Anesthesia induces stress in Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis>), Atlantic cod (<Emphasis Type="Italic">Gadus morhua</Emphasis>) and Atlantic halibut (<Emphasis Type="Italic">Hippoglossus hippoglossus</Emphasis>)

Stress in response to anesthesia with benzocaine, MS-222, metomidate and isoeugenol was studied in Atlantic salmon (Salmo salar), Atlantic halibut (Hippoglossus hippoglossus), and Atlantic cod (Gadus morhua) with no concomitant stress from handling or confinement in association with anesthesia or sampling. All of the anesthetics tested induced a stress response in all species, displayed by a release of cortisol to the water. MS-222 anesthesia elicited the highest cortisol release rates, reaching maximum levels 0.5 h post-exposure and returning to basal levels after 3–4 h. Benzocaine anesthesia caused a bimodal response where the initial peak in cortisol release rate was followed by a second increase lasting towards the end of the trial (6 h). This bimodality was more profound in Atlantic salmon than in Atlantic halibut and Atlantic cod. Metomidate anesthesia induced the lowest release of cortisol of the agents tested in both Atlantic halibut and Atlantic cod, but resulted in a bimodal response in Atlantic salmon where the initial increase in cortisol release was followed by a larger increase peaking at 2–2.5 h post exposure before returning to basal levels after 5 h. The stress induced in Atlantic salmon by isoeugenol anesthesia resembled that of MS-222, but did not reach the same elevated level. Overall, the cortisol release was most profound in Atlantic salmon followed by Atlantic halibut and Atlantic cod.     

Age and growth of two gizzard shads, <Emphasis Type="Italic">Nematalosa come</Emphasis> and <Emphasis Type="Italic">N. japonica</Emphasis>, in coastal waters around Okinawa Island,southwestern Japan

The age and growth of two Nematalosa species around Okinawa Island were examined using sectioned otoliths collected from September 2003 to April 2006. Monthly changes in the frequency of the appearance of a translucent band on the outer margin of the otoliths indicated that ring formation occurred once a year from January to July for Nematalosa come and from January to March for Nematalosa japonica. The von Bertalanffy growth equations for both species were as follows: N. come: L _t  = 365.5{1 − exp[−0.111 × (t + 0.288)]} for females and L _t  = 214.7{1 − exp[−0.700 × (t – 1.110)]} for males; N. japonica: L _t  = 205.1{1 − exp[−1.068 × (t − 1.180)]} for females and L _t  = 195.5 {1 − exp[−1.293 × (t − 1.269)]} for males. The maximum ages observed for N. come and N. japonica were 11 and 6 years old, respectively. The growth of these species was characterized by the slow growth of N. come over many years, resulting in a larger size than N. japonica.     

Econometric model of viability/profitability of octopus (<Emphasis Type="Italic">Octopus vulgaris</Emphasis>) ongrowing in sea cages

The interest in diversifying aquacultural production with new species is evident, preferably with species with a high commercial value and whose consumption is not geographically limited. In this sense, octopus would be a good choice. The present work, therefore, presents an economic-financial analysis of the commercial viability of octopus ongrowing in the Mediterranean, paying special attention to the one- or two-cycle approach. Viability–profitability equations are developed for analysing economic parameters associated with production. To estimate the investment, an offshore ongrowing installation was designed comprising 150 cages containing 30,000 individuals. Growth was estimated for the two strategies: (A) Two consecutive cycles per year (2CY), each lasting 3.5 months from October to June. Initial weight was 0.7 kg, and the mean final weight was 2.7 kg. (B) One growth cycle per year (1CY) beginning in November or December and finishing in April or May, starting with the same weight individuals (0.7 kg) and giving individuals with a final weight of 3.65 kg. The highest costs, the most important from an economic point of view, are feed (38.57 and 40.03%, respectively), fixed assets (25.26 and 17.47%, respectively), juveniles (16.65 and 23.02%, respectively), and in fourth place salaries (14.34 and 15.60%, respectively). The equations obtained for the variables NPV (Net Present Value) and IRR (Internal Ratio of Return) are the following: 1 cycle per year (1CY), NPV = −489,088 − 1.45 K − 1,439,823 C _F − 1,477,890 C _J − 1,460,627 C _O + 1,432,386 SP, IRR = 0.1328 − 7.82 × 10⁻⁸ K − 0.0416 C _F − 0.0437 C _J − 0.0427 C _O + 0.0412 SP. 2 cycles per year (2CY), NPV = −404,431 − 1.46 K − 2,118,410 C _F − 2,121,221 C _J − 2,144,755 C _O + 2,129,223 SP, and IRR = 0.0952 − 6.95 × 10⁻⁸ K − 0.0586 C _F − 0.0588 C _J − 0.0588 C _O + 0.0613 SP. The NPV and IRR values estimated with the econometric equations for each option using the initial variables confirm that 1CY has a higher NPV (3,013,569 €) and IRR (12.27%) than 2CY, with an NPV of 2,396,708 € and IRR of 10.39%. In both cases, octopus ongrowing is economically viable, although 1CY is the most favourable system.     

