Development of a rapid assay for the diagnosis of Myxobolus cerebralis in fish and oligochaetes using loop-mediated isothermal amplification

A loop-mediated isothermal amplification assay was developed for the rapid detection of Myxobolus cerebralis in both fish and oligochaete hosts. The assay was optimized to amplify parasitic DNA by incubation with Bst DNA polymerase and a set of six specially constructed primers at 65 degrees C for 60 min. The amplification products were detected visually using SYBR Green I dye which gave identical results to gel electrophoresis analysis. Parasite DNA was detected from infected oligochaetes, and from the anal fin, caudal fin, dorsal fin and operculum of clinically infected fish. This 'Myxo-LAMP' assay has a detection limit similar to that of a polymerase chain reaction assay (10(-6)), but is more rapid and only requires a water bath for amplification and is therefore practical for simple and rapid diagnosis of infected tissue.     

Effect of the use of the microalga Spirulina maxima as fish meal replacement in diets for tilapia, Oreochromis mossambicus (Peters), fry

The present study addresses the use of the microalga Spirulina maxima as a protein source in diets for tilapia, Oreochromis mossambicus (Peters), fry. Animal protein was replaced with algae protein at ratios of 20%, 40%, 60%, 80% and 100%, and the substitution effect was compared with a control diet in which fish meal was the sole protein. An additional 100% spirulina protein diet was supplemented with phosphorous to test for possible mineral deficiency in the plant-protein-based diet. The six treatments were tested in triplicate in a closed-recirculating system where the fish were fed by hand at 6% of their body weight. After a 9-week feeding period, the growth rate and protein utilization of fish fed the diet with 20% and 40%Spirulina were elevated and not significantly different (P>0.05) from those fed the control diet. Further increases in the alga protein content significantly decreased the growth and feeding performance. The addition of P to the 100%Spirulina diet slightly improved performance in comparison to the same diet without P. None of the treatments produced any clear adverse effects on carcass composition. It is observed that Spirulina can replace up to 40% of the fish meal protein in tilapia diets.     

Effect of different feed formulations on feed efficiency, gonad yield and gonad quality in the purple sea urchin Heliocidaris erythrogramma

The nutritive values of three pelleted prepared diets, based on animal (AP), vegetable (VP) and yeast protein (YP) were studied for the sea urchin Heliocidaris erythrogramma (Val.). Fresh macroalga Ulva australis was used as a natural control diet. Triplicate groups of five animals were fed one of the four diets ad libitum every second day for 85 days. Sea urchins fed pelleted feeds had significantly higher food consumption rates (dry basis) and significantly lower total and protein absorption efficiencies compared with the algal diet. The gonad yield and gonad production efficiency in sea urchins fed the natural diet were significantly higher compared with initial group (gonad yield only) and urchins fed animal and vegetable diets, but did not differ significantly from those of the animals fed the yeast diet. Percent protein and lipid in the gonads were not affected by the dietary source. The taste and smell of gonads were generally better in sea urchins collected from the wild or fed yeast and natural diets than in sea urchins fed animal and vegetable diets. The animal protein diet was the most stable in seawater while the yeast protein diet had the poorest water stability. The results of this study suggest that development of a more stable, single‐cell, protein‐based diet has a potential to promote gonad production of H. erythrogramma.     

Nutrient digestibility studies in Heteropneustes fossilis (Bloch), Clarias batrachus (Linnaeus) and C. gariepinus (Burchell)

The apparent digestibility coefficient (ADC %) for protein in catfishes, Clarias batrachus (Linnaeus) (16–18 cm; 45–50 g), C. gariepinus (Burchell) (21–22 cm; 60–65 g) and Heteropneustes fossilis (Bloch) (10–12 cm; 10–15 g) was estimated for nine feedstuffs of plant and animal origin. Fat digestibility was tested for C. batrachus and C. gariepinus, wherein the influence of feeding frequency on protein digestibility was also examined. The digestibility trials were conducted with triplicate groups of 25 fish each in 70‐L polyvinyl indoor flow‐through (1–1.5 L min^?1) circular tanks (water volume=55 L). Fish were fed to satiation daily, at 08:00 and 18:00 hours, and faecal matter was collected through slow siphoning. For studying the effect of feeding frequency, fish were fed the experimental diets at 08:00, 11:00, 14:00 and 17:00 hours. The protein digestibility of the feedstuffs varied significantly within each test species. However, the protein digestibility values for most of the ingredients tested were comparable between the three catfish species. The most efficient utilization of protein in these fishes was noted for soybean meal followed by lab‐prepared fishmeal. The lowest protein digestibility was recorded for rice bran. Similarly, the utilization of fat from the feedstuffs was also significantly different within each of the two species, C. batrachus and C. gariepinus. Fat digestibility of feed ingredients was similar between the two species. With respect to the fat, soybean meal, lab‐prepared fishmeal, full‐fat soybean meal and dried fish were more efficiently utilized, while the lowest fat digestibility was obtained for rice bran. A marked decline in protein utilization was observed in the two species where the feeding frequency was increased from two to four times a day.     

