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.     

应用高通量测序技术分析大菱鲆幼鱼肠道及其养殖环境的微生物群落结构

采用基于Illumina测序平台的高通量测序技术,对大菱鲆(Scophthalmus maximus)幼鱼肠道及其养殖水体、生物饵料中细菌种类及丰度进行研究。测序结果显示,养殖水体、生物饵料和大菱鲆幼鱼肠道等19个样品共获得有效序列547621条,可聚类于3771个可分类操作单元(OTUs),归属于养殖水体、生物饵料、健康幼鱼和发病幼鱼的操作分类单元(OTU)个数分别为3038、1090、87和777,其中,健康幼鱼与生物饵料、健康幼鱼与养殖水体特有的OTU个数分别为57和0,发病幼鱼与生物饵料、发病幼鱼与养殖水体特有的OTU个数分别为481和31。表明幼鱼肠道微生物多样性与生物饵料密切相关。根据细菌注释结果,拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)和变形菌门(Proteobacteria)在大菱鲆幼鱼肠道中占优势地位,其中,健康幼鱼肠道微生物共聚类为8个门,发病幼鱼的肠道微生物可聚类为19个门。与健康幼鱼相比,发病幼鱼肠道门水平上的3种主要优势菌群落结构出现失衡。此外,对各样品中丰度最高的100位OTU分析显示,幼鱼肠道优势菌种类与生物饵料中的优势菌种类密切相关,而每个发病幼鱼肠道优势菌种类具有一定的独立性。本研究旨在为大菱鲆健康养殖和微生态调控提供实验依据。     

Susceptibility of cultured juveniles of several marine fish to the sevenband grouper nervous necrosis virus

Piscine nodaviruses (betanodaviruses) have been tentatively divided into four genotypes (SJNNV, RGNNV, TPNNV and BFNNV) and it is suggested that host specificity is different among these genotypes. In the present study, a betanodavirus [sevenband grouper nervous necrosis virus (SGNNV)] belonging to the redspotted grouper nervous necrosis virus (RGNNV) genotype, to which most betanodaviruses from warm water fish are identified, was evaluated for its pathogenicity to hatchery-reared juveniles of several marine fish species. When challenged with the virus by a bath method (10(5.1) TCID50 mL(-1)), sevenband grouper, Epinephelus septemfasciatus, Japanese flounder, Paralichthys olivaceus, and tiger puffer, Takifugu rubripes, displayed behavioural abnormalities and mortalities with distinct histopathological signs of viral nervous necrosis and heavily immunostained cells were observed in the central nervous tissues and retina. Bath-challenged rock fish, Sebastiscus marmoratus, and a hybrid of sevenband grouper and kelp grouper, E. moara, did not display any behavioural abnormality or mortality during the experimental period, although many fish showed slight signs of viral infection in nerve cells. Kelp grouper and red sea bream, Pagrus major, showed no behavioural abnormality, mortality or immunohistopathological changes after the virus challenge. These results are, in part, consistent with the natural host range of RGNNV, indicating the complexity in the host specificity of betanodaviruses.     

Vibrio harveyi and other bacterial pathogens in cultured summer flounder, Paralichthys dentatus

A monitoring program for Vibrio harveyi and other potential bacterial pathogens of summer flounder was conducted at two facilities in the Northeast United States. Bacterial samples were collected from larval and juvenile fish and live-feed, and identified using API 20E biochemical profiles and 16S rDNA sequencing. Histopathological examinations were conducted in order to relate histological changes with the presence of potential bacterial pathogens. V. harveyi, Vibrio ichthyoenteri, and Photobacterium damselae subsp. damselae, three known pathogens of flatfish, were isolated from diseased summer flounder. Although a high proportion of juvenile summer flounder showed microscopic signs of disease, the presence of these potential bacterial pathogens in fish was not associated with large-scale mortalities. An outbreak of flounder infectious necrotizing enteritis with 30% cumulative mortality occurred when juveniles were transported to a new facility. Isolates of V. harveyi from the disease outbreak were genetically different from the isolates from the commercial hatchery. V. harveyi isolates from both facilities were pathogenic to summer flounder by intraperitoneal injection. P. damselae subsp. damselae, V. ichthyoenteri, and Vibrio scophthalmi, also found in fish with gross lesions, were not pathogenic to juvenile summer flounder by intraperitoneal injection. Our research shows that several potential bacterial pathogens are associated with morbidity and mortality in summer flounder larvae and juveniles, especially in situations of stress. Increased knowledge about the environmental conditions that lead to disease, as well as the interactions of V. harveyi with other microbial species, could lead to the development of management strategies in summer flounder farms.     

