Morphology of the pig cornea in normal conditions and after incubation in a perfusion apparatus

The fine structure of the pig cornea in normal conditions and after being used in a perfusion apparatus, for 4 h, is described. Earlier reports on the normal morphology of the pig cornea were partly not confirmed. Thus the number of cell layers in the epithelium was found to be 19-23 (a basal cell layer, 4–5 polyhedral cell layers and 14–17 squamous cell layers) compared to earlier reported 6–9 layers.The mean thickness of normal and perfused corneas were 722 μm and 752 μm respectively. Normal corneas had a hydration level of 77.2 % and after perfusion 78.5 %. The normal morphology and morphological changes due to exposure to perfusion were studied by light and electron microscopy. The differences observed between normal and perfused corneas have to be considered limited, and restricted mainly to the anterior squamous epithelium and the endothelium.Taken together our results indicate that the corneas used in the apparatus still had functional integrity.     

Evaluation of Cassia fistula meal as a replacement for soybean meal in practical diets of Oreochromis niloticus fingerlings

This study was undertaken to determine the replacement value of Cassia fistula seed meal (CFM) for soybean meal (SBM) in practical diets of Oreochromis niloticus fingerlings. Five practical diets (350 g kg^?1 crude protein) containing 0 g kg^?1 (control), 170 g kg^?1 (diet II), 340 g kg^?1 (diet III), 509 g kg^?1 (diet IV) and 670 g kg^?1 (diet V) substitution levels of CFM for SBM were formulated and fed to triplicate groups of O. niloticus fingerlings (mean initial weight of 10.22 ± 0.03 g) for 70 days. Fish mortality increased linearly with increase in inclusion levels of CFM in the diet. Growth and diet utilization efficiency were depressed in fish fed diets containing CFM at varying inclusion levels. Feed conversion ratio, specific growth rate and protein efficiency ratio of O. niloticus fed on diet containing 170 g kg^?1 substitution level of CFM were similar (P > 0.05) to the control diet. Digestibility of the different diets decreased with increase in inclusion levels of CFM. Fish fed diet containing 670 g kg^?1 CFM had significantly lower carcass protein. However, no significant differences were observed in carcass protein and lipid contents between fish fed the control diets and diet containing 170 g kg^?1 CFM. The most efficient diet in terms of cost per unit weight gain of fish was obtained in 170 g kg^?1 CFM dietary substitution.     

Characteristics of monoclonal antibodies against Piscirickettsia salmonis

A panel of 28 monoclonal antibodies against Piscirickettsia salmonis was produced using a purified fraction of the bacterium. To determine their specificity to the pathogen, the antibodies were assayed by ELISA and indirect immunofluorescence microscopy. Six monoclonal antibodies were selected based on their strong reaction against P. salmonis and absence of cross-reactivity with other common fish pathogens. Western blot analysis showed that the antibodies reacted to several antigens of P. salmonis . Immunofluorescence assays revealed that these antibodies reacted with the same specificity to different isolates of P. salmonis obtained from the south of Chile. This panel of monoclonal antibodies represents an important tool to develop simple, rapid, sensitive and highly specific methods for the detection of the pathogen and diagnosis of the disease.     

Effects of oxygen supersaturation and temperature on juvenile greenlip, Haliotis laevigata Donovan, and blacklip, Haliotis rubra Leach, abalone

Growth and survival of juvenile greenlip (39.03 (SD 3.80) mm (n=524)) and blacklip (31.92 (SD 4.19) mm (n=531) abalone were investigated at high dissolved oxygen levels (95–120% saturation) between 17 and 19°C. Abalone were fed the same artificial diet and each species was contained in groups of approximately 30 individuals within triplicate tanks for each of six treatments and were exposed to flow through water adjusted to give experimental conditions for up to 75 days. Blacklip abalone held at 16.9°C and 97% oxygen saturation grew in shell length significantly faster than all other treatments of blacklip abalone held at 19°C, and significantly faster than blacklip abalone maintained at 111% oxygen saturation and 17.5°C. Both temperature and oxygen saturation significantly affected the survival of this species. Blacklip abalone held at 19°C had significantly lower survival for both 96% oxygen saturation and 120% oxygen saturation, compared with blacklip abalone maintained at either 110% oxygen saturation and 19°C, or for any 17°C treatment. No significant differences were noted for greenlip abalone within the range tested in terms of growth rate, food consumption rate or survival, indicating that greenlip abalone tolerated these conditions better than did blacklip abalone.     

Kurokura Solution as Immobilizing Medium for Spermatozoa of Tench (Tinca tinca L.)

This study tested KUROKURA solution (Kurokura et al., 1984, Aquaculture 37, 267–274) and its modifications (by increasing NaCl content to 160, 180 and 200 mM) on immobilizing properties for sampling and short-term preservation of potential motility of tench spermatozoa. The immobilizing solution is used because, when collected, the sperm of most samples is contaminated by urine, causing spermatozoa to be of poor quality, with low motilities and velocities (almost 0), thus resulting in a worsened fertilization and hatching rate. Sperm was sampled with a syringe containing an immobilizing solution (IS), allowing an IS:sperm ratio of 2:1, under aerobic conditions at 0–4°C. This sperm solution was stored for 10 h and untreated sperm was collected prior to fertilization as a control. Spermatozoa quality was evaluated for the cell motility and velocity parameters and also for fertilization ability and hatching rate. Results obtained for tench sperm motility, velocity, fertilization and hatching rate showed that only sperm collected in the various immobilizing solutions can be successfully used for artificial insemination and preservation after 10 h at 0–4°C. The best immobilizing solution was found to be KUROKURA 180 (180 mM NaCl, 2.68 mM KCl, 1.36 mM CaCl₂· 2H₂O and 2.38 mM NaHCO₃), giving a fertility and hatching rate of 41%, with no change in rates after 10 h storage of sperm. Control sperm without immobilizing solution showed a fertilization and hatching rate of only 6–7%.     

