Quantification of Flavobacterium psychrophilum in rainbow trout, Oncorhynchus mykiss (Walbaum), tissues by qPCR

A qPCR assay was developed for rapid and sensitive detection of Flavobacterium psychrophilum, the aetiological agent of bacterial cold-water disease and rainbow trout fry syndrome in salmonid fish worldwide. A set of F. psychrophilum-specific primers based on 16S rRNA gene sequences was designed and validated for specific detection and quantification of DNA isolated from representative strains of F. psychrophilum. The qPCR assay exhibited a high specificity for the 16S rRNA gene of F. psychrophilum (from 4 × 10(8) down to 11 copies per reaction) but not for other Flavobacterium species or other bacteria including fish pathogens. This qPCR-based method proved to be useful in the quantification of the F. psychrophilum titre present within organs dissected out from diseased fish. As the F. psychrophilum genome contains six copies of the 16S rRNA gene, we could infer a limit of detection corresponding to two bacteria per reaction, corresponding to 800 bacteria per fish tissue sample, and therefore 20 F. psychrophilum cells mg(-1) of tissue (for sample weighing 40 mg). The qPCR assay reported here could be a useful tool for veterinary diagnostic laboratories to monitor the F. psychrophilum infection level in fish farms.     

Validation of diagnostic assays to screen broodstock for Flavobacterium psychrophilum infections

It is hypothesized that the frequency of bacterial coldwater disease outbreaks can be reduced through the detection of the aetiologic agent, Flavobacterium psychrophilum, in broodstock followed by culling of eggs from heavily infected broodstock. Before a culling programme can be instituted, however, it is necessary to determine the sensitivity and specificity of existing assays for the detection of F. psychrophilum. In this study, tissue and ovarian fluid samples were collected from 224 fish at five hatcheries and screened using an enzyme‐linked immunosorbent assay (ELISA), a membrane‐filtration fluorescent antibody test (MF‐FAT), bacteriological culture and nested PCR. Latent class analysis was used to estimate sensitivity and specificity of kidney culture, kidney ELISA, nested PCR and MF‐FAT. Analytical sensitivity of the ELISA varied but was greatest when bacteria were cultured under iron‐limiting conditions. Diagnostic sensitivity estimates ranged from 0.02 (kidney culture) to 0.97 (kidney ELISA). Specificity estimates ranged from 0.02 (MF‐FAT) to 0.98 (kidney ELISA). In a separate challenge experiment, the ELISA confirmed the presence of F. psychrophilum in sub‐clinically infected fish. Results from this study demonstrate that the ELISA is an appropriate tool to screen broodstock and provides an indication of infection severity, which is crucial for implementation of a screening/culling programme.     

饲料营养与鱼类的健康

营养状态是决定鱼类抵抗疾病能力的一个重要因子.优质饲料对改善养殖鱼类的健康与提高抗病能力至关重要,这一点已被大量实验所证实.饲料中蛋白质、脂肪、碳水化合物等主要营养成分含量不足,会影响鱼类的正常生长,甚至出现缺乏症;现越来越多的研究发现某些必需营养成分(必需氨基酸、必需脂肪酸、维生素和矿物质)含量不足的饲料会导致鱼类营养不良和抗病力弱,微量营养元素与鱼类抗病力的关系研究日益得到重视.     

Use of ongrownArtemia in nursery culturing of the tiger shrimp

Juvenile and adultArtemia produced in a semi flow-through culture system were used as food for postlarval shrimp. The growth performance of shrimp reared on such ongrownArtemia live prey is identical to the growth obtained when feeding newly hatchedArtemia. However, a significantly better stress resistance is obtained when the postlarvae are exposed to a low salinity in a stress test. Besides nutritional and energetic advantages, the use ofArtemia biomass for feeding postlarval shrimp also results in improved economics as expenses for cysts and weaning diets can be reduced.     

Mycoplasma contamination in fish cell lines: an evaluation of detection by differential incorporation of 3H-uridine and 14C-uracil

Abstract. Differential incorporation of uridine and uracil was used to assay for mycoplasma contamination in five fish cell lines: bluegill fry (BF-2), chinook salmon embryo (CHSE-214), epithelioma papillosum cyprini (EPC), fathead minnow (FHM) and rainbow trout gonad (RTG-2). The method was not suitable for monitoring BF-2, CHSE-214, FHM, and RTG-2 cell lines because they incorporated uracil. Differential incorporation of uridine and uracil may be applicable for screening EPC cells because only this cell line could distinguish cultures experimentally infected with Mycoplasma orale from cultures known to be free from microbial contaminants.     

Examining changes in participation in recreational fisheries in England and Wales

Abstract Inland fisheries in England and Wales have high economic and social values. Managing participation to maximise fishery performance is key to maintaining this status. The capital value of fishing rights for migratory salmonid fisheries is €165 million. Coarse fisheries contribute €1030 million to the economy. The central tenet to increasing participation in recreational salmonid fisheries is that an increase in stock size will result in more anglers accessing the fishery. This was examined for salmon on the rivers Usk and Lune where exploitation restrictions increased the number of salmon available to anglers. On the River Lune, the number of salmon available post‐intervention increased by 66%. There was no significant increase in catch while the number of anglers decreased by 16.3%. On the River Usk, the closure of the net fishery potentially resulted in an additional ～1200 salmon being available. Following closure, there was no significant change in rod catch or in the number of anglers. Increased participation is dependent less upon stock manipulation for coarse fisheries and more upon facilitating the activity. In recent years, urban fishery development programmes have provided improved access to local fishing opportunity. Also, new anglers have been targeted through campaigns such as Get Hooked on Fishing and the Scout Angler Badge.     

