Diurnal rhythm of steroid biosynthesis in the testis of terminal phase male of protogynous wrasse, Pseudolabrus sieboldi, a daily spawner

The wrasse, Pseudolabrus sieboldi, is a diandric protogynous labrid fish. Spawning is performed by a terminal phase (TP) male and an initial phase (IP) female between 6:00 and 9:00 h daily during two-month-long spawning season. In the present study, to investigate the roles of steroid hormones in the diurnal spermatogenesis of the P. sieboldi TP male, all steroid hormones produced in the testis were identified and the synthetic pathways of these steroids were determined. Furthermore, the circulating levels of the major steroids produced were analyzed throughout a day at 3-hour intervals during spawning season. In the testis, 11-ketotestosterone (11-KT), estradiol-17β (E2), 17,20β-dihydoxy-4-pregnane-3-one (17,20β-P) and 17,20β,21-trihydroxy-4-pregnen-3-one (20β-S) were synthesized as the major metabolites. In vitro steroid biosynthesis experiments showed similar results to the circulation profiles of the major steroids. This study is the first to clarify the complete steroidogenic pathways in the gonads of a diandric protogynous species throughout its life, when combined with the results of the steroidogenesis in the ovarian follicles. This is also the first report of a clear diurnal rhythm of the steroid production corresponding to the spermatogenic process in the testis of a male teleost.     

Isolation, cloning, sequencing of brain type aromatase and its expression in male and female Wrasse, Pseudolabrus sieboldi

Pseudolabrus sieboldi, wrasse being a diurnal spawner provides a good opportunity to study the endocrine mechanism of estrogen formation in brain and gonads. Moreover, an extremely large amount of E2 was produced in serum and testis of wrasse. It is assumed that the presence of E2 may play a major role in diurnal gametogenesis in male fish. In this study brain type aromatase have been isolated, cloned and sequenced from the brain of wrasse. Further, the expression pattern of brain type aromatase in gonads and adult tissue of male and female fish have been analyzed. In addition, the diurnal expression pattern of brain type aromatase in both male and female fish brain during spawning season have been analyzed. The P450arom (br) was isolated, cloned and sequenced from both male and female bamboleaf wrasse. The P450arom (br) gene (1877 sequenced nucleotide) contains an ORF of 1470 bp, a 5′-UTR of 18 bp and at least 407 bp in 3′-UTR. The amino acid sequence homology in the coding region of wrasse P450arom (br) is high compared to that of medaka, Oryzias latipes (80%), rainbow trout type 2, Oncorhynchu mykiss (78.2%), fugu, Takifugu ribripes (78%) rainbow trout type 1, (76%), goldfish, Carassius auratus (66.8%) and zebrafish, Danio rerio (66.2%). Expression study reveals that P450arom (br) mRNA were most abundant in brains of both male and female fish throughout the day during the spawning season. RT-PCR study revealed that P450arom (br) was expressed in skin, anal fin and tail fin of both male and female wrasse. P450arom (br) was not detected at any time of the spawning day in either ovary or testis of wrasse.     

Environmental and fishing effects on the abundance,size and sex ratio of the blue‐throated wrasse,Notolabrus tetricus,on South Australian coastal reefs

Abstract The blue‐throated wrasse, Notolabrus tetricus (Richardson), is a site‐attached protogynous hermaphroditic species, with a small home range of 1000–2000 m², and the most abundant fish on inshore coastal reefs of South Australia. Long‐term studies over 18–24 years were conducted at an island reserve site and a fished mainland reference site. At the reserve site the retention rate of fish (reduced by mortality + emigration) was directly correlated with female mean size; the female:male sex ratio ranged from 10 to 20:1, consistent with an induction‐inhibition model of sex‐change. At the fished reference site, female mean size declined under intense fishing, and the sex ratio became strongly skewed towards females. Additional spatial surveys over >2000 km of coast showed that: (1) juvenile and adult abundance was variously determined by habitat features, such as substratum‐type, depth, bottom relief, wave exposure and algal canopy cover, and by geographical factors, such as distance off‐shore and within gulfs; and (2) that female mean size and the sex ratio were strongly influenced by recreational fishing, and to a small extent by bottom relief and other factors. The results suggest that female mean size and the sex ratio together can be used as an indicator of recreational fishing intensity at local scales. As populations of sequential hermaphroditic species are sensitive to size‐selective harvesting, management measures should aim to prevent sperm limitation, reproductive failure and skewed sex ratios in this species.     

