The first production of glass eel in captivity: fish reproductive physiology facilitates great progress in aquaculture

Weekly injections of salmon pituitary extracts (SPE) were administered to female Japanese eel, Anguilla japonica at a dose of 20 mg/fish. This induced vitellogenesis and caused oocytes to reach the migratory nucleus stage. Later, a majority of the females that received an injection of SPE at a priming dose, followed 24 h later by 17,20β-dihydroxy-4-pregnen-3-one (DHP), ovulated 15 to 18 h after the final injection. In cultivated males, repeated injections of human chorionic gonadotropin (hCG) at a dose of 1 IU/g BW/week induced spermatogenesis and spermiation. Since potassium ions were revealed to be an essential constituent for the maintenance of motility in the eel spermatozoa, artificial seminal plasma containing KCl was designed as a diluent of milt, and enabled the preservation of milt for several weeks in refrigeration. As a result, artificial fertilization performed immediately after ovulation with pre-diluted and stocked milt consistently resulted in the production of high-quality gametes. Recently, a slurry-type diet made from shark egg yolk has been found to be a suitable feed for captive-bred eel larvae. Although preleptocephalus larvae can be reared with this diet beyond the depletion of their yolk and oil droplet stores, it remains inadequate because larvae reared under this way cannot be raised to the following stage. Therefore, the diet was improved by supplements of krill hydrolysate, soybean peptide, vitamins and minerals. Larvae fed on this new diet have grown to 50 to 60 mm in total length (TL), and have begun to metamorphose into glass eels approximately 250 days after hatching.     

Controlled reproduction in Anguilla anguilla (L.): comparison between spontaneous spawning and stripping‐insemination approaches

This study aimed to compare the fertility of eggs between artificially matured female silver eels that spawned spontaneously and those that were spawned by manual stripping. The effects of the two methods of spawning on ovulation and fertilization rate were also investigated. For this purpose, 18 wild female European eels captured in Bonello lagoon (North Adriatic Sea) were carp pituitary extract‐injected to undergo sexual maturation and ovulation; a final injection of 17,20β‐dihydroxy‐4‐pregnen‐3‐one (DHP) was administered when at least 30% of the oocytes were fully transparent. After the DHP‐injection, nine eels were transferred to a new closed recirculating aquaculture system, where they were housed with spermiating males (sex ratio 4/1) to allow spontaneous spawning (SPT‐group); the remaining nine eels were transferred to a 250 L tank and ovulation was checked at four‐hourly intervals by manual stripping (STR‐group). The number of eggs per female in the SPT‐group was significantly greater than that in the STR‐group. Furthermore, fertilization rates in the SPT‐group were notably higher than those observed in the STR‐group. Significantly, the best performances were obtained among eels in which at least 50% of oocytes were fully transparent at the time DHP was administered. We conclude that the fertility of eggs from spontaneously spawning eels is superior to that of eggs acquired by strip‐spawning and artificial fertilization.     

Field-induced breeding of the dorado,Salminus maxillosus Valenciennes

Induced breeding of the dorado was performed using broodfish held at capture sites near the river shore. Prochilodus platensis pituitary glands (Pit) and human chorionic gonadotropin (HCG) were used as stimulators of ovarian maturation and ovulation. Positive results, with the production of viable larvae, were obtained with single injections of 1 Pit/kg and with multiple injections of 0.5, 0.25 or 0.125 Pit/kg at 2.5-h intervals. Females ovulated after receiving between 0.5 and 1 Pit/kg. During the time period covered by the experiments the percentage of ovulatory responses improved from 36% in December to 86% in February.The threshold dose that triggers final maturation and ovulation was found to be between 0.125 and 0.25 Pit/kg. Latency between injection and ovulation varied from 7 to 9 h at 23.5°C. Viability of intraovarian ovulated eggs rapidly decreased to 0% in less than 4 h at 23.5–25°C.Hatching rates ranged from 0 to 63%; the best value was obtained when eggs were stripped immediately after ovulation from a female treated with five injections of 0.125 Pit/kg at 2.5-h intervals.     

