IncK plasmid-mediated tetracycline resistance in Edwardsiella ictaluri isolates from diseased freshwater catfish in Vietnam

Eight tetracycline resistant Edwardsiella ictaluri isolates obtained from diseased freshwater catfish (Pangasianodon hypophthalmus) in Vietnam, and showing different resistance phenotypes to other antimicrobial agents, were studied. The tet genes were determined using PCR. Conjugation experiments were performed to assess transferability of the tetracycline resistance determinant and the size and incompatibility group (Inc) of each tet-carrying plasmid were determined. PCR and sequencing were used for characterization of the co-transferred resistance genes. A tetA gene was demonstrated in the E. ictaluri isolates and for all of them, Escherichia coli transconjugants were obtained. All transconjugants contained high-molecular weight tetA-carrying plasmids (~ 140 kb) belonging to the incK group, as was shown with the PCR-based replicon typing method. The strA–strB, dhfr1 and sul 2 genes were detected on the tetA-carrying plasmids of the transconjugants showing resistance to streptomycin, trimethoprim and sulfonamides, respectively. The dhfr1 gene was found to be located in a class 1 integron as determined by PCR and sequencing. Interestingly, the 3′ CS region of class 1 integrons was not detected by PCR. This study shows the presence of incK plasmid-mediated tetracycline resistance among E. ictaluri isolates from diseased freshwater catfish in Vietnam.     

Preferences for and perception of cultured marine baitfish by recreational saltwater anglers in Florida

There is strong interest in culturing live marine baitfish for anglers. However, there is scant information about saltwater anglers’ perception of cultured baitfish. Thus, a survey was developed and administered to assess recreational saltwater angler’s preferences for baitfish, their attitudes towards cultured baitfish, supply and demand, and specific information about pinfish (Lagodon rhomboides) and pigfish (Orthopristis chrysoptera). An online survey invitation was e-mailed to 227,829 licensed marine recreational anglers in Florida. A total of 14,871 respondents completed some portion of the survey (6.5% response rate). Inshore destinations were the most common fishing trips taken. Live bait was caught by 57.8% of respondents, purchased by 32.1%, and 10.1% did not use live bait. Perceptions of aquaculture as a means of producing bait were 43.8% positive, 20.6% somewhat positive, and 31% unknown. Consistent availability and price were identified by anglers as the two most important characteristics for an aquaculture produced marine baitfish.     

Quantity, motility and fertility of tench Tinca tinca (L.) sperm in relation to LHRH analogue and carp pituitary treatments

The spermiation of tench males was stimulated with Supergestran containing mammalian LHRHa lecireline at the following doses: 5, 10, 20 and 40 g kg⁻¹ b.w.; then with carp pituitary suspension (CPS) at a dose of 2 mg kg⁻¹ b.w. and with a control of saline physiological solution. The following days, meaning 24, 48 and 72 h after injection, sperm was collected to evaluate volume and the number of sperm per male per kg body weight (B.W.) The percentage of motile sperm and velocity of spermatozoa were measured 48 h after hormonal injection, and 72 h after hormonal injection the sperm was evaluated for fertilization and hatching ability. All 42 males in experimental groups were diploid. Live weight did not differ significantly among experimental groups. The strongest stimulation of spermiation was achieved with LHRHa in dosage of 20 and 40 g kg⁻¹ b.w. and CPS compared to males of the control group and lower dosage of LHRHa. Analysis of variance showed no significant influence of the treatment on the velocity and percentage of motile spermatozoa. The effect of different treatment on the fertilization capacity (the number of spermatozoa per egg was equilibrated) was significant. Significantly the highest quality of sperm collected 72 h after injection expressed by percentage of fertilization and hatching (62–65% fertilization and 61–64% hatching rates, respectively) was found for LHRHa in dosage of 20 and 40 g kg⁻¹ b.w. Significantly the lowest parameters of fertilization and hatching were found for the control group, on the 12% level.     

Diet and trophic role of octopus on an abalone sea ranch

Gastric tract (crop and stomach) and stable isotope analyses (SIA) were used to evaluate the dietary composition of resident, commercially important Octopus aff. O. tetricus Gould, and its importance as a predator of Haliotis laevigata Donovan, on artificial abalone habitats. Taxonomic diversity and resolution of diet were higher in crops (13 taxa) than stomachs (10 taxa), with the latter dominated by unidentifiable material (%Volume = 74%). Crops contained mainly molluscs (~54%, including 31% abalone) and crustaceans (33%), suggesting octopus are important predators of abalone on the sea ranch. Dietary composition of crops did not differ with increasing octopus weight (<300 g, 300–999 g, ≥1,000 g wet weight), paralleling the findings of SIA and reflecting the high stocking densities of abalone that are able to be consumed by octopus of all sizes. The SIA also revealed that octopus occupy a mid‐trophic level, slightly below teleosts and loliginid squids.     

