基于InDel标记的斑节对虾早期性别鉴定方法的建立

为分辨斑节对虾(Penaeus monodon)发育早期的性别,利用改良的插入/缺失(InDel)标记追踪了从受精卵、无节幼体、溞状幼体、糠虾幼体、仔虾后1 d(PL1)、仔虾后30 d(PL30)及亚成虾个体的性别,建立了测序和荧光定量PCR熔解曲线的检测方法。结果表明,InDel标记可扩增获得雌雄特异性序列,用测序法判定亚成虾性别的结果与外部观察结果一致,准确率达100%,对受精卵、无节幼体、溞状幼体及糠虾幼体均可得到雌雄特异性序列;进而建立了基于SYBR Green实时荧光定量PCR(Quantitative real-time polymerase chain reaction,q RT-PCR)熔解曲线的快速鉴定斑节对虾性别的方法,其中雄性特异序列的熔解温度(Melting temperature,Tm)为(79.10±0.10)℃,雌性为(78.45±0.20)℃,通过特异的熔解曲线可准确区分PL30、亚成虾和无节幼体Ⅵ期个体,准确率可达96.3%以上。     

Genetic variation in resistance of the tiger pufferfish Takifugu rubripes to a host-specific monogenean parasite Heterobothrium okamotoi

Fisheries Science - The tiger pufferfish Takifugu rubripes is one of the most valuable aquaculture species in Japan. In this study, we investigated the genetic variation in resistance to...     

Inducement of sex reversal in Sarotherodon niloticus (Linnaeus)

Two androgens, ethynyltestosterone (ET) and methyltestosterone (MT) and two estrogens, diethylstilbestrol (DES) and estrone (E) were orally administered to gonadally undifferentiated Sarotherodon niloticus fry. Diets with hormones were prepared at the following levels: ET and MT at 30 and 60 mg kg^?1 of feed, DES at 25 and 100 mg kg^?1 of feed, and E at 100 and 200 mg kg^?1 of feed. The fish were fed at 10% body weight per day for 25-, 35- and 59-day periods.All-male populations were produced with ET-60 and MT-60 in all three treatment periods, and with ET-30 and MT-30 during the 35- and 59-day treatments.No estrogen treatment resulted in 100% female populations but only about 10% of the DES-100 treatment populations at the three durations were males.Sex ratios were significantly different (P < 0.01) from the expected 1:1 ratio in all fish treated with androgens and estrogens. No ovotestis condition was found. Total mortalities in all treatments and controls were uniformly low, ranging from 1.5 to 8.8% for the treatment period.     

An intensive hatchery rearing protocol for larvae of the bullseye puffer,Sphoeroides annulatus (Jenyns)

In this study bullseye puffer, Sphoeroides annulatus larvae were reared from hatching through to 1 or 2 months after weaning on an experimental scale in three replicate 600 L tanks and on three occasions during the spawning season (nine tanks in total). The rearing protocol used was green water (Nannochloropsis oculata and Isochrysis sp.) 100 000 cells mL^?1 from 0 to 11 days after hatch (DAH), 5–10 rotifers, Brachionus rotundiformis mL^?1 from 2 to 26 DAH, 0.1–1 Artemia mL^?1 from 21 to 34 DAH and weaning from 29 to 34 DAH. Survival to a month after weaning was 1%, a total of 3153 juveniles were produced with an average wet weight of 0.29 ± 0.07 g and a length of 27.5 ± 0.82 mm.     

Genetic diversity in a Tasmanian hatchery population of Atlantic salmon (Salmo salar L.) compared with its Canadian progenitor population

The level of genetic diversity in a cultured Atlantic salmon (Salmo salar) population from Tasmania, Australia was examined at 11 microsatellite loci and compared with that in its progenitor population from the River Philip in Nova Scotia, Canada. The reference progenitor population consisted of archived scales collected from wild River Philip salmon in 1971 and 1972, not long after salmon from this river were imported into Australia in the mid‐1960s. The Tasmanian hatchery stock had a significant reduction in the mean number of alleles (31–43%) and mean allelic richness (28–39%) across all microsatellite loci compared with the wild Canadian population. Mean heterozygosity levels remained unchanged. Estimates of per‐generation effective population sizes for the Tasmanian population, based on allele frequency temporal variance with the wild progenitor population, ranged from 102–207 individuals and reflected hatchery records.     