Influence of Hatchery Protocols on Mitochondrial DNA Variation in Japanese Flounder Juveniles

The Japanese flounder Paralichthys olivaceus is a commercially important fish that is stocked extensively from hatchery rearing programs in Japan. To examine the genetic variability of hatchery-raised juveniles of the Japanese flounder that are used for stocking into natural waters, we analyzed a portion of the mitochondrial genome. The mtDNA region extending from the 3'half of the cytochrome b gene to the central domain of the control region was PCR amplified and analyzed using 11 restriction endonucleases. We identified 34 polymorphic cleavage sites out of a total of 61 sites, which resulted in 67 different haplotypes in a total of 265 offspring, examined from eight hatchery stocks. Haplotype diversity of offspring at each of the eight hatcheries ranged from 0.49 ± 0.09 (SE) to 0.94 ± 0.03 (SE). Also, we observed 40 polymorphic sites out of a total of 59 sites, which resulted in a total of 50 haplotypes in 60 wild flounder. Haplotype diversity of the wild population was 0.98 ± 0.01 (SE). The use of subcultured fishes as broodstock appears to be one of the most important causes of reduced genetic diversity in hatchery-raised flounder juvenile. Our results suggest that the use of wild fish for broodstock is an effective way to maintain genetic variability in Japanese flounder offspring.     

Molecular tracing confirms that infection with infectious pancreatic necrosis virus follows the smolt from hatchery to grow‐out farm

Infectious pancreatic necrosis (IPN) is an important restraint to production of salmonids in aquaculture globally. In order to implement efficacious mitigation strategies for control of this disease, it is important to understand infection routes under current production systems. IPN virus has been shown to be transmitted vertically in Rainbow trout, from broodstock to fingerlings in hatcheries, and there is circumstantial evidence suggesting that vertical transmission can also occur in Atlantic salmon, in addition to horizontal transmission between grow‐out fish in farms. In this study, we show that the smolt carries infection with IPN from hatchery to the marine farm. We do this by comparing sequences from fish groups taken both in hatcheries and on corresponding marine grow‐out farms. We use statistical analysis to prove that sequences obtained from the same fish group in both hatchery and marine farm are more similar than sequences obtained from random fish groups on hatcheries and marine farms.     

Feeding habits of skipjack tuna Katsuwonus pelamis and other tuna Thunnus spp. juveniles in the tropical western Pacific

ABSTRACT: The feeding habits of skipjack tuna Katsuwonus pelamis juveniles (8.5–66.8 mm standard length) were examined, collected from the tropical western Pacific in October to December 1994, and their habits were compared with those of the other tuna Thunnus spp. juveniles (9.8–55.3 mm standard length). The indices, frequency of occurrence of each food item in the total number of stomachs examined (% F ), percentage of number of each food item to the total number of all food items identified (% N ), percentage of wet weight of each food item to the total wet weight of all food items identified (% W ), and relative importance of each food item ( IRI ) were estimated in the north equatorial current (NEC) and the north equatorial countercurrent (NECC) areas. The most dominant prey item of skipjack juveniles in the two areas was fish larvae. Other major prey items in the NEC area were Euphausiacea, Amphipoda, and Copepoda; whereas those in the NECC area were Copepoda, Cephalopoda, Euphausiacea, and Amphipoda. In the other tuna juveniles, the IRI of fish larvae in the two areas was remarkably high. Other prey, Euphausiacea and Cephalopoda in the NEC area only and Cephalopoda in the NECC area, were also found. These results indicate that the skipjack juvenile is primarily a piscivorous feeder although they also depend on various other prey organisms, whereas the other tuna juveniles are stronger piscivorous feeders.     