Gonadal steroidogenesis and the possible role of steroid glucuronides as sex pheromones in two species of teleosts

In general, female zebrafish,Brachydanio rerio, ovulate only in the presence of males. The stimulant must be pheromonal as even male holding water is capable of inducing ovulation. After ovulation the mating phase begins. During this phase the male follows the female and oviposition as well as fertilization takes place. Both the ovulation and the mating are controlled by pheromones synthesized by the gonads. Ovulation can be induced by testicular homogenates. After the lipid material has been extracted from the testicular homogenates, the remaining aqueous phase can still induce ovulation. However, when the aqueous phase is treated with the enzyme-glucuronidase, it loses the ability to induce ovulation. This is an indication that glucuronides, probably steroid glucuronides, are the compounds responsible.During the mating phase, ovulated female zebrafish become attractive to males. It was found that, after ovulation, ovarian extracts contain the compounds responsible for attracting males. The attractant consists of a mixture of steroid glucuronides.After incubation of the gonads with³H-precursors seven steroid glucuronides have been identified in the testis and five in the ovary.Under fish culture conditions the African catfish,Clarias gariepinus, can produce postivitellogenic oocytes throughout the year. However, in capitivity neither males nor females spawn. In female catfish maturation and ovulation can be induced by treatment with gonadotropins. It might be possible that, analogous to the zebrafish, some reproductive processes in the catfish have to be induced by pheromones. It has been demonstrated that pheromonal compounds released by the seminal vesicles are involved in the attraction of female conspecifics. The steroid glucuronide synthesizing capability of the testes and the seminal vesicles of the male catfish are examined, as well as that of the ovary before and after ovulation of the female catfish. Both testes and seminal vesicles appear to be capable of steroid biosynthesis but only the latter synthesizes steroid glucuronides. Six of these conjugates have been isolated and identified. In the female catfish the ovaries are capable of synthesizing seven steroid glucuronides, but only after ovulation.     

Susceptibility of corkwing wrasse Symphodus melops, goldsinny wrasse Ctenolabrus rupestis, and Atlantic salmon Salmo salar smolt, to experimental challenge with Vibrio tapetis and Vibrio splendidus isolated from corkwing wrasse

The virulence of two Vibrio strains, previously isolated from diseased corkwing wrasse Symphodus melops and identified as V. tapetis and V. splendidus, to corkwing and goldsinny wrasse Ctenolabrus rupestris and to Atlantic salmon Salmo salar, was studied under laboratory conditions. Both bacteria were shown to be opportunistically pathogenic to corkwing wrasse, causing significantly higher mortality in the challenged groups than in the controls. Bacterial cultivation of kidney samples and re-isolation of V. tapetis and V. splendidus from most mortalities confirmed the two strains as the probable cause of mortality in the challenged groups. The control group also suffered relatively high mortality, but no specific pathogens that were suspected to be the main cause of death were isolated, other than a mixture of Vibrio spp. and, in the case of one individual, atypical Aeromonas salmonicida. Following injection challenge with both bacterial strains, no mortality was recorded in Atlantic salmon. In bath challenge trials with goldsinny wrasse, only slight mortality was observed in the challenged groups and the unchallenged control group. Bacterial examination showed that atypical Aeromonas salmonicida was the probable cause of death in both bath challenged and control groups of goldsinny wrasse, and no indication of infection by any Vibrio sp. was found.     

Aeromonas salmonicida infectivity studies in goldsinny wrasse, Ctenolabrus rupestris (L.)

Goldsinny wrasse, Ctenolabrus rupestris (L.). were experimentally and naturally infected with Aeromonas salmonicida. Goldsinny wrasse were found to be susceptible to infection with A. salmonicida but significantly more resistant to infection than Atlantic salmon, Salmo salar L. The pathology of the acute infection is described. Following infection significantly higher levels of agglutinating antibody were detected in the sera of recovered wrasse than those observed in the Atlantic salmon controls. However, a large proportion of the recovered wrasse were culture positive for A. salmonicida and appeared to have entered a chronic stage of infection quite distinct from the carrier status that can develop in Atlantic salmon. This study indicates that, although goldsinny wrasse are susceptible to A. salmonicida, these fish are unlikely to become primarily infected, but contract furunculosis from infected Atlantic salmon. However, goldsinny wrasse may act as a reservoir for subsequent infections of cultivated Atlantic salmon because of the development of a chronic form of the disease. The potential for the vaccination of goldsinny wrasse against furunculosis is discussed.     

