A non‐paradoxical dose response to 17α‐methyltestosterone by Nile tilapia Oreochromis niloticus (L.): effects on the sex ratio,growth and gonadal development

Commercial tilapia production is dependent on monosex culture, commonly obtained through the inclusion of an androgen in the diet for a brief period soon after hatch. To determine a minimum effective dose and identify the problems associated with over‐dosing, Nile tilapia Oreochromis niloticus fry were fed diets containing methyltestosterone (MT) at rates up to 1200 mg kg^?1 of diet for 28 days. The minimum effective dose for ≥95% males was 14 mg MT kg^?1 diet. Percent phenotypic males increased as the rate increased from 3.75 (80%) to 30 mg kg^?1 MT (99%). Methyltestosterone given at rates of 120 mg kg^?1 or more reduced efficacy but did not result in a reduced frequency of males relative to that of non‐treated fish. The term ‘paradoxical feminization’ does not adequately describe the observed sex ratios, where no fish were feminized but the efficacy of MT at high doses to masculinize females was reduced. At 1200 mg MT kg^?1, the frequency of females (48 ± 1%) was not different from that in the non‐treated population. The mechanism for the reduced efficacy is not clear and is not adequately explained as an aromatization of androgen to oestrogen response.     

Hormonal sex reversal of tilapias: implications of hormone treatment application in closed water systems

The phenomenon of incidental sex reversal was observed in experiments originally designed to determine the importance of feeding duration and feed quality in the optimization of masculinization and feminization treatments of Oreochromis niloticus (L.) fry by oral application of hormones. Four separate experiments were conducted: (1) masculinization and (2) feminization using three feeding durations and (3) masculinization and (4) feminization using three alternative feed mixtures. Masculinization experiments incorporating 17 α-methyltestosterone (MT) into the feed at 40 mg kg^?1 were applied to putative monosex male progeny whilst feminization treatments were applied to putative all female fry using diethylstilboestrol (DES) at 1000 mg kg^?1. Treatments were applied outdoors in small cages suspended in 900-l circular concrete tanks with a flow through rate of 1-l min^?1. Replicate cages for all treatments and the controls in each experiment were reared together in the same tank. Only the first experiment produced significant differences in sex ratio, with the control treatment producing a slightly lower proportion of males than the three MT treatments at different durations. The most significant finding was that in all experiments the sex ratio of the control differed significantly from expectations and in all but experiment 1, did not differ from those of the treated fish in which sex reversal had clearly been effected. It appears likely that incidental sex reversal occurred in the control fish as a result of the accumulation of active metabolites from the treated fish and/or leachates from uneaten food given to fish in the same water body. The results are discussed in the context of improving the efficiency and safety of hormonal sex reversal treatments.     

Efficacy of 17 α‐methyl testosterone and letrozole on sex reversal of protogynous grouper,Epinephelus tauvina (Forskal, 1775) and spawning performance of sex‐reversed males

This study aimed to test the efficacy of 17 α‐methyl testosterone (17 α‐MT) alone and in combination with letrozole, an aromatase inhibitor, for the induction of sex reversal in protogynous greasy grouper, Epinephelus tauvina. Further, the long‐lasting effects of these treatments and spawning performance of sex‐reversed males were also investigated. Greasy grouper with oocytes in the perinucleolus stage were implanted with 5 mg 17 α‐MT kg^?1 body weight (T₁), 5 mg 17 α‐MT and 0.2 mg letrozole kg^?1 body weight (T₂) and 5 mg 17 α‐MT with 0.4 mg letrozole kg^?1 body weight (T₃) and no androgens/enzyme inhibitor implanted (C). The 17 α‐MT alone and in combination of letrozole‐induced sex reversal in greasy grouper, whereas untreated control fish (C) showed normal ovarian development. However, T₂ and T₃ group showed 100% sex reversal and completion of spermatogenesis up to functional male phase in 2 and 3 months, respectively, whereas T₁ group resulted in only 66.67% functional male with motile spermatozoa after 4 months. Sex‐reversed males successfully fertilized the eggs during induced spawning. There were significant differences on fertilization and hatching rates between T₂ group (79.00 ± 4.36%; 77.67 ± 2.87%, respectively) and T₁ group (57.67 ± 3.17%; 63.87 ± 2.91%, respectively). The result suggested that 17 α‐MT (5.0 mg kg^?1 BW) in combination with letrozole (0.2 mg kg^?1 BW) has the potential to produce 100% sex‐reversed male in short period in greasy grouper, which might greatly help in seed production of greasy grouper.     

