The effects of starvation on fast-start escape and constant acceleration swimming performance in rose bitterling (<Emphasis Type="Italic">Rhodeus ocellatus</Emphasis>) at two acclimation temperatures

To investigate the effects of starvation and acclimation temperature on the escape ability of juvenile rose bitterling (Rhodeus ocellatus), we measured the fast-start escape and constant acceleration swimming performance of fish fasted for 0 (control), 1 and 2 weeks and half-lethal periods (6 or 4 weeks) at two temperatures (15 and 25 °C). Fish acclimated at a high temperature exhibited shorter response latency (R), higher maximum linear velocity (V _max) and longer escape distance during escape movement (D _120ms) than those at the low temperature. Starvation resulted in a significant decrease in V _max and D _120ms at either low or high temperature and a significant increase in R at only the high temperature in the half-lethal period groups (P < 0.05). The relationship between V _max (Y, m s^?1) and starvation time (X, week) was Y ₁₅ = ?0.062X + 1.568 (r = ?0.665, n = 36, P < 0.001) at low temperature and Y ₂₅ = ?0.091X + 1.755 (r = ?0.391, n = 40, P = 0.013) at high temperature. The relationship between U _cat (Y, cm s^?1) and starvation time (X, week) was Y ₁₅ = ?1.649X + 55.418 (r = ?0.398, n = 34, P = 0.020) at low temperature and Y ₂₅ = ?4.917X + 62.916 (r = ?0.793, n = 33, P < 0.001) at high temperature. The slopes of equations showed a significant difference between low and high temperature (F _1,63 = 9.688, P = 0.003), which may be due to the different energy substrate utilization when faced with food deprivation at different temperatures.     

The effects of starvation and re-feeding on growth and swimming performance of juvenile black carp (<Emphasis Type="Italic">Mylopharyngodon piceus</Emphasis>)

We investigated the effects of starvation and re-feeding on growth and swimming performance and their relationship in juvenile black carp (Mylopharyngodon piceus). We measured the specific growth rate (SGR), resting metabolic rate (RMR) and constant acceleration test speed (U _CAT, the maximum swimming speed at exhaustion by constant acceleration test with 0.1667 cm s^?2 rate) in a treatment group (21 days of starvation then 21 days of re-feeding) and control group (routine feeding) (n = 20). Starvation resulted in a 17 % decrease in body mass of black carp (P < 0.05). After 21 days of re-feeding, body mass was greater than that of pre-starvation but still less than that of the control group at 42 days. During the re-feeding phase, the SGR of the treatment group was higher than that of the control group (P < 0.05). Starvation resulted in a significant decrease in the RMR and U _CAT. After 21 days of re-feeding, both the RMR and U _CAT recovered to the pre-starvation levels. In the control group, individual juvenile black carp displayed strong repeatability of the RMR and U _CAT across the measurement periods (P ≤ 0.002). In the treatment group, RMR showed significant repeatability between pre-starvation and re-feeding (P = 0.007), but not between pre-starvation and starvation or between starvation and re-feeding. U _CAT showed significant repeatability between pre-starvation and starvation (P = 0.006) and between pre-starvation and re-feeding (P = 0.001), but not between starvation and re-feeding. No correlation or only a weak correlation was found between any two variables of RMR, U _CAT and SGR, whereas the increment of the U _CAT (ΔU _CAT) was negatively correlated with that of SGR during the starvation phase (r = ?0.581, n = 20, P = 0.007) and re-feeding phase (r = ?0.568, n = 20, P = 0.009). This suggested that within individual black carp, there is a trade-off between growth and maintenance (or development) of swimming performance under food-limited conditions.     