Precocious sex change and spermatogenesis in the underyearling Malabar grouper Epinephelus malabaricus by androgen treatment

The Malabar grouper Epinephelus malabaricus is an important candidate species for commercial aquaculture in tropical and subtropical areas. In nature, this species requires more than 10 years to change sex from female to male and have active spermatogenic tissues in the testis. Thus, it is essential to find a means of producing sperm for seed production. This is the first report of artificial sex change in underyearling E. malabaricus . Female E. malabaricus with immature ovaries at 144 days post-hatch (DPH) were fed a diet with 17α-methyltestosterone (MT) at 50 μg g⁻¹ diet for 6 months. Sex change occurred in most of the treated fish, which had testis with all stages of spermatogenic germ cells including spermatozoa. In contrast, most of the control fish had immature ovaries. These results, which reveal that germ cells in the underyearling grouper have the ability to produce spermatozoa in response to exogenous androgen, demonstrate that sex change can be artificially induced during ovarian development.     

养殖牙鲆性类固醇激素的周年变化

通过放射性免疫方法测定和比较了养殖牙鲆(Paralichthys olivaceus)成鱼血清中两种重要的性类固醇激素—睾酮(T)和雌二醇(E2)的周年变化规律。在雄性个体中,从4月份开始,随着水温的升高及繁殖期的开始,T和E2含量显著上升并在6月份达到最高峰;之后,T和E2含量逐渐下降并在第二年3月左右降低至最低水平。雌性牙鲆个体的E2含量于3月份开始慢慢增高,于6月份达到最高峰;随后,E2的含量逐渐降低并在1月份达到最小值;而雌性个体中的T的含量在2月和6月份出现两次高峰。结果表明,T和E2对牙鲆的性腺发育具有十分重要的作用,在雌雄个体中T和E2均呈现出周期性的变化。     

Gonadal sex differentiation and sex control in red sea bream, Pagrus major

The gonadal sex differentiation in red sea bream, Pagrus major, which is one of the most important species for aquaculture in Japan, was revealed histologically. The suitable conditions for induction of all-male groups in the fish were investigated, and functional males were induced by the conditions of oral administration of 17α-methyltestosterone. The sex determination of this fish was also discussed.     

In vitro steroidogenesis by gonads of the Russian sturgeon, Acipenser gueldenstaedti Brandt

Ovaries and testes from the Russian sturgeon at different stages of sexual maturity were incubated with ;sup3;H-androstenedione or ³H-pregnenolone. The major metabolites of androstenedione in both sexes were testosterone and 5- and 5ß-androstanediols, but no evidence was found for the gonadal production of 11-oxygenated androgens. Pregnenolone was converted to 17-hydroxyprogesterone, androstenedione and testosterone together with reduced metabolites. 11-Oxygenated androgens were found in female serum by gas chromatography-mass spectrometry (GC-MS), in serum of both sexes by radioimmunoassay (RIA), and was detectable by RIA in interrenal but not gonadal tissue. The results suggest that sturgeon may differ from teleosts in that 11-oxygenation may take place in extragonadal tissues.     

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.     

Effect of in vitro xenoestrogens on steroidogenesis in mature female fish, Chasmichthys dolichognathus

In this study, fully vitellogenic oocytes of the longchin goby (Chasmichthys dolichognathus) were exposed to in vitro xenoestrogens such as diethylstilbestrol (DES), bisphenol A (BPA) and nonylphenol (NP) at concentrations of 100 ng/ml in the presence of [³H]17α-hydroxyprogesterone as precursor. The major metabolites produced in vitro are progestogens [17α-hydroxy,20α-dihydroprogesterone (17α20αOHP) and 17α-hydroxy,20β-dihydroprogesterone (17α20βOHP)], androgens [androstenedione (A4) and testosterone (T)] and estrogens [estrone (E1) and estradiol-17β (E2)]. Comparative activities of these chemicals in oocyte steroid biosynthesis showed as follows: NP treatment resulted in clear stimulation of estrogen production, while the opposite occurred in DES treated incubation. Treatment with DES resulted in a higher synthesis of 17α20αOHP. BPA treatment increased the androgen, while estrogen synthesis was slightly inhibited. These results suggest that NP exhibited clearly estrogenic activity on the three chemicals.     

The development of the Norwegian wrasse fishery and the use of wrasses as cleaner fish in the salmon aquaculture industry

Norway leads the world aquaculture production of Atlantic salmon Salmo salar and farmed Norwegian Atlantic salmon is currently consumed around the globe. However, sea lice infestation is a major problem faced by the salmon aquaculture industry in Norway and elsewhere. The use of wild-caught cleaner fish, mainly wrasses, has been recommended over the other available methods as the most economical and environmentally friendly option to control sea lice infestation in salmon farming. Here, we review the development of the Norwegian wrasse fishery and the use of wrasses as cleaner fish. In this document, we address the sea lice problem and introduce the main wrasse species employed as cleaner fish, document the cleaning behaviour of wrasses, present the development of a new wrasse fishery associated with the salmon aquaculture industry, and finally, we identify the main challenges associated with the intensive use of wild-caught cleaner wrasses and provide some insight for future directions of the wrasse fishery and further development of aquaculture techniques to supply salmon facilities with domesticated cleaner fish.     