Effects of dietary carotenoid on the gut and the gonad of the sea urchin Paracentrotus lividus

Gonad colour, determined by accumulated carotenoids, is an important marketability factor in sea urchin products. In this study, dietary carotenoids were fed to Paracentrotus lividus in prepared diets to test their effects on the sea urchin's gonad colour. All diets had the same basic content of protein, lipid, carbohydrate, ash and energy. Four diets were enriched with: (I) all-trans β-carotene; (II) astaxanthin; (III) zeaxanthin/lutein/β-carotene; and (IV) capsanthin/zeaxanthin/lutein/cryptoxanthin/β-carotene. A fifth diet had Dunaliella bardawil algal powder containing all-trans and 9-cis β-carotene added, and a sixth one was a control diet with no added carotenoids. Sea urchins were fed the control diet (no pigments) for 4 weeks, then the six experimental diets for 8 weeks. Carotenoid concentration in the gut and gonad was determined by tri-dimensional photo-diode array high performance liquid chromatography. Four qualitative colour categories were established: brown, pale yellow, medium-orange, and mango-orange.Gonad development and test diameter increased equally in all diet treatments. Carotenoid profile and concentration in the gut were higher than in the gonad or in the feed. Total carotenoid concentration was four- to ten-fold greater in the gut than in the gonad, and carotenoid profile in the gut was different from that of the diet. Gut and gonad total carotenoids, β-carotene, and echinenone concentration were not dependent on total dietary carotenoid concentration. Total carotenoid and β-carotene concentration in the gut, and total carotenoid and echinenone concentration in the gonad were significantly dependent on dietary β-carotene concentration. The percentage of gonads with acceptable colour was positively correlated with dietary and gut β-carotene concentration. Carotenoids not normally contained in the diet of P. lividus (astaxanthin, capsanthin and capsorbin), did not accumulate in the gonad. Echinenone was found in all gut and gonad samples. Over 90% of the gonads from P. lividus fed Dunaliella algal powder diet developed the optimal mango-orange colour and this diet led to the greatest echinenone concentration.     

养殖水电化学同步脱氮响应面优化与验证

本研究先通过单因子实验分析了电流密度、极板面积比、极板间距和初始pH对总氨氮(TAN)和硝态氮(NO3–-N)去除率的影响。采用Design-Expert软件中Box-Behnken的中心组合原则设计四因素三水平响应面实验,考察不同影响因子对脱氮效率的影响,并建立响应面模型优化反应条件,最后对优化的反应条件进行验证。结果显示,电流密度、极板面积比、极板间距和初始pH的变化对TAN去除影响不大,在所选反应条件下,TAN去除率均高于80%,但反应条件的改变显著影响硝酸盐(NO3?)的去除,NO3?去除率在29.8%~80.9%范围内变化。响应面模型的回归系数R2为0.9340,校正系数R2为0.8681,说明该模型具有较好的准确性。NO3?去除最优反应条件：电流密度为25.6 mA/cm2,阴阳极板面积比为1.6∶1,极板间距为2.5 cm,初始pH为6.6,对该反应条件下的脱氮效果展开实验验证发现,TAN去除率为87.3%,NO3?去除率为81.5%。研究表明,电化学处理可实现对TAN和NO3–-N的同步去除,同时,响应面模型的运用有助于优化电化学法在养殖水处理中的脱氮效率。     

养殖水体耐盐高效降亚硝酸盐氮和氨氮芽孢杆菌的筛选与鉴定

亚硝酸盐氮和氨氮是养殖水体恶化的主要成分。从对虾养殖水体中,分离筛选出2株分别对亚硝酸盐氮和氨氮具有较高降解能力的耐盐芽孢杆菌菌株T905和T301。在模拟淡水和海水条件下,当亚硝酸盐氮和氨氮初始浓度分别为44 mg/L和20 mg/L时,3 d后菌株T905对亚硝酸盐氮降解率分别达到72.10%和92.10%,T301对氨氮降解率分别达到55.18%和52.00%。根据形态学特征和生理生化试验结果,鉴定2株菌为枯草芽孢杆菌。     

Antibiofilm potential of a tropical marine Bacillus licheniformis isolate: role in disruption of aquaculture associated biofilms

Microbial biofilms are important in aquaculture industries as they resist antibiotic treatments. In this study, we have investigated the antibiofilm potential of a tropical marine culture Bacillus licheniformis D1 (containing an antimicrobial protein BLDZ1) against two aquaculture associated pathogens namely, Vibrio harveyi and Pseudomonas aeruginosa. Both the test cultures formed biofilms on polystyrene and glass surfaces. The cell free supernatant (CFS) of B. licheniformis inhibited V. harveyi and P. aeruginosa biofilms on polystyrene surfaces up to around 80% and 78% respectively. In addition, the CFS disrupted pre‐formed biofilms of test cultures by about 73%. Fluorescence and scanning electron microscope analysis confirmed the antibiofilm potential of the CFS. The cell free supernatant displayed antiadhesive activity that inhibited the initial attachment of the bacteria during the process of biofilm formation. In addition, the CFS exhibited antimicrobial activity and mediated cell death via cytoplasmic membrane disruption.     