Genetic diversity status of Pacific white shrimp (<Emphasis Type="Italic">Litopenaeus vannamei</Emphasis>) using SSR markers in Iran

Genetic diversity is vital for the maintenance of genetic pool in cultured shrimps. In order to estimate the current status of genetic diversity in Litopenaeus vannamei shrimp in Iran, as an exotic species, a total of 45 individuals from Amiri and Gorgeaj farms in Jask port of Hormozgan province and one hatchery in Gomishan city of Golestan province, were detected using four microsatellite loci. The number of alleles per locus was 5–10, and the mean effective number of alleles (N _e) across populations and loci ranged from 4.834 to 5.148. The overall mean observed heterozygosity (H _o) ranged from 0.450 to 0.479, which was lower than the expected one (0.789–0.794). There was nothing remarkable about any of the allele frequencies across populations or loci. The mean inbreeding coefficient (F _IS) and pairwise genetic differentiation (F _ST) among populations were 41.6 % and 0.133, respectively. The three studied populations departed from Hardy–Weinberg equilibrium (HWE). Analysis of molecular variance revealed 14 % variability among and 86 % within populations. However, considering departing from HWE and the high F _IS and F, the moderate pairwise F _ST values, importance of introducing genetically diverse broodstock and monitoring to control inbreeding is discussed.     

Effect of the early temperature on the growth of larvae and postlarvae turbot, <Emphasis Type="Italic">Scophthalmus maximus</Emphasis> L.: muscle structural and ultrastructural study

Turbot specimens were kept at three temperatures (T _s ): warm (W) (21–22 °C), ambient (A) (17–18 °C) and cold (C) (13–14 °C) during the larval and early postlarval stages. At 90 days posthatching (dph), all of them were transferred to ambient T until 190 dph. At 2–3 dph, the specimens showed a monolayer of red muscle and immature white fibres; external or dermomyotome cells (presumptive myogenic cells) were observed on the surface of the red muscle. In the following stages, many myogenic cells and presumptive myogenic precursors were observed within the myotome, presumably derived of the dermomyotome. When comparing the growth at the same age (2, 10, 25, 37 dph), the body length and the muscle growth were positively influenced by the warm T, being the hyperplasia the muscle parameter more significantly influenced. The development rate was also positively correlated with the high T: the beginning of the metamorphosis took place at 15, 23 and 25 dph at W, A and C temperatures, respectively, with the highest body length values at ambient temperature. The metamorphosis finished at 25, 30 and 37 dph at W, A and C temperatures, respectively, with the highest body length values at warm temperature. However, the muscle cellularity was similar in all the groups at the end of the metamorphosis. At 90 and 190 dph, the largest body length was observed at W temperature. However, the muscle cellularity was similar between A and W; the number of fibres was similar in all the groups at 190 dph, which shows the beginning of a compensatory muscle growth in A and C, mainly in A.     

Molecular endocrine changes of Gh/Igf1 axis in gilthead sea bream (<Emphasis Type="Italic">Sparus aurata</Emphasis> L.) exposed to different environmental salinities during larvae to post-larvae stages

The influence of acclimation of the euryhaline gilthead sea bream (Sparus aurata) larvae/post-larvae to brackish water on growth, energetic contents, and mRNA levels of selected hormones and growth-regulating hypothalamic neurohormones was assessed. Specimens from 49 days post-hatching were acclimated during 28 days to two different environmental salinities: 38 and 20 psu (as brackish water). Both groups were then transferred to 38 psu and acclimated for an additional week. Early juveniles were sampled after 28 days of acclimation to both salinities and one week after transfer to 38 psu. Pituitary adenylate cyclase-activating peptide (adcyap1; pacap), somatostatin-I (sst1), growth hormone (gh1), insulin-like growth factor-I (igf1), and prolactin (prl) mRNA expression were all studied by QPCR. Post-larvae acclimated to 20 psu showed better growth performance and body energetic content than post-larvae maintained at 38 psu. prl, adcyap1, and igf1 mRNA expression levels increased in 20-psu-acclimated post-larvae but decreased upon transfer to 38 psu. GH1 expression did not show significant changes under both experimental conditions. Our results suggested an enhanced general performance for post-larvae in brackish water, supported by the actions of adcyap1, igf1, and prl.     

