Population genetic structure and gene flow in the Japanese sea cucumber Apostichopus japonicus across Toyama Bay, Japan

The Japanese sea cucumber Apostichopus japonicus has a pelagic-lecithotrophic larva. Here, we clarify larval dispersal among Japanese sea cucumber populations by describing the levels of distinctiveness and gene flow among red and green variant populations of A. japonicus across Toyama Bay using 11 polymorphic microsatellite markers. Samples of Japanese sea cucumber populations were collected from three localities (Noto, Shinminato, and Uozu) around Toyama Bay. Geographically separated populations of red and green variants were determined to be genetically distinct, supported by phylogenetic analysis and the distinctiveness range values of pairwise multilocus estimates of F _ST. Contemporary migration analyses indicated that the majority of estimated migration events occurred within A. japonicus populations comprised of only either red or green variants. In a historical gene flow analysis, two best-fit models (n-island and stepping stone models) showed circulation for the gene migration in Toyama Bay among red variant populations. Our results provide useful information on the genetic structure of Japanese sea cucumber populations and will be helpful for genetic conservation and fisheries management of sea cucumber populations in Toyama Bay.     

Effect of salinity on growth and energy budget of red and green colour variant sea cucumber Apostichopus japonicus (Selenca)

Sea cucumber, Apostichopus japonicus (Selenca), tolerates salinity fluctuations inhabiting intertide zone. This study deals with growth, food intake, food conversion and the bioenergetic responses of the red variant (wet weight of 2.60 ± 0.11g) and green variant (wet weight of 2.56 ± 0.08 g) A. japonicus to different salinities of 22, 26, 30, 34, and 38 psu at 16.5 ± 0.5°C. The results showed that salinity had a significant effect on specific growth rate (SGR) of both green and red variants A. japonicus (P < 0.05). Both colour variants of sea cucumber had highest SGR at 30 psu, and then decreased when salinity below or above this point. Maximum SGR (the green 1.07 ± 0.08% day^?1, the red 1.14 ± 0.09% day^?1 respectively) is related with maximum food intake (FI) and highest food conversion efficiency (FCE) (P < 0.05) occurring at 30 psu. Only under 22 psu, the green variant grew faster than the red variant (P < 0.05), and under other four salinity treatments there was no significant difference between SGR of two colour variant holothruians (P > 0.05). Values of adaptable salinity scope for green and red variants sea cucumber survival are 18.5~39 psu and 20.9~38.6 psu respectively. The average energy budget formula of sea cucumber at 30 psu was: 100C = 6G +42F +3U+49R (C, energy ingested; G, energy for growth; F, energy loss as faeces; U, energy used for ammonia excretion; R, energy loss for respiration). The sea cucumber had maximum energy ingested (C) and highest proportion of energy for growth (G) at 30 psu, and then decreased when salinity is above or below this salinity. Both red and green variants of A. japonicus deposited for growth were very low, and the energy loss in faeces and energy for respiration accounted for the majority of assimilation energy. The result clearly showed that the optimum condition for farming green and red variants A. japonicus, both with respect growth and energy allocation, is the salinity scope of 26 ~ 30 psu.     

Completion of the Pacific bluefin tuna Thunnus orientalis (Temminck et Schlegel) life cycle

Tuna aquaculture is currently dependent on the wild capture of juveniles for production. The development of hatchery technology for bluefin and other tunas would be a major step forward in improving sustainability of their aquaculture. The present study overviews the technology in the life cycle completion of the Pacific bluefin tuna (PBT) Thunnus orientalis (Temminck et Schlegel) under aquaculture conditions in Kinki University, and the problems to be solved for the establishment of tuna hatchery technology. On 23 June 2002, broodstock of PBT that were artificially hatched and reared spontaneously spawned in captivity. The resulting eggs hatched and were subsequently reared to the juvenile stage. The spawning fish were the result of a research project started in 1987 to rear wild‐caught juvenile PBT that were several months old. Fertilized eggs were obtained from these fish in 1995 and 1996. Resulting juveniles (the artificially hatched first generation) were reared to maturity and spawned in 2002. Over the summer of 2002, 1.63 million eggs from these fish were used for a mass rearing experiment, and 17 307 juveniles were produced and transferred to an open sea net cage. Of these artificially hatched second‐generation PBT, 1100 grew to approximately 95 cm total length and 14 kg body weight in 22 months. This procedure means the completion of PBT life cycle under aquaculture conditions, which was first attained among large tuna species. The problems awaiting solution in PBT hatchery production are their unpredictable spawning in captivity, to improve survival during the first 10 days post hatch, to reduce cannibalism in larval and juvenile stages, and to solve collision problem causing high mortality during the juvenile stage.     