The effect of temperature on physiological energetics of a fast‐growing selective strain and a hatchery population of the Pacific oyster (Crassostrea gigas)

Growth is an important aquaculture trait, which has been one of the most important traits for genetic improvement in aquatic species. Numerous fast‐growing domesticated aquaculture organisms are being cultured, but the physiological processes underlying growth remain largely unexplored. The Pacific oyster (Crassostrea gigas) is one of the most widely farmed bivalves. In order to improve the productivity traits, a fast‐growing selective strain of C. gigas was established. In this study, we compared physiological energetics of the fast‐growing selective strain and an unselected hatchery population of C. gigas under different conditions of temperature. Components of energy gain (clearance rates, ingestion rates, absorption efficiency and absorption rates), energy loss (metabolic rates) and resulting energy budget (scope for growth) were analysed at four temperatures (15, 20, 25 and 30°C). The results indicated that selective breeding oysters had a higher energy gain compared to unselected oysters, but the selected strain and hatchery population of oysters did not significantly differ in metabolic rates at all tested temperatures (p > 0.05). The enhanced energy gain resulted in almost two times higher scope for growth in selective breeding oysters compared with unselected oysters. Physiological characteristics of C. gigas were greatly affected by temperature and showed a fluctuation with the increase in temperatures. The selective breeding oysters outperformed those of hatchery population in growth performance by an increased energy gain along with a similar energy loss. This study provided insight into physiological energetics accounting for growth improvements in fast‐growing selective strain of C. gigas.     

Growth and feeding responses of a hatchery population of brown trout (Salmo trutta L.) at low temperatures

Abstract— Feed intake, specific growth rate and changes in body composition were studied in age 1+ (140-190 g) brown trout ( Salmo trutta L.) reared at three temperatures (2, 4 and 6°C) under continuous light conditions. Feed in take increased from 35.7 kJ-kg⁻¹.day⁻¹ at 2°C to 95.7 kJ-kg⁻¹-day⁻¹ at 6°C, and the growth rate increased from 0.19%-day⁻¹ to 0.42%-day⁻¹ over the same temperature range. The estimated lower temperature limits for feeding and growth were slightly above 0°C. For all groups of fish, the majority (about 75%) of the weight gain comprised water and protein, but lipid deposition tended to increase with increasing temperature. The deposition of lipid accounted for about 50% of the body energy gain at all temperatures.     

An evaluation of the contribution of hatchery stocking on population density and biomass: a lesson from masu salmon juveniles within a Japanese river system

Stocking is an important management tool for enhancing fisheries resources, but its actual contribution to fisheries resources is controversial, taking into consideration both the positive and negative effects. This study compared density and biomass of hatchery (otolith thermal marked) and wild masu salmon parr between stocked and unstocked rivers to evaluate the contribution of stocking with hatchery‐reared fish. Density and biomass of all fish did not differ between stocked and unstocked rivers. Moreover, density and biomass of wild fish in the stocked rivers were lower than those of the unstocked rivers. Density and biomass of hatchery fish in a non‐natural reproducing river were similar with those of all fish in natural reproducing rivers. These results indicate that hatchery stocking does not have positive effects on population density or biomass but replaces wild fish with hatchery fish and that non‐natural reproducing areas are more suitable as stocking sites.     