Influence of β-hydroxy-β-methylbutyrate on nonspecific humoral defense mechanisms and protection against furunculosis in pikeperch (Sander lucioperca)

Studies have shown that in both in vitro and in vivo tests, β‐hydroxy‐β‐methylbutyrate (HMB) increases the nonspecific cellular and humoral immune response and protection against diseases in animals. The present study examines the influence of HMB on nonspecific humoral defense mechanisms and protection against furunculosis in pikeperch (Sander lucioperca). β‐hydroxy‐β‐methylbutyrate was fed in a pelleted ration of 50 mg kg^?1 feed day^?1 for 4 weeks. Blood was drawn from 12 HMB‐fed and control‐fed pikeperch. The lysozyme and ceruloplasmin activities in the plasma, total immunoglobulin (Ig) levels, and total serum protein were analysed prior to and then after 2 and 4 weeks of HMB ingestion. After 4 weeks of HMB ingestion, a challenge test was performed by injecting the fish with live pathogenic Aeromonas salmonicida bacteria. β‐hydroxy‐β‐methylbutyrate at a dose of 50 mg kg^?1 feed resulted in a statistically significant (P<0.05) increase in the lysozyme activity of the plasma, total Ig, and serum protein levels. Additionally, reduced mortality (40%) after the in vivo challenge with pathogenic A. salmonicida suggested that HMB‐activated nonspecific protection against furunculosis in pikeperch.     

Maximum limits of organic and inorganic mercury in fish feed

The relatively high levels of mercury found in fish feeds might form a fish health and food safety risk. The present study aims to establish sublethal toxic threshold levels in fish and assess feed‐fillet transfer of dietary mercury. Atlantic salmon (Salmo salar L.) parr were fed for 4 months on fish meal‐based diets supplemented with mercuric chloride (0, 0.1, 1, 10 or 100 mg Hg kg^?1 dry weight (DW)) or methylmercuric chloride (0, 0.1, 0.5, 5 or 10 mg MeHg kg^?1 DW). At the end of the experiment, dietary inorganic mercury mainly accumulated in intestine (80% of body burden) and assimilation was low (6%). In contrast, methylmercury readily accumulated in internal organs and muscle (80% of body burden) and had a relatively high assimilation (23%). Highest accumulation of dietary inorganic mercury was observed in the gut and kidney. Fish fed 10 mg Hg kg^?1 had an early (after 2 months) significant increase in renal metallothionein (MT) level and intestinal cell proliferation, followed by intestinal pathological conditions after 4 months of exposure. At 100 mg Hg kg^?1, intestinal and renal function were reduced as seen from the significantly reduced protein and glycogen digestibility and increased plasma creatinine levels. For dietary methylmercury (MeHg), highest accumulation was found in blood and muscle. Intestinal cell proliferation and liver MT significantly increased at 5 mg MeHg kg^?1 after 2 months of exposure. At the end of the experiment, blood haematology was significantly affected in fish fed 5 mg MeHg kg^?1 and these fish exceeded the current food safety limit for mercury. Tissue MT induction and intestinal cell proliferation appeared to be useful and quantifiable early indicators of toxic mercury exposures. Based on the absence of induction of these early biological markers such as MT and cell proliferation, nonobserved effect levels (NOELs) could be set to 0.5 mg Hg kg^?1 for dietary methylmercury and 1 mg Hg kg^?1 for inorganic mercury. Lowest observed effect levels (LOELs) levels could be set to 5 mg kg^?1 for methylmercury and 10 mg Hg kg^?1 for inorganic mercury.     

Induction of Out-of-Season Spawning of Pikeperch, Sander Lucioperca (L.)