Thermal tolerance of a thermally selected strain of rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis> and the pedigrees of its F1 and F2 generations indicated by their critical thermal maxima

A thermally selected strain of rainbow trout has been established by selective breeding since 1966 in Miyazaki, Japan. In the present study, we compared the critical thermal maxima (CTMs), the temperatures at which organisms reach a predefined sublethal endpoint and lose their equilibrium, between a thermally selected and two normal (Donaldson) strains of rainbow trout. The CTM of one normal strain from Nikko (Nikko strain) acclimated to 20 °C (29.7 °C) was significantly lower than those of the thermally selected strain (30.0 °C) and the other Donaldson strain from Aomori (29.9 °C) (P?<?0.05). The F1 generations, F1T and F1N, were produced by crossing thermally selected strain females with Nikko strain males and Nikko strain females with thermally selected strain males, respectively. No significant difference was observed in the CTM between F1T [30.1?±?0.15 °C (n?=?30)] and F1N [30.1?±?0.16 °C (n?=?30)] (P?>?0.05) for fish acclimated to 20 °C, suggesting that the F1 offspring inherited the thermal tolerance trait from one thermally selected strain parent irrespective of whether it was the male or female. F2 offspring of F1T or F1N also showed the thermal tolerance trait. The coefficients of variation for CTM were also compared among all the datasets obtained in the present study and their values for F1 hybrids were lower than those of the parental generation of the Nikko strain (P?<?0.05). In contrast, the coefficients of variation of F2s were the same as those of their parental generation. Furthermore, the thermally selected strain and Nikko strain as a reference family provide a F2 generation for segregating phenotypes, which is required for in-depth genetic analysis of the thermally selected rainbow trout strain.     

Ontogenetic habitat shift in <Emphasis Type="Italic">Pugettia quadridens</Emphasis> on the coast of Sagami Bay,Japan

Population dynamics of the kelp crab Pugettia quadridens, a potential predator of juvenile abalone, were investigated on the coast of Nagai, Sagami Bay, to examine its cryptic ontogenetic habitat shift. Monthly quantitative sampling over 2 years, which was carried out in 11 different subtidal habitats (< 0.1–8 m deep), revealed that P. quadridens changes ontogenetically according to its habitat: P. quadridens occurred in shallow lower subtidal (1–4 m) small red algal turfs (mainly Grateoloupia cornea) almost all year around, when the population was dominated by juveniles and stage I (immature) individuals; on the other hand, in the shallow upper subtidal (0.1–1 m) habitat (mainly Sargassum fusiforme beds near the low tidal mark), P. quadridens occurred only during winter to early spring, when the population was dominated by individuals of senior ontogenetic stages. These finding suggest that P. quadridens inhabit subtidal small red algal turfs during their early benthic phase, and then, the majority of larger individuals of stages II and III migrate to Sargassum beds. Our results suggest that the impact of P. quadridens predation on abalone stocks is limited in the present study site, primarily because the habitat of P. quadridens is segregated from that of juvenile abalone.     

Anti-oxidative functions of <Emphasis Type="Italic">mt2</Emphasis> and <Emphasis Type="Italic">smtB</Emphasis> mRNA expression in the gills and brain of zebrafish (<Emphasis Type="Italic">Danio rerio</Emphasis>) upon cadmium exposure

