Masculinization of Convict Cichlid (<Emphasis Type="Italic">Cichlasoma nigrofasciatum</Emphasis>) by immersion in <Emphasis Type="Italic">Tribulus terrestris</Emphasis> extract

We have examined the effects of Tribulus terrestris (TT), a non-toxic herb, on sex reversal in the Convict Cichlid Cichlasoma nigrofasciatum with the aim of introducing a new environmentally friendly method for masculinization in C. nigrofasciatum. TT is a natural plant product that elevates the testosterone levels in humans and animals. Different concentrations (0.0, 0.10, 0.20 and 0.30 g/l) of TT extract were tested for their effect on sex reversal in C. nigrofasciatum by immersing newly hatched offspring once weekly for 2 months in TT extract. Of the dosages used in the present study, 0.30 g/l TT was the most effective in terms of masculinization, resulting in a maximum male ratio of 87.23% (P < 0.001). Sex ratios of 79 and 85% at 0.10 and 0.20 g/l TT, respectively, were also significantly different from the expected 1:1 ratio (P < 0.001). Histological examinations revealed that the testes of fish treated with TT extract contained all stages of spermatogenesis, clearly demonstrating that the administration of TT extract to C. nigrofasciatum stimulated spermatogenesis. Total survival rates in all treatments and the control were uniformly high, ranging from 88.57 to 90% (P > 0.05). We concluded that TT had no negative effect on the survival rate of C. nigrofasciatum. In addition, all groups of TT-treated fish exhibited successful growth acceleration compared to the control group, but only the 0.30 g/l TT treatment significantly improved the growth rate of C. nigrofasciatum. (P < 0.01). Sex reversal in C. nigrofasciatum demonstrated that TT-treated 0-day-old larvae showed successful sex reversal, spermatogenesis and a better growth rate than untreated progenies.     

Tench (<Emphasis Type="Italic">Tinca tinca</Emphasis> L.) larvae rearing under controlled conditions: density and basic supply of <Emphasis Type="Italic">Artemia</Emphasis> nauplii as the sole food

After artificial reproduction of tench, larvae must be maintained indoors, and studies on rearing conditions are needed, focussing on the reduction of labour and costs. Three experiments on larvae (5th day post-hatch) were conducted for 25 days using Artemia nauplii as the sole food in order to determine basic feeding and density conditions during the first rearing period. Tench were maintained in 25 l fibreglass tanks, supplied with an artesian water flow throughout of 0.2 l min⁻¹. Water temperature was 22.5 ± 1°C, and the photoperiod was natural. Larvae fed on a restricted amount of nauplii reached high survival rates, even with the minimum of 50 nauplii larva⁻¹ day⁻¹. This amount of food may be sufficient at least for the first 25 days of exogenous feeding if fast growth is not the priority, and high densities can be maintained with good survival rates (over 90% up to 160 larvae l⁻¹ and 77% with 320 larvae l⁻¹). When food was supplied in excess once a day, high survival rates were achieved (91–97%), without differences among the densities tested. Animals at a density of 100 l⁻¹ reached the highest length (15.57 mm) and individual weight (46.8 mg). This growth is greater than those reported in studies feeding several times a day. It could be deduced that, while live food remains available for tench, it is not necessary to feed so frequently. Considering the relationship among the initial number of animals, final survival and growth and ration supplied, the new data reported here are useful to establish suitable stocking densities under both culture and experimental conditions.     

Fortification of <Emphasis Type="Italic">niboshi</Emphasis> with histidine prepared from <Emphasis Type="Italic">katsuobushi</Emphasis> protein digest

ABSTRACT:   Free and total histidine contents in the muscle of various fishes and invertebrates caught in the Seto Inland Sea were estimated. These contents were higher in the muscles from skipjack, yellowfin tuna, yellowtail and mackerel than in those from lizard fish, sea bream and hairtail. Invertebrate muscles, except squid, contained low free and total histidine. Boiled and dried anchovy ( niboshi ) contained lower free histidine than that of dried anchovy (suboshi). To prepare free histidine-rich extracts, the waste residue of soup stock from smoke-dried and shaved skipjack ( katsuobushi ) was digested with proteases. The extract of the enzyme digest contained high free histidine and was used to fortify niboshi with free histidine. If taken, the fortified niboshi may suppress food intake, and prevent obesity.     