Development of an in vitro culture system for producing eel larvae from immature ovarian follicles in Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis>

To standardize conditions during the final maturation and ovulation of ovarian follicles from Japanese eel, we have developed a culture system for the production of fertilizable eggs from post-vitellogenic ovarian follicles in vitro. Post-vitellogenic ovarian follicles were incubated in culture medium supplemented with 17α,20β-dihydroxy-4-pregnen-3-one (DHP) with or without bovine serum albumin (BSA) to assess the effects of protein concentration. Eggs that ovulated during incubation were fertilized, and the remaining follicles were incubated in prostaglandin F_2α (PGF_2α) for a further 3 or 6 h before fertilization. Male eels were injected repeatedly with human chorionic gonadotropin. The quality of eggs obtained under the different culture conditions was evaluated after artificial fertilization in terms of hatching success. Hatching rates tended to decrease with increasing concentrations of BSA in the incubation medium in a dose-dependent manner. The addition of PGF_2α drastically increased the number of eggs that ovulated, but the rate of hatching was greatly decreased compared with eggs obtained earlier by DHP incubation alone. The larvae obtained from artificially fertilized eggs produced in vitro survived for 14 days without feeding. We conclude that in vitro culture systems thus have a great potential for the acquisition of good quality eggs under tightly controlled artificial conditions, culminating in the production of eel larvae.     

Hormone-induced spawning of the Australian freshwater fish Murray cod, Maccullochella peeli (Mitchell) (Percichthyidae)

Murray cod, Maccullochella peeli, originally captured from the wild, underwent normal gonadal development in earthen ponds. Handling of broodfish in the 3 months before a breeding season caused atresia and resorption of oocytes in most females. Cod were removed from the ponds when the water temperature reached 20°C during spring, and final oocyte maturation and ovulation were induced in mature females by injecting 1000 or 2000 IU/kg human chorionic gonadotrophin (HCG) or 2–5 mg/kg of a preparation of the pituitary gland from common carp (CPG). Control treatments and dosages of 100–750 IU/kg HCG did not induce ovulation. Broodfish were held at 21 ± 1°C in 2000-l tanks after injection. The time of stripping and fertilization of Murray cod eggs was an important factor determining their hatchability. There was generally high post-fertilization mortality of eggs stripped within 1 h or between 4 and 6 h of ovulation, but high hatchability of eggs stripped 2–3 h after ovulation. The mean hatchability of eggs stripped 48.5–49.5 h after the injection of 1000 IU/kg HCG was 79.8%, but there were significantly lower mean hatchabilities of eggs stripped after 46–48 h and 50–52 h, as well as after the injection of 2000 IU/kg HCG. Results using CPG were variable. Possible reasons for the high post-fertilization mortality of Murray cod eggs are discussed, and techniques for broodfish handling, injection, stripping and the fertilization and incubation of eggs are presented.     

Induced spawning in perch, Perca fluviatilis L., using FSH + LH with pimozide or metoclopramide

Induced spawning in perch, Perca fluviatilis L., was studied using follicle stimulating hormone and lutenizing hormone (FSH + LH) with the addition of pimozide or metoclopramide. The doses administered to fish were: 75 IU kg^?1 of FSH + LH with 5 or 10mgkg^?1 of pimozide (P) or metoclopramide (M) in females, and 25 IU kg^?1 of FSH + LH with 2.5 or 5 mg kg^?1 of P or M in males. Hormonal injections in males (except the injection of FSH + LH with 10 mg kg^?1 of M) did not influence the quantity of milt obtained. All females treated in this experiment had oocytes at the same stage of nucleus migration, so the time of ovulation was very synchronous, i.e. 16-24 h after injection. The relationship between the quality of the eggs, expressed as a survival to eyed-egg stage, and latency was inverse. Spawning success, expressed as a spawning effectiveness coefficient (S_e), was highest in the fish that had been treated with FSH + LH with a higher dose of M.     

Improvement of ovulation induction by additive injection of 17,20β‐Dihydroxy‐4‐pregnen‐3‐one after human chorionic gonadotropin administration in a pelagic egg spawning marine teleost,nibe croaker Nibea mitsukurii (Jordan & Snyder)

This study aimed to develop the consistent ovulation induction method in a pelagic egg spawning marine teleost, nibe croaker Nibea mitsukurii. Attempts to induce oocyte maturation and ovulation in nibe croaker using human chorionic gonadotropin (hCG; 0.5 IU g^?1) resulted in the normal progression of oocyte maturation and hydration, but a failure to induce ovulation in many individuals. This ovulation disorder was similarly observed even when the dose of hCG was increased 10 times (5 IU g^?1) or decreased to one tenth (0.05 IU g^?1), indicating that it cannot be completely overcome solely by hCG administration. However, this ovulation disorder could be completely overcome by subsequent administration of 17,20β‐dihydroxy‐4‐pregnen‐3‐one (DHP) at the appropriate dose (0.5 μg g^?1) and time (20 h after hCG administration). An increase in the number of individuals that ovulated due to DHP administration led to an increase in individuals producing larvae, resulting in an approximately threefold increase in the estimated number of larvae produced compared with the group of fish administered hCG alone. Thus, this ovulation induction method using DHP administration after hCG was demonstrated to overcome the ovulation disorder in nibe croaker and could be applicable to commercially important species with similar ovulation problems.     