There were not any past studies about metallothionein isoforms (smtB and mt2) having anti-oxidative functions on zebrafish after Cd²⁺ exposure. On the other hand, the anti-oxidative enzymatic factors such as superoxide dismutase (sod), glutathione peroxidase (gpx1a), and catalase (cat) are used as references to investigate whether the smtB and mt2 have anti-oxidative responses on the gills and brain of zebrafish after 1–6 h of 0 and 1.78 μM Cd²⁺ exposure. The anti-oxidative system such as sod, cat, and gpx1a mRNA expressions demonstrated a cascade response upon Cd²⁺-induced oxidative stress in the present study. Interestingly, the smtB mRNA expression levels increased by 3.2- to 6.1-fold, and mt2 raised by 4.1- to 11.3-fold in gills at 1 and 3 h after exposure to Cd²⁺, respectively. On the other hand, the smtB mRNA levels increased by 10.6- to 58.6-fold, but mt2 mRNA levels increased by 2.3- to 11.1-fold in brain at 1 and 3 h after exposure to Cd²⁺, respectively. In addition, both tissues showed increased apoptosis levels at 3 h, and recovery after 6 h of Cd²⁺ exposure. From the results, we suggest that both mt2 and smtB play a role in anti-oxidation responses within 6 h after exposure to Cd²⁺. In conclusion, the smtB mRNA levels have a higher response than mt2 in the brain, but both mRNA expressions appear to have a similar pattern in the gill. We suggest that smtB plays an important role to defend oxidative stress in the brain of adult zebrafish upon acute Cd²⁺ exposure.     

Long-term effects of stocking density on survival,growth performance and marketable production of the sea urchin <Emphasis Type="Italic">Strongylocentrotus intermedius</Emphasis>

Sea urchins were stocked at a density of 15 (D15), 30 (D30), 45 (D45) and 60 (D60) urchins/cage (0.3 m long × 0.2 m wide × 0.4 m high) in a laboratory culture environment for 16 months. The wet body weight (BW) and test diameter growth were monitored at 2-month intervals during the experiment. At the conclusion of the experiment, the surviving sea urchins were counted and the gonad wet weight (GW) and gonad color were measured. Specific growth rate (SGR) of body weight, survival rate (SR), gonad index (GI), gonad color difference (ΔE ₀₀), coefficients of variation (CV) of BW, GW, GI and ΔE ₀₀, total biomass yield (TY) and total gonad yield (TGY) per cage were calculated. Two marketable yield variables, graded according to gonad index, i.e., marketable biomass yield (MY) and marketable gonad yield (MGY), were also calculated. Coefficient of variation of final body weight (FW) and final test diameter (FTD) of sea urchins increased as the stocking density increased, indicating the existence of adverse social interactions. These adverse social interactions detrimentally affected FW, FTD, SGR, GW and GI (P < 0.01). Although SR decreased with the increasing densities, no statistical significant difference was detected. Sea urchins at D15 had the lowest gonad color difference (ΔE ₀₀) (P < 0.05). However, statistically equal CV of ΔE ₀₀ indicates this density effect was not a result of adverse social interactions. TY and TGY increased with increased density and can be described by the following equations: TY = 84.18X ^0.64, R ² = 0.999 and TGY = 24.16X ^0.38, R ² = 0.979. However, the MY and MGY were not significantly different among stocking densities. The results of this study demonstrate that in intensive culture S. intermedius at low stocking density can achieve high growth rate, gonad index and desirable color without decreasing the marketable yield. Farmers should choose to culture S. intermedius at low stocking densities.     

Toxicological and hematological effect of <Emphasis Type="Italic">Terminalia arjuna</Emphasis> bark extract on a freshwater catfish, <Emphasis Type="Italic">Heteropneustes fossilis</Emphasis>