Microsatellite and mitochondrial DNA analyses reveal no genetic difference between two pufferfish species torafugu <Emphasis Type="Italic">Takifugu rubripes</Emphasis> and karasu <Emphasis Type="Italic">T. chinensis</Emphasis>

It has been reported that the nucleotide sequences of the mitochondrial and nuclear genes of Takifugu pufferfish torafugu T. rubripes and karasu T. chinensis show 99–100% sequence identity, indicating a very close relationship between the two species. To further investigate this genetic relationship, we compared genetic variation at four microsatellite loci and at the mitochondrial control region (CR) (561 bp) between groups of T. rubripes caught at two locations [TrG, caught in the Genkai Sea off Tsushima Island in 2003 (n = 50); TrS, caught in the Suwo Sea off Kita-Kyushu in 2008 (n = 50)] and T. chinensis caught at one location (TcK, caught off the east coast of Korea in 2004; n = 50). Analyses using microsatellite loci showed that genetic diversity index values of the TrG, TrS and TcK groups were 0.9505, 0.9350 and 0.9335, respectively, while values of genetic distance and genetic differentiation between TrG and TcK (0.0543 and 0.0189, respectively) were smaller than those between TrG and TrS (0.0857 and 0.0194, respectively). Analyses using CR for the same specimens showed that genetic distances were consistent with those obtained using microsatellite loci. These results, together with our previous observations, suggest that T. rubripes and T. chinensis are very closely related and possibly can be regarded as the same species.     

Lack of arterial PO<Subscript>2</Subscript> downregulation in Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis> L.) during long-term normoxia and hyperoxia

Regulation of arterial partial pressure of O₂ (P_aO₂) in Atlantic salmon (Salmo salar) was investigated during resting conditions in normoxic and hyperoxic water. Dorsal aorta cannulated adult Atlantic salmon (1.2–1.6 kg, n = 8) were exposed to 2 week sequential periods of normoxia [16.7 ± 1.1 kPa (mean ± SD)] and hyperoxia (34.1 ± 4.9 kPa) in individual tanks containing seawater (33.7 ± 0.2 ppt) at stable temperature conditions (8.7 ± 0.7°C) and a light regime of L:D = 12:12. Tank design and sampling procedures were optimized to provide suitable shelter and current for the fish, and to allow repeated, undisturbed sampling of blood from free-swimming fish. Fish were sampled regularly through the experimental period. P_wO₂, P_aO₂, blood ion composition (Na⁺, K⁺, Cl⁻), acid–base status (pH, PCO₂, HCO₃ ⁻), haematocrit and glucose were measured. The most frequently observed P_aO₂ values were in the range of 60–80% of P_wO₂, both during normoxia and hyperoxia, and P_aO₂ values were significantly lower during normoxia than during hyperoxia. Blood pH, PCO₂ and HCO₃ ⁻ were significantly elevated during hyperoxia, while, Na⁺, Cl⁻ and Hct were significantly lower. K⁺ and glucose showed no significant differences. This study demonstrates a lack P_aO₂ regulation in Atlantic salmon to low partial pressures, in contrast to previous reports for many aquatic gill breathing animals. Both during normoxia and hyperoxia, P_aO₂ reflects P_wO₂, and alterations in external PO₂ consequently result in proportional arterial PO₂ changes. Physiological adaptation to hyperoxia, as illustrated by changes in several blood parameters, does not include down-regulation of P_aO₂ in Atlantic salmon. The lack of P_aO₂ regulation may make Atlantic salmon vulnerable to the oxidative stress caused by increased free radical formation in hyperoxic conditions.     