Use of Nannochloropsis sp. isolated from the East China Sea in larval rearing of sea cucumber (Apostichopus japonicus)

The supply of microalgae to hatcheries is a limiting factor for the mass larval production of sea cucumber (Apostichopus japonicus) in Fujian Province, China. In this study, Nannochloropsis sp. isolated from the East China Sea was tested as food for A. japonicus larvae. The first trial compared the effect of mono‐, bi‐ and trialgal diets comprising three microalgae (Chaetoceros muelleri, Dunaliella tertiolecta and Nannochloropsis sp.) on A. japonicus larval growth, survival, settlement and juvenile growth. The results showed that there were no significant differences in survival and settlement between larvae fed with Nannochloropsis sp. and other diets. All diet treatments yielded similar juvenile sea cucumber output. In the second trial, A. japonicus larvae were fed equally four times daily at three different rations (5000, 20 000 and 40 000 cells mL^?1 day^?1). Larvae fed 20 000 cells mL^?1 day^?1 were significantly larger than larvae in other groups and experienced the highest survival rate. In the third trial, A. japonicus larvae were fed 20 000 cells mL^?1 day^?1 in three different frequency (2, 3 and 4 meals day^?1). The greatest body length was observed in larvae that received 3 meals day^?1. Survival and settlement of larvae fed 2 meals day^?1 were significantly lower than other two groups. These results suggest that Nannochloropsis sp. can be used as a diet for the large‐scale production of A. japonicus seed, and larvae fed three times daily at a ration of 20 000 cell mL^?1 day^?1 are recommended for hatchery production of A. japonicus.     

Reproductive performance in induced and spontaneous spawning of the mangrove red snapper, Lutjanus argentimaculatus: a potential candidate species for sustainable aquaculture

A reliable breeding technique was developed for the mangrove red snapper, Lutjanus argentimaculatus (Forsskal 1775), to help sustain the aquaculture of this immensely popular species in Southeast Asia. Using standardized indices of female maturity (based on mean oocyte diameter of ≥0.40 mm), time of injection (1000–1130) and sex ratio (one female to two males), a single injection of 100 μg kg^?1 luteinizing hormone‐releasing hormone analogue (LHRHa) (n=16 fish), but not 50 μg kg^?1 (n=five fish), successfully induced egg (62.5% success rate) and larval (43.8%) production. Human chorionic gonadotropin (hCG) at 500 IU kg^?1 (n=five fish) also failed to induce spawning, but doses of 1000 (n=22 fish) and 1500 IU kg^?1 (n=15 fish) gave spawning (77.3% and 80.0% respectively) and hatching success rates (72.7% and 60.0% respectively) that were not significantly different from those of 100 μg kg^?1 LHRHa. No spawning was observed in saline‐injected controls (n=seven fish). While mean spawning latency, egg diameter, egg production per spawn, percent egg viability, hatching rate, percent of normal larvae and cumulative survival of eggs to normal larvae did not differ significantly among the effective hormones and doses, 1000 IU kg^?1 hCG had a higher percentage (76.5%) of total spawns with egg production per spawn in excess of one million than those of 1500 IU kg^?1 hCG (50.0%) and 100 μg kg^?1 LHRHa (40.0%). Mangrove red snapper spontaneously spawned from March–April to November–December with a peak of egg collection and spawning in May–June. Egg collection per spawn ranged from 0.05 to 6.35 million. Spontaneous spawning of mangrove red snapper exhibited lunar periodicity with spawns mostly occurring 3 days before or after the last quarter and new moon phases and occurred consistently between 02:00 and 04:00 hours. High fecundity and good egg quality, coupled with the ability to respond to induce spawning or natural spawning in captivity, provide a sound basis for improving the sustainability of red snapper aquaculture in Southeast Asia.     

