Production of polyclonal antiserum against recombinant VP28 protein and its application for the detection of white spot syndrome virus in crustaceans

The VP28 gene of white spot syndrome virus (WSSV) was cloned into pRSET B expression vector. The VP28 protein was expressed as a protein with a 6-histidine taq in Escherichia coli GJ1158 with NaCl induction. Antiserum was raised against this recombinant-VP28 protein in rabbits and it recognized VP28 protein in naturally and experimentally WSSV-infected shrimp, marine crabs, freshwater prawns and freshwater crabs. The antiserum did not recognize any of the other known WSSV structural proteins. Various organs such as eyestalks, head muscle, gill tissue, heart tissue, haemolymph, tail tissue and appendages were found to be good materials for detection of WSSV using the antiserum and detection of WSSV was successful in experimentally infected Penaeus monodon and P. indicus at 12 and 24 h post-infection (p.i.), respectively. The antiserum was capable of detecting WSSV in 5 ng of total haemolymph protein from WSSV-infected shrimp.     

Several micronutrients in the rotifer Brachionus sp. may not fulfil the nutritional requirements of marine fish larvae

The current best practice intensive culture of larval Atlantic cod includes feeding rotifers from onset of exogenous feeding until 25–30 days after hatching. These larvae grow considerably slower and develop higher frequencies of deformities than larvae reared in semi‐extensive systems, using copepods as feed. The present study compares the micronutrient concentrations in rotifers with those of copepods, with the aim of identifying nutrients that may be limiting for normal growth and development of cod larvae. An additional criterion used is the nutrient requirements given for fish in general, by NRC (1993) , as nutrient requirements of cod remains to be determined. Rotifers were fed on four different diets, consisting of baker's yeast with cod liver oil (3.3 : 1 dry weight (DW)/v), baker's yeast with Algamac 2000^TM (3.5 : 1 DW), baker's yeast with live algae Chlorella (4.1 : 1 DW), and Culture Selco 3000^TM (CS). CS was a complete commercial diet for rotifers while the other diets are considered as based on raw ingredients. Compared with copepod nutrient levels, rotifers grown on yeast‐based diets supplemented with either cod liver oil, Algamac 2000 or Chlorella were apparently sufficient for covering the requirements in cod larvae for all the B‐vitamins, except thiamine. Rotifers cultured on the CS diet also had sufficient amounts of thiamine. Of the minerals, only calcium and magnesium were sufficient, using this criterion while iron was on the borderline. However, with reference to the requirements given for larger fish ( NRC 1993 ), only thiamine, vitamin A, manganese, selenium and perhaps copper, appear too low in the rotifers cultured without extra micronutrient supplementation. The other nutrients were present at levels intermediate between copepod and fish requirement levels. This study suggests that it is necessary to develop enrichment techniques to produce rotifers with sufficient amounts of all micronutrients. Such techniques will also be important tools for determining which nutrients are present at levels below the actual requirements in cod larvae.     

The maturation of the salmon GnRH system and its regulation by gonadal steroids in masu salmon

Pituitary gonadotropin (GTH) secreting cells and brain gonadotropin-releasing hormone (GnRH) secreting neurons are known to be subjected to feedback control by gonadal steroid in teleosts. In masu salmon, Oncorhynchus masou, salmon GnRH (sGnRH) neurons in the ventral telencephalon (VT) and the preoptic area (POA) are involved in the control of GTH cells because sGnRH synthesis in these areas is activated with gonadal maturation. In this study, we attempted to clarify mechanisms of feedback control of sGnRH neurons by gonadal steroids. We examined the effects of 17-methyltestosterone (MT) on sGnRH synthesis in yearling and 2-year-old female fish (which were immature during experimentation in May), and the effects of castration on sGnRH synthesis in underyearling precocious male fish in August. sGnRH synthesis in the POA, but not in the VT, was increased by MT administration in 2-year-old females only, indicating higher sensitivity to MT in the preoptic sGnRH neurons. Castration increased sGnRH synthesis in the VT but not in the POA. These results suggest that sGnRH neurons in the VT and those in the POA are differentially regulated by gonadal steroids.     

A comparison of fatty acid composition and quality aspects of eggs and larvae from cultured and wild broodstock of common sole (Solea solea L.)

