks of 9000 psi applied at a TI of 192 min for durations of 16 or 24 min. Ultraviolet radiation of perch sperm with doses of 3240–6480 ergs/mm² resulted in 100% inactivation of paternal chromosomes, and perch eggs fertilized with inactivated sperm had survival rates of > 50%, thereby establishing methods for producing gynogenetic perch. Studies comparing the growth and performance of diploid vs. triploid perch are underway. Tetraploid perch are being reared to sexual maturity to evaluate their potential as brood fish.     

Physiological changes in rainbow trout held under crowded conditions and fed diets with different levels of vitamins E and C and highly unsaturated fatty acids (HUFA)

Rainbow trout (Oncorhynchus mykiss) maintained in crowded (100 kg m^− 3) and uncrowded (20 kg m^− 3) conditions were fed 42 days with five experimental diets having different levels of vitamin E (25.6 and 275.6 mg kg diet^− 1), C (0 and 1000 mg kg diet^− 1) and HUFA (highly unsaturated fatty acids, 12.5 and 320.5 g kg diet^− 1): −E−HUFA, −E+HUFA, +E−HUFA, +E+HUFA, −C+E+HUFA. Cortisol, plasma metabolites, tissue glycogen, fish composition, and tissue fatty-acid profile were evaluated at the end of the experimental period. In general, no changes in cortisol levels were associated with crowding, although +E+HUFA and −C+E+HUFA fish showed higher levels (mean ± SE, 55.5 ± 11.1 and 78.0 ± 11.3 ng ml^− 1) as a consequence of a possible interaction between chronic crowding and diet composition. Protein and glucose con-centration in plasma displayed no effect of crowding, but liver glycogen showed a general tendency to decrease in −E−HUFA, −E+HUFA, +E−HUFA, +E+HUFA, −C+E+HUFA crowded groups (70.2 ± 2.1, 52.1 ± 2.5, 73.4 ± 7.4, 91.7 ± 3.3, 74.2 ± 8.4 mg g^− 1 tissue, respectively) compared to uncrowded groups (108.9 ± 14.2, 82.7 ± 8.8, 92.4 ± 10.7, 99.1 ± 10.0, 103.5 ± 15.6 mg g^− 1 tissue, respectively), thus proving significant in −E+HUFA fish. Variations in total lipids, triglycerides, total cholesterol and HDL as well as LDL cholesterol in plasma were manifested under crowding conditions, displaying a certain influence of vitamin E and HUFA dietary content. Final body composition, in general, showed no change attributable to fish density, but some differences associated with diet composition were found in lipid and moisture percentages of crowded fish. Liver and muscle fatty-acid profile revealed a clear effect of the dietary lipid source that was more evident in muscle than in liver at normal fish density, and in some cases this effect was modulated by dietary vitamin E and C content and fish-culture conditions.     

Effect of dietary vitamin C supplementation on the oxygen consumption, ammonia-N excretion and Na/K ATPase of Macrobrachium nipponense exposed to ambient ammonia

The 96-h LC₅₀ of ammonia-N and the effects of dietary vitamin C on oxygen consumption, ammonia-N excretion and Na⁺/K⁺ ATPase activity of Macrobrachium nipponense exposed to ambient ammonia were investigated. The results showed that the 96-h LC₅₀ of ammonia-N was 36.6 mg l⁻¹ for the freshwater prawn, M. nipponense, at pH 8.0. When prawns were exposed to high ambient ammonia-N concentrations, the oxygen consumption rate increased and ammonia excretion decreased. Dietary vitamin C supplementation led to higher oxygen consumption and lower ammonia excretion. Na⁺/K⁺ ATPase activity increased with increased ambient ammonia-N exposure in the range of 0–18.3 mg l⁻¹, and then was reduced at ambient ammonia-N 36.6 mg l⁻¹. Na⁺/K⁺ ATPase activities of prawns fed a vitamin C-supplemented diet were significantly lower than those of prawns fed a diet which was not supplemented with vitamin C.     

Induced spermiation in 3-year-old sterlet, Acipenser ruthenus L.

Spermiation in 3‐year‐old sterlet, Acipenser ruthenus (L.), males maintained under warm water conditions was induced by intramuscular injection of either (i) (D‐Ala⁶)‐GnRH‐ProNHEt (Kobarelin); (ii) mammalian GnRH analogue+metoclopramide (Ovopel); or (iii) human chorionic gonadotropin (Biogonadyl). The volume of milt, sperm concentration and motility were measured. A higher percentage of spermiating males was obtained after Kobarelin or Ovopel treatment (81.8% and 77.7% respectively) in comparison with fish treated with Biogonadyl (40.0%). However, only stimulation with Ovopel guaranteed motile spermatozoa in all spermiating males. Moreover, treatment with Ovopel resulted in the highest average milt volume, sperm concentration and motility. The average total number of motile spermatozoa (milt volume×sperm concentration×sperm motility) per individual was 0.99×10⁹, 5.31×10⁹ and 0.02×10⁹ after stimulation with Kobarelin, Ovopel or Biogonadyl respectively. Our data indicate that Ovopel is a good stimulator of spermation in sturgeons. Moreover, the time of sexual maturation could be significantly reduced in sturgeons maintained in cages under warm water conditions.     

Dopamine modulates the physiological response of the tiger shrimp Penaeus monodon

Levels of glucose, lactate, pO₂, pCO₂, HCO₃⁻, TCO₂, Na⁺, K⁺, Cl⁻, protein, and oxyhemocyanin in the hemolymph and its osmolality and pH were measured when tiger shrimp, Penaeus monodon (13.5 ± 1.5 g body weight), were individually injected with saline or dopamine at 10^− 8, 10^− 7, or 10^− 6 mol shrimp^− 1. Results showed that hemolymph glucose, lactate, pCO₂, HCO₃⁻, and TCO₂ values increased from 2 to 4 h; hemolymph osmolality, Na⁺, and total protein had increased at 2 h; and hemolymph K⁺ decreased from 2 to 8 h after the dopamine injection. All physiological parameters returned to the control values 4–16 h after receiving dopamine. The dopamine injection also significantly decreased the oxyhemocyanin/protein ratio of P. monodon which occurred at 2 h, resulting from an elevation of hemolymph protein and a slight decrease of oxyhemocyanin. These results suggest that stress-inducing dopamine caused a transient period of modulation of energy metabolism, osmoregulation, respiration, and the acid–base balance in P. monodon in adapting to this environmental stress.