Sexual patterns and protogynous sex reversal in the rusty parrotfish, Scarus ferrugineus (Scaridae): histological and physiological studies

Gonadal histology confirmed that Scarus ferrugineus is a diandric protogynous fish. The process of protogynous sex reversal was investigated through histological observations on the gonads of females changing sex to male. This process was divided into three stages on the basis of changes in the structure of the germinal and somatic elements. Ovaries of functional females (stages IV-V) were filled with vitellogenic oocytes during the breeding season but contained no trace of spermatogenic tissue. During post-spawning period, the remaining vitellogenic oocytes began to degenerate and accompanied by a drop in plasma levels of estradiol-17β. At the commencement of sex change, previtellogenic oocytes began to degenerate and stromal cell aggregation was observed in the central region of the lamellae. At mid-reversal stage, steroid-producing cells (Leydig cells) developed at the border of the stromal aggregate and spermatogonial cysts appear at the periphery of lamellae. Finally, sex change to secondary males was considered complete, with the beginning of active spermatogenesis and spermiation. Plasma levels of testosterone remained low throughout the sex change, but II-KT increased rapidly parallel to the increased number of Leydig cells while the level of estradiol-17β decreased. The results indicate also that the sex-changed males had higher level of II-KT than primary males, while primary males had higher level of testosterone. Histological examination revealed that testes of primary and secondary males are almost identical in organization of the spermatogenic cysts, association of sertoli cells, and developing germ cells but differ in clustering and development of Leydig cells.     

Induction of sex reversal in blue drum (Nibea mitsukurii) and gynogenetic yellow drum (Nibea albiflora) by oral administration of letrozole

The objective of this study was to test the efficiency of an aromatase inhibitor, letrozole (LZ), to induce masculinization in blue drum (Nibea mitsukurii) and gynogenetic yellow drum (N. albiflora). Two experiments (Exp‐1 and Exp‐2) were performed to investigate the effect of LZ on the growth, sex ratio and gonad development in these two fishes. In Exp‐1, blue drum were treated with oral administration of letrozole at different doses (0, 1, 10, 100 mg/kg). In Exp‐2, gynogenetic yellow drum were orally administered different doses of LZ (0, 10, 100 mg/kg) and a dose of 10 mg/kg 17α‐methyltestosterone (MT). The treatments in both experiments were performed from 25 to 85 days post hatch (dph). As a result, all the LZ‐treated fish were phenotypic males despite of slower growth than the control during the treatment period. Free spermatozoa were observed from 150 dph onwards and the gonadosomatic index of the LZ‐treated fish did not differ significantly from that in the control at maturity. However, the sex‐reversed fish induced by MT exhibited a high proportion (33.4%) of testicular abnormalities. Our results showed that oral administration of LZ was efficient in inducing sex reversal in drums, and thereby facilitated their mono‐sex culture.     

Advances in hatchery and grow-out technology of cobia Rachycentron canadum (Linnaeus)

This paper describes advances in hatchery and grow‐out technology of cobia (Rachycentron canadum, Linnaeus). In 2007, methods for capture, transport, acclimation, sampling, conditioned spawning, larval rearing, fingerling production, nursery, shipping and grow‐out have been perfected. Survival rates ranging from 17.5% to 35% were achieved from egg to shipping size fingerlings (1.0 g) in 2007 at the University of Miami Experimental Fish Hatchery, with production of approximately 20 000 fingerlings per 12 000 L tank. Wild and F1 broodstock cobia have been conditioned to spawn through temperature manipulation producing viable eggs for experimental and production level larval rearing trials in several hatcheries. Brood fish have also been induced to spawn using hormones. Cobia appear to be susceptible to infestations by parasitic protozoa such as Amyloodinium ocellatum and to infections caused by deleterious bacteria such as Photobacterium spp. and Vibrio spp. Prophylactic methods used to prevent and control epizootic diseases at the hatchery are summarized. Improved techniques for cage management were implemented, and both novel designs of submerged cages deployed in exposed areas and traditional gravity cages in protected areas have been used for commercial ongrowing of cobia in the Americas and the Caribbean region.     