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).     

2-methylisoborneol implicated as a cause of off-flavour in channel catfish, Ictalums punctatus (Rafinesque), from commercial culture ponds in Mississippi

Abstract. Samples of water, sediment and fish were collected from six commercial channel catfish, Ictalurus punctalus (Rafinesque), ponds from May to October and analysed for muddy-odour producing compounds. Three episodes of off-flavour in the fish occurred in late summer. The episodes were of 2–3 months' duration and correlated with the levels of 2-methylisoborneol in water, sediment and fish. Oscillatoria agardliii was the dominant phytoplankton in ponds with off-flavour fish. Other known muddy odour-producing compounds such as geosmin, 2-isopropyl-3-mcthoxypyrazinc, 2-isobutyl-3-methoxypyrazine and 2-isobutyl-6-methoxypyrazinc were not detected in the water sediment or at concentrations greater than 5·0 ng/l.     

Direct actions of serotonin on gonadotropin-II and growth hormone release from goldfish pituitary cells: interactions with gonadotropin-releasing hormone and dopamine and further evaluation of serotonin receptor specificity

In this study, the direct actions of serotonin (5HT) on gonadotropin (GTH)-II and growth hormone (GH) release in the goldfish were tested at the pituitary cell level. 5HT (10 nM - 10 µM) stimulated GTH-II but inhibited GH release from perifused goldfish pituitary cells in a dose-dependent manner. The minimal effective dose of 5HT tested to suppress basal GH secretion (10 nM) was 10-fold lower than that to stimulate GTH-II release (100 nM). The GTH-II releasing effect of 5HT was abolished by repeated 5HT treatment (10 µM) whereas the corresponding inhibition on GH release was unaffected. These results suggest that 5HT receptors on goldfish gonadotrophs and somatotrophs exhibit intrinsic differences in terms of sensitivity to stimulation and resistance to desensitization. Salmon GTH-releasing hormone (sGnRH, 100 nM) stimulated GTH-II and GH release from goldfish pituitary cells. The GTH-II releasing action of sGnRH was unaffected by simultaneous treatment of 5HT (1 µM). However, the corresponding GH response to sGnRH (100 nM) was inhibited. In the goldfish, dopamine is known to stimulate GH release through activation of pituitary D1 receptors. In the present study, the GH-releasing action of dopamine (1 µM) and the D1 agonist SKF38393 (1 µM) was significantly reduced by 5HT (1 µM). To examine the receptor specificity of 5HT action, the effects of 5HT1 and 5HT2 analogs on GTH-II and GH release were tested in goldfish pituitary cells. The 5HT1 agonist 8OH DPAT (0.1 and 1µM) and 5HT2 agonist methyl 5HT (0.1 1µM) mimicked the GTH-II releasing effect of 5HT. The 5HT1 agonist 8OH DPAT (0.1 and 1µM) also stimulated GH release but the 5HT2 agonist methyl 5HT (0.1 and 1µM) was inhibitory to basal GH secretion. In addition, 5HT (1µM) -stimulated GTH-II release was abolished by the 5HT1 antagonist methiothepin (10µM) and 5HT2 antagonist mianserin (10µM). Similarly, the inhibitory action of 5HT (1µM) on basal GH release was blocked by the 5HT2 antagonist mianserin (10µM). The 5HT1 antagonist methiothepin (10µM) was not effective in this regard. These results, taken together, indicate that 5HT exerts its regulatory actions on GTH-II and GH release in the goldfish directly at the pituitary cell level, probably through interactions with other regulators including sGnRH and dopamine. The GTH-II releasing action of 5HT is mediated through 5HT2 and possibly 5HT1 receptors. The inhibition of 5HT on basal GH release is mediated through 5HT2 receptors only. Apparently, 5HT1 receptors are not involved in this inhibitory action. In this study, a paradoxical stimulatory component of 5HT on GH release by activating 5HT1 receptors is also implicated.     

Dietary carotenoids and skin melanin content influence the coloration of farmed red porgy (Pagrus pagrus)

The present study investigates the effects of dietary carotenoid sources on the coloration of the red porgy, Pagrus pagrus. Red porgies (131.9 ± 16.2 g; mean ± SD) were fed for 12 weeks on five different diets supplemented with red carotenoids (mainly astaxanthin esters) supplied from Haematococcus pluvialis algae and yellow carotenoids (mainly β‐carotene, lutein, and zeaxanthin) supplied from Alfalfa, Medicago sativa L. and Spirulina, Spirulina pacifica. The carotenoid‐supplemented diets did not have any marked effect on the growth rate, the feed conversion ratio, the daily feeding rate, or the hepatosomatic index of red porgy. The biochemical indices measured in plasma including cholesterol, total proteins, glucose, lactate, phospholipids, non‐esterified fatty acids, triglycerides, and thyroid hormones did not differ significantly among groups. Diet did not affect significantly the melanophore‐area coverage, the melanin skin concentration and skin lightness. Carotenoid‐supplemented diets affected significantly the carotenoid deposition in the skin, the presence and distribution of erythorphores and xanthophores, and skin hue and chroma. Overall, data have shown the efficacy of Haematococcus algae in promoting a reddish coloration in red porgy.