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.     

Loss of genetic variability in the captive stocks of tambaqui,Colossoma macropomum (Cuvier, 1818), at breeding centres in Brazil,and their divergence from wild populations

The loss of variability in farmed populations and the risks of interactions with wild populations support the need for the genetic monitoring of species farmed throughout the world. In Brazil, the tambaqui is the most widely farmed native fish species. Despite this, there are no data on the pedigree of the farmed stocks, and the potential for interactions with wild populations in the Amazon basin has raised concerns with regard to the genetic variability of these stocks. The present study analysed sequences of the mitochondrial Control Region and 12 microsatellites to characterize the genetic variability of seven historically important commercial tambaqui breeding centres located in four different regions of Brazil, and compared these sequences with those obtained from individuals collected from a wild population. High levels of genetic diversity were found in the wild population, whereas genetic diversity was reduced in both markers in most captive populations, except for the broodstock located near the Amazon River. High F_ST and D_EST indices were recorded between the wild population and most of the captive stocks analysed. The drastic reduction in genetic diversity found in most captive stocks and the difference between these stocks and the wild population may have been the result of the small size of the founding populations and the absence of breeding management. The renewal of the broodstocks and the application of breeding management techniques are recommended. In the Amazon region, in addition, the use of broodstocks that are genetically very different from local wild populations should be avoided.     

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.     

Essential oils to control ichthyophthiriasis in pacu,Piaractus mesopotamicus (Holmberg): special emphasis on treatment with Melaleuca alternifolia

In vitro effect of the Melaleuca alternifolia, Lavandula angustifolia and Mentha piperita essential oils (EOs) against Ichthyophthirius multifiliis and in vivo effect of M. alternifolia for treating ichthyophthiriasis in one of the most important South American fish, Piaractus mesopotamicus (Holmberg), were evaluated. The in vitro test consisted of three EOs, each at concentrations of 57 μL L^?1, 114 μL L ^?1, 227 μL L^?1 and 455 μL L ^?1, which were assessed once an hour for 4 h in microtitre plates (96 wells). The in vitro results demonstrated that all tested EOs showed a cytotoxic effect against I. multifiliis compared to control groups (P < 0.05). The in vivo treatment for white spot disease was performed in a bath for 2 h day^?1 for 5 days using the M. alternifolia EO (50 μL L ^?1). In this study, 53.33% of the fish severely infected by I. multifiliis survived after the treatment with M. alternifolia (50 μL L ^?1) and the parasitological analysis has shown an efficacy of nearly 100% in the skin and gills, while all the fish in the control group died. Furthermore, the potential positive effect of M. alternifolia EO against two emergent opportunistic bacteria in South America Edwardsiella tarda and Citrobacter freundii was discussed.     

Swimming performance of brown trout and grayling show species‐specific responses to changes in temperature

Fishways have historically been constructed to restore and preserve the ecological connectivity for fish in fragmented rivers. However, the fishways are often selective on species due to different size and swimming capacity. As the proportion of dammed rivers is still increasing, there is a growing need for more information on wild fish and their migration potential. In this study, we compared the swimming capacity of wild caught brown trout (Salmo trutta) and European grayling (Thymallus thymallus) until the fish were exhausted in a critical swimming speed (U_crit) test, under three different naturally occurring stream temperatures in Norway: 1.7, 5.5 and 10°C. The results indicated that trout swim better at the warmer temperatures than at colder temperatures. The grayling showed consistent swimming patterns with little variation across all tested temperatures. The results therefore signify the need to have operational fishways already early in the spring when the grayling migration starts and highlight the need for more studies on fish migration abilities across a wider range of species and seasons.     

Associations between morphometric variables and weight and yields carcass in Pirapitinga Piaractus brachypomus

The objective of this work was to evaluate the association between morphometric variables and carcass characteristics in Pirapitinga. We used a thousand specimens of Pirapitinga with an average weight of 1,200 g, which were stunned, slaughtered, weighed, measured, and processed for morphometric and processing yield analysis, to obtain weights, carcass and fillet yields. Initially, the linearity of the variables was verified. Pearson's simple and partial correlation tests were performed between all metrics. Track analysis was performed considering the weights and yields of carcass and filet as dependent variables and the others as independent variables. Ridge regression models were used to eliminate the effects of multicollinearity among the independent variables. Observations showed that the simple correlations between body weights and yields were superior to the partial ones in terms of magnitude. The length and circumference of the specimens were the measures most linearly associated with weight, carcass weight and fillet with rib weight. As for carcass yield and fillet yield, linear correlation coefficients were low and not significant when associated with body weights. With the path analysis, we could observe significant positive correlations between the morphometric measurements for weight, carcass weight and fillet with rib weight. The standard length, body circumference and body circumference/body height ratio are the most correlated. The latter are the most important variables in the evaluation of body weights in Pirapitinga fish. They can serve as criteria for indirect selection in searching for fish with better carcass characteristics. As for fillet and carcass yields, the morphometric variables have not shown to be efficient for indirect selection.     