Eggs from a F₁ cultured broodstock of sole were compared with eggs from wild‐caught breeders throughout one spawning season, to evaluate if egg quality may be affected by culture‐related conditions. Fourteen batches of eggs from cultured broodstock and 17 batches from wild‐caught sole were compared with respect to fatty acid (FA) composition, egg size, fertilization rate and hatching rate. Based on a multivariate analysis of the FA profiles, it was possible to discriminate between culture and wild inheritance. Eggs from cultured broodstock had high levels of C20:1(n‐9), C18:2(n‐6) and C18:3(n‐3), whereas eggs from wild fish had high levels of C16:1(n‐7), C20:4(n‐6) and C20:5(n‐3). Differences in FA profiles were most likely related to dietary differences. Fertilization and hatching rates were generally low and lowest in eggs from cultured broodstock, but not related to FA composition. Larval growth of one batch from each group was compared. Larval growth was not correlated to broodstock origin, FA composition or egg or larval size. However, larval survival was significantly lower for larvae from cultured broodstock.     

The pelagic habitat analysis module for ecosystem‐based fisheries science and management

We have developed a set of tools that operate within an aquatic geographic information system to improve the accessibility, and usability of remote‐sensed satellite and computer‐modeled oceanographic data for marine science and ecosystem‐based management. The tools form the Pelagic Habitat Analysis Module (PHAM), which can be applied as a modeling platform, an investigative aid in scientific research, or utilized as a decision support system for marine ecological management. Applications include fisheries, marine biology, physical and biological oceanography, and marine spatial management. The GIS provides a home for diverse data types and automated tools for downloading remote sensed and global circulation model data. Within the GIS environment, PHAM provides a framework for seamless interactive four‐dimensional visualization, for matching between disparate data types, for flexible statistic or mechanistic model development, and for dynamic application of user developed models for habitat, density, and probability predictions. Here we describe PHAM in the context of ecosystem‐based fisheries management, and present results from case study projects which guided development. In the first, an analysis of the purse seine fishery for tropical tuna in the eastern Pacific Ocean revealed oceanographic drivers of the catch distribution and the influence of climate‐driven circulation patterns on the location of fishing grounds. To support management of the Common Thresher Shark (Alopias vulpinus) in the California Current Ecosystem, a simple empirical habitat utilization model was developed and used to dynamically predict the seasonal range expansion of common thresher shark based on oceanographic conditions.     

Detection and molecular characterization of infectious spleen and kidney necrosis virus from major ornamental fish breeding states in Peninsular Malaysia

‘Gold standard’ OIE reference PCR assay was utilized to detect the presence of infectious spleen and kidney necrosis virus (ISKNV) in freshwater ornamental fish from Malaysia. From total of 210 ornamental fish samples representing 14 species, ISKNV was detected in 36 samples representing 5 fish species. All positive cases did not show any clinical signs of ISKNV. Three restriction enzymes analyses showed that the fish were infected by identical strains of the same virus species within Megalocytivirus genus. Major capsid protein (MCP) genes of 10 ISKNV strains were sequenced and compared with 9 other reference nucleotide sequences acquired from GenBank. Sequence analysis of MCP gene showed that all strains detected in this study were closely related to the reference ISKNV with nucleotide sequence identity that was ranging from 99.8% to 100%. In addition, phylogenetic analysis of MCP gene revealed that viruses from genus Megalocytivirus can be divided into three genotypes: genotype 1 include reference ISKNV and all other strains that were detected in this study, genotype 2 include viruses closely related to red sea bream iridovirus (RSIV), and genotype 3 include viruses closely related turbot reddish body iridovirus (TRBIV).     

Urine properties in carp, Cyprinus carpio L., with sekoke disease

Abstract. Changes in the properties of urine and blood of carp with sekoke disease were studied. With the progress of this disease, the parameters of blood and particularly urine changed significantly. After 120 days, urine flow, osmolarity and concentrations of inorganic ions (K⁺, Ca²⁺, Mg²⁺, Cl⁻) and organic compounds (ammonium, creatine and creatinine, protein, glucose) in the experimental group showed higher values than those of the controls. Even after 60 days, when clinical signs of sekoke disease were not evident, abnormally high concentrations of ammonium, protein and glucose in the urine were found in the experimental group.     

