Response of juvenile pacu (Piaractus mesopotamicus Holmberg, 1887) to balanced digestible protein

This 45‐day work aimed to determine the response of juvenile pacu (Piaractus mesopotamicus) to balanced digestible protein (BDP) and to use these responses to determine whether the optimum economic levels of BDP would differ depending on the form in which the fish is sold. Six isoenergetic diets containing 163, 201, 238, 272, 315 and 348 g/kg BDP (dry‐matter basis) were prepared through the serial dilutions of a high‐protein diet with the low‐protein diet. Fish (initial average body weight, 10.82 ± 0.14 g) were fed with respective experimental diets three times a day until apparent satiation. The optimum biological level of BDP was calculated as 326 g/kg (dry‐matter basis) by the quadratic regression model for maximum body weight gain of juvenile pacu. To maximize economic returns (US$/kg) for different end products, the optimum economic levels of BDP were calculated as 311, 317 and 319 g/kg (dry‐matter basis) by an economic model for whole‐body, eviscerated and sliced juvenile pacu respectively. This finding revealed that obtaining maximum biological performance of fish in the case of high feed costs or low prices of the end product in consumer market would substantially reduce the economic returns.     

Acute toxicity of ammonia in Pacu fish (Piaractus mesopotamicus,Holmberg, 1887) at different temperatures levels

Piaractus mesopotamicus juveniles (total length 12 ± 0.5 mm) were exposed to different concentrations of ammonia‐N (un‐ionized plus ionized ammonia as nitrogen), using the static renewal method at different temperature levels (15, 20 and 25°C) at pH 7. The 24, 48, 72, 96 h LC₅₀ values of ammonia‐N in P. mesopotamicus juveniles were 5.32, 4.19, 3.79 and 2.85 mg L^?1 at 15°C; 4.81, 3.97, 3.25 and 2.50 mg L^?1 at 20°C; and 4.16, 3.79, 2.58 and 1.97 mg L^?1 at 25°C respectively. The 24, 48, 72, 96 h LC₅₀ values of NH₃‐N (un‐ionized ammonia as nitrogen) were 0.018, 0.014, 0.013, 0.009 mg L^?1 at 15°C temperature; 0.023, 0.019, 0.016 and 0.012 mg L^?1 at 20°C; 0.029, 0.026, 0.018 and 0.014 mg L^?1 at 25°C. The temperature increase from 15 to 25°C caused an increase of ammonia‐N susceptibility by 21.80%, 9.55%, 31.92% and 30.87%, after 24, 48, 72 and 96 h exposure respectively. Furthermore, we found that exposure of fish to ammonia‐N caused an elevation in total haemoglobin and blood glucose with an increase of 2 mg L^?1 concentration. Ammonia levels tolerated, especially in different temperatures levels, have important implications for the management of aquaculture.     

Depletion study and estimation of withdrawal periods for florfenicol and florfenicol amine in pacu (Piaractus mesopotamicus)

The intensive production of farmed fish is at a global all‐time high, and the control of bacteria proliferation in fish farms requires the frequent use of antimicrobials. This practice raises important environmental concerns related to the emergence of antimicrobial‐resistant bacteria strains. Only a few antimicrobial drugs have been approved for use in aquaculture, one of which is florfenicol. This work studies the depletion and withdrawal period of florfenicol and its main metabolite, florfenicol amine, in pacu (Piaractus mesopotamicus), a neotropical characin widely farmed in the southern hemisphere. Juvenile pacu (average weight of 724 g) were stocked in a closed‐loop laboratory system with controlled water temperature (25.8°C), and were fed for 10 consecutive days with a diet containing an intended dose of 10 mg/florfenicol per kg bw. Muscle and skin tissues were collected at 1, 3, 6, 8, 10, 12 and 16 days post‐treatment, and florfenicol and florfenicol amine were quantified using a validated ultra‐high performance liquid chromatography‐tandem mass spectrometry (UPLC‐MS/MS) method. The limits of quantitation for florfenicol and florfenicol amine were 10 ng/g in muscle and 50 ng/g in skin. Considering a maximum residue limit of 1000 ng/g for the sum of florfenicol and florfenicol amine in muscle with skin in natural proportions a withdrawal period of 5 days (water temperature 25.8°C) or 129 degree days was calculated on the basis of the upper limit of the one‐sided 95% confidence interval for the 99^th percentile derived from the residue depletion study.     