Increasing demand for eco-friendly botanical piscicides and pesticides as replacements for harmful synthetic chemicals has led to investigation of new sources of plant materials. Stem bark of Terminalia arjuna, which has been used as a popular folk medicine since ancient time, was examined for its piscicidal activity. This study aims to determine toxicity of ethanol extract of T. arjuna bark on fresh water stinging catfish (Heteropneustes fossilis), along with evaluation of changes in hematological parameters of the fishes exposed to a lethal concentration. The percent mortality of fishes varied significantly in response to concentrations of the extract and exposure times (between exposure time F = 36.57, p < 0.001; between concentrations F = 39.93, p < 0.001). The lethal concentrations (LC₅₀) of ethanol extract were found to be 12.7, 8.94, 5.63 and 4.71 mg/l for 24, 48, 72 and 96 h, respectively. During acute toxicity test, blood samples of treatment fishes showed significant decreases in the red blood cells count, hematocrit content, hemoglobin concentration, mean corpuscular hemoglobin concentration and plasma protein level when compared to those of the control group, while there were significant increases in the mean corpuscular volume, mean corpuscular hemoglobin, white blood cells count and plasma glucose concentration. These results suggest that T. arjuna bark extract could be considered as a potent piscicide due to its toxic effect on fish, particularly fish hematology.     

<Superscript>1</Superscript>H-NMR metabolite profiling of abalone digestive gland in response to short-term starvation

The effect of a short-term feeding and starvation experiment on juvenile abalone (Haliotis rubra × H. laevigata) was investigated (average length = 67 mm; average weight = 48 g). All aquaculture experiments were conducted at The University of Melbourne, Australia. Artificial feed was supplied ad libitum to the fed group, and no feed was supplied to the starved group. A modified metabolite extraction protocol using deuterated solvents was developed for ¹H-NMR-based metabolite profiling of digestive gland in response to the short-term feeding/starvation experiment, to avoid lyophilisation prior to biochemical analysis. PLS-DA revealed that fed and starved abalone are metabolically distinct from each other after 28 and 56 days. After 28 days, the fed group was defined by an increase in arginine, glucose, glutamate, glycine, inosine and uracil (P < 0.05), and the starved group was defined by an increase in N,N-dimethylglycine. After 56 days, the fed group still displayed increased glucose (P < 0.05), while N,N-dimethylglycine remained elevated in the starved group (P < 0.05). Arginine and glycogen were all higher at 28 days compared to 56 days, suggesting decreased anaerobic energy production at the later time point. Only glucose and N,N-dimethylglycine were significantly different between the fed and starved groups after 56 days, suggesting that abalone had not acclimatised to the starvation treatment after 28 days. These results infer N,N-dimethylglycine is a robust marker for short-term starvation in abalone. ¹H-NMR was also conducted on the artificial feed and starved abalone faecal matter, revealing the biochemical differences between them and digestive gland tissue. These methodology and results will facilitate a deeper understanding of the nutritional and physiological requirements of abalone in an aquaculture setting.     

<Emphasis Type="Italic">Virgibacillus proomii</Emphasis> and <Emphasis Type="Italic">Bacillus mojavensis</Emphasis> as probiotics in sea bass (<Emphasis Type="Italic">Dicentrarchus labrax</Emphasis>) larvae: effects on growth performance and digestive enzyme activities

This study examined the effects of two probiotics (Virgibacillus proomii and Bacillus mojavensis) on the digestive enzyme activity, survival and growth of Dicentrarchus labrax at various ontogenetic stages in three separate experiments. These probiotics were incorporated as single or mixed into fish feed for a period of 60 days. The growth parameters, proximate composition of whole body, digestive enzymes and gut microbiology were monitored at regular. The increments in length and weight and the survival were significantly higher (P < 0.05), and the values of food conversions were significantly lower (P < 0.05) in fishes fed the probiotic. The administration of V. proomii and B. mojavensis in diet resulted in an increase (P > 0.05) in body ash and protein content and in the specific activity of phosphatase alkaline and amylase in the digestive tract of all the fishes. V. proomii and B. mojavensis persisted in the fish intestine and in the feed in high numbers during the feeding period (group 1: 5.8 × 10⁴ CFU/ml, group 2: 9.6 × 10⁴ CFU/ml, and group 3: 9.8 × 10⁴ CFU/ml day 60). The two probiotics V. proomii and B. mojavensis were adequate for improved growth performance and survival and for healthy gut microenvironment of the host.     