Optimum temperature and salinity conditions for growth of green algae <Emphasis Type="Italic">Chlorella ellipsoidea</Emphasis> and <Emphasis Type="Italic">Nannochloris oculata</Emphasis>

ABSTRACT:   The effects of temperature and salinity on growth of green algae Chlorella ellipsoidea and Nannochloris oculata were determined to compare the optimum culture conditions. A four-temperature (15, 20, 25, and 30°C) × three-salinity (10, 20, and 30) factorial design with triplicates was applied. Specific growth rate (SGR), maximum density, and duration to reach maximum density of C. ellipsoidea were significantly affected by both temperature and salinity. The highest SGR was observed in C. ellipsoidea at 25°C and salinity 10, but the maximum density was very low. The highest maximum density was achieved in C. ellipsoidea at 15°C and 10. The slope constant of the linear relationship between semilogarithmic growth of C. ellipsoidea and day of culture was highest at 15°C and 10. The SGR and duration to reach maximum density of N. oculata were significantly affected by both temperature and salinity. However, maximum density of N. oculata was significantly affected by temperature, but not salinity. The highest maximum density was achieved in N. oculata at 25°C and 30, but SGR was significantly lower than that of N. oculata at 25°C and 10. The slope constant of the linear relationship between semilogarithmic growth of N. oculata and day of culture was highest at 25°C and 30. Based on these results, the condition of 15°C and salinity 10 seemed to be optimal for maximum density of C. ellipsoidea , and the condition of 25°C and 10 and 30 for SGR and maximum density for N. oculata , respectively.     

Chemical composition of dried eyeballs from <Emphasis Type="Italic">Euphausia superba</Emphasis> and <Emphasis Type="Italic">Euphausia pacifica</Emphasis>

ABSTRACT:   Dried krill eyeballs were obtained from Euphausia superba and Euphausia pacifica by using a patented industry method and their chemical compositions were examined. Crude protein content was 77.7% and 80.8% of the dry matter of E. superba and E. pacifica , respectively. The dominating amino acids in both krill were glutamic acid, aspartic acid, glycine, arginine, leucine and tyrosine. Crude fat content was 10.9% and 5.4% of dry matter of E. superba and E. pacifica , respectively. The main lipid class of the extracted lipids was phospholipids at 88.5% in E. superba and 96.4% in E. pacifica . The dominating fatty acids in both krill were 22:6 ( n- 3), 20:5 ( n- 3), 16:0 and 18:1 ( n- 9). Astaxanthin (3, 3'-dihydroxy-β, β-carotene-4, 4'-dione) content of E. superba and E. pacifica was 566 mg/100 g and 252 mg/100 g of dry matter, respectively. Retinol of E. superba and E. pacifica was 153.0 mg/100 g (510 000 IU/100 g) and 57.6 mg/100 g (192 000 IU/100 g) of dry matter, respectively. The lipophilic extract of E. superba by using n -hexane contained 1923 mg/100 g of astaxanthin at approximately four times higher than the dried eyeballs.     

Dietary amino acid <Emphasis Type="SmallCaps">l</Emphasis>-threonine requirement of fingerling Indian catfish, <Emphasis Type="Italic">Heteropneustes fossilis</Emphasis> (Bloch) estimated by growth and biochemical parameters