Growth and metabolism of pacu (Piaractus mesopotamicus Holmberg 1887) juveniles fed diets containing different protein, lipid and carbohydrate levels

To verify the potential of lipids and carbohydrates to spare dietary protein and to understand the intermediary metabolism of interaction of these nutrients in pacu juveniles, an experiment was carried out to evaluate pacu physiological and performance parameters. The experimental design was completely randomized with 12 treatments in a 2 × 2 × 3 factorial arrangement, consisting of diets containing two digestible protein levels (200 and 230 g kg⁻¹ PD), two lipid levels (40 and 80 g kg⁻¹) and three carbohydrate levels (410, 460 and 500 g kg⁻¹). Fish‐fed 230 g kg⁻¹ digestable protein (DP) showed increased glycaemia, decreased hepatic glycogen, as well as a smaller intake index and better feed conversion ratio. The higher dietary lipid level (80 g kg⁻¹) reduced protein intake and serum protein concentration, increased liver and body fat content, but did not affect growth. At a lipid level of 80 g kg⁻¹, the increase in dietary carbohydrate levels promoted greater weight gain (WG), crude protein intake (CPI) and better feed conversion ratio (FCR). For fish fed diets containing 40 g kg⁻¹ lipid, the best energy‐productive values (EPV) were obtained at 460 g kg⁻¹ carbohydrate. Increased levels of the main nutrients in the diets reduced the levels of serum triglycerides, while the increase in energy concentration increased the hepatosomatic (HSI) and glycaemia index values. Pacu used lipids as effectively as carbohydrates in the maximization of protein usage, as long as dietary protein was at a level of 230 g kg⁻¹ DP. The physiological parameters indicated that the best balance between the DP, dietary lipid and carbohydrate levels within the ranged this trial was obtained at 230, 40 and 460 g kg⁻¹, respectively, without lower growth.     

Growth performance and body composition of pacu Piaractus mesopotamicus (Holmberg 1887) in response to dietary protein and energy levels

Improper dietary protein and energy levels and their ratio will lead to increased fish production cost. This work evaluated effects of dietary protein : energy ratio on growth and body composition of pacu, Piaractus mesopotamicus. Fingerling pacu (15.5 ± 0.4 g) were fed twice a day for 10 weeks until apparent satiation with diets containing 220, 260, 300, 340 or 380 g kg^?1 crude protein (CP) and 10.9, 11.7, 12.6, 13.4 or 14.2 MJ kg^?1 digestible energy (DE) in a totally randomized experimental design, 5 × 5 factorial scheme (n = 3). Weight gain, specific growth rate increased and feed conversion ratio (FCR) decreased significantly (P < 0.05) when CP increased from 220 to 271, 268 and 281 g kg^?1 respectively. Pacu was able to adjust feed consumption in a wide range of dietary DE concentration. Fish fed 260 CP diets showed best (P < 0.05) protein efficiency ratio and FCR with 11.7–12.6 MJ kg^?1; but for the 380 CP‐diets group, significant differences were observed only at 14.2 MJ kg^?1 dietary energy level, suggesting that pacu favours protein as energy source. DE was the chief influence on whole body chemical composition. Minimum dietary protein requirement of pacu is 270 g kg^?1, with an optimum CP : DE of 22.2 g MJ^?1.     

Growth,nitrogen gain and indispensable amino acid retention of pacu (Piaractus mesopotamicus,Holmberg 1887) fed different brewers yeast (Saccharomyces cerevisiae) levels

A feeding‐and‐digestibility trials were carried out to evaluate the efficacy of replacing fishmeal with brewers yeast Saccharomyces cerevisiae in diets of pacu, Piaractus mesopotamicus, juveniles. The feeding trial was conducted during 54 days with 450 fish (26.6 ± 1.7 g) testing six isonitrogenous (270 g kg^?1 crude protein) and isoenergetic (19 MJ kg^?1 crude energy) diets, with increasing yeast level to replace 0 (control), 30, 35, 50, 70 or 100% of dietary fishmeal. Growth performance and feed utilization increased with increasing dietary yeast level until 50% fishmeal replacement. Protein retention efficiency was higher in fish fed 35 and 50%. Protein digestibility and the fillet hue (the red/green chromaticity) were not significantly different among all treatments. Nitrogen gains were significantly improved in fish fed 35% replacement diet compared to fish fed the control diet. The retentions of indispensable amino acids tended to increase with increasing dietary yeast levels, with maximum retention at 35–50%. On the basis of our results, replacing 50% fishmeal by yeast in pacu diets successfully improved feed efficiency and growth performance, and reduced nitrogen losses, thereby reducing the nitrogen outputs from fish farms.     

