The effect of temperature on the energy budget of the Manila clam, <Emphasis Type="Italic">Ruditapes philippinarum</Emphasis>

In this study, the energy budget of the Manila clam, Ruditapes philippinarum, was evaluated after one-week acclimation periods at 5, 10, 15, 20, and 25°C. Small clams (151 ± 12 mg DW) and large clams (353 ± 16 mg DW) were fed with the microalgae, Isochrysis galbana. Filtration rate, ingestion rate, assimilation efficiency, oxygen-consumption rate, and ammonia excretion rate were measured. Both filtration rate and ingestion rate of small and large clams were found to be related to temperature. The highest Q ₁₀ values were measured in the range 15–20°C for both small and large clams. Assimilation efficiency of both small and large clams was not significantly influenced by temperature, although the maximum mean values were detected at 20°C. Oxygen consumption rate and ammonia excretion rate of small and large clams were found to be related directly to temperature over the entire range, with a maximum being detected at 25°C. The highest Q ₁₀ value was estimated in the range 10–15°C with regard to oxygen consumption rate, and in the range of 15–20°C with regard to ammonia excretion rate. Scope for growth (SFG) was positive at all temperatures, achieving a maximum value at 20°C in both small and large clams, primarily as a consequence of the enhanced ingestion rate which offset the concomitant elevation in the metabolic rate. In this study we have estimated the thermal optimum for this species at 20°C.     

Detection of quinolone-resistance genes in Photobacterium damselae subsp. piscicida strains by targeting-induced local lesions in genomes

Quinolone-resistant strains of the fish-pathogenic bacterium, Photobacterium damselae subsp. piscicida are distributed widely in cultured yellowtail, Seriola quinqueradiata (Temminck & Schlegel), in Japan. The quinolone resistance-determining region (QRDR) was amplified with degenerate primers, followed by cassette ligation-mediated PCR. Open reading frames encoding proteins of 875 and 755 amino acid residues were detected in the gyrA and parC genes, respectively. Resistant strains of P. damselae subsp. piscicida carried a point mutation only in the gyrA QRDR leading to a Ser-to-Ile substitution at residue position 83. No amino acid alterations were discovered in the ParC sequence. A mutation in the gyrA gene was also detected in nalidixic acid-resistant mutants of strain SP96002 obtained from agar medium containing increased levels of quinolone. These results suggest that GyrA, as in other Gram-negative bacteria, is a target of quinolone in P. damselae subsp. piscicida. Furthermore, we attempted to detect a point mutation using targeting-induced local lesions in genomes (TILLING), which is a general strategy used for the detection of a variety of induced point mutations and naturally occurring polymorphisms. We developed a new detection method for the rapid and large-scale identification of quinolone-resistant strains of P. damselae subsp. piscicida using TILLING.     

Estimation of the mode of inheritance of thermal tolerance in the guppy <Emphasis Type="Italic">Poecilia reticulata</Emphasis>

High water temperature influences the survival, growth, and maturation of fish. Genetically characterizing thermal tolerance is one of the most important subjects in fish culture. To identify the genetic characterization of thermal tolerance, this characteristic was compared among strains, and among parents and their offspring, in the guppy Poecilia reticulata. In the strain comparison, significant differences in survival rate were observed among the five strains examined, and between females and males. Females exhibited greater tolerance than males in four of five strains examined. In the comparison between parents and their offspring, stronger influence of female parent than of male parent was observed. Offspring obtained from surviving females exhibited greater tolerance than those from dead females. This tendency was typically observed in male offspring. The survival rate in male offspring obtained from dead female parents was lower than that of those from surviving females. The high-temperature tolerance of male parents did not influence this characteristic in offspring as strongly as that of female parents. These results suggest that the major gene or genes, which has a dominant resistant allele and a recessive sensitive allele, are probably passed on by sex-linked inheritance, located on the X chromosome.     

