First isolation of Tenacibaculum maritimum from wedge sole, Dicologoglossa cuneata (Moreau)

The first isolation of Tenacibaculum maritimum from wedge sole, Dicologoglossa cuneata, is reported. The pathogen was recovered from ulcers of cultured fish, from three different outbreaks. The six isolates obtained were biochemically and serologically characterized and diagnosis was confirmed by polymerase chain reaction using specific primers and partial 16S rRNA gene sequencing. The isolates constituted a homogeneous phenotypic group; however, they belong to two of the different serotypes described within this species. A virulence evaluation of the isolates using Wedge sole fry was also performed.     

Comparison of procedures to determine protein content of developing bean seeds (Phaseolus vulgaris)

Micro-Kjeldahl, Lowry and Bradford procedures were compared for determining the protein content ofPhaseolus vulgaris seeds during their development. Micro-Kjeldahl and biuret techniques were also compared with mature seeds of a normal and a genetically-improved bean cultivar. The protein contents of casein and soy protein isolate were as well estimated by these four methods. For many samples of both bean and food protein products large disagreements were found between micro-Kjeldahl and the other three procedures.     

Characterization of hepatic and extrahepatic glutathione S-transferases in rainbow trout (Oncorhynchus mykiss) and their induction by 3,3′,4,4′-tetrachlorobiphenyl

Liver, kidney, gill and olfactory epithelium cytosolic fractions of rainbow trout (Oncorhynchus mykiss) were examined for glutathione S-transferase (GST) contents. Proteins retained on a glutathione (GSH)-affinity matrix were separated as monomers by reversed-phase HPLC and characterized by immunoblotting, mass spectrometry and partial amino acid sequence. For each organ concerned, a specific pattern of these proteins was determined and appeared similar for liver and kidney on one hand, and for gill and olfactory epithelium on the other hand. It was confirmed that the prominent hepatic GST is a class enzyme, also constitutively expressed as a major isoform in the four organs studied. Moreover, a class variant and two new class GST subunits were characterized in minor fractions. An unknown protein, which was found major in gills and olfactory epithelium, exhibited some characteristics of class GSTs. Occurrence of possible GSH-adduct formation observed on two distinct monomers in specific experimental conditions is discussed. These results and methods were used to investigate the effect of 3,3,4,4-tetrachlorobiphenyl (TCB), a polychlorinated biphenyl (PCB), on GST expression in trout liver. From HPLC-profiling, significant co-induction of the major class and the two minor class GST subunits was observed in trout after waterborne exposure to TCB which was followed by a slight increase in 1-chloro-2,4-dinitrobenzene (CDNB) activity. The present work allows qualitative evaluation of the specific detoxification potential of rainbow trout. The use of HPLC-profiling of GSTs as a possible tool for the biomonitoring of polluted aquatic environment is suggested.     

The effects of long-day photoperiod on growth, body composition and skin colour in immature gilthead sea bream (Sparus aurata L.)

Immature gilthead sea bream (Sparus aurata L.) with a mean initial weight of 25.6 g were reared over 11 months to market size under different photoperiods: 16L:8D; 24L:0D and a control. Differences in final mean weight were significant between the three treatments (P<0.001): 16L:8D, 465.0 g; 24L:0D, 445.9 g; control, 402.6 g. Fish from the 24L:0D consumed most while 16L:8D fish converted it most efficiently. The lipid content of fillets was lower (P<0.05) for the 24L:0D (5.37±0.16 g 100 g^?1 of fillet) than for the control (6.02±0.12 g), and the moisture content lower for the control than for either treatment groups. Skin luminosity (L^*) was directly related to the number of hours of light exposure.     

Immunogene expression in head kidney and spleen of common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.) following thermal stress and challenge with Gram-negative bacterium, <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

To evaluate the effect of thermal and microbial stress on the immune response of common carp (Cyprinus carpio L.), relative mRNA expression level of pro-inflammatory cytokines [tumor necrosis factor alpha (TNF-α) and interleukin (IL)-1β] and other genes related to immune or stress response [inducible nitric oxide synthase (iNOS), heat shock protein 70 (Hsp70), superoxide dismutase one (SOD1), and glucocorticoid receptor (GR)] was measured by quantitative PCR (qPCR). In addition, total protein and total immunoglobulin level in blood plasma of experimental common carp was also assayed. All the above parameters were estimated 24 h post-challenge with Gram-negative bacterium, Aeromonas hydrophila. Common carp (54.89?±?6.90 g) were initially exposed to 20 °C (control group) and 30 °C (thermal stress group) water temperature for 30 days, followed by experimental challenge with 2.29?×?10⁸ colony forming unit/mL (CFU/mL; LD₅₀ dose) of A. hydrophila. Exposure of fish to thermal stress and subsequently challenge with A. hydrophila significantly (P?<?0.05) increases the IL-1β mRNA expression in head kidney and spleen of common carp by ~?39.94 and ~?4.11-fold, respectively. However, TNF-α mRNA expression in spleen decreased ~?5.63-fold in control fish challenged with A. hydrophila. Thermal stress and challenge with bacterium suppresses the iNOS and GR mRNA expression in spleen of common carp. Moreover, significant (P?<?0.05) increase in total protein content of blood plasma (~?43 mg/g) was evident in fish exposed to thermal stress and challenged with A. hydrophila. In conclusion, our study highlights the importance of elevated temperature stress and microbial infection in differential regulation of expression of several immunogenes in common carp.     