Characterization of molecular variants of GnRH, induction of spermiation and sex reversal using salmon GnRH-A and domperidone in the protogynous diandric fish, Synbranchus marmoratus Bloch, (Teleostei, Synbranchidae)

This paper studies the molecular variants of gonadotropin-releasing hormone (GnRH) present in the brain of the protogynous swamp eel, Synbranchus marmoratus, and the effects of the administration of salmon GnRH analogue (sGnRH-A) and the dopamine receptor antagonist, domperidone (DOM) on final maturation and gamete release in this species. Evidence for the presence of two GnRH variants, sGnRH and cIIGnRH were obtained by reverse phase high-pressure liquid chromatography (RP-HPLC) and radioimmunoassay with different antisera. The effects of treatment with sGnRH-A+DOM were checked by three ways: oocyte and milt release by stripping, histological analysis of the gonadal tissue, and androgen serum levels at different times throughout the experiment. In males, spermiation was induced after three weeks of treatment. In the female group, sGnRH-A+DOM did not induce ovulation at the end of the experiment. Histological analysis of the gonads from the female group showed evidence of sex reversal. All the treated fish had elevated androgen serum levels from the third week, with respect to control fish. In all cases, serum estradiol levels were undetectable. These results suggest that treatment with sGnRH analog and DOM induce sex reversal in female and spermiation in males. This revised version was published online in July 2006 with corrections to the Cover Date.     

Stringed bed suspended bioreactors (SBSBR) for in situ nitrification in penaeid and non-penaeid hatchery systems

For establishing nitrification in prawn (non-penaeid, salinity 10–15 ppt) and shrimp (penaeid, salinity 30–35 ppt) larval production systems, a stringed bed suspended bioreactor (SBSBR) was designed, fabricated, and validated. It was fabricated with 5 mm polystyrene and low density polyethylene beads as the substrata for ammonia and nitrite oxidizing bacterial consortia, respectively, with an overall surface area of 684 cm². The reactors were activated in a prototype activator and were transported in polythene bags to the site of testing. Performance of the reactors activated with the nitrifying bacterial consortia AMONPCU-1 (ammonia oxidizers for non-penaeid culture) and NIONPCU-1 (nitrite oxidizers for non-penaeid culture) was evaluated in a Macrobrachium rosenbergii larval rearing system and those activated with AMOPCU-1 (ammonia oxidizers for penaeid culture) and NIOPCU-1 (nitrite oxidizers for penaeid culture) in a Penaeus monodon seed production system. Rapid setting up of nitrification could be observed in both the static systems which resulted in a higher relative per cent survival of larvae.     

A comparison of aromatase inhibitors for the sex reversal of female Atlantic salmon (Salmo salar L.)

Immersion treatment of all female Atlantic salmon alevins with 1,4,6-androstatriene-3, 17-dione (ATD) (Steraloids Inc.) at doses of 0.5 and 5 mg l^?1 resulted in up to 54% masculinisation and fertility was the same as normal males.     

Vibrios in hatchery cultures of the razor clam,Solen marginatus (Pulteney)

Hatchery culture of the razor clam, Solen marginatus (Pulteney), has recently been developed in Galicia (NW Spain). However, recurrent episodes of mortalities of larval and post‐larval cultures have been recorded during the course of various studies. The disease signs were similar to those described for other bivalve species in outbreaks caused by bacteria of the genus Vibrio. In this article, we present the results of microbiological monitoring of two batches of razor clams with different survival rates. All fermentative isolates were identified as members of the Splendidus clade within the genus Vibrio. Some of these isolates, identified as Vibrio splendidus‐like, were clearly associated with the batch suffering mortalities, indicating their possible role as pathogens. Similar strains were found in the broodstock, suggesting vertical transmission of these bacteria. This is the first study of the microbiota associated with hatchery culture of S. marginatus, and the results will provide useful information for the optimization of a protocol for hatchery culture of this bivalve species.     