Pikeperch were induced to spawn 3 months prior to the natural spawning period through photothermal and hormonal stimulation. Females (five specimens in each group) were stimulated with injection of human chorionic gonadotropin (hCG) once (200 IU kg^–1), twice (200 IU kg^–1, second dose after 48 h–400 IU kg^–1) or three times (200 IU kg^–1, after 24 h–200 IU kg^–1 and after another 24 h–200 IU kg^–1). The control group was injected once with 0.9% NaCl. The males were stimulated with a single hormone dose of 200 IU kg^–1. Eggs were obtained from all the hormonally treated fish. None of the control group females, which were only stimulated photothermally, ovulated any eggs. The time of ovulation was 66–71 h following the first injection, and the eggs viability until the eyed stage (from 71.5 to 77.5%) did not depend on the number of hormone doses (P > 0.05). The out-of-season spawning method described in this paper could be used to provide pikeperch larvae for intensive culture systems (recirculating water systems) before natural spawning season and to produce larger-sized pikeperch fingerlings for stocking.     

The effects of different proportions of the 17β‐estradiol and 17α‐methyltestosterone on growth,sex reversal and skin colouration of the electric blue hap (Sciaenochromis ahli Trewavas, 1935)

This study was performed to investigate the effects of 17β‐estradiol (ES) and 17α‐methyltestosterone (MT) on growth, development, survival, sex ratio and colour change in the electric blue hap (Sciaenochromis ahli Trewavas, 1935). The hormones were not supplemented to the control feed, while six other feeds were prepared by adding 20, 40 and 60 mg kg^?1 17β‐ES or 20, 40 and 60 mg kg^?1 17α‐MT to each, resulting in seven different feed treatments. Average live weight of the fish supplemented with these diets was 0.42 ± 0.04 g. At the end of the study, the highest weight gain was observed in fish fed 60 mg kg^?1 17α‐MT group (2.62 ± 0.11 g) and the difference with the groups fed with 17β‐ES was found to be significant. All fish fed 17α‐MT were male, while the rates of feminization in fish fed 17β‐ES at 20, 40, 60 mg kg^?1 were 91.11%, 88.88% and 93.33% respectively. Survival rates were respectively determined as 80%, 95.56%, 84.44%, 93.33%, 77.78%, 84.44% and 84.44% for the control, 20, 40, 60 mg kg^?1 17β‐ES and 20, 40, 60 mg kg^?1 17α‐MT treatments. The best colouration was achieved in the 17α‐MT groups (P < 0.05). The L* values varied between 32.98 ± 4.44 and 61.35 ± 2.19, a* values between ?7.06 ± 0.22 and ?3.42 ± 0.11, and b* values between ?7.74 ± 0.10 and 11.65 ± 0.03, while Chroma (C*) and Hue (H°_ab) angle values varied between 7.54 ± 0.22 and 13.60 ± 0.01 and between 119.76 ± 0.05 and 239.73 ± 4.86. In conclusion, the 17α‐MT feeding was found to have a greater effect on the growth, feed conversion ratio, masculunization and pigmentation of the electric blue haps than the 17β‐ES treatment.     

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.     

Effects of a nonsteroidal aromatase inhibitor on gonadal differentiation of bluegill sunfish Lepomis macrochirus

In the present study, the efficacy of Letrozole, a potent nonsteroidal aromatase inhibitor (AI), on gonadal sex differentiation and sex reversal was examined in bluegill sunfish (Lepomis macrochirus). In Experiment 1, using AI diet treatments (50, 150, 250 and 500 mg kg⁻¹) from 30 to 90 days posthatch (dph), AI interrupted ovarian cavity formation at a dose of 500 mg kg¹ diet and one intersex fish was identified in this group. The proportions of males in all the treated groups were significantly higher than those in the control group. In Experiment 2, using AI immersion treatments (250, 500 and 1000 μg L⁻¹) during 30–50 dph, the treated groups of 500 and 1000 μg L⁻¹ produced significantly more males than the control and 250 μg L⁻¹ groups. Histological examination revealed no differences in ovary or testis tissue between control and AI‐treated fish. There were no significant differences detected in body weight and length among the AI treated and control groups (P>0.05) for both experiments. The results from these two experiments suggest that inhibition of aromatase activity by AI could influence sex differentiation in bluegill sunfish.     