A comparison of human chorionic gonadotropin and luteinizing hormone releasing hormone analogue for ovulation induction in black sea bass Centropristis striata (Linnaeus, 1758)

Mature black sea bass, Centropristis striata L. (200–800 g), were captured in coastal South Carolina during the spawning season and administered hormones for ovulation induction and strip spawning. During both study years, control groups of females were incorporated into the study design and administered sham injections containing physiological saline solution. In 2004, females received a single intramuscular injection of human chorionic gonadotropin (hCG) (330 IU kg⁻¹) (n=8) or two injections of hCG at 24‐h intervals (n=8). In 2005, females received a single injection of hCG (n=10) or an analogue of luteinizing hormone releasing hormone (LHRHa) (n=10). In 2004, all fish administered a single dose of hCG ovulated at least once. Six fish ovulated on two consecutive days and one fish ovulated on 3 days consecutively. In contrast, six of eight fish receiving two doses of hCG ovulated once, five ovulated on 2 days successively and three fish ovulated 3 days in succession. Of the fish that spawned, no differences were found in any reproductive parameters. In 2005, all fish administered hCG or LHRHa ovulated at least once. Three fish administered hCG ovulated twice, four fish ovulated on three consecutive days and one fish 4 days successively. All fish administered LHRHa spawned at least twice, six fish ovulated thrice and three fish ovulated 4 days, successively. A significant difference in fertility was found between hCG (75.6±11.4%) and LHRHa (55.6±27.4%). The results of this study indicate that both hCG and LHRHa are effective for ovulation induction in prespawning black sea bass.     

Induced Spawning of Nassau Grouper Epinephelus striatus

Nassau grouper Epinephelus striurus females ovulated 48–51 h after the first of two intramuscular injections of human chorionic gonadotropin given 24 h apart (usually 0.7 IU/gram body weight). Typical spawns contained 400,000–600,000 eggs. With fresh milt and clean water, fertilization rate was 85 and 86%. Survival from fertilization to first feeding for six spawns was 73–94%.     

Augmentation of free amino acids in eggs of red snapper,Lutjanus campechanus as part of the induced spawning protocol

An attempt was made to increase free amino acids (FAA) concentrations in eggs of red snapper Lutjanus campechanus as part of an induced spawning protocol. Mature female red snapper were given intramuscular injections of human chorionic gonadotropin (HCG) at a 1100 IU kg^?1 of female body weight. Immediately after the HCG injection, one set of females received an intramuscular injection of FAA in a buffered saline solution at a dose of 20 mg FAA cocktail kg^?1 of body weight. The FAA cocktail was 25% isoleucine, 25% leucine, 25% lysine and 25% valine on a molecular weight basis. A second set of fish did not receive the FAA supplement. Both sets of fish were equally successful in spawning with ovulation occurring 21–32.5 h post injection. Injection of FAA into brooders as eggs began final maturation and hydration resulted in an increase in leucine (nmoles mg egg^?1) and a slight increase in isoleucine (nmoles egg^?1) in recently ovulated eggs. FAA supplementation via brood injection altered FAA utilization rates by embryos and during larval development, resulted in a greater yolk sac and oil globule reserve in larvae as they approached the time of first feeding.     

Induced Ovulation of Southern Flounder Paralichthys lethostigma Using Gonadotropin Releasing Hormone Analogue Implants

Implanted pellets that provide a sustained release of [D-Ala⁶ Des-Gly¹⁰] LHRH-ethylamide (GnRHa) were used to induce maturation and ovulation of Southern flounder Paralichthys lethostigma . Of the 12 females whose ovaries contained follicles with a maximum diameter ≥500 μm, 11 ovulated for the first time within 90 h of hormone implantation. Only 1 fish with a maximum follicle diameter less than 500 μm ovulated within 2 wk after implantation. Ovulated eggs were manually stripped from the females and mixed with sperm from several males. Most females were spawned 1 to 3 times on consecutive days with variable fertility. One female was spawned 11 times producing 668,000 eggs. Fertility was evaluated by examining the incubated eggs for early stages of embryonic cleavage. The percentage of fertile eggs in subsamples of incubated eggs ranged from 7–95%. The results indicate that GnRHa implants can be used to induce repeated ovulation in this species. The variability in fertility is discussed in relation to egg quality.     