Effects of processing on physical characteristics of diets with distinct levels of carbohydrates and lipids: the outcomes on the growth of pacu (Piaractus mesopotamicus)

The use efficiency and feed conversion of extruded and pelletized diets were compared. Eight isoproteic diets (220 g kg⁻¹ digestible protein) were assayed for 90 days in a 2 × 2 × 2 multifactorial design with two carbohydrate levels (400 and 500 g kg⁻¹), two lipids levels (40 and 80 g kg⁻¹) and two diet processing (pelletization and extrusion) with three repetitions. The growth of Piaractus mesopotamicus fed with these diets and the quality control indices of diets were gauged. The density of extruded diets was lower as carbohydrate level was 400 g kg⁻¹ and lipid 40 g kg⁻¹. The interaction carbohydrate and diet processing presented higher leaching value for low carbohydrate level in extruded diet. Fish fed with extruded diets presented the best feed conversion and protein efficiency ratio. When high levels of carbohydrate and lipid are combined, the weight gain is impaired. The interaction between diet processing diet and lipid levels resulted in the best fish performance when pelletized diets with 40 g kg⁻¹ lipid or extruded diets with 80 g kg⁻¹ lipid were considered. The protein efficiency ratio increased with the increment of carbohydrates in the pelletized diets. The fish show low tolerance to lipids and a preference for carbohydrate when the lipid productive values are taken into account.     

Efficacy of garlic (Allium sativum) extract applied as a therapeutic immersion treatment for Neobenedenia sp. management in aquaculture

Garlic, Allium sativum L., extract administered as a therapeutic bath was shown to have antiparasitic properties towards Neobenedenia sp. (MacCallum) (Platyhelminthes: Monogenea) infecting farmed barramundi, Lates calcarifer (Bloch). The effect of garlic extract (active component allicin) immersion on Neobenedenia sp. egg development, hatching success, oncomiracidia (larvae) longevity, infection success and juvenile Neobenedenia survival was examined and compared with freshwater and formalin immersion. Garlic extract was found to significantly impede hatching success (5% ± 5%) and oncomiracidia longevity (<2 h) at allicin concentrations of 15.2 μL L^?1, while eggs in the seawater control had >95% hatching success and mean oncomiracidia longevity of 37 ± 3 h. At much lower allicin concentrations (0.76 and 1.52 μL L^?1), garlic extract also significantly reduced Neobenedenia infection success of L. calcarifer to 25% ± 4% and 11% ± 4%, respectively, compared with 55% ± 7% in the seawater control. Juvenile Neobenedenia attached to host fish proved to be highly resistant to allicin with 96% surviving 1‐h immersion in 10 mL L^?1 (15.2 μL L^?1 allicin) of garlic extract. Allicin‐containing garlic extracts show potential for development as a therapy to manage monogenean infections in intensive aquaculture with the greatest impact at the egg and larval stages.     

Use of neem oil (Azadirachta indica) to control caligid copepod infestation on Asian seabass (Lates calcarifer)

Asian seabass (Lates calcarifer) culture in marine cages have been developed rapidly in Malaysia. The high intensity of culture might facilitate several disease infection including caligid infestation. This study was conducted to evaluate the effect of neem oil (Azadirachta indica) against caligid parasites on seabass. The prevalence and clinical pathogen of Caligus infection has been described through a sampling of 150 fish from floating cage at different cultured periods. Acute toxic tests with different concentrations of neem oil were carried out on the parasite and fish host. Results revealed that the 96‐hr median lethal concentration (LC₅₀) of Caligus and fish (10.3 ± 2.5 g of body weight) were 2 and 20 ppm respectively. The in vivo test indicated that 10 ppm of neem oil could result in 100% antiparasitic efficacy within 96 hr. These results therapeutically demonstrated the efficacy of neem oil in caligid control.     