Comparative performance of hatchery-reared and wild Scylla paramamosain (Estampador, 1949) in pond culture

The performance of hatchery‐reared juveniles either in aquaculture grow‐out systems or stock enhancement is likely to be dependent on a range of factors during the hatchery phase of production. With recent progress in the development of hatchery systems for the mud crabs Scylla spp., there is growing interest in evaluation of the quality of hatchery‐reared juveniles relative to wild seed crabs as currently used in aquaculture. Hatchery‐reared and wild‐collected Scylla paramamosain juveniles were stocked either together in ponds or separately. All crabs were tagged with microwire‐coded tags, so that origin could be determined in the mixed groups. Preliminary validation demonstrated that tagging did not affect survival or growth, with a tag retention of 94%. After 106 days of culture, there was no significant difference in survival at harvest between the two sources of crabs. The wild juvenile crabs had a significantly higher initial weight:carapace width (CW) ratio compared with those from the hatchery, indicating a difference in condition. However, where crabs were stocked separately, the hatchery‐reared animals exhibited significantly faster growth than those collected from the wild, both in terms of specific growth rate and CW increase per month. However, in the mixed ponds, where there was competition with wild crabs, there was no significant difference in growth rate between crabs from the two sources. Overall, the results demonstrate that the growth performance of hatchery‐reared S. paramamosain can at least equal that of wild‐collected seed crabs in ponds culture.     

Mortality of Northern Bluefin Tuna Thunnus thynnus Due to Trauma Caused by Collision During Growout Culture

Abstract.— Indoor and outdoor experiments were conducted to improve the rearing techniques for artificially hatched northern bluefin tuna Thunnus thynnus during growout culture. Collisions with the walls of tanks or nets caused mass mortality that occurs during growout. The period when collisions frequently occur and the types of injury caused by collision were examined in this study. Juveniles were reared in indoor tanks from 30 to 120 d after hatching, and in an open sea net cage from 42 to 150 d after hatching. Dead fish were collected and counted daily in both of the experiments. In the indoor experiment, the sampled fish were preserved in 10% formalin solution, and each of 10 specimens of about 30, 50, 70, 85, 100, 130, 160 and 225 mm in body length (BL) were examined using x‐rays to detect injury of the bones. Juvenile and young adult bluefin tuna showed a reduction in numbers caused by collision with the tank or the net wall during the experiments. In the indoor tank, there were 1,200 fish on day 30 but only eight on day 120. The daily mortality increased from day 30 after hatching, when juveniles reached 50‐mm BL and remained over 4%/d until day 60 when juveniles grew to 300‐mm BL. The proportion of dead fish with injuries of bone, especially of the vertebral column and the parasphenoid, increased after fish reached 50‐mm BL, and exceeded 60% in fish with BL 85 mm or greater. In the open sea net cage, there were 3,841 fish at the start of the experiment on day 42 and only 65 on day 150. In this experiment, the reduction was greatest from the start of the experiment until day 80, when fish grew to approximately 25 cm in total length. Significant bacterial, viral or parasitic diseases were not observed in these fish; the only findings were dislocations of the vertebral column and injuries to the upper and lower jaws. These results show that the loss of juvenile and young adult bluefin tuna was caused by collision with the tank or net wall that fatally damaged the bones of the vertebral columns and the parasphenoid.     

High incidence of death due to collision of hatchery-reared Pacific bluefin tuna Thunnus orientalis juveniles in sea cages, as revealed by head and spinal injuries

High mortality of hatchery-reared Pacific bluefin tuna (PBT) juveniles is frequently observed in sea cages after transfer from indoor tanks, and is likely due to collisions of the fish with the net wall of the sea cages. Because collisions frequently cause bone injuries due to physical impact, such injuries can be considered an index of obvious collisions. In the present study, bone injuries of dead juveniles were investigated in order to examine the prevalence of net collision as a cause of death among hatchery-reared PBT juveniles in sea cages. Juvenile PBT were reared in sea cages for 90 days after transfer, and dead fish were collected and examined by the clearing and staining method or by dissection to detect injury to the vertebral column and parasphenoid. The prevalence of injuries to the vertebral column and parasphenoid until day 30 after transfer was low (0.0–12.0 %) when the fish were 5.5–15.2 cm in total length (TL). However, it drastically increased to 17.8–78.0 % for fish 21.0–39.2 cm TL from day 31 onward after transfer. These results suggest that the mortalities of juvenile PBT larger than 20 cm TL in cages were induced by net collisions.     

Association of early juvenile yellowfin tuna <Emphasis Type="Italic">Thunnus albacares</Emphasis> with a network of payaos in the Philippines

To understand how early juvenile yellowfin tuna use the habitat and environment provided by fish aggregating devices (FADs), fish (19–31 cm FL) implanted with ultrasonic transmitters into their abdominal cavities were released in a network of payaos in Panay Gulf, the Philippines. Self-recording receivers were attached to the anchor ropes of the payaos to detect the presence of the fish. Some aspects of the behavior of juveniles were similar to those reported in adults. One juvenile showed a diurnal vertical swimming pattern, swam within a limited shallow range during the nighttime, and dived to deeper waters during the daytime. Two juveniles performed deep dives over 100 m during payao-to-payao excursion. Three juveniles showed a diurnal horizontal swimming pattern that was synchronized. In contrast, juveniles stayed <6 days in the network, shorter than adults. No juveniles returned to the same payao after an interruption of over 24 h. It is suggested that juveniles in this area are just starting to migrate and are temporarily staying around a payao for a few days to forage before continuing their migration.     