In vitro steroidogenesis by testicular fragments and ovarian follicles in a hybrid sturgeon, Bester

In this study, developmental changes in the steroidogenic capacity of testicular fragments and isolated ovarian follicles of a hybrid sturgeon, Bester, at a variety stage of developments were examined. Testicular fragments or isolated ovarian follicles were incubated in L-15 medium in the presence or absence of different concentrations of five preparations; forskolin, human chorionic gonadotropin (HCG), pregnenolone (P₅), 17-hydroxyprogesterone (17OHP) and testosterone (T) for 18 h at 15 °C. After incubation, concentrations of 11-ketotestosterone (11 KT) (testis) and, 17-estradiol (E₂) (ovarian follicles) and 17,20-dihydroxy-4-pregnen-3-one (DHP) (testis and ovarian follicles) were measured. 11KT was detected in the media following incubation with P₅, 17OHP and T. Its concentration was higher during late spermatogenesis and prespermiation and lower at the degeneration stage. Both P₅ and 17OHP were converted to DHP during the prespermiation stage. Forskolin had little stimulatory effect on the synthesis of 11KT and DHP and HCG did not induce the production of these steroids.E₂ was detected in the medium following incubation of follicles with P₅, 17OHP and T at all stages of oocyte development. The concentration of E₂ in the medium increased during vitellogenesis with the peak production occurring at the tertiary yolk stage. In contrast, the potencies of follicles to produce steroids shifted to the production of DHP during migratory nucleus stage. Forskolin and HCG had little effect on the synthesis of E₂ and DHP. These results demonstrated that the failure of spontaneous spermiation or ovulation is not due to the insufficient synthesis of DHP, but may due to the lack of availability of precursors.     

Application of controlled-release,GnRHa-delivery systems in commercial production of white bass X striped bass hybrids (sunshine bass), using captive broodstocks

Hatchery-produced white bass (Morone chrysops) and striped bass (M. saxatilis) reared to maturity in a commercial aquaculture facility, were successfully spawned using controlled-release delivery systems containing the gonadotropin-releasing hormone analog DAla⁶, Pro⁹[NEt]-GnRH (GnRHa). Two-year-old white bass females (mean weight, 0.81 kg) were implanted with different polymer-based, GnRHa delivery systems at doses ranging from 40 to 89 μg GnRHa kg⁻¹ body weight. GnRHa treatment on 20 February 1994, when females contained oocytes up to 720 μm in diameter, induced ovulation of all fish between 35 to 82 h after treatment. The white bass eggs produced were fertilized with sperm from striped bass for the production of sunshine bass. An average of 294500 eggs kg⁻¹ were produced, with a mean fertility of 81.2%, 24 h survival of 46.5%, and overall hatching success of 45%. Survival from hatch to 30 days post-hatch was 78% and the fry weighed between 0.07 and 0.1 g. Overripening of eggs began within 1 h from ovulation and maximum fertilization (60%) was observed when eggs were stripped 0.5 h after ovulation. Fertilization success decreased thereafter to 31% and 10% by 1 h and 3 h after ovulation, respectively. Control fish not treated with GnRHa did not show any signs of final oocyte maturation during the period of the study. GnRHa administration via controlled-release delivery systems appears to be a very effective method for inducing high fecundity ovulation of captive white bass broodstocks, and producing eggs of high fertility and hatching success.     

Induced ovulation using LHRHa in anadromous obscure puffer Takifugu obscurus cultured entirely in freshwater