Stable carbon (δ13C) and nitrogen (δ15N) isotopes as natural indicators of live and dry food in Piaractus mesopotamicus (Holmberg, 1887) larval tissue

This study proposed the use of the stable isotope technique to track the type of food utilized by pacu Piaractus mesopotamicus larvae during their development, and to identify the moment when the larvae start using nutrients from the dry diet by retaining its carbon and nitrogen atoms in their body tissues. Five‐day‐old pacu larvae at the onset of exogenous feeding were fed Artemia nauplii or formulated diet exclusively; nauplii+formulated diet during the entire period; or were weaned from nauplii to a dry diet after 3, 6 or 12 days after the first feeding. δ¹³C and δ¹⁵N values for Artemia nauplii were ?15.1‰ and 4.7‰, respectively, and ?25.0‰ and 7.4‰ for the dry diet. The initial isotopic composition of the larval tissue was ?20.2‰ and 9.5‰ for δ¹³C and δ¹⁵N respectively. Later, at the end of a 42‐day feeding period, larvae fed Artemia nauplii alone reached values of ?12.7‰ and 7.0‰ for δ¹³C and δ¹⁵N respectively. Larvae that received the formulated diet alone showed values of ?22.7‰ for δ¹³C and 9.6‰ for δ¹⁵N. The stable isotope technique was precise, and the time at which the larvae utilized Artemia nauplii, and later dry diet as a food source could be clearly defined.     

Preliminary success using hydrogen peroxide to treat Atlantic salmon,Salmo salar L., affected with experimentally induced amoebic gill disease (AGD)

Currently, the only effective and commercially used treatment for amoebic gill disease (AGD) in farmed Tasmanian Atlantic salmon is freshwater bathing. Hydrogen peroxide (H₂O₂), commonly used throughout the aquaculture industry for a range of topical skin and gill infections, was trialled in vitro and in vivo to ascertain its potential as an alternative treatment against AGD. Under in vitro conditions, trophozoites of Neoparamoeba perurans were exposed to three concentrations of H₂O₂ in sea water (500, 1000 and 1500 mg L^?1) over four durations (10, 20, 30 and 60 min) each at two temperatures (12 and 18 °C). Trophozoite viability was assessed immediately post‐exposure and after 24 h. A concentration/duration combination of 1000 mg L^?1 for >10 min demonstrated potent amoebicidal activity. Subsequently, Atlantic salmon mildly affected with experimentally induced AGD were treated with H₂O₂ at 12 and 18 °C for 15 min at 1250 mg L^?1 and their re‐infection rate was compared to freshwater‐treated fish over 21 days. Significant differences in the percentage of filaments affected with hyperplastic lesions (in association with amoebae) and plasma osmolality were noted between treatment groups immediately post‐bath. However, the results were largely equivocal in terms of disease resolution over a 3‐week period following treatment. These data suggest that H₂O₂ treatment in sea water successfully ameliorated a clinically light case of AGD under laboratory conditions.     

Reproductive cycle,collection and early growth of Aulacomya ater,Molina 1782 (Bivalvia: Mytilidae) in northern Chile

We evaluated the reproductive cycle of Aulacomya ater at Caleta Punta Arenas (Antofagasta, Chile), finding two important spawning periods (August–September and October–November) associated with declining water temperatures. Mytilid collectors were installed at Caleta Punta Arenas and Caleta Errázuriz (250 km further south) in November and then extracted in consecutive time periods. The results showed settlements of A. ater along with Choromytilus chorus (Molina) and Semimytilus algosus (Gould). An analysis of the spat size structure from each period revealed different micro‐cohorts for each species; the micro‐cohorts of C. chorus were predominant. Daily growth rates were estimated using the average sizes of the micro‐cohorts on two consecutive dates. Although the daily growth rates did not vary significantly between sites for a given species, these rates were lowest for A. ater and highest for S. algosus. The abundance and greater growth of C. chorus caused interspecific competition with A. ater that was detrimental to the survival of the latter. We hypothesize the existence of a metapopulation of A. ater in northern Chile that would explain the settlement of four micro‐cohorts on collectors after 28 days at Caleta Punta Arenas, and the settlement recorded after 80 days at Caleta Errázuriz.     