Potential of cinnamon (<Emphasis Type="Italic">Cinnamomum verum</Emphasis>) oil to control <Emphasis Type="Italic">Streptococcus iniae</Emphasis> infection in tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis>)

In this study, four essential oils—cinnamon oil, leech lime oil, lemongrass oil, and turmeric oil—were examined for their antimicrobial activities against Streptococcus iniae, a bacterium that is pathogenic in fish, in which it causes streptococcosis. Cinnamon oil was the most potent antimicrobial agent among these oils, with a minimal inhibitory concentration (MIC) of 40 μg/ml. By using gas chromatography–mass spectrometry (GC–MS), it was found that the major components of cinnamon oil were cinnamaldehyde (90.24), limonene (2.42%), cinnamyl acetate (2.03%), linalool (1.16%), and α-terpineol (0.87%). Of these compounds, only cinnamaldehyde exhibited antimicrobial activity against S. iniae, with an MIC of 20 μg/ml. In an in vivo trial, no mortality was apparent in fish fed on fish diets supplemented with 0.4% (w/w) of cinnamon oil and with 0.1% (w/w) of oxytetracycline 5 days prior to infection with S. iniae. These results indicate that cinnamon oil had a protective effect on experimental S. iniae infection in tilapia, and thus has the potential to replace the antibiotics used to control this disease.     

Age and growth of the lizardfish <Emphasis Type="Italic">Saurida elongata</Emphasis> from the Tsushima/Korea Strait

We studied the age and growth of the lizardfish Saurida elongata using ground thin sections of otoliths (sagittae) from specimens collected in the Tsushima/Korea Strait between May 1999 and June 2001. A total of 695 individuals with fork lengths (FL) ranging from 189 to 478 mm were examined. The frequency of translucent zone occurrence at the outer margin of the otoliths indicated that translucent zones formed once a year between November and February. Most of the males examined were estimated to be 2–7 years old and the females, 3–9 years old. The maximum estimated age of a male specimen was 10 years and that of a female, 11 years. The estimated von Bertalanffy growth curves were FL _t  = 451{1 − exp[−0.172(t + 2.50)]} and FL _t  = 515{1 − exp[−0.151(t + 2.47)]} for males and females, respectively. At all ages, the FL at a specific age for females was greater than that for males, suggesting that females of this species grow faster than males.     

Optimisation of larval culture of the mussel <Emphasis Type="Italic">Mytilus</Emphasis><Emphasis Type="Italic">edulis</Emphasis> (L.)

The blue mussel Mytilus edulis is a commercially important species whose fishery and culture generally relies on natural spat collection. Hatchery-production could provide an alternative source of seed, enabling reliable expansion of the industry. Mussel spawning and larval rearing trials were carried out to optimise elements of hatchery production. Culturing fertilised eggs at low density (20–200 eggs cm⁻²) rather than high density (400–720 eggs cm⁻²) significantly improved the quality of first veliger larvae and differences in this improvement were evident between the eggs from different females (maternal effects). Veliger larval growth at 17 or 21°C was significantly faster than growth at 14°C. Feeding veliger larvae an identical total ration either daily or at 2–3 day intervals did not significantly affect their growth. Different microalgal diets (1: Isochrysis sp. (clone T-ISO), 2: Chaetoceros calcitrans forma pumilus, 3: C. muelleri, 4: mixed Isochrysis sp. (clone T-ISO) and C. calcitrans f. pumilus, and 5: mixed Isochrysis sp. (clone T-ISO) and C. muelleri) were tested on veliger larval growth and mixed diets outperformed single-species diets.     

Ex situ and in situ measurements of juvenile yellowfin tuna <Emphasis Type="Italic">Thunnus albacares</Emphasis> target strength

To provide target strength (TS) information for estimating the body length of yellowfin tuna Thunnus albacares and its abundance around fish aggregating devices, TS was measured ex situ and in situ. In the ex situ TS measurements, two cameras synchronized with a 200 kHz echosounder were used to obtain the precise orientation of the yellowfin tuna under free swimming conditions. The ex situ TS (dB re 1 m²)–fork length (FL, cm) regression was: TS = 27.06 log (FL) − 85.04. Ex situ TS was found to reach its maximum in the tilt angle range of −15° to −20° after excluding TS samples with insignificant correlation to the tilt angle. The angle between the vertebra and the swim bladder was approximately 25° according to X-ray images, supporting the above tilt range. The relationship between the swim bladder volume (V _SB, ml) and the fork length was: V _SB = 0.000213 FL³. The results from the in situ TS measurements indicated that the tilt angle was highly concentrated between −10° and 15°. The results from a calculation using the ex situ TS–FL equation with the fork length from biological sampling agreed strongly with the average in situ TS.     