Puffer fishes of the order Tetradontidae exist in both anadromous and non-anadromous seawater resident forms. The obscure puffer akifugu obscurus is an anadromous species whose intensive culture in freshwater has developed rapidly in China. To mass produce larvae for intensive culture, induction of ovulation of 3-year-old obscure puffer cultured entirely in freshwater was attempted by giving the fish multiple injections of LHRHa. Twenty 3-yearold cultured obscure puffers were treated with multiple injections of LHRHa at a constant dosage of 30 g of LHRHa kg-1 of body weight with 36 h interval between injections. Beginning two days after hormonal treatment, abdominal palpation was performed every day to check expansion and hardening of the abdomen of the fish due to hydration of oocytes that indicates the completion of final oocyte maturation. The first four fish ovulated after the second LHRHa injection on day 3, but most females ovulated after the fourth hormone injection. A total of 18 fish (90%) ovulated over a period of 7 days, while no fish ovulated in the control group. The mean fertilization rate and the mean hatch rate was 67.1% and 87.6%, respectively. The viability of newly hatched larvae was very good, with a high survival rate of 98.6% 24 h post-hatch. These results indicate that cultured obscure puffer, which are not allowed to undergo diadromous migration and are entirely cultured in freshwater, could be induced to complete maturation and ovulate by simple multiple injections of LHRHa and that the eggs could be successfully hatched into viable larvae.     

Induced breeding of the sabalo,Prochilodus platensis Holmberg

Maturation and ovulation were induced in female Prochilodus platensis with multiple homologous pituitary extract injections (total dose 0.9 to 1.4 hypophysis/kg) or single injections of pituitary extract combined with human chorionic gonadotrophin ((1 hypophysis + 500 I.U.)/kg). One of the females given the latter treatment produced viable eggs. Latency time from injection to ovulation was 9 h (at 28°C). Hatching occurred 12 h after fertilization. Survival after 30 days of rearing was 96%.     

LHRHa‐induced ovulation of the endangered‐Caspian brown trout (Salmo trutta caspius) and its effect on egg quality and two sex steroids: testosterone and 17α‐hydroxyprogestrone

To induce synchronized ovulation, migrating wild Caspian brown trout (Salmo trutta caspius) females were treated with two interperitoneal injections of Des‐Gly¹⁰, d ‐Ala⁶ LHRH (LHRHa), given 3 days apart. Two injections of 100 μg kg^?1 body weight of this hormone effectively induced ovulation. Within 27 days from the second injection, all fish injected with 100 μg kg^?1 LHRHa had ovulated compared with 54.5% of the controls. The mean time to ovulation was reduced significantly (P<0.05) from 31.67±4.84 days in control fish and 28.83±7.31 days in sham‐treated fish to 16.36±1.61 days in fish injected with 100 μg kg^?1 LHRHa. The fertilization rate in 50 and 100 μg kg^?1 LHRHa‐injected fish was significantly lower than that in the control fish (P<0.05). In fish injected with 50 and 100 μg kg^?1 LHRHa, significant (P<0.05) changes in testosterone (T) and 17α‐hydroxyprogestrone (OHP) levels were observed. After the second LHRHa injection, the fish injected with 100 μg kg^?1 showed the highest serum levels of testosterone and OHP. These results demonstrate that the use of LHRHa can effectively reduce the mean time to ovulation and induce synchronized ovulation in Caspian brown trout.     

Hormone Induced Spawning of Summer Flounder Paralichthys dentatus

During their first year in captivity, summer flounder Paralicthys denratus were induced to spawn with gonadotropin releasing hormone analogue (GnRHa) implants, injected carp pituitary extract (CPE) or human chorionic gonadotropin (hCG) injections. The percentage of fertile eggs was greatest (69%) in CPE-treated females. CPE, but not GnRHa or hCG, was capable of stimulating oocyte growth (increased follicle diameter during vitellogenesis) followed by ovulation. Fish with maximum ovarian follicle diameters between 180 and 435 μm at the initiation of CPE injections produced the greatest percentage of fertile eggs. For most females, fertilization rate was greatest for the first batch of eggs ovulated. The mean fertilization rate for the first spawn of CPE-treated fish was 42% compared with 14% for the second spawn from the same fish. Fish with maximum initial follicle diameters of 585 40 μm that were implanted with GnRHa ovulated the greatest number of eggs, but fertility was low and variable. Approximately 35% of females injected with hCG ovulated a limited number of eggs, but only one hCG-treated female produced fertile eggs. Only a limited number of spermiating males were available for spawning trials. Hormone treatments used on females were ineffective for inducing or maintaining spermiation in male summer flounder.     