An eight-week feeding experiment was conducted to quantify the dietary threonine requirement of young catfish, Heteropneustes fossilis (9.20 ± 0.85 cm, 3.60 ± 0.45 g) using isonitrogenous and isoenergetic diets [40% crude protein (CP); 4.28 kcal g/100 g, gross energy (GE)] containing casein, gelatin and l-crystalline amino acids. Six dietary treatments supplemented with graded levels of l-threonine (0.50, 0.75, 1.00, 1.25, 1.50 and 1.75 g per 100 g, dry diet), in gradations of 0.25 g per 100 g dry diet were formulated. Fish were randomly stocked, in triplicate groups, in 55-l indoor polyvinyl flow-through circular tanks and fed experimental diets at 4% of their body weight divided over two equal feedings at 08:00 and 16:00 hours. Feeding schedule and ration size were worked out prior to the start of the feeding trial. Live weight gain (263%), feed conversion ratio (FCR) (1.35) and protein efficiency ratio (PER) (1.85) were significantly higher (P < 0.05) in fish fed a diet containing 1.25% dietary threonine. However, second-degree polynomial regression analysis of live weight gain, FCR, PER and body protein deposition data indicated the dietary threonine requirement to be 1.37, 1.26, 1.23 and 1.24 g per 100 g of dry diet, respectively. Whole-body moisture decreased significantly (P < 0.05) with the increase of dietary concentration up to 1.25%. A significantly (P < 0.05) higher protein content was evident in fish fed a diet containing 1.25% threonine. Body fat increased significantly (P < 0.05) with the increase of dietary concentration and was found to be highest at a 1.75% threonine concentration. A significantly (P < 0.05) higher ash content was reported at the 0.50 and 0.75% threonine levels. Body protein deposition was also found to be significantly (P < 0.05) higher at the 1.25% threonine level, followed by the 1.50% threonine level. Based on these results, it is recommended that the diet for fingerling H. fossilis should contain threonine at a level of 1.27 g per 100 g of dry diet, corresponding to 3.17 g per 100 g of dietary protein for optimum growth and efficient feed utilization. No mortality was observed during the experiment.     

Sterols from bivalves <Emphasis Type="Italic">Calyptogena soyoae</Emphasis> and <Emphasis Type="Italic">Bathymodiolus septemdierum</Emphasis> living in deep sea

From the two species of bivalves, Calyptogena soyoae around a cold seep and Bathymodiolus septemdierum near hydrothermal vents in the sea, sterols were isolated using high-pressure liquid chromatography analysis of the lipid fraction guided by characteristic ¹H NMR signals of the sterol skeleton. The minor sterol composition of C. soyoae included 24-methylenecycloartanal, cycloeucalenol, and obutusifoliol, which are known phytosterols. From B. Septemdierum, lathosterol and cholesterol as main sterols together with more diverse sterols were obtained. The difference between these species and their sterol contents is most likely because of feeding modes and metabolism of nutrients from their habitat.     

Suspension feeding and growth of ark shell <Emphasis Type="Italic">Anadara granosa</Emphasis>: comparison with ubiquitous species <Emphasis Type="Italic">Scapharca subcrenata</Emphasis>

The ark shell Anadara granosa is a species peculiar to the Ariake Sound. To determine why distribution of A. granosa in Japan is largely confined to this area, we examined feeding and growth of A. granosa as functions of environmental and biological variables. The results were compared with those of another ark shell Scapharca subcrenata, which is ubiquitous in Japan. Feeding experiments indicated that A. granosa is eurythermal and euryhaline, as is S. subcrenata, but is adapted to temperature slightly higher than S. subcrenata. Weight-specific clearance rate (CR) of A. granosa as a function of soft-body dry-weight (w) followed the power function of w (CR=2.7×w ^−0.37), with coefficient and exponent very close to those for S. subcrenata. Growth rate of A. granosa increased linearly with daily ration, similar to S. subcrenata. Thus, feeding and growth characteristics of A. granosa were comparable to those of S. subcrenata and no ‘peculiarities’ of the former were detected. Therefore, the factors that make A. granosa a species restricted to the sound are probably not directly related to feeding or growth characteristics.     

A comparative account of the structure of the growth hormone encoding gene and genetic interrelationship in six species of the genus <Emphasis Type="Italic">Labeo</Emphasis>