Induction of Spawning in the Sea Cucumber Holothuria scabra (Echinodermata: Holothuroidea)

With the advent of sea cucumber aquaculture in the South Pacific region, a reliable method is needed to induce large numbers of animals to spawn in captivity. Broodstock of the sea cucumber Holothuria scabra , collected from Stradbroke Island, Moreton Bay (27°90'N, 153°24'E) Australia, during the reproductive season from October to January, were used in spawning trials. During the 1997–1998 summer between one to five weeks of captivity, 100% of animals were induced to spawn in four trials at dusk on or close to a new or full moon, using nine males and nine females contained in a Reln tank and 30 cm of filtered sea water, using a 3–5 C temperature shock. H. scabra was induced to spawn in small numbers during the 1996–1997 summer despite a marked degree of weight loss, and all animals spawned during 1997–1998 with minimal loss of weight. The difference in the number of spawned eggs between animals of similar size and mean numbers of spawned eggs in consecutive trials decreased the longer animals were held in captivity before spawning. The hatch rate of eggs was reduced significantly for broodstock held for more than one month. Hatch rate and numbers of spawned eggs are important indicators of egg viability of broodstock maintained in captivity for an extended period.     

Different responses between orange variant and cultured population of the Pacific oyster Crassostrea gigas at early life stage to temperature‐salinity combinations

Although breeding of rare shell colour variants has drawn widespread attention from shellfish breeders, the potential disadvantages of their adaptive capacity have been ignored in practice. To explore the difference in adaptive capacity between orange shell variant (OSO) and commercially cultured population (CPO) of the Pacific oyster Crassostrea gigas at early life stage, the development to D‐larvae and larval survival and growth (just 23 and 30°C for larval experiment) of them were compared under different temperature (16, 23 and 30°C) and salinity (17, 25 and 33 psu) combinations. In this study, at 23°C and 25 psu, for both OSO and CPO there was no difference in fertilization rates and survival (p > .05) (mean percentages of D‐larvae after fertilized 40 hr ≥ 95.00%; mean larval survival rates on day 10 > 80.00%). However, the percentage of D‐larvae of CPO at 40 hr was significantly (p < .05) higher than OSO at temperatures of 16 and 30°C and 25–33 psu and 17 psu at 23°C. Similarly, CPO has a better larval survival on day 10 and growth than OSO at salinities of 17 and 33 psu at 23°C. Overall, our results indicate that OSO can have an equally good performance like CPO at early life stage under optimal condition (23°C; 25 psu), but the potential disadvantages in adaptive capacity will be shown at suboptimal conditions. These findings can guide future hatchery breeding of OSO, and suggest the potential disadvantages in adaptive capacity in rare colour variants need more attention in further breeding.     

Suspended culture of the sea cucumber Apostichopus japonicus below a Pacific oyster raft – potential for integrated multi‐trophic aquaculture

To discuss the possibility of co‐culturing Pacific oyster with the sea cucumber Apostichopus japonicus, a field experiment was conducted in an oyster farm. Apostichopus japonicus juveniles (mean wet weight, 0.08 g) were cultured below an oyster raft and at a control station for 216 days, and the wet weight and stable isotope ratios (δ¹³C, δ¹⁵N) were analysed together with settling organic matter (OM) collected using sediment traps. All sea cucumbers cultured below the raft survived (survival rate, 100%), while at the control station one individual disappeared (96%). During 216 days, the juveniles at the oyster and control stations grew to a mean weight of 5.5 and 2.6 g, attaining respective specific growth rates of 2.0% and 1.6% (paired t‐test, P < 0.001). Settlement rates of carbon and nitrogen at the oyster station were ~5 times larger than those at the control station. The stable isotope analysis showed that settling OM at both stations originated from coastal phytoplankton and that phytoplankton represented the primary food source for A. japonicus. The rapid growth of A. japonicus at the oyster station was concluded to be due to the abundant supply of oyster biodeposits, which could be ingested by this species.     

Effects of stocking density,temperature, and salinity on larval survival and growth of the red race of the sea cucumber <Emphasis Type="Italic">Apostichopus japonicus</Emphasis> (Selenka)

The red race of the sea cucumber Apostichopus japonicus was introduced into China from Japan for large-scale seed production because of its economic value. This paper reports the effects of stocking density, temperature, and salinity on survival and growth of early larvae before and after feeding, in order to establish conditions for optimal larval growth and production. To maximize the yield per unit of space, densities of 0.5–1 larvae/ml are recommended for non-feeding larvae, while 0.1–0.2 larvae/ml are best for feeding larvae. Higher survival and growth values were obtained for both non-feeding and feeding larvae at temperature ranges from 21 to 24°C. Larvae reared at a salinity of 30‰ always showed maximum growth and survival. Based on results of this study, a temperature range from 21 to 24°C and a salinity of 30 are considered optimal for early development of the red A. japonicus.     