Enhancement of <Emphasis Type="Italic">Penaeus monodon</Emphasis> shrimp postlarvae growth and survival without water exchange using marine <Emphasis Type="Italic">Bacillus pumilus</Emphasis> and periphytic microalgae

We have investigated the possibility of using a consortium of marine bacterium and periphytic microalgae to improve the water quality and increase the growth and survival of the shrimp Penaeus monodon in a hatchery system. Three treatments were evaluated for their effect on P. monodon postlarvae (PL) when the culture water was not changed: Bacillus pumilus alone (B); periphytic microalgae alone (M); B. pumilus + periphytic microalgae (BM). P. monodon PL raised in a tank of unchanged water without bacterium and periphytic microalgae served as the control. The water in tanks of the M and BM treatments had significantly low levels of total ammonia-nitrogen (TAN) (0.03 and 0.01 mg l⁻¹, respectively) and nitrite-nitrogen (NO₂-N) (0.03, 0.01 mg l⁻¹, respectively) than that in the B (TAN 0.80, NO₂-N 0.68 mg l⁻¹) and control (TAN 1.11, NO₂-N 1.12 mg l⁻¹) tanks. Moreover, PL cultured in tanks M and BM had significantly higher survival and specific growth rates and a significantly higher resistance to the reverse salinity stress test than those in the B and control tanks. Compared to the control PL, the PL cultured in the BM tanks had significantly higher levels of protein, lipid, polyunsaturated fatty acids, ecosapentaenoic acid, and docosahexaenoic acid. The culture water in tanks BM also contained significantly less Vibrio than the control water. Our results illustrate the beneficial effects of a B. pumilus and periphytic microalgae consortium on improving the water quality and the growth and survival of shrimp PL grown in a hatchery system.     

Alterations in blood electrolytes of a freshwater catfish <Emphasis Type="Italic">Heteropneustes fossilis</Emphasis> in response to treatment with a botanical pesticide, <Emphasis Type="Italic">Nerium indicum</Emphasis> leaf extract

The present study aimed at investigating the effects of Nerium indicum leaf extract on the blood electrolytes of Heteropneustes fossilis for short- and long term. Fish were subjected to Nerium indicum leaf extract for short term (11.27 mg/L i.e. 0.8 of 96 h LC₅₀) and long term (2.81 mg/L i.e. 0.2 of 96 h LC₅₀). Fish were killed on each time intervals from control and experimental (Nerium indicum) groups after 24, 48, 72, and 96 h in short-term exposure and after 7, 14, 21, and 28 days in long-term experiment. Blood samples were analyzed for calcium and inorganic phosphate levels. Acute exposure of Nerium indicum leaf extract caused a progressive decrease in the serum calcium levels after 48 h in fish H. fossilis, which persists till the close of the experiment (96 h). The serum inorganic phosphate levels remain unaffected till 48 h in the Nerium indicum leaf extract–exposed fish. After 72 and 96 h, the levels exhibit a decrease. Chronic Nerium indicum leaf extract treatment provoked a decrease in serum calcium levels at day 14. This decrease continues till 28 days. The serum phosphate level of the Nerium indicum leaf extract–treated fish decreases on day 14 and 21. However, on day 28, the levels become close to the normal values. We conclude that Nerium indicum leaf extract exposure alters the blood electrolytes of the fish, thus causing physiological disturbances which might affect seriously the normal vital functions, growth rate, reproduction, and their survival in nature.     

Growth,diet, and reproduction of Eurasian perch <Emphasis Type="Italic">Perca fluviatilis</Emphasis> L. in Lake Varese,northwestern Italy

