Epizootiology of Renibacterium salmoninarum, the causal agent of bacterial kidney disease in salmonid fish

Abstract. By use of a selective medium, Renibacterium salmoninarum was recovered from rainbow trout, Salmo gairdneri Richardson. With asymptomatic animals, small numbers of Renibacterium colonies were recovered only from the anterior part of the kidney; whereas in clinically diseased fish, dense pure culture growth was obtained from kidney, spleen, heart, blood, ascitic fluid and faeces. There was no evidence for the presence of R. salmoninarum in the water or sediment of 56 freshwater fish farms in England and Wales. Nevertheless, laboratory–based experiments showed that the pathogen was excreted in the faeces of clinically diseased fish. Moreover, there was survival of the bacterial cells for up to 21 days in sediment/faecal material.     

Relative potencies of natural estrogens on vitellogenin and choriogenin levels in the Indian freshwater spotted snakehead, <Emphasis Type="Italic">Channa punctata</Emphasis>: in vivo and in vitro studies

The relative efficacies of three natural estrogens viz., estrone (E₁), estradiol-17β (E₂) and estriol (E₃) to induce synthesis of vitellogenin (Vg) and choriogenin (Chg) were assessed in primary hepatocyte cultures of the Indian freshwater spotted snakehead, Channa punctata. Hepatocytes were isolated from the spotted snakehead liver by a non-enzymatic protocol. Optimum culture conditions were standardized for ensuring their viability and functioning. Isolated hepatocytes were cultured for 48 h for monolayer formation and then exposed to various concentrations (0.001–10 μM) of the three estrogens. Competitive homologous ELISAs, developed and validated for spotted snakehead Vg and Chg were employed to determine the amounts of these two proteins secreted into the culture medium after 48 h of incubation. The results reveal that although all the three estrogens were effective in inducing the production of Vg and Chg in a dose-dependent manner, there were differences in their relative potencies. Of three estrogens, E₁ was the least potent and could induce synthesis of Vg and Chg only at a minimum concentration of 0.5 μM; whereas significant levels of both the proteins were quantified in culture medium by exposing the hepatocytes to E₂ or E₃ even at a concentration of 0.001 μM. All three estrogens were effective in inducing synthesis of Vg and Chg in vivo also. These results suggest the possibility of employing the above in vitro experimental design to monitor the presence of estrogens/estrogen-like chemicals in natural waters, which could interfere with the estrogen receptor system of fish. This study further points to the possibility of using Chg, in addition to Vg, as a parameter for screening various chemicals for their estrogenic activity.     

Regulation of growth, fatty acid composition and delta 6 desaturase expression by dietary lipids in gilthead seabream larvae (Sparus aurata)

The Δ6 and Δ5 desaturases and elongases show only very limited activity in marine fish, and little is known of the possibility of enhancing Δ6 desaturase gene expression in these fish. The use of plant oils in marine fish diets is limited by their lack of n−3 highly unsaturated fatty acids (HUFA) despite an abundant content of the 18C fatty acid precursor linoleic and α-linolenic acids. The objective of the present study was to determine the ability of larval gilthead seabream to utilize vegetable oils and assess the nutritional regulation of Δ6 desaturase gene expression. Seventeen-day-old gilthead seabream larvae were fed during a 17-day period with one of four different microdiets formulated with either sardine fish oil (FO), soybean, rapeseed or linseed oils, respectively, or a fifth diet containing defatted squid meal and linseed oil. Good larval survival and growth, both in terms of total length and body weight, were obtained by feeding the larvae either rapeseed, soybean or linseed oils. The presence of vegetable oils in the diet increased the levels of 20:2n−9 and 20:2n−6, 18:2n−9, 18:3n−6, 20:3n−6 and 20:4n−6, in larvae fed rapeseed and soybean oils in comparison to those fed FO. In addition, a sixfold increase in the relative expression of Δ6 desaturase-like gene was found in larvae fed rapeseed and soybean oils, denoting the nutritional regulation of desaturase activity through its gene expression in this fish species. However, feeding linseed oil did not increase the expression of the Δ6 desaturase gene to such a high extent.     

