First isolation of Tenacibaculum maritimum from wedge sole, Dicologoglossa cuneata (Moreau)

The first isolation of Tenacibaculum maritimum from wedge sole, Dicologoglossa cuneata, is reported. The pathogen was recovered from ulcers of cultured fish, from three different outbreaks. The six isolates obtained were biochemically and serologically characterized and diagnosis was confirmed by polymerase chain reaction using specific primers and partial 16S rRNA gene sequencing. The isolates constituted a homogeneous phenotypic group; however, they belong to two of the different serotypes described within this species. A virulence evaluation of the isolates using Wedge sole fry was also performed.     

Histopathological effects of cadmium on the gills of the freshwater fish, Macropsobrycon uruguayanae Eigenmann (Pisces, Atherinidae)

Evaluations of histopathological lesions in gill tissue were carried out in the freshwater fish Macropsobrycon uruguayanae following 30 and 60 days of exposure to 1.5 mg 1^-1 of cadmium. The study was conducted on both fed and starved animals in order to determine the influence of feeding condition on cadmium toxicity. The main lesions observed and quantified were: (1) hyperplasia of primary lamellar epithelium; (2) hyperplasia of secondary lamellar epithelium; (3) separation of respiratory epithelium; (4) shortening of secondary lamellae; (5) epithelial necrosis; (6) fusion of adjacent secondary lamellae; (7) hypertrophy of respiratory epithelium; (8) lamellar telangiectasis; (9) hyperplasia of chloride cells; (10) mucinous metaplasia; and (11) inflammatory infiltration. Lesions 6, 8, 9 and 11 were only induced by exposure to cadmium, while lesion 4 could be produced only by starvation. Starved fish also showed a reduction in total body weight and length. Lesion 2 was shown to be non-specific, and produced by either cadmium, starvation or even exposure time. The possible mode of action of the experimental factors are discussed in relation to the observed pathologies.     

Comparison of dot blot and PCR diagnostic techniques for detection of white spot syndrome virus in different tissues of Penaeus monodon

Comparison of PCR and dot blot diagnostic techniques for detection of white spot syndrome virus (WSSV) was made on different tissues of infected Penaeus monodon including eye stalk, eye stalk with eye, gills, cuticle, pleopod, periopods, uropods and telson. Dot blots of crude DNA extracted from infected tissue samples showed positive reactions with all the samples; however, the sensitivity of the dot blot was reduced with the purification of DNA samples extracted from pleopod, telson and uropod. PCR was found to be more sensitive when compared to dot blot. Both crude DNA and purified DNA samples extracted from all the tissues except for eye stalk with eye showed single step nested PCR positive reaction. The amplification of all or either of the three bands of 941 bp, 525 bp and 204 bp size varied with the tissues analysed. The severity of infection assessed by PCR amplification was found to be maximum in cuticle and telson followed by gill. Other tissues such as eye stalk, pleopod, periopods and uropod were observed to have mild infection. The maximum intensity of the PCR product was for the smallest amplified product of 204 bp followed by 525 bp and the weakest intensity was observed for the 941 bp size. The limitation of PCR due to inhibiting factors present in tissues could be overcome with the use of dot blot which gave positive reaction from the DNA extracted from eye stalk including the eye but yielded no amplification by PCR.     

Toxicity of aqueous aluminium to the ectoparasitic monogenean Gyrodactylus salaris

Both laboratory and field experiments have demonstrated that aqueous aluminium can act as a paraciticide to the monogenean Gyrodactylus salaris Malmberg, 1957 infecting freshwater salmonids. However, the reproductive conditions of gyrodactylids surviving to cessation of an aluminium exposure is unknown. Therefore, Atlantic salmon (Salmo salar L.) parr infected with G. salaris was experimentally exposed to elevated concentrations of aqueous aluminium for more than 1 month. During this period, the infection increased the first week before it peaked and started to steadily decline approaching elimination. When almost all parasite specimens were eliminated, the water quality was adjusted to normal aluminium-poor freshwater. During the next 3 weeks, the surviving G. salaris were shown to clearly resume their reproduction. The experiment demonstrates the potential of the gyrodactylids to reproduce after cessation of an aluminium exposure. Thus, the as-yet unknown mechanism behind the toxic effects of aluminium on G. salaris survival and reproduction might to some extent be reversible. The present study gives further support for this metal as a parasiticide to G. salaris but demonstrates at the same time that in order to eliminate the parasites totally, the aluminium treatment must eradicate all G. salaris from the skin of the infected hosts. Furthermore, the potential development of resistance to aluminium should also be studied.     

High yield and rapid growth of Neoparamoeba pemaquidensis in co-culture with a rainbow trout gill-derived cell line RTgill-W1

Neoparamoeba pemaquidensis is an ubiquitous amphizoic marine protozoan and has been implicated as the causative agent for several diseases in marine organisms, most notably amoebic gill disease (AGD) in Atlantic salmon. Despite several reports on the pathology of AGD, relatively little is known about the protozoan and its relationship to host cells. In this study, an in vitro approach using monolayers of a rainbow trout gill cell line (RTgill-W1, ATCC CRL-2523) was used to rapidly grow large numbers of N. pemaquidensis (ATCC 50172) and investigate cell-pathogen interactions. Established cell lines derived from other tissues of rainbow trout and other fish species were also evaluated for amoeba growth support. The amoebae showed preference and highest yield when grown with RTgill-W1 over nine other tested fish cell lines. Amoeba yields could reach as high as 5 x 10(5) cells mL(-1) within 3 days of growth on the gill cell monolayers. The amoebae caused visible focal lesions in RTgill-W1 monolayers within 24 h of exposure and rapidly proliferated and spread with cytopathic effects destroying the neighbouring pavement-like cells within 48-72 h after initial exposure in media above 700 mOsm kg(-1). Disruption of the integrity of the gill cell monolayers could be noted within 30 min of exposure to the amoeba suspensions by changes in transepithelial resistance (TER) compared with control cell monolayers maintained in the exposure media. This was significantly different by 2 h (P < 0.05) compared with control cells and remained significantly different (P < 0.01) for the remaining 72 h that the TER was monitored. The RTgill-W1 cell line is thus a convenient model for growing N. pemaquidensis and for studying host-pathogen interactions in AGD.     