No data have previously been reported on Eurasian perch Perca fluviatilis L. in Lake Varese. In this study, the growth, diet, and reproductive biology of the Eurasian perch population were investigated with the aim of providing information that may serve as a basis for efficient resource management. A total of 240 specimens were caught during the monthly sampling campaign from November 2006 through October 2008. The length-to-weight relationships were W _t = 8.4 × 10⁻³ L _t^3.10 (males) and W _t = 4.1 × 10⁻³ L _t^3.36 (females). The parameters for the von Bertalanffy growth function for pooled sexes were L _∞  = 33.17 cm, k = 0.20 year⁻¹, and t ₀ = −1.34 year. Perch in Lake Varese spawn from April through May. Sexual maturity is reached when males are 2 years old, in females mostly when they are 3 years old. Relative fecundity (F _rel) and absolute fecundity (F _abs) were assessed for females. Fecundity values were similar to data reported for other European populations: females of age 2+ F _rel = 102,457 ± 12,275, age 3+ F _rel = 131,767 ± 5,891, and age 4+ F _rel = 131,252 ± 15,555. Perch diet spectrum was wide and somewhat characterized by season. Perch in Lake Varese feed on macroinvertebrates, mainly Chironomidae and Chaoborus, zooplankton, and juvenile rudd Scardinius erythrophthalmus.     

Domestication of the mud crab <Emphasis Type="Italic">Scylla serrata</Emphasis>

The significant decrease in wild mud crab population highlights the need to manage the resources and domesticate crabs. This paper presents the initial results of the domestication of mud crab Scylla serrata aimed at producing good-quality captive broodstock. The analysis of the genetic structure of the base population was done as a prerequisite for domestication. Adult S. serrata from the northern to southern parts of the Philippines (Cagayan, Camarines, Samar, and Surigao) were obtained for genetic diversity analysis and domestication. Analysis of molecular variance showed that differences in the genetic variability between the four populations were not significant. Moreover, no significant deviation from Hardy–Weinberg Equilibrium was observed in each sample population and even in pooled populations. Body weight was positively correlated with the carapace width. Second spawning occurred 41–46 days after the first spawning and 34 days from second to third spawning. However, there was a decrease in the number of zoea in repeat spawnings. Twenty-four first-generation (F₁) families were produced from the four sites. The duration from spawning of the base population (P₀) to attainment of broodstock size F₁ was 10–14 months. Four second-generation (F₂) families were produced after 11–12 months. Up to the F₂, crabs tested negative for six viruses: white spot syndrome virus, infectious hypodermal and hematopoietic necrosis virus, gill-associated virus, yellow head virus, Taura syndrome virus, and infectious myonecrosis virus. The reproductive performance of P₀ was comparable to the succeeding generations. Several families were obtained from one population in a year. However, due to the cannibalistic behavior of crabs, more space is required for the nursery and grow-out phase. The domestication of S. serrata is the first study done on any mud crab species in the Indo-west Pacific region. The initial results would serve as guide to understand and eliminate the barriers to mud crab domestication. The breeding technology developed from this study will support the production of good-quality seedstock for farming.     

The use of alternative prey (crayfish, <Emphasis Type="Italic">Procambarus clarki</Emphasis>, and hake, <Emphasis Type="Italic">Merlucius gayi</Emphasis>) to culture <Emphasis Type="Italic">Octopus vulgaris</Emphasis> (Cuvier 1797)

The effects of two alternative prey (crayfish and hake) were tested on growth and survival of both juveniles and adults of Octopus vulgaris in two experiments. Octopuses fed the control (squid) were larger (3.0 ± 0.7 kg) than those fed crayfish (2.4 ± 0.6 kg) at the end of experiment I. Similarly, overall growth rates were higher for octopuses fed squid (1.7 ± 0.3 and 1.2 ± 0.2 %BW day⁻¹, respectively). Average feeding rates for the experiment were not different, being 6.5 ± 0.9 and 7.5 ± 0.9 %BW day⁻¹, respectively, for octopuses fed either squid or crayfish. Nevertheless, food conversions for the experiment were higher (42.4 ± 2.7%) for octopuses fed squid compared to the ones fed crayfish (23.9 ± 1.9 g). For experiment II, hake and crayfish were compared to squid; the final weight of octopuses fed squid, hake or crayfish was 1,183.0 ± 242.7 g, 1,175.6 ± 240.1 g and 922.3 ± 160.1 g, respectively. Overall growth rates for the experiment were 1.9 ± 0.2 %BW day⁻¹, 1.9 ± 0.3 %BW day⁻¹ and 1.1 ± 0.3 g, respectively. Final weight and growth rates were never different (P > 0.05) between octopuses fed squid and hake, but were always higher (P < 0.05) compared to the ones fed crayfish. Average feeding rates for experiment II were similar for the three diets, and of 4.6 ± 1.5, 4.2 ± 1.3 %BW day⁻¹ and 5.1 ± 0.9 %BW day⁻¹, respectively, for octopuses fed squid, hake or crayfish. Food conversions for experiment II were of 41.0 ± 9.6%, 40.5 ± 9.9% and 21.3 ± 7.4 g, respectively, for octopuses fed squid, hake or crayfish, and were always higher for octopuses fed squid and hake compared to crayfish. The results indicate that crayfish is not an adequate replacement for the usual prey to fatten octopus, even considering its much lower market price.     