The growth hormone (GH) gene isolated and cloned from various Labeo species (L. rohita, L. calbasu, L. fimbriatus, L. gonius, L. bata, and L. kontius) is shown to contain a single copy in the haploid genome, with an overall size of ∼2.5 kb. The GH gene in all the Labeo species studied has five exons and four introns of various sizes with the exon/intron boundary sequence of GT/AG. The length variation of the GH gene between the species is found to be due to length variation in the form of several deletions in the third intron. The length of individual exons is the same in all the species with an open reading frame (ORF) of 630 bp (210 amino acids) except in L. rohita, which has a 9 bp deletion in the fourth exon, resulting in a shorter GH of 621 bp (207 amino acids). The similarity in the nucleotide and amino acid sequences between the different Labeo species is greater than 97%, in spite of eight amino acids being altered in the GH protein of Labeo that reside outside the conserved domain sequence required for its function. Nucleotide substitutions are seen in the form of 20 transitions and three transversions in the ORF of the GH gene. Both types of transitions (A–G; T–C) and only one type of transversion (A–C) are detected in the GH gene. Codon preference in GH gene shows a strong preference for G and C in the wobble position of the codons. Genetic interrelationships determined between Labeo and other species of fishes using nucleotide sequence of GH cDNA supports the overall teleost classification of Nelson (Fishes of the World. Wiley, New York, 1984) with separate clades for Ostariophysi, Protacanthopterygii, and Acanthopterygii. Besides, the unweighted pair group method with arithmetic means (UPGMA) analysis clearly distinguishes between the species having five exons and four introns in the GH gene from the species having six exons and five introns in the same gene. The Labeo species analyzed in the present study could be clustered into two groups using the maximum-parsimony method on the intron sequences data of the GH gene.     

Hybridization success of three common European cyprinid species, <Emphasis Type="Italic">Rutilus rutilus,Blicca bjoerkna</Emphasis> and <Emphasis Type="Italic">Abramis brama</Emphasis> and larval resistance to stress tests

ABSTRACT:   Hybridization success at early developmental stages and larval resistance to osmotic, thermal and fasting tests in roach Rutilus rutilus , silver bream Blicca bjoerkna , common bream Abramis brama and their F1 hybrids were investigated. Results revealed that hybrid survival rates were similar to parents. At the eyed embryo stage, however, a maternal effect was observed as a general trend during hatchling and larval stages. After these stages, hybrids displayed a higher survival rate than their parents. Under stress tests, no survival was observed after 40 min for osmotic and thermal shocks and after 24 days for the prolonged fasting test in these species and their F1 hybrids. The median survivals of hybrids were intermediate between the two parents. For total mortality, hybrids were also affected by a maternal effect but to the advantage of the hybrids.     

The effect of different diets on the growth and mortality of juvenile pikeperch (<Emphasis Type="Italic">Sander lucioperca</Emphasis>) in the transition from live food to formulated feed

The efficiencies of four feeding methods were compared in the transition period from live food to dry feed in pikeperch (Sander lucioperca). Pond-nursed fry were trained to artificial feed without a transition (Group P) and with a transition, using a combination of dry feed and different kinds of live food—Chironomus larvae (Group C), Tubifex (Group T) and Daphnia (Group D)—during a 12-day-long trial. The ratio of dry feed for the transition groups was increased from 0 to 100% in 3-day intervals. The highest specific growth rates and condition factors were detected for Group C (4.2%, 1.46 ± 0.5) and Group T (4.2%, 1.48 ± 0.6), respectively. The survival percentages of the groups were as follows: 86.7 ± 9% (Group C), 78 ± 4% (Group T), 52 ± 1.4% (Group D) and 41 ± 9% (Group P). The rate of cannibalism ranged between 6.3 and 13.7% during the transition period. Larger-sized fish seemed to learn more rapidly to accept dry feed than their smaller counterparts.     

Effects of heavy metals in river waters in Japan on immobility and mortality of <Emphasis Type="Italic">Daphnia magna</Emphasis> and <Emphasis Type="Italic">Oryzias latipes</Emphasis> larvae

ABSTRACT:   Samples of river waters containing high concentrations of zinc and other heavy metals but low concentrations of other anthropogenic contaminants were collected to investigate the relationship between toxicity of heavy metals and naturally present organic matters or hardness, as well as the effects of heavy metals on aquatic organisms. Acute toxicity tests were conducted for the water samples using Daphnia magna and medaka Oryzias latipes . Almost all the D. magna died in river waters containing high concentrations of zinc, but O. latipes in the same waters were hardly affected. Since the test organisms were not only exposed to zinc but also other heavy metals in the river waters, we examined the toxicity using toxic units composed of zinc, copper, lead, and cadmium. The results of a bioassay with the river waters showed that the mortality of D. magna did not depend solely on the total number of toxic units of heavy metals. The organic matters and the hardness of the river waters could decrease the acute toxicity of zinc and other heavy metals to D. magna .     