Expression and characterization of the periplasmic cobalamin-binding protein of Photobacterium damselae subsp. piscicida

Cobalamin (vitamin B₁₂) is an essential cofactor in a variety of enzymatic reactions and most prokaryotes contain transport systems to import vitamin B₁₂. A gene coding for a periplasmic cobalamin-binding protein of Photobacterium damselae subsp. piscicida was identified by in silico analysis of sequences from a genomic library. The open reading frame was composed of 834 bp encoding a protein of 277 amino acids. The protein showed 61% identity with the vitamin B₁₂-binding protein precursor of P. profundum , 53% identity with the corresponding protein of Vibrio parahaemolyticus and 43% identity with the periplasmic binding protein BtuF of Escherichia coli. The expression of the native protein was investigated in P. damselae subsp. piscicida , but BtuF was weakly expressed under normal conditions. To characterize the BtuF of P. damselae subsp. piscicida , the recombinant protein was expressed with a C-terminal His₆-tag and purified; the molecular weight was estimated to be approximately 30 kDa. The protein does not contain any free thiol group, consistent with the view that the two cysteine residues are involved in a disulphide bond. The purified BtuF binds cyanocobalamin with an affinity constant of 6 ± 2 μ m .     

Blood chemistry and organ nutrient composition in Atlantic salmon, Salmo salar L., fed graded amounts of wheat starch

After feeding Atlantic salmon, Salmo salar L., five graded amounts of wheat starch from 0 to 310 g kg^?1, low but increased levels of glycogen in kidney, heart and gills were determined. No variations were found in proximate or glycogen compositions of muscle, whereas the liver composition reflected the diet composition. Whole-body homogenates varied in dry matter and lipid levels; the variation was according to g lipid eaten per fish. Plasma glucose levels ranged above average levels only when the starch level in the feed was higher than 220 g kg^?1. Although feed intake and thereby lipid intake increased as dietary carbohydrate increased in the present experiment, no differences in plasma concentrations of cholesterol and only small changes in triglycerides were determined. The increase in dietary carbohydrate was balanced with protein, and total plasma protein concentrations followed the decrease in feed protein content. Haematocrit, haemoglobin, mean cell volume and mean cell haemoglobin all showed significantly reduced levels as dietary starch increased, indicating a fibre effect from ‘left-over’ starch in the intestine followed by reduced absorption of divalent ions, such as iron.     

Morphological studies of peripheral blood cells of the Chinese sturgeon, <Emphasis Type="Italic">Acipenser sinensis</Emphasis>

The peripheral blood cells of one-year-old Chinese sturgeon (Acipenser sinensis) have been studied by light microscopy and transmission electron microscopy. The erythrocyte count was 84.86 × 10⁴ cell mm⁻³ in the peripheral blood of the fish and that of leukocytes was 2.24 × 10⁴ cell mm⁻³. The erythrocytes and four main types of leucocyte—thrombocytes, lymphocytes, granulocytes (including neutrophils and eosinophils), and monocytes, were identified in the peripheral blood. In addition to normal erythrocytes, reticulocytes and division of erythrocytes were observed. Thrombocytes were the most numerous among the leukocytes, and the number of neutrophils with lobated nuclei was larger than for other fish. The structures of the erythrocytes, lymphocytes, monocytes, granulocytes, and thrombocytes of the fish were studied. The erythrocytes were almost completely devoid of organelles, except for some mitochondria and granules. A large number of vacuoles and a few organelles were observed in cytoplasm of the monocytes. There were many microvilli on the membrane and pseudopodia-like cytoplasm bulge in the lymphocytes. The neutrophils were round or oval in shape with bilobed, trilobed, or multilobed nuclei whereas the eosinophils had big special granules, dark stained. There were many vesicles in some thrombocytes, which were related to its phagocytosis; some thrombocytes had almost no cytoplasm or organelles.     

Parentage assignment of stocked black sea bream <Emphasis Type="Italic">Acanthopagrus schlegelii</Emphasis> in Hiroshima Bay using microsatellite DNA markers

The genetic contribution of 51 broodstock, comprising 29 females and 22 males, reared at Hiroshima City Marine Products Promotion Center for the production of stocked black sea bream was monitored during two consecutive years using seven microsatellite DNA loci. The high discrimination ability of these markers was reflected in the polymorphic identification content (PIC=0.831), the exclusion probability (Q≈1), and the low probability of identity index (I=3.635⁻¹⁰). The total number of breeders contributing to the mating process was estimated at 32 (62.7%) in 2000 and 30 (58.8%) in 2001. On pedigree reconstruction, 69.3% of the offspring were successfully assigned to a single broodstock pair. Loss of alleles accounted for 16.9% during seed production; nevertheless, 90.9% of males and 69.0% of females participated in the mating process. Based on microsatellite genetic tagging, 58.9% of the fish sampled during the two months after release were identified as hatchery stock, presenting no significant differences from wild conspecifics in either fork length or body weight.     