Growth and survival of grouper <Emphasis Type="Italic">Epinephelus</Emphasis><Emphasis Type="Italic">coioides</Emphasis> (Hamilton) larvae fed free-living nematode <Emphasis Type="Italic">Panagrellus redivivus</Emphasis> at first feeding

The free-living nematode, Panagrellus redivivus, was tested as live food for grouper Epinephelus coioides larvae during the first feeding stage. A series of experiments were conducted to determine the acceptability of the free-living nematodes in grouper larvae at first feeding, the optimum nematode density and the response of the larvae to nutritionally enriched nematode. All experiments were conducted in 200-L conical tanks filled with 150-L filtered seawater and stocked at 15 larvae L⁻¹. Duration of feeding experiments was up to day 21 (experiment 1) and 14 days (experiment 2 and 3). Brachionus plicatilis and Artemia (experiment 1) and Brachionus plicatilis alone (experiment 2 & 3) was used as the control treatment. Observations indicated that the grouper larvae readily fed on free-living nematodes as early as 3 days posthatching, the start of exogenous feeding. Optimum feeding density for the larvae was 75 nematodes ml⁻¹. The enrichment of cod liver oil or sunflower oil influenced the total lipids and n-3 highly unsaturated fatty acids of P. redivivus, which in turn influenced those of the grouper larvae, however, growth and survival of the larvae were not affected (P > 0.05). The results from this investigation showed that the nematode, P. redivivus, can be used as first live food for grouper larvae from the onset of exogenous feeding until they could feed on Artemia nauplii.     

Spatial learning-induced <Emphasis Type="Italic">egr</Emphasis>-<Emphasis Type="Italic">1</Emphasis> expression in telencephalon of gold fish <Emphasis Type="Italic">Carassius auratus</Emphasis>

The immediate-early gene (egr-1) expression was used to examine the neuron’s response in telencephalon of goldfish during spatial learning in small space. Fishes were pre-exposed in the experimental apparatus and trained to pick food from the tray in a rectangular-shaped arena. The apparatus was divided into identical compartments comprising three gates to provide different spatial tasks. After the fish learned to pass through the gate one, two more gates were introduced one by one. Fish made more number of attempts and took longer time (P < 0.05) to pass through the first gate than the gate two or three. This active learning induces the expression of egr-1 in telencephalon as established by western blot analysis. Subsequently, the fish learn quickly to cross the similar type of second and third gate and make fewer errors with a corresponding decline in the level of egr-1 expression. As the fish learned to pass through all the three gates, third gate was replaced by modified gate three. Interestingly, the level of egr-1 expression increased again, when the fish exhibit a high exploratory behavior to cross the modified gate three. The present study shows that egr-1 expression is induced in the telencephalon of goldfish while intensively acquiring geometric spatial information to pass through the gates.     

Probiotic bacteria <Emphasis Type="Italic">Lactobacillus rhamnosus</Emphasis> influences the blood profile in rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis> (Walbaum)

This paper reports the effect of feeding probiotic diets on blood profiles in rainbow trout. Two experiments were performed: in the first, fish of average weight 75 g were offered either a commercial feed or the same incorporated with 10⁹ CFU g⁻¹ of lactic acid bacteria Lactobacillus rhamnosus for 30 days; in the second study performed for a similar duration, fish of average weight 126 g were offered formulated diets that either contained the same bacteria in heat-killed or freeze-dried form (nearly 10¹¹ CFU g⁻¹), or the basal diet without the bacteria. Blood samples were collected at different times after commencement of probiotic feeding to determine the total cholesterol, triglyceride contents, the plasma alkaline phosphatase activity, plasma protein and hematocrit value. The plasma cholesterol significantly increased upon probiotic feeding in the first experiment. A significant elevation (P < 0.05) of plasma cholesterol and triglyceride and alkaline phosphatase activity level was found in the freeze-dried probiotic fed groups at 20 and 30 days postfeeding. This was concomitant with the increased plasma protein and hematocrit values in FD group at 20 and 30 days. Likewise, the heat-killed probiotic fed group registered significantly high values of triglycerides, alkaline phosphatase activity, and plasma protein compared to the control diet fed groups after 20 days of feeding. Thus, alterations in the blood profiles could serve as supplementary information when examining the benefits of probiotics for fish.     