Inherent bias in using aggregate CPUE to characterize abundance of fish species assemblages

We have analyzed the practice of assessing an assemblage of fish species in a multispecies fishery on the basis of aggregate catch per unit effort (CPUE), which is the summed catch of all species per unit of effort. We show that at the onset of fishing or of a large positive or negative change in fishing effort, aggregate CPUE will be hyper-responsive, that is, relative change of aggregate CPUE will be greater than that of aggregate abundance. We also show that as the fishery reaches equilibrium, the aggregate CPUE in most circumstances will continue to be hyper-responsive, with a greater relative change from its value at the start than the aggregate abundance. However, there are less likely circumstances in which the aggregate CPUE will be hyper-stable compared to aggregate abundance. The circumstances leading to hyper-responsiveness or hyper-stability depend on the distribution of productivity and fishery vulnerability parameters among the species in the aggregation.     

Utilization of monounsaturated fatty acid (18:1n-9, oleic acid) by freshwater prawn Macrobrachium rosenbergii (de Man) juveniles

The utilization of oleic acid as an energy source and the effects of oleic acid levels and/or dietary soy bean lecithin (SBL) on oleic acid utilization, growth and survival, and lipid class and fatty acid compositions of freshwater prawn, Macrobrachium rosenbergii (de Man), juveniles were determined.
Increase in levels of dietary oleic acid from 10 to 80 g kg⁻¹ significantly ( P ≤ 0.05) reduced growth, survival and feed conversion efficiency of M. rosenbergii juveniles during the 40-day feeding period. Inclusion of 20 g kg⁻¹ SBL had no significant effect ( P ≥ 0.05) on growth and survival, nor was there any interactive effect between dietary SBL and oleic acid levels.
Body fatty acid profile of prawns reflected dietary fatty acid quantity. The fatty acid composition of prawns fed diets containing 80 g kg⁻¹ oleic acid had tremendously high proportions of oleic acid. Polar lipids, mostly phosphatidylcholine (PC), constituted the bulk of the extracted total lipids. Prawns fed with SBL had significantly ( P ≥ 0.05) higher PC content.
Oleic acid was metabolized for energy by M. rosenbergii juveniles at the same rate regardless of dietary level of SBL and/or oleic acid. Expired ¹⁴CO₂ accounted for half of the ingested radioactivity 48 h after feeding with labelled diets. No significant difference in the amount of ¹⁴CO₂ expired by prawns fed the labelled test diets was found. Per cent radioactivity ingested and absorbed into the body was also not significantly different in prawns of the different dietary treatments.     

Expression of HSP70 in response to heat-shock and its cDNA cloning from Mediterranean blue mussel

Expression of HSP70 in response to heat-shock was investigated at the protein and mRNA levels in Mediterranean blue mussel. Western and Northern blot analyses revealed that HSP70 was expressed following heat-shock in the mantle at both protein and mRNA levels, suggesting that gene expression of HSP70 is implicated in the cellular response to heat-shock stress in mussel. It was then attempted to clone HSP70 cDNA in order to determine the primary structure of mussel HSP70. As a result, two full-length cDNA encoding HSP70 were isolated from a cDNA library prepared from the heat-shocked mantle. The isolated cDNA consist of single open reading frames of 2067 bp and 1911 bp which encode proteins of 689 amino acids and 637 amino acids, respectively. Both HSP70 cDNA encode an ATPase do main, and a substrate-binding do main in addition to a Glu-Glu-Val-Asp (EEVD) peptide motif that is specific for cytosolic HSP70. These findings suggest that the cDNA clones obtained in the present study encode cytosolic HSP70.     