Inhibitory neurotransmitter serotonin and excitatory neurotransmitter dopamine both decrease food intake in Chinese perch (<Emphasis Type="Italic">Siniperca chuatsi</Emphasis>)

Aminergic neurotransmitters play important roles in the regulation of food intake. However, their effects on feeding in fish have been less explored and still unclear. In the present study, the effects of serotonin (5-HT) and dopamine (DA) on food intake were evaluated through intraventricular (ICV) administration in Chinese perch (Siniperca chuatsi) and the mRNA expression levels of neuropeptide Y (NPY), agouti gene-related protein (AgRP), and pro-opiomelanocortin (POMC) were detected. At 1 h post-injection, 5-HT significantly decreased food intake in a dose-dependent manner. The mRNA expression of NPY and AgRP were significantly decreased (p < 0.05), whereas the mRNA expression of POMC was significantly increased (p < 0.05), suggesting the involvement of NPY, AgRP, and POMC in inhibitory action of 5-HT on food intake in Chinese perch. DA significantly decreased (p < 0.05) food intake and AgRP mRNA expression at 1 h post-injection, indicating the inhibitory effect of DA on food intake might be mediated through AgRP. This might shed new light on the regulation of food intake in Chinese perch.     

Effect of stocking density on growth performance,serum biochemical parameters,and muscle texture properties of genetically improved farm tilapia, <Emphasis Type="Italic">Oreochromis niloticus</Emphasis>

The objective of this study was to assess the effects of stocking density on growth performance, serum biochemical parameters, and muscle texture properties of genetically improved farmed tilapia (Oreochromis niloticus, GIFT). Juvenile GIFT with an average initial weight of 12.54?±?0.45 g (mean?±?SD) were randomly stocked in 16 tanks (80 L) in a recirculation aquaculture system at four densities of 10 (D1), 20 (D2), 30 (D3), and 40 (D4) fish per tank for 56 days, with quadruplicate for each density. There were no significant differences in water temperature among the four treatments (P?>?0.05). D4 had the significantly lowest dissolved oxygen content (5.52 vs 5.69–6.09 mg L^?1) (P?>?0.05) and pH (6.63 vs 6.87–7.20) (P?<?0.05). NO₂-N and NH₄-N concentrations significantly increased with increasing stocking density (P?<?0.05). Weight gain (WG) and specific growth rates (SGR) decreased with increasing stocking density. The lowest WG (617.20 vs 660.45–747.06%), SGR (3.52 vs 3.62–3.81% day^?1), and highest feed conversion ratio (1.68 vs 1.53–1.58) were observed in D4. Fish at D4 had significantly lower condition factor (3.11 vs 3.29–3.37%) and survival rate (91.25 vs 97.50%) than those from D1 and D2 (P?<?0.05). With increasing stocking density, serum total cholesterol, triglyceride, and total protein concentrations decreased (P <?0.05) and aspartate aminotransferase and alanine aminotransferase activities increased (P <?0.05). D4 fish had higher moisture content (78.80 vs 76.97%) and lower crude protein content (18.14 vs 19.39%) in muscle than D1 fish (P?<?0.05). Compared to D1 and D2, D3 and D4 had lower muscle hardness (1271.54–1294.07 vs 1465.12–1485.65 g), springiness (0.62–0.65 vs 0.70–0.72), gumminess (857.33–885.32 vs 1058.82–1079.28 g), and chewiness (533.04–577.09 vs 757.53–775.69 g) (P <?0.05). High stocking density resulted in growth inhibition, declines in flesh quality, and disturbance to several serum biochemical parameters.     