The use of preserved copepods in sea bream small‐scale culture: biometric,biochemical and molecular implications

Considering the well‐known problems arising from the use of rotifers and Artemia as live prey in larval rearing in terms of fatty acid deficiencies, the aim of this study was to evaluate a partial or complete replacement of traditional live prey with preserved copepods during the larviculture of gilthead sea bream (Sparus aurata). Sea bream larvae were randomly divided into 4 experimental groups in triplicates: group A larvae (control) fed rotifers followed by Artemia nauplii; group B fed a combined diet (50%) of rotifers–Artemia and preserved copepods; group C fed rotifers followed by preserved copepods; and group D fed preserved copepods solely. Survival and biometric data were analysed together with major molecular biomarkers involved in growth, lipid metabolism and appetite. Moreover, fatty acid content of prey and larvae was also analysed. At the end of 40 days treatment, a stress test, on the remaining larvae, was performed to evaluate the effects of different diets on stress response. Data obtained evidenced a positive effect of cofeeding preserved copepods during sea bream larviculture. Higher survival and growth were achieved in group B (fed combined diet) larvae respect to control. In addition, preserved copepods cofeeding was able to positively modulate genes involved in fish growth, lipid metabolism, stress response and appetite regulation.     

Technical and economic feasibility of integrating seahorse culture in shrimp/oyster farms

Integrated multi‐trophic aquaculture systems (IMTA) are building ecosystems designed to produce aquatic organisms with less environmental impact. We have developed a simple system and a management strategy for introducing seahorse (Hippocampus spp.) culture in shrimp/oyster farms to increase economic sustainability. The system includes broodstock, nursery and grow‐out phases of Hippocampus reidi G. culture. We performed a test in a 42.4‐ha marine shrimp/oyster farm, which showed that this system is technically feasible. Then, we performed a partial budget analysis including cost‐return, cash flow and financial feasibility analysis. More than 12 000 marketable size seahorses may be produced in 402 m² annually. Initial investment was only US$ 21 103.00. The benefit–cost ratio was ~US$ 20.00 for each US$ 1.00 spent, internal rate of return was 131% and payback period <2 years. In conclusion, the introduction of seahorse in shrimp/oyster IMTA system is technically feasible, profitable, resilient and shows high liquidity. This system may be easily established in developing countries and has potential to provide seahorses to supply different markets (live specimens for the marine aquarium trade and dried specimens for traditional Chinese medicine and the curio trade). In addition, it can indirectly contribute to preserve natural populations.     

Replacement of fish oil with a DHA‐rich Schizochytrium meal on growth performance,activities of digestive enzyme and fatty acid profile of Pacific white shrimp (Litopenaeus vannamei) larvae

This trial was conducted to evaluate the effects of replacing dietary fish oil with Schizochytrium meal for Pacific white shrimp (Litopenaeus vannamei) larvae (initial body weight 4.21 ± 0.10 mg). Six test microdiets were formulated using Schizochytrium meal to replace 0 g/kg, 250 g/kg, 500 g/kg, 750 g/kg, 1000 g/kg or 1500 g/kg fish oil DHA. No significant differences were observed in survival, growth, final body length and activities of digestive enzyme among shrimp fed different diets (p > .05). No significant differences were observed in C20:5n‐3 (EPA) in muscle samples (p > .05). C18:3n‐3 and C20:4n‐6 in muscle increased as Schizochytrium meal replacement level increased (p < .05). No significant differences were observed in C22:6n‐3 (DHA) and n‐3 fatty acids among shrimp fed diets that algae meal replaced 0 g/kg ‐ 1000 g/kg of fish oil. Shrimp fed diet R150 had higher DHA content than other groups and had higher n‐3 fatty acids than that of shrimp fed diets R50, R75 and R100 (p < .05). C18:2n‐6, PUFA and n‐6 fatty acids in muscle increased, while n‐3/n‐6 ratio decreased with increasing algae meal replacement level from 0 g/kg to 1000 g/kg (p < .05). In conclusion, Schizochytrium meal could replace 1500 g/kg fish oil DHA in the microdiets without negatively affecting shrimp larvae survival, growth and activities of digestive enzyme.     

Probiotic characteristics and aflatoxin B1 binding ability of Debaryomyces hansenii and Kazaschtania exigua from rainbow trout environment

The aim of this study was to evaluate probiotic properties and the aflatoxin B₁ adsorption ability of yeasts isolated from rainbow trout intestine and fish feed to assess their use in the formulation of feed additives. Growth at pH 2, bacterial pathogens inhibition, bacterial pathogens co‐aggregation, autoaggregation, homologous and heterologous inhibition against lactic acid bacteria were evaluated. Moreover, aflatoxin B₁ adsorption was tested. All strains were able to maintain viable (10⁷ cells/ml) at pH 2. All strains isolated from intestine were identified as Kazaschtania exigua, while strains isolated from feed were all identified as Debaryomyces hansenii. Kazaschtania exigua RC035 and RC037 showed the strongest antimicrobial activity while K. exigua RC037 and RC038 were the most efficient co‐aggregating bacterial pathogens. All strains exhibited strong autoaggregation. None of the tested yeast strains showed homologous inhibition towards other yeasts and heterologous inhibition towards lactic acid bacteria strains. Debaryomyces hansenii RC031 demonstrated aflatoxin B₁ adsorption capacity (21%). The results of the present study indicate that select strains of Kazaschtania exigua and D. hansenii showed potential to improve the health of rainbow trout by inhibiting pathogens and binding AFB₁ and their use as probiotics may improve the production of rainbow trout in aquaculture systems.     