Retinomotor responses of juvenile bluefin tuna Thunnus thynnus

SUMMARY: In bluefin tuna culture, a high mortality of juveniles is caused by bumping into the tank and net-pen walls at dawn. This bumping can possibly be attributed to visually disoriented behavior of the fish. To examine this possibility, the authors carried out retinomotor response experiments with juvenile bluefin tuna (50.7–96.8 mm total length, at which they were transferred from a indoor tank to a sea net-pen) and measured ambient light intensity at the culture site at dawn. The light intensity at which the transition from scotopic to photopic vision takes place was 7.52 lx and the time taken by the transition was 15 min. At dawn, the ambient light intensity rapidly increased from scotopic light intensity level and attained photopic light intensity level in 10 min. This incompatibility of the retinal adaptation with the change in the ambient light intensity could cause the visual disorientation of the fish. It is therefore possible that the visually disoriented juveniles cannot control their high power swimming and thus bump into the walls at dawn.     

Evaluation of the growth and survival rate of the Caribbean Sea cucumber,Isostichopus badionotus (Selenka, 1867), early juveniles produced in captivity

Isostichopus badionotus is a new commercially important sea cucumber in the Caribbean that has rapidly shown signs of overexploitation. There is no information on the growth and survival rates for wild or hatchery‐produced early juveniles. Hatchery production and determination of juvenile growth rates are essential for future aquaculture efforts for this species. In this study, daily growth rate (GR), specific growth rate (SGR), weight gain (WG), survival rate (SR), and weight–length relation (WLR) were evaluated using small (0.01 g) and large (0.25 g) hatchery‐produced early juveniles under laboratory conditions. Growth was rapid compared to other cultured sea cucumber species; maximal values of SGR and WG were obtained for small juveniles (4.58%/day and 2,191%, respectively), while maximal GR and SR were obtained for large juveniles (0.29 mm/day and 99.17%, respectively). A wide range of sizes was evident at the end of each trial. The WLR of small and large juveniles fit with a negative allometric growth trend. This study highlights the potential production of I. badionotus as early juvenile stages withstand hatchery handling well and exhibit a high SR, reaching commonly released mean sizes (>0.7 g and 25 mm) after almost 3 months (120 days postfertilization).     

Detection and molecular characterization of betanodaviruses retrieved from bivalve molluscs

Betanodaviruses are small ssRNA viruses responsible for viral encephalopathy and retinopathy, otherwise known as viral nervous necrosis, in marine fish worldwide. These viruses can be either horizontally or vertically transmitted and have been sporadically detected in invertebrates, which seem to be one of the possible viral sources. Twenty‐eight new betanodavirus strains were retrieved in three molluscs species collected from different European countries between 2008 and 2015. The phylogenetic analyses revealed that strains retrieved from bivalve molluscs are closely related to viruses detected in finfish in Southern Europe in the period 2000–2009. Nevertheless, a new betanodavirus strain, markedly different from the other members of the RGNNV genotype, was detected. Such a massive and varied presence of betanodaviruses in bivalve molluscs greatly stresses the risks of transmission previously feared for other invertebrates. Bivalve molluscs reared in the same area as farmed and wild finfish could act as a reservoir of the virus. Furthermore, current European regulations allow relaying activities and the sale of live bivalve molluscs, which could pose a real risk of spreading betanodaviruses across different geographic regions. To our knowledge, this is the first study, which focuses on the detection and genetic characterization of betanodaviruses in bivalve molluscs.     

Identification of hatchery-reared and naturally produced Atlantic salmon, Salmo salar L., juveniles based on examination of otoliths

. Atlantic salmon, Salmo salar L., parr and smolts from three Norwegian rivers were examined with respect to whether or not they had been released from a hatchery. The hatchery background was known for some released fish and could be ascertained for others from their aberrant body morphology and eroded fins. Fish released after one winter in a hatchery had opaque otoliths like the fish examined from two hatcheries. Naturally produced fish showed a distinct, seasonal growth pattern in the otoliths, alternating between opaque summer zones and hyaline winter zones. Fish released as fry showed an otolith pattern similar to that of naturally produced fish. A test revealed little discrepancy between two independent otolith readings even though the test reading was conducted without any information about the fish accompanying the otoliths. The results suggest that examination of otoliths may help distinguish between juveniles that are hatchery-reared and juveniles that are naturally produced in the river, provided that the hatchery-reared fish have a 1-year history in the hatchery prior to release.     