Supplementing the feed of pikeperch [Sander lucioperca (L.)] juveniles with MacroGard and its influence on nonspecific cellular and humoral defense mechanisms

This study examined the influence of β‐1.3/1.6‐glucan (MacroGard) on the innate immune system in healthy pikeperch (Sander lucioperca) juveniles. MacroGard was fed in a pelleted ration of 1 or 2 g kg^?1 feed for 6 weeks. The control group of fish was fed feed with no supplement. Blood, pronephros and spleen samples for the study of nonspecific cellular and humoral defense mechanisms were collected from 10 fish from each group. No changes were observed in either the behaviour or the health of the fish throughout the rearing period. Fish mortality was not noted in any of the groups. Additionally, the supplementation of feed with MacroGard did not have a significant impact on the fish growth rate as expressed in both absolute and relative values (P>0.05). The condition of the fish from the experimental group did not differ from that of the control group specimens, and the feed conversion ratios were also very similar (P>0.05). The results showed that MacroGard administered in two doses significantly (P<0.05) activated the metabolic activity and potential killing activity of spleen phagocytes, compared with the control‐fed pikeperch. The highest phagocyte activity was observed with a dose of 2 g kg^?1. The proliferative response of pronephros lymphocytes indicated a similar pattern. MacroGard significantly increased proliferative response of lymphocytes stimulated by mitogens (P<0.05). MacroGard also significantly increased the lysozyme activity and total immunoglobulin levels in the serum. In this study neither significantly different ceruloplasmine activity or total protein level in serum were observed. The result showed that MacroGard at the optimal dose of 1 g kg^?1 feed and a twofold higher dose does not have a negative influence on the hepatocytes or increased nonspecific cell‐ and humoral‐mediated immunity in healthy pikeperch.     

Induced spawning in perch, Perca fluviatilis L. using carp pituitary extract and HCG

Induced spawning in Eurasian perch, Perca fluviatilis L., was studied using human chorionic gonadotrophs (hCG) or acetone-dried common carp pituitary with or without hCG. The dose administered to fish was 5700 IU hCG kg^-1 or 4.0 mg kg^-1 carp pituitary with or without 500-700 IU hCG kg^-1 for females and 2850 IU hCG kg^-1 or 2.0 mg kg^-1 carp pituitary with or without 250-350 IU hCG kg^-1 for males. There were no statistically significant differences in quantity of milt in treated and control groups, although the best result was observed when males were treated with a triple injection of hCG and carp pituitary extract. Male spermiating success, expressed as a quantity of milt, was negatively correlated with fish weight. All females treated in this experiment had oocytes at the same division, so time of ovulation was highly synchronous. Spawning success, expressed as a spawning effectivity coefficient (S_c), was highest in fish treated with the triple injection. Spawning methods described in this paper were successful, even though hormones from another order of fish were used.     

Effects of dietary chitosan oligosaccharide complex with rare earth on growth performance and innate immune response of turbot,Scophthalmus maximus L.

An 8‐week‐feeding trial was conducted to investigate the effect of dietary chitosan oligosaccharide complex with rare earth (COS‐REE) on growth performance and innate immune response of turbot, Scophthalmus maximus L. (Initial average weight was (12.1 ± 0.1) g) as well as disease resistance against Edwardsiella tarda. Six practical diets (approximately 53.01% protein and 12.57% lipid) were formulated to contain graded levels (0, 75, 150, 300, 600 and 1200 mg kg^?1) of COS‐REE. Results of the present study showed that, compared to the control group (0 mg kg^?1), the specific growth rate (SGR) was significantly higher in fish fed the diet with 300 mg kg^?1 COS‐REE (P < 0.05), while the feed conversion ratio (FCR) significantly decreased (P < 0.05). The phagocytic index (PI) and the activity of super oxide dismutase (SOD) of serum in fish fed the diet with 300 mg kg^?1 COS‐REE was significantly higher than fish fed the control diet (P < 0.05), but no significant differences were observed in malondialdehyde (MDA) and hepatic metallothionein (MT) concentrations. After 8 weeks, fish were challenged by intraperitoneal injection with E. tarda, and COS‐REE‐treated fish demonstrated increased protection capability. These results suggested that COS‐REE could enhance growth, innate immunity and disease resistance in turbot, and the optimum dose was approximately 300 mg kg^?1.     

Effects of brewer’s yeast extract on growth performance and health of juvenile pikeperch Sander lucioperca (L.)