Comparison of iodine and glutaraldehyde as surface disinfectants for red porgy (Pagrus pagrus) and white sea bream (Diplodus sargus sargus) eggs

The efficacy of iodine and glutaraldehyde as fish egg surface disinfectants were assessed in red porgy (Pagrus pagrus) and white sea bream (Diplodus sargus sargus) eggs, two species of interest for Mediterranean aquaculture. Iodine was effective in reducing the bacterial load of the 1-day-old eggs when applied at 50 mg L⁻¹ for 5 min. The same concentration did not cause any significant change in hatching success or survival of the larvae for the first 5 days. Glutaraldehyde failed to reduce the bacterial load of the fish eggs at concentrations that were safe for the eggs (100 mg L⁻¹ for 5 min), as it had a significant effect in preventing hatching of the developed embryo. Disinfecting 0-day-old eggs with iodine resulted in a significant reduction of hatching percentage, while larval survival thereafter was unaffected. The results of the present study suggest that iodine may be an appropriate egg disinfectant for both red porgy and white sea bream.     

Fertilization and Development of Eggs of the Sea Urchin Lytechinus variegatus Maintained on an Extruded Feed

Sea urchin eggs are used extensively as models for studies in developmental and molecular biology. Developing aquaculture techniques and facilities for sea urchins would facilitate their use for this purpose and for production of young sea urchins for stock enhancement. A basic requirement for the aqua-culture of sea urchins is the availability of a feed that predictably produces eggs of a consistent, high quality. We tested an extruded feed with Lyrechinus variegatus for this purpose. Lytechinus variegatus (mean horizontal diameter 55 mm, 75 g wet weight) were collected in April 1998. Nine individuals were maintained in each of four aquaria with closed, recirculating filtered sea water. They were fed approximately 1 g extruded feedindividual per week. All feed was consumed. The mean weight of the sea urchins changed little with this feeding regime. The sea urchins spawned spontaneously on a monthly basis during water changes. Four females in February and one in March were spawned by injection with potassium chloride. Fertilization was 99–100% successful in each case. Egg diameters from these spawnings and a spontaneous spawning in April ranged from 102 × 6 to 128 × 1 μm. Gastrulation occurred in <20 h and metamorphosis after 22 to 37 d. Newly metamorphosed juveniles ranged in size from 435 × 38 to 473 × 56 μm. Egg size, larval size, the chronology of embryonic and larval development, and size of newly metamorphosed juveniles are similar to those obtained from field individuals reported in the literature. These results indicate that a predictable production of high quality eggs by sea urchins under controlled conditions in the laboratory with a prepared feed is feasible.     

Winter monsoon promotes the transport of Japanese temperate bass Lateolabrax japonicus eggs and larvae toward the innermost part of Tango Bay,the Sea of Japan

Northwesterly cold winds characteristic of the East Asian Winter Monsoon (EAWM) dictate winter climatic conditions over the Japanese Archipelago. Japanese temperate bass Lateolabrax japonicus is a commercially important coastal fish that spawns offshore in winter and uses shallow waters as nursery habitats. To investigate the effects of EAWM on the planktonic period of L. japonicus, eggs, larvae, and juveniles were quantitatively collected in Tango Bay on the Sea of Japan side in winter and spring from 2007 to 2017. Although eggs occurred close to the mouth of the bay, planktonic larvae occurred further inside as they developed. The horizontal distribution of planktonic larvae, combined with water velocity data obtained from mooring observations, indicated that planktonic larvae are transported south‐ to westward through Ekman current and an anticyclonic circulation, which are driven by northwesterly winds. To evaluate survival during the planktonic period in each year class, the abundance of benthic larvae/juveniles was divided by winter total landings of Lateolabrax spp. (proxy of the spawning stock size). This survival index exhibited a positive correlation with the northwesterly component of winter winds, and a negative correlation with winter air temperature (average from December to February, Spearman's correlation, p < .05). There was, however, no significant correlation with winter water temperature or winter freshwater discharge in the bay. We conclude that northwesterly cold winds of EAWM play a critical role in transporting L. japonicus eggs and larvae toward nursery habitats, specifically beaches and estuaries fringing the innermost part of Tango Bay.     