Ecology and conservation of the American eel in the Caribbean region

The majority of American eel, Anguilla rostrata LeSueur, knowledge is derived from temperate regions in the United States and Canada, with little known from its tropical Caribbean distribution. Findings of original research on American eel distribution, abundance, population biology, habitat ecology and threats from the Caribbean island of Puerto Rico were synthesised. American eel were captured from 48 of 116 sites (41.4%) in 26 of 49 river basins (53.1%) during 2005–2016, and it was extirpated upstream of dams and migration barriers >3.0 m high (38.9% of habitat). Mean density and biomass were 438.9 fish/ha and 23.44 kg/ha, respectively. Upstream habitats favoured larger individuals, and females were larger than males. The swim‐bladder parasite Anguillicoloides crassus Kuwahara, Niimi & Hagaki was not found in 120 eels examined. Realised threats include dams and other migratory barriers, habitat loss and alteration and pollution; exotic species and commercial fishing are impending threats; and the least understood is climate change.     

Survival and growth of reciprocal crosses between two stocks of the Hong Kong oyster Crassostrea hongkongensis (Lam & Morton, 2003) in southern China

The Hong Kong oyster, Crassostrea hongkongensis, is one of the most economically important and well‐known oyster species in southern China. To explore the possibility of improving the performance of the Hong Kong oyster, complete diallel crosses between two oyster stocks, the Zhuhai stock (Z: fast growth line F4) and the Maowei Sea stock (M: resistant line F3), were conducted using pooled gametes. Three replicates, each consisting of two pure stocks (ZZ and MM) and two reciprocal crosses (ZM and MZ), were successfully generated. High fertilization rate and hatching level were observed among all the experimental groups, suggesting that there was no sperm–egg recognition barrier between geographic populations. Reciprocal crosses had higher survival rate compared to the pure stock crosses and the rate of survival increased with progeny growth. Growth heterosis became obvious both in larval and adult stages, and was primarily influenced by the egg origin and mating strategy at the larval stage. Also, the phenotypic traits of all progeny differed amongst the culture sites, suggesting a significant environmental effect. The Zhuhai site was more suitable for oyster aquaculture than the Maowei Sea site. Thus, our results demonstrated that crossbreeding between Zhuhai and Maowei Sea stocks of Hong Kong oyster to produce superior heterosis represents a promising means to improve the fishery yield of this species in southern China.     

“吉富”系列罗非鱼生长性状的配合力、杂种优势与遗传相关分析

"吉富"系列罗非鱼是中国当前养殖的主要罗非鱼种类。该研究以3个品系"吉富"尼罗罗非鱼(Oreochromis niloticus)(新吉富、吉富、吉诺玛)为研究材料,应用3×3完全双列杂交设计构建了9个组合,采用数量遗传学方法对各组合子一代的体质量、全长、体长、体高和体宽5个生长性状的配合力、杂种优势与遗传相关进行了分析。结果表明:5个生长性状均未检测到一般配合力,而特殊配合力效应达62%以上,表明这些性状主要受到特殊配合力的影响;进一步的杂种优势分析发现吉诺玛♀×新吉富♂、新吉富♀×吉诺玛♂、吉富♀×新吉富♂这3个组合的杂种效果明显,其中以吉诺玛♀×新吉富♂为最佳组合;5个生长性状之间均存在显著的遗传相关与表型相关(P0.05),可进行多性状的聚合利用。该研究结果为"吉富"系列罗非鱼的配套利用提供了有益的资料和数据,同时为开展罗非鱼配套系育种提供了依据。     

Exophiala angulospora infection in hatchery‐reared lumpfish (Cyclopterus lumpus) broodstock

Samples from moribund lumpfish were collected in a marine hatchery in Scotland in 2015. Black nodules were noted on the skin, and gills and fungal hyphae were extensively distributed in musculature and internal organs. Multifocal chronic inflammatory lesions displaced structures in all affected organs. Mortalities commenced on completion of spawning in May and were evenly distributed over the second year in the temperature range 11–15°C. The main systemic infection causing agent was initially identified based on morphological characteristics as an Exophiala species. Ribosomal DNA (rDNA) ITS regions of the isolates were subsequently sequenced confirming the isolates belonged to Exophiala genus. All isolates fell in a single phylogenetic cluster, which is represented by Exophiala angulospora. Fish were treated with either formalin or Bronopol or a combination of both, but there was no effect on the pattern or numbers of mortalities. Isolates were also tested against three different concentrations of Latrunculin A, Amphotericin B and Itraconazole with no success. It is of utmost importance to increase the knowledge on pathogen–host interactions to successfully develop sustainable control methods.