Development of the use of wrasse to control sea lice on salmon farms in Ireland

Abstract. Developments in the use of wrasse as cleaner-fish in the control of sea lice ( Lepeophtheirus salmonis, Caligus elongatus ) in Ireland are reviewed. Corkwing, Symphodus melops , and goldsinny, Ctenolabrus rupestris , have controlled L. salmonis (<1 louse per salmon) on commercial farms. C. rupestris and rockcook, Centrolabrus exoletus , removed both adult and chalimus C. elongatus in aquarium trials. No egg-bearing lice were ever found on salmon in cages with wrasse, and numbers of mobile lice were maintained at <1 per salmon smolt.
Wrasse are fished using shrimp pots, optimally set for about 2 h. The fishery is seasonal and limited to >10cm wrasse occurring close to the farm, and distant from other farms. Localized reductions in the body length and numbers of wrasse have been detected in the second year of fishing in two areas respectively. Due to the fishery limitations and high fecundity of wrasse, impacts on wrasse populations should remain localized. All wrasse populations fished are screened for microbial diseases, and protozoan and metazoan parasites. Results are typical for wild marine fish but metazoan skin parasites are rare. No pathogens, including tests with an Aeromonas sp. found in wrasse, have been found to be transferable to salmon.
Wrasse are the only effective and continuous method of sea lice control legally available. Research efforts should be directed towards improving the availability, pathogen-free status, and husbandry of wrasse, to enable cleaner-fish technology to be more widely and efficiently utilized.     

Diseases of north European wrasse (Labridae) and possible interactions with cohabited farmed salmon, Salmo salar L

There have been several reported studies of wrasse health but none of these has shown transmission of wrasse diseases when stocked with farmed Atlantic salmon. Most of the studies have focussed on bacterial and parasite issues, including treatment of bacterial diseases with antibiotics and vaccination of wrasse. Classical and atypical furunculosis have been reported in wrasse following stress, and wrasse have been susceptible to vibrio infection. Further study is required on the vaccination of wrasse for furunculosis with latent carrier status to maximize survival. There are studies on viral diseases such as infectious pancreatic necrosis, infectious salmon anaemia and pancreas disease and although these did not give any undue concern for salmon health, there is also scope for further study in this area. Resident parasite communities of wrasse are largely host-specific and do not appear to be a threat to salmon. Given that wrasse have not, to date, been a vector of disease in salmon, attention should be placed on maintaining best practice in cohabiting wrasse with salmon. Other issues that should be addressed are good welfare of wrasse in pens and identifying measures of this, the identification of losses of wrasse in pens, being alert to potential emerging diseases through health screening of mortalities and assessing the risks associated with carrying forward wrasse from one salmon production cycle to the next. Issues of exploitation by fishing on wild wrasse stocks and improved biosecurity may be addressed by the increased movement by the industry to the stocking of farmed wrasse.     

Correction to: The development of the Norwegian wrasse fishery and the use of wrasses as cleaner fish in the salmon aquaculture industry

Fisheries Science - A correction to this paper has been published: https://doi.org/10.1007/s12562-021-01502-z     

Growth performance of mixed sex, hormonally sex reversed and progeny of YY male tilapia of the GIFT strain, Oreochromis niloticus

The growth performance of three experimental groups consisting of mixed sex fish (control), hormone-treated fish and progeny of YY male tilapia, all originated from the genetically improved farmed tilapia (GIFT) strain was evaluated. Masculinization of sexually undifferentiated fry was achieved by providing a supplement of 5 mg of 17-α-methyltestosterone per kg of feed over a period of 21 days (after sac absorption). Both mixed sex and progeny of YY male groups were fed a standard commercial ration. Mixed sex fish did not deviate significantly ( P >0.05) from the 1:1 male to female ratio. Percentages of male averaged 75% in hormone-treated fish and 95% in YY male group over the sampling periods and at final harvest. The effect of sex on weight and length was statistically significant ( P <0.001). The model used to analyse weight and length included experimental group and sex in each culture period as the fixed effects, and replicate cages as the random effect. Over the culture period of 141 days, there were no statistical differences ( P >0.05) in body weight and length between mixed sex, hormonally treated and progeny of YY males. There were also no significant differences in level of variability in harvest weight between three groups of fish when the data were classified into five categories (=<100, 100 to <150, 150 to <200, 200 to <250 and ≥250 g). It is concluded that monosex culture of all male tilapia would be of no advantage over mixed sex culture for the GIFT strain under conditions of cages suspended in earthen ponds.