Melanomacrophages in three species of free-ranging sharks from the northwestern Atlantic, the blue shark Prionacae glauca (L.), the shortfin mako, Isurus oxyrhinchus Rafinesque, and the thresher, Alopias vulpinus (Bonnaterre)

The melanomacrophage aggregates or cells (MMC) are commonly used as biomarkers of exposure to pollution in fish, albeit their numbers and morphological characteristics can be influenced not only by environmental toxins but also by a range of physiological parameters and pathological conditions. Accordingly, before we can use MMC as biomarkers in any fish species, their normal, 'background' characteristics have to be established in apparently healthy fish. The knowledge of MMC in sharks is minimal. The aim of this study was to characterize MMC from 51 free-ranging, large pelagic sharks from the northwestern Atlantic, including shortfin mako, Isurus oxyrhinchus , thresher, Alopias vulpinus and blue shark, Prionacae glauca . The rationale of this study was twofold. First, because, sharks have life histories predisposing them to the accumulation of environmental toxins they constitute good sentinel species of the health of the global marine ecosystem. Second, because many shark populations are in decline or threatened by extinction, we need to expand our understanding of their health status in order to develop appropriate protective measures. All sharks were collected by sports fishing gear between June and August in 2007. Their health condition was assessed by necropsy, morphometrics, and by microscopic examination of gonads, livers, spleens and kidneys. Routine, haematoxylin and eosin and/or Pearl's reagent-stained paraffin embedded sections were studied by light microscopy. Our results provide the first data on the morphometric and morphological characteristics of MMC in viscera of apparently healthy free-ranging sharks from the northwestern Atlantic.     

Effect of dietary protein/carbohydrate ratio on activities of hepatic enzymes involved in the amino acid metabolism of Nile tilapia, Oreochromis niloticus (L.)

This study investigated the influence of different dietary protein/carbohydrate ratios on activities of enzymes involved in the amino acid metabolism as well as on growth performance and body composition of Nile tilapia (Oreo- chromis niloticus). Fish were fed semi-synthetic diets differing in their protein/carbohydrate ratio. The diets were formulated to be isolipidic and isoenergetic (on the basis of metabolizable energy). Fish were reared individually. After a prefeeding phase, tilapia were randomly assigned to three groups and were fed the experimental diets at a relatively low level. The study lasted 9 weeks. Proximate composition, gross energy content, and activities of aspartate aminotransferase (ASAT) and alanine aminotransferase (ALAT) in the liver of fish were determined. While the mean growth rate and feed conversion efficiency did not differ significantly among the feeding groups, the body composition showed significant differences. The dietary protein/carbohydrate ratio had no significant effect on the protein and energy gain, but the lipid gain was significantly higher in tilapia fed the low-protein diet. The activities of ASAT and ALAT per gram of liver and per gram of soluble protein were significantly increased in fish fed the high-protein diet. There was a significant positive correlation between the activities of ASAT and ALAT. The activities of both enzymes in the liver were significantly higher in fish with a higher protein gain. Measuring the activities of enzymes involved in the amino acid metabolism can provide more information about the metabolic utilized dietary protein.     

Incorporation of Maize Gluten in Supplementary Feed and its Impact on Growth and Flesh Quality of Some Carps

Maize gluten (a byproduct from the starch industry) was used at four different levels (5, 10, 15 and 20% w/w) replacing the fish meal, a component of supplementary diet, under a semi-intensive culture system. The impact of different diets on the physico-chemical parameters of water, growth and flesh quality of catla, Catla catla; rohu, Labeo rohita; and mrigal, Cirrhina mrigala was studied. Growth in terms of body weight gain was highest when fed a diet having 5% maize gluten (replacing fish meal at 25% level) for all the fish species. However, incorporation of maize gluten at higher levels led to decline in growth, since maize gluten is deficient in essential amino acids like methionine, lysine and threonine, etc. The flesh quality in terms of total protein, total lipid, total sugar, moisture and total ash was also determined at the termination of the experiment to see whether there was any significant variation with the incorporation of maize gluten or not.