Controlled reproduction in the wild European eel (Anguilla anguilla): two populations compared

This study aimed to describe the response variability of female silver eels in terms of gonad development and eggs production to a standardized gonadotropic treatment (Carp pituitary extracts—CPE), and to relate this variability to population characteristics. For this purpose, sexual maturation, ovulation, and fertilization were induced in two eel populations coming from different locations in Adriatic Sea (Comacchio—CM and Marano-Grado—MG lagoons), and after that, their reproductive capacity was valuated. External (Silver index—SI, Eye index—EI, Pectoral fin length index—PFLI, Condition factor—K) and hormonal (17β-estradiol—E2, testosterone—T) parameters were measured, and some subject/group were killed for histological and lipid analysis and age determination. Morphometric parameters showed the CM-Group to have highest values of Body weight (BW), Body length (BL), and K, while MG-Group presented highest PFLI and Gonadosomatic index (GSI) values. Regarding hormonal analysis, the CM-Group showed significantly higher T and E2 levels than the MG-Group, both groups showed considerably rapid increase at T5 (5th injection). A positive trend in gonadal development was found through histological evaluation; a more regular maturation was observed in the MG-Group, whereas the CM-Group presented an exponential oocytes development starting from T10 (10th week), which led to an anticipated spawning. Lipid content showed significant differences in T0 (start study), post-ovulation, and Control (30th week) between CM and MG eels. As to zootechnical performances, while MG eels released spontaneously into the water, the CMs were stripped in order to check ovulation. The MG eels were statistically the most productive with 40.1 ± 6.33 % BW of eggs released. Furthermore, CM females ovulated mainly between the 19th and 22nd week (77.8 % spawned eels) instead in the MG’s ovulation goes from the 24th to the 28th week (100 % spawned eels). As fertilization is of concern, in both groups fertilized eggs were obtained with no difference in larvae production. These results seem to indicate that bigger dimensions, higher K, and larger lipid content (Comacchio eels) could fasten gonadic maturation without positively influencing reproductive performance of animals, both in term of quantity and quality of produced eggs. Smaller females with a highest SI (Marano-Grado eels) presented a more regular gonadic development, leading the animals to spontaneous spawning.     

Reproductive parameters of the chub mackerel <Emphasis Type="Italic">Scomber japonicus</Emphasis> estimated from human chorionic gonadotropin-induced final oocyte maturation and ovulation in captivity

ABSTRACT:   Final oocyte maturation and ovulation of captive chub mackerel Scomber japonicus with fully yolk-accumulated oocytes were induced by a single injection of human chorionic gonadotropin. Reproductive parameters, including spawning frequency and batch fecundity, which are required to estimate spawning biomass in pelagic fish by the daily egg production method, were analyzed. Germinal vesicle migration (GVM) occurred at 18–24 h post-injection, and the hydration and ovulation of oocytes were completed at 30 and 36 h post-injection, respectively. The results of the maturation process suggest that fish with GVM-stage ovaries captured in the daytime from the field are capable of spawning on the night following their capture. The oocytes used in the oocyte size-frequency distribution method for batch fecundity estimates should be at late GVM and more advanced stages. The results of sequential artificial insemination showed that the quality of ovulated eggs held in the ovarian lumen rapidly deteriorated as time progressed after ovulation. This indicates that the fertilization window for the ovulated eggs of chub mackerel lasts only a few hours, and spawning behavior should be performed within a few hours after ovulation in the wild population.     

Carbon nanotube enhanced label-free immunosensor for amperometric determination of oocyte maturation-inducing hormone in fish

Maintaining high-quality fish eggs stably and efficiently is important for aquaculture. We developed a label-free immunosensor system for measuring 17,20β-dihydroxy-4-pregnen-3-one (DHP). DHP is suddenly secreted before ovulation as a maturation-inducing hormone in fish, and therefore, DHP levels are an indicator for predicting ovulation. The method is based on immunologic reactions and amperometric measurement using cyclic voltammetry (CV). For biomolecular immobilization on the surface of sensing electrode, Au electrode, we used self-assembled monolayers of thiol-containing compounds to fix anti-DHP immunoglobulin. In addition, we used a single-walled carbon nanotube to improve sensitivity. Using this electrode, we were able to determine the CV signal change caused by the antigen–antibody complex. The proposed immunosensor system showed a linear correlation (correlation coefficient: 0.9827) between the anodic peak current of the CV and the DHP level in range from 15.6 to 50,000 pg ml^?1. The sensor system was then applied to monitor DHP of goldfish (Carassius auratus). Blood plasma of fish was collected every 3 h after administering a DHP inducer. In the measurement, the anodic peak current of the CV showed distinct changes depending on DHP levels in the blood plasma. A good relationship was observed between DHP levels determined by our proposed system and the conventional method (correlation coefficient: 0.9351).