Juvenile European pikeperch, Sander lucioperca, were fed commercial feed (group C) or experimental feed supplemented with NuPro^® nucleotide‐rich Saccharomyces cerevisiae yeast protein (extract obtained through a cell wall removal process) in doses of 20, 40 or 60 g kg^?1 feed (groups N2, N4, N6) for 8 weeks. Growth, non‐specific immunity parameters, histological structure of the liver and intestine, proximate whole‐body composition and blood biochemical parameters were assayed. It was noted that brewer’s yeast extract has immunomodulatory proprieties. NuPro^® in doses of 40 and 60 g kg^?1 feed strongly stimulated non‐specific (innate) cellular and humoral immunity in pikeperch. The experimental feed did not have a significant impact on pikeperch growth (P > 0.05). The proximate composition of the fish bodies and the hepatosomatic and viscerosomatic (VSI) indices were also not affected, which indicated that the tested diets had no negative impact on the metabolism or deposition of nutrients in fish tissues. The lower levels of transaminases AST and ALT, which were noted in the groups with the two highest doses of NuPro^® (P < 0.05), might indicate improved liver function. It was also demonstrated that the brewer’s yeast extract stimulates the absorption activity of intestinal epithelial cells.     

Induced sex reversal of dusky grouper, Epinephelus marginatus (Lowe)

The present paper describes sex reversal in dusky grouper, Epinephelus marginatus (Lowe). Over a 4-month feeding period, 5 mg kg^?1 of the male hormone 17-α methyltestosterone was combined with frozen fish as feed. After 2.5 months of treatment, the gonads of all six fish treated in the experiment presented tissues with oocytes and spermatogonia, and after 3 months, there were only a few remnants of oogonial tissue left in the gonads. All fish in the experiment gave sperm of varying quality after 4 months of the trial. The quantity of the hormone administered to the fish over these 4 months was 89-113 mg kg^?1. Spermatozoid motility varied from 95% to 40%, and lasted for about one hour in sea water. Using the combined sperm of three males, fertilization ranged from 50% to 95%. Fertilized eggs and hatched larvae continued to develop normally.     

Induced spawning in bream, Abramis brama (L.), using carp and bream pituitary extract and hCG

Induced spawning in bream, Abramis brama (L), was studied using acetone-dried common carp pituitary (CP) and bream pituitary (BP) with or without the addition of human chorionic gonadotrophin (hCG). The total dose administered to fish was of 5.0 mg kg^?1 BP or 4.0 mg kg^?1 CP with or without 2000-2200 IU hCG kg^?1 for females and 2.5 mg kg^?1 BP or 2.0 mg kg^?1 CP with or without 1000–1100 IU HCG kg^?1 for males. In all male treated groups 100% of spermiation was observed: in females the most effective method was a triple injection with hCG and carp pituitary, resulting in 79% of females ovulated (over 68% of eyed eggs). Biological quality of eggs, expressed as a percentage of eyed eggs, was negatively correlated with time elapsing between resolving (final) injection and ovulation. Spawning success, expressed as a value of S_e (spawning effectiveness coefficient), was higher in fish treated with triple injection.     

The effect of 11 β-hydroxyandrostenedione on pikeperch Stizostedion lucioperca (L.)

Oral administration of 11 β-hydroxyandrostenedione was used to sex inverse juvenile Stizostedion lucioperca (L.) reared under intensive culture conditions. Androgen was mixed into the commercial trout food in the following doses: 0, 30, 60 and 90 mg kg^?1 diet. Administration (for 21 days) of 60 mg 11 β-hydroxyandrostenedione kg^?1 of the diet produced 93% males and 7% bisexual fish. In hormone-treated groups, no morphological changes were observed in the liver. No significant differences were observed in growth or in the hepatosomatic index between the androgen-treated groups and the control group.     

Spawning induction in Kutum Rutilus frisii kutum (Kamenskii, 1901) using carp pituitary extract or GnRH analogue combined with metoclopramide

Kutum Rutilus frisii kutum (Kamenskii, 1901), Cyprinidae is an endemic fish of the Caspian Sea. Iranian Fisheries Organization (Shilat) produce up to 200 million fry (1–2 g body weight (b.w.)) to restock the Caspian Sea population annually. Some of these fry are produced by spawning induction in broodfish by carp pituitary extract (CPE). The objective of this study was to assay the effectiveness of the gonadotropin releasing hormone analogue (d ‐Ala⁶, Pro⁹‐Net GnRH) alone or in combination with metoclopramide (MET), a dopamine antagonist, on the percentage of ovulated females, latency period, ovulation index and fertilization success. The following hormone treatments were tested: single injection of 2 mg kg^?1 b.w. of CPE as a positive control, GnRHa alone 20 and 40 μg kg^?1 b.w. and combination of GnRHa and MET as follows: 5 μg+2.5 mg, 10 μg+ 5 mg and 20 μg+10 mg kg^?1 b.w. Negative control group was injected with 0.7% saline. The percentage of ovulated females, ovulation index and fertilization success were 90%, 71.3±1.24%, 68.4±2.3%, respectively, in the group treated with GnRHa+MET at a dose of 20 μg+10 mg kg^?1 b.w. and were significantly higher than those in the positive control (60%, 64.5±0.23%, 69.1±4.5%) (P<0.05). However, the latency period in this group was longer than that in the positive control (P<0.05). Only 20% and 40% fish ovulated in groups that received 20 or 40 μg kg^?1 b.w. GnRHa. No fish ovulated in the negative control.     

