Nitrate removal efficiency and bacterial community of polycaprolactone-packed bioreactors treating water from a recirculating aquaculture system

A denitrification reactor packed with polycaprolactone (PCL) as a carbon source and biofilm carrier was developed to remove nitrate nitrogen (\( {\mathrm{NO}}_3^{-}-\mathrm{N} \)) from the water of a recirculating aquaculture system for 115 days. The hydraulic retention time was set to 6 h, and the water flow rate was 0.95 L h^?1. The removal rates of \( {\mathrm{NO}}_3^{-}-\mathrm{N} \) ranged from 34.67 to 155.7 g \( {\mathrm{NO}}_3^{-}-\mathrm{N} \) m^?3 day^?1 when the effluent \( {\mathrm{NO}}_3^{-}-\mathrm{N} \) concentrations ranged between 1.62 and 72.25 mg L^?1. No obvious changes in the Fourier transform infrared spectra of the PCL before and after use were observed. Bacterial community structure in the biofilm of the PCL granules was analyzed using Illumina MiSeq sequencing technology. Proteobacteria and Bacteroidetes were predominant in the biofilm, with relative abundances of 63.9 and 27.4%, respectively. Three genera, Acidovorax, Azospira, and Diaphorobacter, were capable of both denitrification and PCL degradation. This study indicates that PCL-packed reactors may be used and optimized for removing nitrate from aquaculture effluents.     

Apparent digestible protein,energy and amino acid availability of three plant proteins in Florida pompano,Trachinotus carolinus L. in seawater and low‐salinity water

Two experiments were conducted with Florida pompano, Trachinotus carolinus L. at 3 and 28 g L^?1 salinity to determine apparent crude protein digestibility (ACPD), energy digestibility (AED) and amino acid availability (AAAA) from soybean meal (SBM), soy protein isolate (SPI) and corn gluten meal (CGM). Mean AAAA was similar to ACPD. In fish adapted to 3 g L^?1 salinity, they were 81.2% and 81.9% (CGM), 93.6% and 92.2% (SBM), 93.8% and 93.1% (SPI) for AAAA and ACPD respectively. In fish adapted to 28 g L^?1, they were 84.5% and 83.4% (CGM), 86.5% and 87.1% (SBM), and 83.4% and 85.0% (SPI) for AAAA and ACPD respectively. The AED was highest for SPI and lowest for SBM and inversely related to carbohydrate. The ACPD, AED and AAAA of soy products appeared to be lower in high salinity, whereas CGM was unaffected. The data suggest that SBM, SPI and CGM should be further evaluated as partial fishmeal replacements in Florida pompano diets. Application of the generated coefficients can be used to develop well‐balanced, low‐cost diets for Florida pompano reared in low salinity or seawater.     

Plasma levels of immune factors and sex steroids in the male seahorse <Emphasis Type="Italic">Hippocampus erectus</Emphasis> during a breeding cycle

To better understand the endocrine- and immune-response pattern during reproduction in a fish species having parental care behaviors and also to accumulate the endocrine- and immune-related data for future explanations of the low reproductive efficiency in seahorse species, the variations of immune factors and sex steroids in the plasma of the male lined seahorse Hippocampus erectus at different breeding stages, i.e., pre-pregnancy, pregnancy (early, middle, and late periods), and post-pregnancy, were investigated in the present study. The immune factors included monocytes/leucocytes (M/L), leucocyte phagocytic rate (LPR), immunoglobulin M (Ig M), interleukin-2 (IL-2), interferon-α (IFN-α), and lysozyme (LZM). The sex steroids included testosterone (T), 11-ketotestosterone (11-KT), 11β-hydroxytestosterone (11β-OHT), 17α-methyltestosterone (17α-MT), 17β-estradiol (E2), and 17α-hydroxy-20β-dihydroprogesterone (17α-20β-P). Moreover, the immune metabolic activity of epithelium cells in the brood pouch at different breeding stages was also analyzed through ultrastructural observations of the abundance of cytoplasmic granules, mitochondria, endoplasmic reticulum, lysosomes, and exocytosis. The results show that a higher immune level was observed during pregnancy, particularly in the early and middle periods, and a lower immune level was noted during pre-pregnancy. Correspondingly, the epithelium cells in the brood pouch also showed a stronger immune metabolic activity during pregnancy and weaker activity during pre-pregnancy. Four sex steroids of T, 11β-OHT, 17α-MT, and E2 were higher during pre-pregnancy and lower during post-pregnancy, whereas 11-KT and 17α-20β-P, which were positively correlated with part immune factors, were higher during pregnancy. No negative correlations between sex steroids and immune factors were observed. In conclusion, the higher immune competence during pregnancy may indicate that parental care could improve immunity, which may be the major factor for no immunosuppressive effect of sex steroids during reproduction in the seahorse H. erectus, unlike noncaregiving fishes in which inhibitions of sex steroids on immunity are frequently observed. Moreover, higher 11-KT and 17α-20β-P during pregnancy than during pre-pregnancy and post-pregnancy may suggest that these two steroids are also involved in parental care regulation.     

Aeromonas hydrophila B-haemolysin: purification and examination of its role in virulence in 0-group channel catfish, Ictalurus punctatus (Rafinesque)

Abstract. A single step procedure for purification of B-haemolysin from Aeromonas hydrophila is described which recovered 94% of the haemolytic activity with a sixfold increase in specific activity. Toxicity of haemolysin in 0-group channel catfish was 0–6 mg of purified protein per gram offish. Of six strains of A. hydrophila evaluated for differential toxin production and virulence in 0-group channel catfish, three B-haemolysin-deficient strains were virulent while three B-haemolysin-producing strains were avirulent.     