南海北部黄鳍金枪鱼幼鱼和成鱼鳃的组织学比较研究

本研究运用光学显微镜、扫描电镜以及透射电镜技术研究了南海北部黄鳍金枪鱼(Thunnus albacares)幼鱼和成鱼鳃的显微组织结构、鳃表面超微结构和鳃小片内部超微结构特点。结果表明, 黄鳍金枪鱼鳃丝顶端弯曲, 鳃弓、 鳃丝和鳃耙表面具有不同类型的细胞。表面超微结构研究显示黄鳍金枪鱼的鳃具有高的片层密度、独特的斜向血流模式以及鳃的融合特性, 幼鱼和成鱼鳃结构之间存在显著不同。鳃小片内部超微结构研究显示鳃扁平上皮细胞覆盖于鳃丝和鳃小片表面, 顶端存在微绒毛或微脊结构, 相对幼鱼, 成鱼具有高的细胞质密度; 离子细胞主要分布于鳃小片以及鳃小片基部, 幼鱼离子细胞顶端开口为微绒毛, 成鱼为小坑状。本研究阐明了黄鳍金枪鱼幼鱼和成鱼鳃的组织结构, 丰富了黄鳍金枪鱼鳃的基础生物学资料, 为研究高速游泳鱼类鳃的形态特征与其高速游泳习性之间的关系提供了参考。     

Lumpfish (Cyclopterus lumpus,Linnaeus) is susceptible to viral nervous necrosis: Result of an experimental infection with different genotypes of Betanodavirus

In recent years, the use of cleaner fish for biological control of sea lice has increased considerably. Along with this, a number of infectious diseases have emerged. The aim of this study was to investigate the susceptibility of lumpfish (Cyclopterus lumpus) to Betanodavirus since it was detected in asymptomatic wild wrasses in Norway and Sweden. Three betanodaviruses were used to challenge lumpfish: one RGNNV genotype and two BFNNV genotypes. Fish were injected and monitored for 4 weeks. Brain samples from clinically affected specimens, from weekly randomly selected fish and survivors were subjected to molecular testing, viral isolation, histopathology and immunohistochemistry. Reduced survival was observed but was attributed to tail‐biting behaviour, since no nervous signs were observed throughout the study. Betanodavirus RNA was detected in all samples, additionally suggesting an active replication of the virus in the brain. Viral isolation confirmed molecular biology results and revealed a high viral titre in BFNNV‐infected groups associated with typical lesions in brains and eyes of survivor fish. We concluded that lumpfish are susceptible to Betanodavirus, as proven by the high viral titre and brain lesions detected, but further studies are necessary to understand if Betanodavirus can cause clinical disease in this species.     

Size‐dependent stress response in juvenile Arctic charr (Salvelinus alpinus) under prolonged predator conditioning

Predator conditioning can be used to improve post‐release antipredator recognition of hatchery‐reared salmonids. However, possible negative stress‐related effects of prolonged predator conditioning on juvenile fish physiology are poorly understood. We studied the effects of prolonged (91 days) predator odour exposure on whole‐body cortisol level and spleen size in six full‐sib families of juvenile hatchery‐bred Arctic charr (Salvelinus alpinus). Chemical cues from water containing charr‐fed pikeperch (Sander lucioperca) were used as the predator exposure stimuli and lakewater was used as a chemical control. Our study revealed that juvenile body cortisol levels post‐predator conditioning were affected by treatment, fish size and their interaction. Importantly, among the smaller (i.e. slowest growing) charr, the predator‐exposed fish had higher cortisol levels than control fish, while the opposite pattern was true for the larger fish. These results suggest that chemical cues from charr‐fed predators induce a prolonged stress response in juvenile charr. As prolonged predation exposure seems to elevate stress levels in a size‐dependent manner, the larger, faster growing fish could possibly have intrinsically lower stress responses to predation threats than smaller, slower growing fish. Possible coupling between stress sensitivity and growth requires further attention due to the likely implications for the management of unintended domestication among captive‐reared salmonids.