Microalga <Emphasis Type="Italic">Isochrysis galbana</Emphasis> in feed for <Emphasis Type="Italic">Trachinotus ovatus</Emphasis>: effect on growth performance and fatty acid composition of fish fillet and liver

Seeking alternatives to the depleting fish oil are crucial for marine fish aquaculture, which is currently dependent on fish oil as the primary source of n-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFAs). Five isonitrogenous (46% crude protein) and isolipidic (16% crude lipid) feed diets (FO, ISO2.9, ISO4.8, ISO6.7, ISO8.6) were formulated by partially replacing fish oil with microalgae Isochrysis galbana. These diets were fed to triplicate tanks of Trachinotus ovatus (mean initial weight 1.92 g) for 80 days. This work demonstrates that a moderate inclusion (around 4.5–5.0 wt%, equivalent to the replacement of 24–26 wt% fish oil) of I. galbana biomass in fish diet improves fish growth performance, lipid deposition and enhances total n-3 fatty acids, DHA, and EPA contents in neutral and polar lipids (PLs) of fish muscle and liver of T. ovatus. The results disclosed in this study suggest that I. galbana microalgae represents a potential high-quality substitute for fish-based ingredients in aquaculture feeds, which can be a promising sustainable solution to resolve the depleting fish oil resource in a cost-effective manner.     

Feeding,somatic condition and survival of sand lance <Emphasis Type="Italic">Ammodytes</Emphasis> sp. larvae in Mutsu Bay,Japan

To clarify the recruitment process of sand lance Ammodytes sp., we investigated larval condition factor, relative gut fullness (%GF), prey abundance and oceanographic structure in Mutsu Bay, Japan, during 1999–2001. Ammodytes sp. larvae, which were collected by horizontal hauls of Motoda nets and a ring net at depths of 1, 10, 20, 30 and 40 m, were mainly distributed at 10–30 m. Larvae at the first feeding time until 12 mm in body length (BL) fed predominantly on copepod nauplii, whereas large larvae with BL of 12.1–14.0 mm fed on a mixture of copepod nauplii, copepodites and appendicularians from late February to April. A path analysis showed that difference in water density between 35- and 5-m depths negatively affected naupliar abundance at 10–30-m depth (standardised path coefficient β = ?0.71, p = 0.005 for 3.3–8.0-mm BL larvae and β = ?0.78, p < 0.001 for 8.1–12.0-mm BL larvae). Naupliar abundance positively affected the %GF of Ammodytes sp. larvae (β = 0.75, p < 0.001 for 3.3–8.0-mm BL larvae and β = 0.66, p < 0.001 for 8.1–12.0-mm BL larvae), whereas it was negatively affected by water temperature (β = ?0.45, p = 0.008 for 3.3–8.0-mm BL larvae and β = ?0.56, p = 0.002 for 8.1–12.0-mm BL larvae), and the temperature effect was weak compared with that of naupliar abundance. In turn, %GF positively affected larval somatic weight (β = 0.91, p < 0.001 for 6.0-mm BL larvae and β = 0.70, p = 0.005 for 10.0-mm BL larvae). The recruitment failure in 1999 was likely caused by a reduced condition factor, which resulted from low naupliar abundance. In contrast, the abundance of nauplii and Oithona similis copepodites was high in 2000 and 2001. It is possible that the higher recruitment success in 2001 was because of the higher water temperatures in Mutsu Bay, sustaining faster growth of the larvae than in 2000 under the high-prey abundance conditions.     

Rapid measurements of fat content in live and slaughtered common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.)