Combined effects of temperature and diet on growth and biochemical composition of the Pacific oyster Crassostrea gigas (Thunberg) spat

Temperature and quality of the available food are important factors that influence the physiology of oysters; however, the combined effects have not been well studied. We evaluated the impacts of the temperature and diet on the growth, survival and biochemical composition in the Pacific oyster Crassostrea gigas spat, cultured in the laboratory for 8 weeks at 23, 26, 29 and 32°C and fed Isochrysis sp.‐Pavlova lutheri (IP) and Dunaliella tertiolecta (Dt). The growth and biochemical composition showed a pattern, which changed in response to rising temperature. The shell length was significantly longer, in spat fed the IP diet, except at 32°C, where both diets produced poor growth results. The survival was <50% after 5 weeks at 32°C, whereas at all other temperatures it was >88%. High temperatures directly increased lipids and saturated fatty acids, while the proteins, carbohydrates and unsaturated fatty acids decreased. High temperatures achieved in the environment, as those reached on clear summer days during low tides, are an important stressor in oyster spat, especially when the quality of the available food is poor.     

Colouring ornamental fish (Cyprinus carpio and Carassius auratus) with microalgal biomass

Koi carp and goldfish value increases with intensity of skin colour, which is an important quality criterion. Fish cannot fully synthesize their own carotenoid colourings and these must therefore be included in their diet. Two trials were undertaken to investigate skin colour enhancement in ornamental species (i.e. three chromatic varieties of koi carp (Cyprinus carpio), namely Kawari (red), Showa (black and red) and Bekko (black and white) and goldfish (Carassius auratus)) by feeding a dietary carotenoid supplement of freshwater microalgal biomass [Chlorella vulgaris, Haematococcus pluvialis, and also the cyanobacterium Arthrospira maxima (Spirulina)], using a diet containing synthetic astaxanthin and a control diet with no colouring added for comparison. In the first trial, five homogeneous duplicate groups of 25 juvenile koi carp (C. carpio) (initial mean body weight 24.6 ± 0.7 g) were fed, for 10 weeks, one of the four diets containing 80 mg colouring/kg diet. In the second trial, this procedure was repeated for five homogeneous duplicate groups of 25 goldfish (C. auratus) (initial mean body weight of 0.9 ± 0.1 g). Initial and final samples of skin along the dorsal fin were withdrawn, from five fish per group, for subsequent analysis of total carotenoid content (spectrophotometric analysis), and red hue (colorimetric analysis, CIE (1976) L* a* b* colour system). Growth and feed efficiency were not significantly different between groups administered by the various dietary treatments. In both trials, dietary carotenoid supplementation increased total skin carotenoid content. The more efficient colouring for koi carps was found to be C. vulgaris biomass, providing both maximum total carotenoid deposition and red hue for the three chromatic koi carp varieties studied, and particularly for the kawari variety. For goldfish the best colouring obtained, as ascertained by total carotenoid content, was also achieved using C. vulgaris biomass, and red hue was maximum when using H. pluvialis biomass.     

Host preference of adult Caligus elongatus Nordmann in the laboratory and its implications for Atlantic cod aquaculture

The sea louse Caligus elongatus utilizes a range of marine fish species as hosts, and it has been suggested that fish farms are infected with adult C. elongatus from wild fish hosts. We assessed experimentally whether dislodged lice return to their original host species. Adult lice from wild lumpfish and saithe were presented to lumpfish, sea trout, Atlantic cod, saithe and European plaice. All fish species became infected. Lumpfish and cod were strongly preferred, followed by saithe, trout and plaice. Lice from wild saithe showed a preference for saithe and lumpfish, followed by cod, sea trout and plaice. The mtCO1 gene was used to determine the genotypes of the lice. Lice from lumpfish were genotype 1. Lice from saithe were mainly genotype 2. The two genotypes of C. elongatus differed slightly in their host preferences: lice from saithe were less discriminating. The data support the hypothesis that adult C. elongatus may transfer between different host species. Cod is an attractive host for lice from both lumpfish and saithe, which suggests that C. elongatus may become a serious pest of farmed cod.     

Identification of Gyrodactylus (Monogenea) species parasitizing salmonid fish using DNA probes

Abstract. Oligonucleolides GsV4B, GdV4 and GtV4, complementary to sequences from Gyrodactylus Salaris, G. derjavini and G. truttae , respectively, were designed following examination of the sequence of the small subunit ribosomal RNA (srRNA) gene from each species. The oligonucleotides were end-labelled with digoxigenin for use us species-specific probes. The V4 region of the srRNA gene was amplified from individual gyrodactylids using the polymerase chain reaction (PCR) and aliquots of the PCR products werespotted onto nylon membrane. Optimum conditions for hybridization of the oligonucleotide probes to these samples were determined. The use of these three probes against aliquots of PCR product provides an objective, sensitive and specific method suitable for routine diagnostic use in identifying Gyrodactylus species found on salmonid fish, but cannot be used to distinguish G. salaris and G. thymulli .