Year‐round natural spawning,early development,and the effects of temperature,salinity and prey density on captive ornate goby Istigobius ornatus (Rüppell, 1830) larval survival

Natural spawning, early development and larviculture of the ornate goby Istigobius ornatus in captivity were studied for the first time. I. ornatus spawned 46 times from 31 October 2013 to 31 October 2014. Fecundity ranged from 246 to 10,214 eggs per clutch, with an average hatching rate of 77.8% ± 9.9% (M ± SEM). Fertilized eggs (1.31–1.54 × 0.46–0.50 mm in diameter) were adhesive demersal and oval‐shaped. Embryonic development lasted 84 hr at 27.5 ± 0.5°C. Newly hatched larvae [2.12 ± 0.04 mm in total length (TL)] transformed to the juvenile stage completely when TL was 7.79 mm. Effects of different water temperatures (24, 28 and 32°C) and salinities (10, 15, 20, 25, 30, 35 and 40 g/L) on per cent survivals (%) and survival activity indices (SAIs) were tested. Survival was not significantly different under different temperatures; SAIs was significantly higher at 28°C. Larvae showed the significantly higher survival and SAIs at salinities 10–30 g/L than at 35 and 40 g/L. Effect of different prey densities on survival was significantly higher in 7 days post hatch larvae fed 20 and 30 rotifers/ml. These findings could guide future programs in captive breeding technology development and commercial production of other marine ornamental gobies.     

Quality evaluation of indoor‐ and outdoor‐cultured sea cucumber (Apostichopus japonicus) seedlings: Insight from survival and immune performance in response to combined stress of hyperthermia and hyposalinity

The mass mortalities of sea cucumber Apostichpous japonicus have prevailed in northern China, mainly attributing to the emergence of extreme environmental conditions, that is hyperthermia and hyposalinity. The high‐quality sea cucumber seedlings appear to possess more robust resistance to adverse conditions. There are usually indoor‐ and outdoor‐cultured seedlings in industrial production of sea cucumbers. Although the outdoor‐cultured sea cucumbers are practically considered to be more strong and robust, the effective evaluation approach to distinguish these seedlings has been scarce. The current study compared survival and immune performances of indoor‐ and outdoor‐cultured A. japonicus under combined exogenous stressors, that is hyperthermia and hyposalinity. Results based on secondary stress induction revealed that the activities of immune enzymes and levels of catecholamines in body wall of outdoor‐cultured seedlings were prominently higher than those of indoor‐cultured seedlings recovered for 0–72 hr following sublethal stress (30°C and 25 psu of salinity). The opposite case occurred on immune enzymes in coelomic fluid of the two sources of seedlings except for myeloperoxidase. Importantly, the outdoor‐cultured seedlings, which were recovered for 72 hr after sublethal stress, exhibited a 93% of cumulative survival rate following 7 days of recovery after lethal stress (33°C and 20 psu of salinity), 27% higher than the indoor‐cultured seedlings. Collectively, the outdoor‐cultured A. japonicus seedlings showed more superior quality than the hatchery‐produced seedlings in terms of survival and immune performance. These findings provide practically useful information towards quality distinction of the indoor‐ and outdoor‐cultured sea cucumbers, which could benefit the aquaculture industry to obtain high‐quality seedlings.     

Reproduction,growth and stress response in adult red sea bream,Pagrus major (Temminck & Schlegel) exposed to different photoperiods at spawning season

Adult red sea bream, Pagrus major (body weight, 1.0–2.0 kg) was exposed to three photoperiods [12 h light:12 h dark (12L:12D), 16 h light:8 h dark (16L:8D) and 24 h light:0 h dark (24L:0D)] from 2 months before spawning till the end of the spawning season to investigate growth, spawning and stress response. During the spawning season, tanks were checked every morning for spawned eggs. The growth performance in fish under 24L:0D was stimulated with significantly higher feed intake than those under other photoperiods (P<0.05). The number of eggs and gonadal histology confirmed that three and five females out of six in each of duplicate tanks of the 16L:8D treatment spawned. In contrast, only two out of six females in one tank of the 24L:0D treatment spawned, and no spawns were observed in the 12L:12D treatment. At the end of the spawning period, both 17β‐estradiol and testosterone levels were significantly higher in fish exposed to 16L:8D followed by 12L:12D and 24L:0D photoperiods (P<0.05). Photoperiod manipulation did not cause significant stress response in fish (P>0.05). The results suggest that stimulating the growth performance of red sea bream at reproductive stage with a 24L:0D photoperiod is possible if the fish are subjected to this photoperiod long before the onset of the spawning season.     