Antifungal effect of Origanum onites essential oil as an alternative to formalin in the artificial incubation of narrow‐clawed crayfish (Astacus leptodactylus Eschscholtz, 1823)

As alternative to formalin, the antifungal effect of a plant product [Origanum onites L. (Lamiaceae) oil] was investigated for use in the artificial incubation of narrow‐clawed crayfish (Astacus leptodactylus Eschscholtz) eggs. For this purpose, this study was conducted as two experiments. In experiment I, the eggs were artificially incubated for 40 days. In experiment II, juveniles were cultured to determine effects of O. onites oil on juveniles for 30 days. The experimental groups were as follows: formalin (3500 ppm for 15 min), O. onites oil (300 ppm for 15 min, 700 ppm for 2 min and 1000 ppm as a dip treatment 15 split‐second) and a control (no treatment). In the experiment I, the highest hatching rate (86%) and survival rate of stage II juveniles (80%) were observed in 1000 ppm dip group. These results were similar to that of formalin group (85% and 79%) respectively. The control group exhibited the lowest hatching rate (49%) and stage II rate (42%) compared with the 1000 ppm dip group and 3500 ppm formalin treatments. However, other concentrations (300 and 700 ppm) of O. onites showed toxic effects on the eggs and there was no hatching. In the experiment II, the survival rate and growth performance of the crayfish juveniles were similar in all groups. This study indicated that the 1000 ppm O. onites dip treatment could be a good alternative to formalin for improved egg hatchability in the artificial incubation of crayfish eggs.     

Larval rearing of the giant Azorean barnacle,Megabalanus azoricus (Pilsbry, 1916): feeding trials,larval development and settlement on artificial substrata

A series of experiments were conducted to obtain an efficient larval rearing protocol for Megabalanus azoricus. The first part of this study investigates the effect of microalgae‐based diets on survival and larval development. Mono and mixed‐diets were tested at 20 ± 1°C, in a sequence of 11‐day feeding experiments. The second part presents a preliminary study on the influence of a biofilm on recruitment and use of oyster spat collectors in a mass rearing system. A photographic record of larval development and a brief reference to the diagnostic features that enable quick larval staging are also presented, along with morphometric measurements. Of the microalgae tested (Chaetoceros sp., Chloromonas sp., Dunaliella sp., T‐Isochrysis sp. and Skeletonema sp.) the mixed‐diet Skeletonema sp. with T‐Isocrysis sp. showed the highest survival percentages: total survival ranged from 79.7 to 85.7% and 69.7–80.0% of nauplii were in stage VI after 11 days of rearing. Cypris were also present, but only represented 5.3% of the survivors at most. In the mass rearing system juveniles were found settled in the collectors after 25 days, at 20 ± 1°C. However recruitment was less than 1%. Preliminary results showed no settlement preference towards collectors with biofilm. Nevertheless, this study provides the first record of M. azoricus settlement under laboratorial conditions and represent a starting point for future larval rearing studies.     

Total nitrogen,carbon and phosphorus digestibility in gilthead seabream (Sparus aurata) and European seabass (Dicentrarchus labrax) fed with conventional and organic commercial feeds: implications for particulate waste production

Organic fish farming is postulated as a more environmentally friendly activity than conventional activities. However, environmental sustainability as regards waste output has not been addressed. In this respect, the most important environmental problems associated with fish farming are those related with the discharge of particulate wastes. In this study, total nitrogen, carbon and phosphorus digestibility was estimated in a wide size range of gilthead seabream (Sparus aurata) and European seabass (Dicentrarchus labrax) fed a commercially available conventional diet and an organic feed, as a previous step to assessing particulate waste output. Neither fish size nor feed nutrient content was seen to influence nutrient digestibility in either species. The organic diet provided higher carbon but lower nitrogen and phosphorus digestibility than conventional feed in both species. Simulations under real ongrowing conditions resulted in lower values for nitrogen and higher values for carbon particulate wastes when conventional feed was supplied to both species. Particulate phosphorus wastes were 2.6‐ to 3.4‐fold higher for fish fed the organic diet. Restrictions concerning NO the raw materials used for organic diet formulation and differences in nutrient bioavailability between the diets seemed to be the underlying reason for the differences found.     