Effects of fish CYP inducers on difloxacin <Emphasis Type="Italic">N</Emphasis>-demethylation in kidney cell of Chinese idle (<Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis>)

A drug–drug interaction occurs when the effect of one drug is altered by the presence of another drug which is generally associated with the induction of cytochrome P450s (CYPs) activity. Thus, unexpected treatment failures often happen resulting from inappropriate coadministration in fisheries. However, little information is available about CYP induction in fish. The reaction of difloxacin (DIF) biotransformation to sarafloxacin (SAR) belongs to N-demethylation catalyzed mainly by CYP(s). In order to supply useful information on CYP induction, the present study assessed the effects of fish-specific CYP inducers on DIF N-demethylation and enzyme kinetics in kidney cell of Chinese idle (CIK; grass carp (Ctenopharyngodon idellus)) by RP-HPLC. Results demonstrated that the amounts of SAR formation and enzymatic parameters Clint and Vmax were significantly increased due to β-naphthoflavone (BNF) pretreatment. Therefore, we suggest that CYP1A may be involved in DIF N-demethylation in CIK. This study provides instructive information to ensure treatment success via avoiding CYP induction in fisheries.     

Induction of (<Emphasis Type="Italic">Z</Emphasis>)-8-heptadecene in marine green alga <Emphasis Type="Italic">Bryopsis maxima</Emphasis> by mechanical wounding

ABSTRACT:   After mechanical wounding of the marine green alga Bryopsis maxima in a phosphate buffer, one main organic component was released to the solution. The organic compound was characterized by a molecular ion at 238  m/z by gas chromatography–mass spectrometry (GC-MS) and was deduced as 8-heptadecene by the fragment pattern. To identify the stereochemistry of 8-heptadecene, synthetic ( Z / E )-8-heptadecene was coinjected to the solution of wounded B. maxima . Comparing retention times and mass spectra with the synthetic sample, the target hydrocarbon obtained from wounded B. maxima was coincident with ( Z )-8-heptadecene.     

Effects of stocking density on survival and growth of juvenile tench (<Emphasis Type="Italic">Tinca tinca</Emphasis> L.)

In two 120-day experiments, performed in the laboratory at 22°C, the effects of stocking density on the survival and growth of juvenile tench (Tinca tinca L.) were evaluated. Fish were kept in fibreglass tanks, supplied throughout with flow of artesian water, and fed a dry diet for salmonids, in excess, supplemented with restricted amounts of Artemia nauplii. In the first experiment four-month-old juveniles (0.31 ± 0.04 g and 32.00 ± 1.17 mm TL) were stocked at four densities—0.18, 0.88, 1.05, and 2.10 g l⁻¹. Survival was high (>89%) for all treatments. Final densities ranged between 1.10 g l⁻¹ (significantly lowest) and 10.46 g l⁻¹ (significantly highest). The density increase was significantly higher (611%) for fish stocked at the lowest initial density (0.18 g l⁻¹) than for fish stocked at 0.88, 1.05, and 2.10 g l⁻¹, for which the density increase averaged 457%. In the second experiment, 4.5-month-old juveniles (0.58 ± 0.17 g and 39.54 ± 0.83 mm TL) were stocked at three densities—1.05, 3.00, and 4.00 g l⁻¹. Survival was high (>96%) for all treatments. Final densities ranged between 4.08 and 16.53 g l⁻¹ and were significantly higher for greater initial densities. The density increase was greatest (413%) for fish stocked at the highest density (4 g l⁻¹) and was not significantly different for fish stocked at 1.05 and 3 g l⁻¹. Considering all the densities in the two experiments, for stocking at 4 g l⁻¹ the final density was 15 times higher than that reached after stocking at 0.18 g l⁻¹, without harmful effects on survival and growth. This final density (equivalent to 16.53 kg m⁻³) is in the range recommended for other fish species in this period under intensive conditions.     