Effects of light intensity on larval development and juvenile growth of sea cucumber Apostichopus japonicus

The growth rate, survival rate, development and setting rate of larval sea cucumber Apostichopus japonicus were measured under four light intensities (0, 50, 500 and 2,000 lx), and the growth rate and metabolism of the juvenile sea cucumbers were investigated under four light intensity treatments (0, 100, 1,000 and 2,500 lx). The light requirements (i.e. intensity) of the sea cucumber changed as they grew. Better growth performance, development and survival rate were observed in embryos and larvae under 500 lx treatment. However, reduced light intensity (50 lx) increased the settlement rate of the larvae. The highest specific growth rate occurred in juvenile sea cucumbers under 1,000 lx. The sea cucumbers in this group also had the lowest oxygen consumption rate and ammonia excretion rate, but the highest O:N ratio, indicating that the optimal light intensity for the juvenile sea cucumber culture was 1,000 lx. Therefore, appropriate light intensities were suggested to provide larval and juvenile sea cucumbers with better growth and development conditions.     

Breeding and larval development of Holothuria mammata,a new target species for aquaculture

Sea cucumber commercial fisheries in the North‐eastern Atlantic and Mediterranean Sea started in Turkey in the 1990s. Due to ineffective management practices, the sea cucumber resource was overexploited. Holothuria mammata is one of the sea cucumber species exploited from this geographical area, showing high potential for its aquaculture. This research was undertaken to develop the breeding, larval and juveniles rearing techniques for development of H. mammata aquaculture. Broodstocks collected from Ria Formosa (Faro, S Portugal) were successfully induced to spawn by thermal stimulation from July to October, yielding up to 6.95 million eggs/female. The eggs and larvae were reared and their development described. H. mammata showed the five typical larval stages of most aspidochirote holoturians. Juveniles stage was reached after 21 days post‐fertilization. This is the first work focused on the aquaculture biotechnology of H. mammata, however, further research is need to improve the survival of juveniles to ensure the future production of this species.     

Mortality event involving larvae of the carpet shell clam Ruditapes decussatus in a hatchery: isolation of the pathogen Vibrio tubiashii subsp. europaeus

Diseases caused by bacteria belonging to the genus Vibrio are a common, as yet unresolved, cause of mortality in shellfish hatcheries. In this study, we report the results of routine microbiological monitoring of larval cultures of the carpet shell clam Ruditapes decussatus in a hatchery in Galicia (NW Spain). Previous episodes of mortality with signs similar to those of vibriosis affecting other species in the installation indicated the possibility of bacterial infection and led to division of the culture at the early D‐veliger larval stage. One batch was cultured under routine conditions, and the other was experimentally treated with antibiotic (chloramphenicol). Differences in larval survival were assessed, and culturable bacterial population in clams and sea water was evaluated, with particular attention given to vibrios. Severe mortalities were recorded from the first stages of culture onwards. The pathogen Vibrio tubiashii subsp. europaeus was detected in both batches, mainly associated with larvae. Moreover, initial detection of the pathogen in the eggs suggested the vertical transmission from broodstock as a possible source. Experimental use of antibiotic reduced the presence and diversity of vibrios in sea water, but proved inefficient in controlling vibrios associated with larvae from early stages and it did not stop mortalities.     

A preliminary identification of genomic loci for body colour variation in the sea cucumber Apostichopus japonicus

The sea cucumber Apostichopus japonicus is one of the most economically important aquaculture species in China, which shows a distinct characteristic of body colour variation. A number of purple variants were obtained in the coast of China, which shows great potential to develop a niche market. In the present study, to understand the genetic basis of green‐purple colour variation, a genome‐wide association study based on the 2b‐RAD sequencing was conducted. As a result, 8,795 genome‐wide single nucleotide polymorphism (SNP) markers were obtained and then applied to the association analysis. Ten most associated SNP loci with p‐value <1.0 × 10^–8 were obtained, and they were assigned to 4 chromosomes (chr17, chr21, chr15 and chr05). In genomic regions neighbouring to those SNPs, 33 candidate genes with annotation were obtained and the endothelin‐converting enzyme‐1, which was reported to be involved in pigmentation variation, was identified. To our knowledge, this is the first report of the genetic locus for body colour variation in this species. These findings will be useful for the fine mapping of the determining gene for body colour variation of A. japonicus.