The South American freshwater fish Prochilodus lineatus (Actinopterygii: Characiformes: Prochilodontidae): new species in Vietnamese aquaculture

The freshwater characiform fish Prochilodus lineatus is a detritivorous species that has its native distribution area in South America but has been imported to China for aquaculture purposes. This is the first time that it is being reported in Vietnam, both from aquaculture and captured from a river channel. According to local authorities, the species is becoming increasingly important in local aquaculture and its spread can be expected. Keeping in mind the high biomass production in the rivers of its native distribution area, a successful establishment of P. lineatus into Vietnamese fresh waters may exert serious impacts on the local ecosystems.     

Effects of rearing density on growth,fatty acid profile and bioremediation ability of polychaete Nereis diversicolor in an integrated aquaculture system with rainbow trout (Oncorhynchus mykiss)

The objective of this study was to assess the effects of rearing density on the growth and fatty acid profile of Nereis diversicolor and on its capability to bioremediate wastewater in an integrated culture system with rainbow trout (Oncorhynchus mykiss). To this end, a batch of juvenile N. diversicolor (0.03 ± 0.01 mg) was assigned into four different densities (T₁ = 250, T₂ = 500, T₃ = 1,000, T₄ = 2,000, besides T₀ = with no worm) in three replicates. The worm groups were fed with solid waste that was supplied from tanks containing rainbow trout (107.17 ± 13.5 g; 1.39 ± 0.18 kg/m³). During the experiment (60 days), the water temperature was 17.71 ± 0.6°C. The results revealed that SR%, SGR% and WG% of N. diversicolor in T₁ were significantly higher than those of T₂, T₃ and T₄. Both FER rate and biomass gain in T₄ were significantly higher than those in the other groups. The highest removal rate of NO₂‐N (73.72%), NH₃‐N (65.70%), PO₄‐P (59.32%), BOD₅ (69.60%) and TSS (82.33%) were observed in T₄. The major fatty acids presents were palmitic acid, oleic acid, linoleic acid, stearic acid and alpha‐linolenic acid in all worm‐treated groups, with no difference observed in the concentration of these fatty acids among them. Taken together, these findings suggest that organic wastes from rainbow trout farms could be recycled to achieve a sustainable aquaculture goal, and demonstrate that a high percentage of fatty acids in fish feed is not absorbed by rainbow trout but is done by N. diversicolor.     

Influence of traditional Chinese medicine and Bacillus species (TCMBS) on growth,immune response and disease resistance in Nile tilapia,Oreochromis niloticus

Multispecies herbs and probiotic bacteria offered to fish might enhance their immune response and increase disease resistance, but the dose effects of herbal‐probiotic application remain unclear. Therefore, the effect of herbal‐probiotic mixtures of traditional Chinese medicine (TCM) of composition Astragalus membranaceus, Angelica sinensis, Crataegus hupehensis and probiotic Bacillus species (BS) of composition Bacillus subtilis and Bacillus lincheniformis as natural immunostimulants in tilapia Oreochromis niloticus have been investigated. Fish were randomly divided into four groups: control diet (CT), TCMBS1 [TCM at 3 g/kg and BS at 7 g/kg], TCMBS2 [TCM at 5 g/kg and BS at 5 g/kg], TCMBS3 [TCM at 7 g/kg and BS at 3 g/kg]. Tilapia in the TCMBS3 group showed significant improvement in weight gain, specific growth rate, and lowered feed conversion ratio compared with other treated groups and the control. Concerning immune indexes, all treated groups significantly enhanced lysozyme, superoxide dismutase, catalase, protease and antiprotease activities, with highest values in catalase and antiprotease activities in TCMBS3 compared with control. TCMBS3 demonstrated higher expression of βeta‐defensin, lysozyme, heat shock protein 70, catalase and transforming growth factor‐βeta compared with other treated groups or the control group in both mid‐intestines and head‐kidney. After challenge with Streptococcus agalactiae, the best survival was found in TCMBS3 (97%), followed by TCMBS2 (73%), and TCMBS1 (69%) compared with the CT (35%). Collectively the present results suggest that TCMBS3 dose might potentiate a higher immune response and disease resistance in fish.