Mechanical control of Ichthyophthirius multifiliis Fouquet, 1876 (Ciliophora) in a rainbow trout hatchery

Ichthyophthirius multifiliis, more commonly known as ‘white spot’ is recognised as one of the most pathogenic diseases of wild and cultured freshwater fish. In trout hatchery systems infections can quickly establish and result in high mortality if left unregulated. Current control centres around the use of regular bath treatments with formalin, however, treatments are not always effective and are labour intensive. Environmentally safe, non-chemical alternatives are required. This paper describes the performance of a mechanical system developed to remove cysts from commercial trout raceways. The system consists of two parts: a specially designed suction head connected to a pump that is used to vacuum the bottom of hatchery raceways, and a low-adhesion polymer raceway lining. Following tests on a series of potential linings, the most efficacious product in preventing settlement of cysts was used to line three raceways in a commercial rainbow trout (Oncorhynchus mykiss) hatchery. Over a period of three months, lined raceways were vacuumed on a daily basis. The three control raceways were maintained according to the farms normal husbandry procedures. Approximately thirty fish were sampled from each raceway at fortnightly intervals, and the abundance of the parasite in each raceway determined. The mechanical system led to a significant reduction in the abundance of the parasite in the test raceways, with a mean abundance of 361.88 ± 260.75 parasites per fish in control raceways compared with 1.64 ± 1.59 parasites per fish in the test raceways at the peak of the infection. Additionally fish survival was significantly higher in test raceways over control, with a mean of 84.5% of the stock surviving in the test raceways compared with only 70.6% in the controls by trial end.     

Mechanical control of Ichthyophthirius multifiliis Fouquet, 1876 (Ciliophora) in a rainbow trout hatchery

Ichthyophthirius multifiliis, more commonly known as ‘white spot’ is recognised as one of the most pathogenic diseases of wild and cultured freshwater fish. In trout hatchery systems infections can quickly establish and result in high mortality if left unregulated. Current control centres around the use of regular bath treatments with formalin, however, treatments are not always effective and are labour intensive. Environmentally safe, non-chemical alternatives are required. This paper describes the performance of a mechanical system developed to remove cysts from commercial trout raceways. The system consists of two parts: a specially designed suction head connected to a pump that is used to vacuum the bottom of hatchery raceways, and a low-adhesion polymer raceway lining. Following tests on a series of potential linings, the most efficacious product in preventing settlement of cysts was used to line three raceways in a commercial rainbow trout (Oncorhynchus mykiss) hatchery. Over a period of three months, lined raceways were vacuumed on a daily basis. The three control raceways were maintained according to the farms normal husbandry procedures. Approximately thirty fish were sampled from each raceway at fortnightly intervals, and the abundance of the parasite in each raceway determined. The mechanical system led to a significant reduction in the abundance of the parasite in the test raceways, with a mean abundance of 361.88 ± 260.75 parasites per fish in control raceways compared with 1.64 ± 1.59 parasites per fish in the test raceways at the peak of the infection. Additionally fish survival was significantly higher in test raceways over control, with a mean of 84.5% of the stock surviving in the test raceways compared with only 70.6% in the controls by trial end.     

Brook trout (Salvelinus fontinalis) population dynamics and mottled sculpin (Cottus bairdi) occurrence in relation to acidic episodes in streams

We monitored water chemistry and populations of brook trout (Salvelinus fontinalis) and mottled sculpins (Cottus bairdi) in 5 streams and some tributaries to determine how the severity of acidic episodes (low pH and high Al) influenced density and distribution of these species. Among streams, Linn Run had the lowest pH (4.8) and highest concentrations of total dissolved Al (>400 μg-l^?1) during periods of high discharge. Densities of brook trout and mottled sculpins were low in Linn Run, but high in tributaries, where pH (6.5) and Al (30 μg-l^?1) remained well below lethal levels. Among the other streams, brook trout density was highest where the acidic episodes were least severe and the density of age-0 brook trout was lowest when precipitation during the early months of the year was above normal. Immigration of brook trout from downstream areas maintained populations in the stream segments subjected to severe episodes. Mottled sculpins were not found in streams with severe episodes.