Immune protection in carp, Cyprinus carpio L., after immunization with Aeromonas hydrophila crude lipopolysaccharide

Abstract. Vaccination with crude lipopolysaccharide (LPS) induced better protection against infection with Aeromonas hydrophila in carp than vaccination with formalin killed vaccine. Dipping fish in vaccine for 2 h at 25°C was more effective than intraperitoneal injection of the vaccine in procedural simplicity, lower stress loading and the degree of protection acquired. In carp immunized with crude LPS by the dip method, antibodies were not detected by bacteriai agglutination, passive haemagglutination and the agar diffusion tests. The results indicate that the protection against A. hydrophila infection in carp is not dependent on humoral immunity.     

Comparative analysis of using cereal grains and compound feed in semi-intensive common carp pond production

The semi-intensive system of carp farming is dominant in inland aquaculture. Although it appears to be simple, this system has a whole range of options, from those based on traditional rearing methods with use of plants from the region where the fish are reared to production based on the use of high-quality concentrated feeds. In this paper, we conduct a comparative analysis of the effect of using cereal grains as opposed to compound feed (pelleted and extruded) on the quantity of fish produced, flesh quality, histological changes in the digestive tract, water quality, and structure of the biocoenosis in the pond ecosystem. When the fish are fed cereal grains, production ranges from 500 to 1500 kg/ha, while using compound feed gives production that is twice as great. The proportions of protein and water are higher and that of lipids lower in flesh of common carp reared on compound feed in relation to fish fed cereal grains. Use of extruded feed gives better content of n-3 polyunsaturated fatty acids and a better n-3/n-6 ratio than when pelleted feed and cereal grains are used. Comparison of water quality in carp ponds under conditions of using various types of feed indicates that there are no significant differences. Although it was expected that feed with a higher percentage of proteins would cause disturbances in the aquatic ecosystem, their continual incorporation into the cycling of matter in the water contributes to maintenance of ecological equilibrium and even indirectly promotes growth of the common carp.     

Distribution and induction of cytochrome P450 1A1 in the rainbow trout brain

Cytochrome P450 (CYP) 1A1 participates in the activation as well as detoxification of environmental pollutants such as aromatic hydrocarbons. This CYP form is also efficiently induced by aromatic hydrocarbons. The presence of CYP 1A1 in the brain might thus be of physiological and toxicological importance. In the present investigation on rainbow trout, the distribution of 7-ethoxyresorufin-O-deethylase (EROD) activity, a cytochrome CYP 1A1 catalyzed reaction, was measured in whole tissue homogenates from brain parts. In control fish, a relatively high activity was found in the rainbow trout olfactory bulb compared to the other brain parts. Although an EROD induction (3 to 7-fold) by β-naphthoflavone (BNF) was recorded in all brain parts from the rainbow trout, the highest induced activity was measured in the olfactory bulbs. To ascertain the distribution of EROD activity in cells, whole brain tissue was subfractionated by differential centrifugation. The fractionation scheme separated mitochondria (P2 fraction) and microsomes (P3 fraction) as determined by marker enzymes and electron microscopy. In control rainbow trout, a low EROD activity could be measured in the P2 fraction. BNF induced the EROD activity in both P2 and P3 fractions. Western blotting showed the induction by BNF of a protein band in the P2 and P3 fractions with a molecular mass around 58,000 when highly specific anti-cod CYP 1A1 antibodies were used. ELISA measurements confirmed the induction of CYP 1A1 protein in the rainbow trout brain subcellular fractions.     

Protective immunity of sevenband grouper, Epinephelus septemfasciatus Thunberg, against experimental viral nervous necrosis

This paper describes the protective immune responses of sevenband grouper, Epinephelus septemfasciatus Thunberg, immunized with live piscine nodavirus, the causative agent of viral nervous necrosis (VNN), or the Escherichia coli – expressed recombinant coat protein. Nodavirus-neutralizing antibodies were detected at titres ranging from 1:158 to 1:1257 in serum of sevenband grouper which survived intramuscular injection with the virus, by a cell culture assay system. The virus-neutralizing ability of immune serum was also confirmed by injecting virus previously treated with serum into fish. This indicates establishment of acquired immunity in survivors and thus explains why survivors from natural infection are resistant to recurrence of the disease. Young sevenband grouper were immunized twice by intramuscular injections with the recombinant coat protein. Immunized fish produced neutralizing antibodies at high titres for at least 110 days and showed significantly lower mortalities in virus challenge tests. These results suggest the potential for vaccination against VNN in sevenband grouper, which is susceptible to piscine nodavirus at all life-stages.     

Post-molt processes of cuticle formation and calcification in the Japanese mitten crab <Emphasis Type="Italic">Eriocheir japonicus</Emphasis>

This study examines the following in the Japanese mitten crab: (1) the structure of the exoskeleton with special reference to its calcification; (2) the progression of post-molt cuticle formation and calcification. In the crab, the structure and calcification state of the exoskeleton at the molt and during the inter-molt stage were similar to those of other crustaceans. During the inter-molt, the exoskeleton consisted of four cuticle layers; the outermost epicuticle, the exocuticle, the endocuticle and the innermost membrane layer. Intense calcification was observed in the exo- and endocuticle. At the molt, the synthesis of the epi- and exocuticle was already complete, and the addition of the endocuticle began after the molt. Calcification of the exocuticle initiated soon after the molt, but there was a delay between endocuticle matrix synthesis and calcification. Histology showed that the process of calcification was similar to that in other crustaceans. However, calcium concentrations within the exoskeleton continued to increase and never reached the levels of the inter-molt stage at the end of the experiment. This suggests that the Japanese mitten crab is relatively slow to calcify compared to other crustaceans.