The persistence of infectivity of Tetracapsulabryosalmonae-infected water for rainbow trout, Oncorhynchus mykiss (Walbaum)

Proliferative kidney disease (PKD) is caused by the infection of susceptible salmonid fish with spores of the myxozoan Tetracapsula bryosalmonae , a parasite harboured and released by several species of bryozoans. Under natural conditions, PKD is a water-borne infection of fish, whose outcome and spatio-temporal dissemination depend on the viability of spores present in the water. In order to evaluate the duration of parasite infectivity, juvenile rainbow trout, Oncorhynchus mykiss , were exposed for 20 h to T. bryosalmonae -infected water at various times post-water collection or after different filtration procedures. When infected water was held in a temperature range of 14.5–17 °C for up to 14 days, PKD was transmitted to the fish only between 0 and 12 h post-water collection and its infectivity vanished between 12 and 24 h. Similarly, the infectivity of water passed through 25 μm but not through 1 μm mesh filters, and was lost in the material eluted from the 1 μm filtration membrane although the parasite's DNA was amplified from this material. The parasitic infectivity in water appears to be fragile and this may offer opportunities to decrease the impact of PKD in trout farms by the implementation of management procedures aimed at reducing the number of the bryozoan-holding surfaces located in the river, immediately upstream from these farms.     

Nutrient composition and metamorphosis success of Atlantic halibut (Hippoglossus hippoglossus, L.) larvae fed natural zooplankton or Artemia

Atlantic halibut larvae were fed docosohexanoic acid- (DHA) selco enriched Artemia (RH-cysts) or wild zooplankton in duplicate tanks from first-feeding and 60 days onward. The zooplankton were collected from a fertilized sea water pond and consisted mainly of different stages of Eurytemora affinis and Centropages hamatus . There were no differences in survival, or in growth during the first 45 days of feeding, between larvae fed the two prey items, but the larvae fed Artemia showed much higher incidence of malpigmentation and impaired eye migration than larvae fed zooplankton. The prey organisms contained similar amounts of dry matter and protein, but Artemia was higher in lipid and glycogen than the zooplankton. Larvae fed Artemia were higher in both glycogen and lipid than the zooplankton-fed larvae towards the end of the feeding period. There were large differences between the prey organisms in the concentrations of essential fatty acids (% of total fatty acids) which was reflected in the fatty acid composition of the larval body. It is concluded that the macronutrient composition of Artemia in the present study was probably within the optimal range for promotion of growth and survival in young Atlantic halibut. The concentration of n-3 HUFA, and especially DHA, is however, very much lower in enriched Artemia than in copepods, and may be one of the factors triggering developmental errors in Atlantic halibut.     

Morphological characterization and notes on the life cycle of a newly discovered variant of Loma salmonae (Putz, Hoffman & Dunbar) from a natural infection of chinook salmon, Oncorhynchus tshawytscha (Walbaum)

Two variants of Loma salmonae occur in net-pen reared chinook salmon, Oncorhynchus tshawytscha. The typical variant (OA) has a host specificity for salmonids of the genus Oncorhynchus whereas the atypical variant (SV) has a host specificity for brook trout, Salvelinus fontinalis, and in this study, the ultrastructure of the two are compared. In fish at 8 weeks post-exposure xenomas of the SV variant have a very high proportion of mature spores compared with other developmental stages, while in xenomas of the OA variant there are fewer spores and many other developmental stages. Spores of the SV variant had up to 20 turns of their polar tube whereas those of the OA variant only had 17. Furthermore, the spores of the SV variant were significantly larger than those of the OA variant. The sporophorous vesicle for both variants appears to form around a sporogonial plasmodia, which results in many spores developing within the vesicle.     

Larval pigmentation, survival and growth of Penaeus indicus fed the nematode Panagrellus redivivus enriched with astaxanthin and various lipids

The highly unsaturated fatty acid (HUFA) content of the free-living nematode Panagrellus redivivus was successfully increased when three different lipid sources (cod-liver oil, marilla oil and capelin oil) from marine animals were used in the nematode culture medium. The lipid enrichment significantly increased the ω3 HUFA content, particularly eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), of the nematodes compared with lipid non-enriched nematodes. Penaeus indicus larvae fed the lipid-enriched nematodes had significantly ( P < 0.05) greater larval survival (69–77%) until metamorphosis than those fed non-enriched nematodes (54%).
This nematode species was also enriched with astaxanthin to determine the effects of this carotenoid on pigmentation, survival and growth during the larval development of P. indicus . The pigment, delivered via nematodes, significantly ( P < 0.05) improved larval coloration and survival (88%) compared with that of placebo-pigmented nematodes (78%). However, there was no strong evidence to show the benefit of either pigment and/or lipid enrichment on larval growth and development of P. indicus .
This study has shown that the nutritional value of the nematodes can be enhanced by the addition of fish oil into the culture medium. Supplementation of EPA and DHA, together with synthetic astaxanthin, allow the nematodes to be used as a sole diet for the larval culture of P. indicus .