Differences in uptake and killing of pathogenic and non‐pathogenic bacteria by haemocyte subpopulations of penaeid shrimp,Litopenaeus vannamei, (Boone)

Phagocytosis is an important function of both invertebrate and vertebrate blood cells. In this study, the phagocytic activity of haemocyte subpopulations of penaeid shrimp, Litopenaeus vannamei, (Boone), against pathogenic and non‐pathogenic particles was investigated in vitro. The haemocytes of penaeid shrimp were firstly separated by centrifugation on a continuous density gradient of iodixanol into four fractions with five subpopulations (sub), of which sub 1 (hyalinocytes) and sub 4 (semi‐granulocytes) have the main function in phagocytosis of both pathogenic and non‐pathogenic bacteria as well as fluorescent polystyrene beads. It was found that these haemocyte subpopulations engulfed virulent Vibrio campbellii and Vibrio harveyi at a higher rate than non‐virulent Escherichia coli and polystyrene beads. When these bacteria were mixed with shrimp haemocyte subpopulations and incubated for 180 min, the percentage of viable intracellular V. campbellii (25.5 ± 6.0%) recovered was significantly higher than the percentage recovered from V. harveyi (13.5 ± 1.1%). No viable intracellular E. coli was observed in this study. In contrast to V. harveyi and E. coli, V. campbellii containing endosomes did not acidify in time. Incubation of haemocyte subpopulations with the most virulent V. campbellii strain resulted in a significant drop in haemocyte viability (41.4 ± 6.3% in sub 1 and 30.2 ± 15.1% in sub 4) after 180 min post‐inoculation in comparison with the less virulent V. harveyi (84.1 ± 5.6% in sub 1 and 83.4 ± 4.1% in sub 4) and non‐virulent E. coli (92.7 ± 2.8% in sub 1 and 92.3 ± 5.6% in sub 4) and polystyrene beads (91.9 ± 1.6% in sub 1 and 84.4 ± 3.4% in sub 4). These findings may be a valuable tool for monitoring shrimp health and immunological studies.     

Effects of various levels of dietary protein on survival,molting frequency and growth of juvenile blue crabs (Callinectes sapidus)

An 18-week study was conducted to examine growth characteristics of the juvenile blue crab (Callinectes sapidus Rathbun) fed various levels of dietary protein. Hatchery raised individuals (157) of identical age (160 days) from the same gravid female were assigned to one of four dietary treatments: adult brine shrimp (Artemia salina Leach) and artificial pelleted diets containing crude protein levels of 23, 37, and 49%.Growth rate, measured as a function of mean weight and mean carapace width, indicated that brine shrimp-fed crabs were larger (P < 0.05) than crabs fed artificial pelleted diets. Also, blue crabs fed either 37 or 49% crude protein were larger than crabs fed the 23% crude protein diet. Molting frequency was greater in the brine shrimp-fed crabs than in those fed the artificial diets. No differences in growth were detected when comparing males vs. females fed the artificial diets. However, females demonstrated greater growth than males within the brine shrimp treatment. Protein concentration of freeze-dried whole blue crab carcasses was greater in brine shrimp-fed crabs than in crabs fed the artificial pelleted diets (P < 0.05).     

Protein and energy nutrition of brook trout (Salvelinus fontinalis) at optimal and elevated temperatures

The digestible protein (DP) and digestible energy (DE) requirements for maintenance and growth of brook trout (Salvelinus fontinalis) were determined using a factorial model at either optimum (15 °C) or elevated temperature (19 °C). Several key parameters of the factorial model were measured using a series of inter‐related studies. The maintenance requirements for DP and DE were 0.10 gDP kg^?0.69 day^?1 (15 °C) and 0.31 gDP kg^?0.78 day^?1 (19 °C), and 34.86 kJDE kg^?0.84 day^?1 (15 °C) and 46.14 kJDE kg^?0.86 day^?1 (19 °C). The total requirements for DP were 0.10 gDP kg^?0.69 day^?1 + 2.14PG (protein gain) (15 °C) and 0.31 gDP kg^?0.78 day^?1 + 1.98PG (19 °C). The total requirements for DE were 36.86 kJDE kg^?0.84 day^?1 + 1.58EG (energy gain) (15 °C) and 46.14 kJDE kg^?0.86 day^?1 + 1.64EG (19 °C). The partial efficiencies for growth were 0.47 (15 °C) and 0.51 (19 °C) for protein, and 0.63 (15 °C) and 0.61 (19 °C) for energy. Nutrient gain was lower at the elevated temperature; however, feed formulation for brook trout should be adjusted to match changes in nutrient requirements at different culture temperatures. The protein and energy requirements model will be useful for developing commercial feeds and feeding charts for brook trout.