The objective of this study on common carp (Cyprinus carpio) was to obtain and predict the first cross-validated data of the fat content on market size carps using a non-invasive or non-destructive method in situ. The carps (1680 ± 388 g; n = 136) used were from a semi-intensive system and were on a different diet (cereal, pelleted and extruded diet). For the evaluation of the fat content, a Fish Fatmeter FM 692 from Distell.com. (FFM) and a manual measurement of back fat height using a digital calliper were used. For the prediction model, the following basic body measurements (variables) were used: total body length, body length, body height, the width of the body, and the circuit of the body. The body weight, weight of intestines, weight of gonads, weight of hepatopancreas, and fillet yield (%) were measured, and the Fulton coefficient was calculated. The study was focussed on evaluating the applicability of these methods and the accuracy of the obtained result, respectively. Results showed that all the rapid methods had a strong correlation. Multiple regression models with forward selection of variables were used throughout. The final prediction model between predicted and observed values for the fat content for FFM and calliper being adjusted index of determination is shown here (R _adj ²  = 0.88; 5 variables and R _adj ²  = 0.91; 7 variables), respectively.     

Medicinal plant extracts modulate respiratory burst and proliferation activity of rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) leukocytes

The present study was designed to investigate the immunomodulatory effects of Aloe vera, Curcuma longa, Echinacea purpurea, Lavandula officinalis, Origanum vulgare, Panax ginseng, and Rheum officinale extracts on leukocytes purified from rainbow trout (Oncorhynchus mykiss) head kidney. The cells were cultured in a medium containing increasing doses of extracts; afterwards, they were tested for reactive oxygen species production after stimulation with phorbol myristate acetate (PMA) and proliferation in the presence or absence of phytohemagglutinin from Phaseolus vulgaris (PHA-P). After a 2-h exposure, the extracts of L. officinalis, O. vulgare, and R. officinale strongly reduced the oxidative burst activity of PMA-stimulated leukocytes, in a dose-dependent manner (P ≤ 0.05). A. vera, C. longa, E. purpurea, and P. ginseng extracts reduced this response with lower efficacy and especially at lower concentrations. On the contrary, the highest concentration of ginseng extract stimulated the respiratory burst of leukocytes compared to untreated control cells. After a 72-h exposure, the extracts of L. officinalis, R. officinale, C. longa, E. purpurea, and P. ginseng had a clear dose-dependent stimulatory effect on leukocyte proliferation (P ≤ 0.05). The results suggest that these medicinal plants can be considered as reliable sources of new antioxidants or immunostimulants to be used in aquaculture.     

Immunomodulatory effects of <Emphasis Type="Italic">Rhodomyrtus tomentosa</Emphasis> leaf extract and its derivative compound,rhodomyrtone, on head kidney macrophages of rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>)

Rhodomyrtus tomentosa is a medicinal plant that shows biological effects including immunomodulatory activity on human and other mammals but not in fish. In this study, we evaluated the in vitro immunomodulatory effects of R. tomentosa leaf extract and its active compound, rhodomyrtone, on the immune responses, using rainbow trout (Oncorhynchus mykiss) head kidney (HK) macrophages as a model. The tested immune functions included the expression of genes involved in innate immune and inflammatory responses and the production of reactive oxygen species (ROS). Gene expression was evaluated after exposure to 10 μg mL^?1 of R. tomentosa and 1 μg mL^?1 of rhodomyrtone for 4 and 24 h. R. tomentosa and rhodomyrtone induced changes in the expression of pro-inflammatory cytokines (il1β, il8, and tnfα), anti-inflammatory cytokines (il10 and tgfβ), inducible enzymes (inos, cox2, and arginase), and an antioxidant enzyme (gpx1). Co-exposure of R. tomentosa with LPS resulted in a prominent reduction in the expression of genes related to an inflammatory process (il1β, il8, tnfα, inos, saa, hepcidin, and gpx1), suggesting anti-inflammatory effects. Similarly, co-exposure of rhodomyrtone with LPS led to a downregulation of inflammation-related genes (il1β, inos, saa, and hepcidin). In addition, exposure to both natural plant products caused a reduction in cellular ROS levels by HK macrophages. The present results indicate that R. tomentosa and rhodomyrtone exerted immunostimulatory and anti-inflammatory effects on fish macrophages, thus opening up the possibility of using these natural products to further develop immunostimulants for health management in aquaculture.