The tetrasporophyte of Asparagopsis armata as a novel seaweed biofilter

The red seaweed Asparagopsis armata (Harvey; Rhodophytae, Bonnemaisoniaceae) produces biologically active secondary metabolites that are valuable natural ingredients for cosmetics and medicine and its cultivation may therefore be a profitable venture. The tetrasporophyte of this species (“Falkenbergia rufolanosa”) was successfully tank-cultivated as a continuous biofilter for the effluent of a commercial fish farm in southern Portugal. Optimal stocking density for highest biomass yield and a low level of other algal species in winter and late spring was 5×g centrifuged fresh weight l^− 1. The effect of total ammonia nitrogen supply (TAN flux) on biofiltration and biomass yield was investigated in winter and spring. Results revealed that A. armata is currently the seaweed-biofilter with the highest TAN removal of up to 90 μmol l^− 1 h^− 1 at a TAN flux of about 500 μmol l^− 1 h^− 1. In the tanks used, this is equivalent to a removal of up to 14.5 g TAN m^− 2 day^− 1. At a lower TAN flux of about 40 μmol l^− 1 h^− 1, TAN removal by A. armata is more than double to what is reported at this flux for another successful seaweed biofilter, the genus Ulva. Monthly variation of A. armata biomass yield peaked in May and was lowest in January. At TAN fluxes between 300 and 400 μmol l^− 1 h^− 1, an average water temperature of 21.7 °C and a total daily photon flux density of 47 Mol m^− 2, seaweed yield was over 100 g DW m^− 2 day^− 1 with a recorded maximum of 119 g. During spring, autumn and early summer, the biomass of A. armata within the experimental tanks doubled every week. A model for the up scaling of this finfish integrated aquaculture of A. armata varies the investment in biofilter surface area and estimates the return in biofiltration and biomass yield. Highest TAN removal efficiencies will only be possible at low TAN fluxes and a very large biofilter area, resulting in a low production of biomass per unit area. To remove 50% of TAN from the effluent (1 mt Sparus aurata; 21 °C), 28 m² of biofilter, designed to support a water turnover rate of 0.8 Vol h^− 1 would be necessary. This system produces 6.1 kg FW (1.5 kg DW) of A. armata per day and has the potential to turn biofiltration into an economically sustained, beneficial side effect.     

The basics of bio-flocs technology: The added value for aquaculture

The expansion of the aquaculture production is restricted due to the pressure it causes on the environment by the discharge of waste products in the water bodies and by its dependence on fish oil and fishmeal. Aquaculture using bio-flocs technology (BFT) offers a solution to both problems. It combines the removal of nutrients from the water with the production of microbial biomass, which can in situ be used by the culture species as additional food source. Understanding the basics of bio-flocculation is essential for optimal practice. Cells in the flocs can profit from advective flow and as a result, exhibit faster substrate uptake than the planktonic cells. The latter mechanisms appear to be valid for low to moderate mixing intensities as those occurring in most aquaculture systems (0.1–10 W m^− 3). Yet, other factors such as dissolved oxygen concentration, choice of organic carbon source and organic loading rate also influence the floc growth. These are all strongly interrelated. It is generally assumed that both ionic binding in accordance with the DLVO theory and Velcro-like molecular binding by means of cellular produced extracellular extensions are playing a role in the aggregation process. Other aggregation factors, such as changing the cell surface charge by extracellular polymers or quorum sensing are also at hand. Physicochemical measurements such as the level of protein, poly-β-hydroxybutyrate and fatty acids can be used to characterize microbial flocs. Molecular methods such as FISH, (real-time) PCR and DGGE allow detecting specific species, evaluating the maturity and stability of the cooperative microbial community and quantifying specific functional genes. Finally, from the practical point of view for aquaculture, it is of interest to have microbial bio-flocs that have a high added value and thus are rich in nutrients. In this respect, the strategy to have a predominance of bacteria which can easily be digested by the aquaculture animals or which contain energy rich storage products such as the poly-β-hydroxybutyrate, appears to be of particular interest.