Effect of stocking density on survival and growth performance of pikeperch, <Emphasis Type="Italic">Sander lucioperca</Emphasis> (L.), larvae under controlled conditions

The effect of stocking density on the survival and growth of pikeperch, Sander lucioperca (L.), larvae was examined in two consecutive experiments. In experiment I, 4-day-old larvae [body wet weight (BW): 0.5 mg; total body length (TL): 5.6 mm] were reared in 200-l cylindro-conical tanks in a closed, recirculating system (20 ± 0.5°C) at three stocking densities (25, 50 and 100 larvae l⁻¹) and fed a mixed feed (Artemia nauplii and Lansy A2 artificial feed) for 14 consecutive days. At densities of 25 and 100 larvae l⁻¹, growth rate and survival ranged from 2.7 to 1.9 mg day⁻¹ and from 79.2 to 72.3%, and fish biomass gain ranged from 0.6 to 2.0 g l⁻¹, respectively. There were two periods of increased larval mortality: the first was at beginning of exogenous feeding and the second during swim bladder inflation. In experiment II, 18-day-old larvae (BW: 35 mg; TL: 15.6 mm) obtained from experiment I were reared under culture conditions similar to those of experiment I, but at lower stocking densities (6, 10 and 15 larvae l⁻¹). The fish were fed exclusively with artificial feed (trout starter) for 21 consecutive days. At densities of 6 and 15 larvae l⁻¹, the growth rate and fish biomass gain ranged from 28.8 to 23.1 mg day⁻¹ and from 2.0 to 3.3 g l⁻¹, respectively. The highest survival (56.5%) was achieved at a density of 6 larvae l⁻¹. Mortality at all densities was mainly caused by cannibalism II type behaviour (27–35% of total). In both experiments, growth and survival were negatively correlated and fish biomass gain positively correlated with stocking densities. The present study suggests that the initial stocking density of pikeperch larvae reared in a recirculating system can be 100 individuals l⁻¹ for the 4- to 18-day period post-hatch and 15 individuals l⁻¹ for the post-19-day period.     

Growth of wild pearl oysters <Emphasis Type="Italic">Pinctada fucata,Pinctada margaritifera</Emphasis> and <Emphasis Type="Italic">Pinctada sugillata</Emphasis> (Bivalvia: Pteriidae) in Taiwan

In order to understand growth features of pearl oysters in the genus Pinctada, i.e. Pinctada fucata, Pinctada margaritifera, and Pinctada sugillata in Taiwan, a total of 3062 wild individuals of these species from juvenile to adult were collected monthyly from March 2001 to April 2002 in Jukeng, Pingtung County, south-west Taiwan. Quantitative measurements of live oysters were conducted for shell height (SH), shell length (SL), shell width (SW), hinge length (HL), and wet weight (WW). Different cohorts were identified through multiple length frequency analysis on SH of P. fucata and P. margaritifera, and growth curves with seasonal variation were estimated for these species. Pinctada fucata in Taiwan had a different seasonal growth pattern from the Japanese population, but had similar growth rates during the high growth period. The growth rate of P. margaritifera in Taiwan was slower than in French Polynesia, the Solomon Islands, and the Red Sea. Comparisons of morphological growth features among the three species show large differences in the SW-related features. Pinctada fucata in Taiwan had larger SW than in Japan and Korea. The differences in growth rates and morphological features suggested that the wild Taiwanese oysters may retain genetically pristine characteristics, thus genetic conservation might be urgently needed.     

Improving growth in juvenile turbot (<Emphasis Type="Italic">Scophthalmus maximus</Emphasis> Rafinesque) by rearing fish in switched temperature regimes

The effect of thermal history (16 and 20°C) on growth of juvenile turbot, Scophthalmus maximus (initial mean weight 72.6 g, n = 157) was studied. Fish were divided into four groups, two groups remaining at constant temperature (C16, C20), while fish in the other groups were transferred from either 16 to 20°C (F16-20) or from 20 to 16°C (F20-16). Between 35 and 42 fish in each tank were individually tagged at the start of the experiment. The final mean weights were significantly higher in the F20-16 group (230 g) than in the C20 (213 g), F16-20 (211 g) and C16 (205 g) groups. The overall growth rate was highest in the F20-16 group (1.17% day⁻¹) but comparable in the three other groups (1.00–1.04% day⁻¹). Our findings indicate that, even at near-optimal temperature for a given size, the temperature history of the fish may influence future growth. Based on these indications, we conclude that as turbot grow larger, the temperature should be reduced to take advantage of the change in optimal temperature for growth with increasing fish size rather than rearing at constant temperatures.