Analysis of exotoxins produced by atypical isolates of Aeromonas salmonicida,by enzymatic and serological methods

In this study, exotoxins produced by 62 Aeromonas salmonicida strains and the bacterium Haemophilus piscium were analysed. Enzymatic assays, zymograms and serological detection were used to monitor secretion by bacterial strains of the previously described exotoxins P1, GCAT and AsaP1 and also the extracellular P2 metallo-gelatinase and a serine caseinase, which is different from the P1 protease and has not yet been characterized. Based on the results, the strains were divided into five groups. One comprised the type strains for A. salmonicida ssp. masoucida, H. piscium and 36% of the atypical isolates, and another, a type strain for A. salmonicida ssp. smithia together with 14% of the atypical isolates. A second type strain of A. salmonicida ssp. smithia was grouped with 8% of the atypical isolates. The largest group contained the type strains for A. salmonicida ssp. achromogenes and 38% of the atypical isolates. The type strains for A. salmonicida ssp. salmonicida were in the last group with all the four typical strains and 4% of the atypical isolates. The combination of zymogram and serological detection used is recommended as the most reliable method for characterizing A. salmonicida strains according to their exotoxin secretion.     

Developments in the nutrition of Menidia estor Jordan 1880

The Mexican silverside, Menidia estor, is a species with great regional importance and with very high prices in local markets. Unfortunately, due to high fisheries pressure, environmental degradation and pollution, the species has become endangered. Recently, there has been much progress in the biotechnology of this species, aimed at its culture, and the present paper describes the advances in feeding and nutrition of those important fish. M. estor is a stomachless, zooplantophagous fish, which also occasionally feeds on small fish and crustaceans in the adult stage. Studies on the digestive enzymatic activities show high proteolytic capacity and a late or different model of digestive maturation from that described for marine stomach fish. Nutritional studies on M. estor have shown that juveniles have dietary requirements of about 400 g kg^?1 protein and 80 mg kg^?1 vitamin C. The upper level of dietary carbohydrate for good growth and survival for juveniles is about 150 g kg^?1. Based on the high docosahexaenoic acid (DHA) content in flesh when fed on diets low in DHA, it is believed that the species has the capacity to biosynthetize >20‐carbon fatty acids from 18‐carbon fatty acids. The high levels of DHA in flesh makes this fish a very significant potential component of human nutrition. Using these findings the first practical diets for commercial culture of the species have been developed.     

Effects of Using Live Grass Shrimp (Palaemonetes varians) as the only Source of Food for the Culture of Cuttlefish, Sepia officinalis (Linnaeus, 1758)

The life cycle of cuttlefish fed ad libitum exclusively on live grass shrimp (Palaemonetes varians) was studied during 5 consecutive generations. Different culture temperatures promoted different (P < 0.05) exponential growth for each life cycle, being summer generations shorter than those of winter. Higher temperatures promoted higher IGR’s and mortality, while lower temperatures promoted increased life span, reproduction stages, total fecundity and total egg biomass. Increased generations also seemed to increase fertility. A “hybrid” generation promoted the best results in terms of hatchling weight, individual fecundity and fertility. Mean egg weight was related to female size and embryonic development took longer at lower temperatures. Brood stock sex ratios seemed to be temperature related. All of these culture aspects were also compared between themselves in order to establish future brood stock methodologies. Grass shrimp proved to be a good diet for the culture of cuttlefish throughout the life cycle. The use of only one species reduces costs and labor associated to cuttlefish culture.     

Litopenaeus vannamei (Boone) post-larval survival related to age, temperature, pH and ammonium concentration

Transport of post‐larvae shrimp used in aquaculture is an important element of successful cultivation because of the potential for stress during stocking procedures. To find optimum transport conditions, several bioassays were performed in the laboratory to evaluate survival of whiteleg shrimp Litopenaeus vannamei 5–30‐day‐old postlarvae under conditions similar to those encountered during transport from the hatchery to nursery and shrimp ponds. Postlarvae were exposed for 4 h to different temperatures and pH levels ammonia concentrations. Survival was significantly reduced after a 4 h exposure to pH 9 and was inversely related to temperature with or without 7 mg L^?1 of ammonia. The 15‐ and 20‐day‐old postlarvae had higher survival rates than other ages. The lowest survival occurred in alkali conditions (pH 9), with 7 mg L^?1ammonia at 30 and 32°C. To assure optimal survival of postlarvae during transfer from the hatchery to the nursery and shrimp ponds, we recommend temperatures below 28°C, pH no higher than 8, no ammonia and post‐larval age at least 15 days.