Flavobacterium psychrophilum in rainbow trout, Oncorhynchus mykiss (Walbaum), hatcheries: studies on broodstock, eggs, fry and environment

The occurrence of Flavobacterium psychrophilum at four rainbow trout hatcheries was investigated to provide more knowledge about the reservoirs and transmission of this bacterium. Broodstock were sampled at stripping (including both unfertilized and fertilized eggs), and the offspring were then sampled at the eyed egg and fry stages. Water and surface samples (e.g. hatchery trays) were also sampled. Flavobacterium psychrophilum was found in ovarian fluid and milt, indicating that broodstock may serve as a reservoir and are latent carriers of the pathogen. Flavobacterium psychrophilum was not found on or inside eggs, but further egg studies will be necessary to elucidate the possibility of vertical transmission of the pathogen. Flavobacterium psychrophilum was isolated from water samples, but only from water that had been in close contact with farmed rainbow trout or eggs. Flavobacterium psychrophilum isolates were characterized and compared with well-characterized strains, using degradation of elastin, serotype and ribotype profiles. Different ribotypes of F. psychrophilum were found between hatcheries, but a common ribotype A was found at all four hatcheries. Different ribotypes were found in broodstock without clinical disease, whereas only a few ribotypes (mostly ribotype A) were found in diseased fry. The same ribotype A was found in broodstock, in water samples from hatchery trays and in fry, which suggests the possibility of transmission of F. psychrophilum between broodstock and offspring.     

The life history of Tenualosa macrura in Sarawak, further notes on protandry in the genus and management strategies

Abstract  The contraction of Tenualosa macrura (Bleeker) to the Sarawak coast and a small part of Sumatra, and the decline in its fishery mirror what has happened to three other species in the genus. Little was known of this species in its last stronghold in Sarawak where it is important for management and conservation reasons to understand its biology and life history. It forms part of a multispecies fishery where effort is uniform, but catches follow a lunar cycle. Tenualosa macrura is protandrous, but low numbers of small females suggest diandry. In Sumatra Tenualosa macrura spawn throughout the year, but in Sarawak have a seasonal peak in the NE monsoon (December). This has management implications because the Sarawak fishery ceases at this time because of adverse weather, hence reducing fishing pressure on spawning T. macrura . Management measures introduced in Sarawak for Tenualosa toli (Valenciennes) may therefore not be required for T. macrura .     

The essential fatty acid requirement of milkfish (Chanos chanos Forsskal)

The essential fatty acid (EFA) requirement of milkfish was examined by a 12-week feeding trial using defined, purified diets at water temperature of 28–29°C and salinity of 32. The test diets contained varying levels of 18:0 (triglyceride form, TG), 18:3(n–3), 18:2(n–6) and (n–3) highly unsaturated fatty acids (n–3 HUFA). Milkfish juveniles were starved for 7 days and were than fed lipid-free diet for 30 days before the initiation of feeding trials. Low growth and feed efficiency together with high mortalities were observed in fish fed the lipid-free diet as well as in the EFA-deficient diet. Supplementation of 2% 18:2(n–6) to the tristearin based diet did not improve growth rate of milkfish as effectively as feeding with (n–3) fatty acids. The highest weight gain was obtained in milkfish fed a combination of 5% 18:0 + 1.0% 18:3(n–3) + 0.5% 20:5(n–3) + 0.5% 22:6(n–3) although the supplementation of 2% 18:3(n–3) alone or combination of 0.5% 20:5(n–3) + 0.5% 22:6(n–3) to the tristearin based diets were also effective for improvement of growth. Thus, (n–3) fatty acids, such as 18:3(n–3) and (n–3)HUFA were nutritionally more important than 18:2(n–6) for milkfish. The fatty acid composition of the polar lipids from whole body of milkfish juveniles fed the various test diets were influenced by the composition of the dietary fatty acids.