Quantifying the dietary niacin requirement of the Indian catfish, Heteropneustes fossilis (Bloch), fingerlings

A growth study was conducted to determine the dietary niacin requirement of the Indian catfish, Heteropneustes fossilis (Bloch), fingerlings (Mean weight 9.41 ± 0.18 g). Semi‐purified diets with five levels (0, 5, 10, 20 and 40 mg kg^?1 diet) of supplemental niacin were fed to H. fossilis for 15 weeks. Each diet was fed to three replicate groups of fish. Results indicated that the highest (P < 0.05) weight gain was for the fish fed the diet supplemented with 20 mg niacin kg^?1, followed by fish fed the diets with 40, 10 and 5 mg niacin kg^?1, and the lowest in fish fed the unsupplemented control diet. Patterns of specific growth rate (SGR) and protein efficiency ratio (PER) were similar to those of the weight gain. Survival of fish fed the control diet and niacin‐supplemented diet was 58% and 91–100% respectively. Niacin deficiency signs such as anaemia, anorexia, lethargy and skin haemorrhage were observed in fish fed the control diet. The haematocrit values (Ht) were higher (P < 0.05) in fish fed the diets supplemented with niacin than in fish fed the control diet. The hepatosomatic indexes (HSI) of fish fed with or without niacin‐supplemented diets were not significantly (P > 0.05) different from each other. Both body protein and lipid content were higher (P < 0.05) in fish fed the diet supplemented with 20 and 40 mg niacin kg^?1, respectively, than those fish fed other diets. The niacin content in liver significantly (P < 0.05) reflected the supplementation level in the diet and ranged from 29.11 to 40.31 mg g^?1 tissue. The associated liver niacin content for growth was about 47 μg g^?1 tissue. Quadratic regression analysis showed that the dietary niacin requirement for maximal growth of H. fossilis under these experimental conditions was about 25 mg kg^?1 diet.     

The effect of feeding the leucine metabolite β-hydroxy-β-methylbutyrate (HMB) on cell-mediated immunity and protection against Yersinia ruckeri in pikeperch (Sander lucioperca)

The present study examined the influence of leucine metabolite β‐hydroxy‐β‐methylbutyrate (HMB), a natural product HMB, on nonspecific cell‐mediated defence mechanisms and protection against enteric redmouth disease (ERM) in pikeperch (Sander lucioperca). β‐hydroxy‐β‐methylbutyrate was fed in a pelletted ration at 50 mg kg^?1 of the feed for 8 weeks. The phagocytic ability and potential killing activity of blood and pronephric phagocytes were examined in HMB‐ and control‐fed fish before and after 8 weeks of feeding HMB. Simultaneously, the proliferative response of blood and pronephric lymphocytes stimulated by mitogens concanavaline A and lipopolisaccharide were examined in experimental and control groups. Following 8 weeks of HMB feeding, a challenge test was performed by injecting the fish with live pathogenic bacteria Yersinia ruckeri. β‐hydroxy‐β‐methylbutyrate applied in the diet for 8 weeks prompted a statistically significant (P<0.05) increase in phagocytic ability and potential killing activity of the blood and pronephric phagocytes and the proliferative response of blood and pronephric lymphocytes. The changes in these mechanisms correlated with protection against Y. ruckeri, the ERM pathogen. The results showed that feeding HMB increased the nonspecific cell‐mediated defence mechanisms and protection against ERM by reducing cumulative mortality (30%) following the challenge with pathogenic bacteria. Future studies will include determination of optimal doses and protocols of oral application of HMB to maximize the immunomodulatory effects and protection against viral diseases in intensive pikeperch culture.     

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.