Quantitative PCR assay for the detection of the parasitic ciliate <Emphasis Type="Italic">Cryptocaryon irritans</Emphasis>

We developed a quantitative PCR assay for detecting the parasitic ciliate Cryptocaryon irritans, which causes “white spot disease” in marine fishes, from the natural environment. A specific primer set for C. irritans was designed and its high specificity was confirmed in silico: almost all of the sequences deposited in the GenBank nucleotide database were covered, 22/23 for the forward primer and 7/7 for the reverse primer. We estimated that there were 3,415.9 rRNA gene copies per genome of C. irritans. In artificial mixture experiments to validate whether the qPCR assay is applicable to natural samples, the estimated copy numbers showed significantly positive correlations with the number of theronts added (p < 0.001). When we applied this qPCR assay to natural samples collected bimonthly from surface and bottom seawaters at an aquaculture site (water depth, 10 m) from May 2009 to March 2010, we only detected C. irritans (112.0 ± 6.3 cells/l) in the surface seawater sample in November. This qPCR assay is a useful tool for detecting C. irritans rapidly and quantitatively in natural environments; it could also help advance our understanding of the ecology of C. irritans, as well as facilitate the diagnosis of the disease.     

Detection of prostaglandin E<Subscript>2</Subscript> in polychaete <Emphasis Type="Italic">Perinereis</Emphasis> sp. and its effect on <Emphasis Type="Italic">Penaeus monodon</Emphasis> oocyte development in vitro

Prostaglandins are involved in the reproductive processes in a variety of animals, including crustaceans. It was found that polychaetes, the best maturation diet for shrimp broodstock, possessed the greatest variation of prostaglandin E₂ (PGE₂) when compared with other live feeds. The level of PGE₂ varied according to sizes, feed intake, sources and type of polychaete. The matured and also larger sand polychaete Perinereis sp. contained higher PGE₂ levels than younger and smaller sand polychaetes (18.16 ± 5.82 ng PGE₂ mg⁻¹ protein for polychaetes at an average length of 10 cm up to 160.8 ± 37.09 ng PGE₂ mg⁻¹ protein for polychaetes at an average length of 17 cm). The PGE₂ levels in ovaries and haemolymph of female shrimp fluctuated with the developmental stage of the ovaries. The highest concentration of PGE₂ in haemolymph was at stage 3 of ovarian development, whereas the highest concentration of PGE₂ in shrimp ovaries was at stage 4. In vitro incubation of Penaeus monodon pre-vitellogenic oocytes with polychaete extract and synthetic PGE₂ demonstrated that both PGE₂s enhanced oocyte development, especially during late development and ovulation. The putative role of PGE₂ from polychaetes or the presence of PGE₂ in polychaetes may be a factor in their role as a dietary constituent required for shrimp oocyte development.     

Behavioural response of carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis>, L.) pond stock upon occurrence of hunting great cormorant (<Emphasis Type="Italic">Phalacrocorax carbo sinensis</Emphasis>) flocks

Besides various physiological and biological indices, stress in fish can be expressed directly by changes in their behaviour. Electrofishing point sampling approach based on CPUE (catch per unit effort) was used to evaluate fish density (escapement) into the littoral zones of four Czech ponds as a reaction to the occurrence and hunting activities of great cormorant (Phalacrocorax carbo sinensis) flocks. The behaviour of two-year-old common carp (Cyprinus carpio) before and after cormorant arrival was compared. Significant differences (P < 0.001) in CPUE were recorded in fish density before and after cormorant arrival on all of four ponds under study—Žofinsky: 0.04 ± 0.18 and 22.38 ± 26.87 (mean ± SD), Stary Hospodář: 0.05 ± 0.29 and 25.49 ± 25.45, Žebrákov: 0.16 ± 0.57 and 28.58 ± 24.75 and Travičny: 0.03 ± 0.19 and 17.18 ± 18.20, respectively. The evaluation of fish wounding in the CPUE fish samples proved the proportion of fish injured by cormorant attacks ranging from 3.33 to 18.19% in the ponds under study.