Effects of dietary carbohydrate on weight gain and gonad production in small sea urchins,Lytechinus variegatus

In experiment 1, juvenile sea urchins (n = 80, 0.088 ± 0.001 g wet weight and 5.72 ± 0.04 mm diameter) were held individually and fed ad libitum one of three semi‐purified formulated diets (n = 16 individuals treatment^?1). In the diets, protein was held constant (310 g kg^?1 dry, as fed) and carbohydrate level varied (190, 260, or 380 g kg^?1 dry, as fed). Wet weights were measured every 2 weeks. Total wet weight gain was inversely proportional to dietary carbohydrate level and energy content of the respective diet. In experiment 2, sea urchins (5.60 ± 0.48 g wet weight, n = 40) fed 190 g kg^?1 carbohydrate consumed significantly more dry feed than those fed 260 g kg^?1, but not more than those fed 380 g kg^?1 carbohydrate. Based on differential feed intake rates, sea urchins that consumed more feed also consumed higher levels of protein and had the highest weight gain. Consequently, protein content and/or protein: energy ratio may be important in determining feed utilization and growth among sea urchins in this study. The average digestible energy intake was approximately 70 kcal kg^?1 body weight day^?1, suggesting daily caloric intake of juvenile Lytechinus variegatus is lower than in shrimp and fish.     

Food consumption and digestive enzyme activity of Clarias batrachus exposed to various temperatures

The effect of temperature on the food consumption rate and the digestive enzyme activities of Clarias batrachus (80.60 ± 5.34 g) were evaluated. Fish were exposed to six different temperatures of 10, 15, 20, 25, 30 and 35 °C following an acclimation temperature of 25 °C. The rate of temperature change was 2 °C day^?1. Highest food consumption was recorded at 25 °C. It gradually reduced with decreasing water temperature. Food consumption rate was significantly (P < 0.05) lower at 10 °C compared with other treatments. Hence, 46.67, 8.20–23.58 and 1.02–6.15% reduced food consumptions were recorded in groups exposed at 10, 15 and 20 °C temperatures, respectively, compared with the 25 °C. The consumption rate was not affected in fish exposed at 30 and 35 °C. Total protease, trypsin and chymotrypsin activities were significantly (P < 0.05) higher in fish exposed at 25 °C compared with others. Lipase activity was significantly (P < 0.05) higher in fish exposed at 30 °C compared with others. Lowest enzyme activities were recorded at 10 °C. Water temperature below 25 °C affected the food consumption and digestive enzyme activities in fish that served as indicators of stress in fish.     

Formulation of cheaper artificial feeds for shrimp culture: Preliminary biochemical,physical and biological evaluation

A compound shrimp diet was prepared making use of the locally available natural food stuffs, slaughter house products and factory by-products. The ingredients, viz: beef liver (discarded for human use on veterinary grounds), coconut oil cake, soya flour, mangrove leaves, and wheat flour, were analysed biochemically for their nutritive value. The formulated diet is in a dry pellet form. It was assessed experimentally for its physical stability and subsequently fed to various size groups of a palaeomonid prawn (Leptocarpus potamiscus) for biological evaluation. The results of the investigations suggest that beef liver meal and coconut oil cake are rich in protein and can be utilized in the formulation of aquatic diets. The diet is highly water stable, consumable and assimilable. It should be tested upon both palaeomonid and penaeid prawns in further experiments.     

Production of polyclonal antiserum against recombinant VP28 protein and its application for the detection of white spot syndrome virus in crustaceans

The VP28 gene of white spot syndrome virus (WSSV) was cloned into pRSET B expression vector. The VP28 protein was expressed as a protein with a 6-histidine taq in Escherichia coli GJ1158 with NaCl induction. Antiserum was raised against this recombinant-VP28 protein in rabbits and it recognized VP28 protein in naturally and experimentally WSSV-infected shrimp, marine crabs, freshwater prawns and freshwater crabs. The antiserum did not recognize any of the other known WSSV structural proteins. Various organs such as eyestalks, head muscle, gill tissue, heart tissue, haemolymph, tail tissue and appendages were found to be good materials for detection of WSSV using the antiserum and detection of WSSV was successful in experimentally infected Penaeus monodon and P. indicus at 12 and 24 h post-infection (p.i.), respectively. The antiserum was capable of detecting WSSV in 5 ng of total haemolymph protein from WSSV-infected shrimp.     

Median Lethal Concentration (LC50) of Piscicidal Plants and Their Utilization in Aquaculture

Aqueous and ethanol extracts of Acacia pennata (bark), Catunaregam uliginosa (fruit), Diospyros lenceofolia (fruit), and Sapindus mukorossi (fruit) were evaluated as piscicides on the freshwater fish Danio dangila, Danio rerio, and Puntius shalynius, and the air-breathing Heteropneustes fossilis. Catunaregam uliginosa (fruit) possessed relatively higher piscicidal potency followed by Sapindus mukorossi (fruit), Acacia pennata (bark), and lastly Diospyros lanceofolia (fruit). Catunaregam uliginosa can thus be used for eradicating unwanted wild fishes in a short time span at low concentrations so that the health of aquatic environment is maintained.