Environmental factors affecting outbreaks of winter saprolegniosis in channel catfish, Ictalurus punctatus (Rafinesque)

Abstract. Previous studies have led to the development of a laboratory model for a disease termed 'winter saprolegniosis', locally referred to as 'winter kill', occurring in channel catfish, Ictalurus punctatus Rafinesque, raised in commercial ponds in the southeastern United States. In the laboratory, the onset of disease was routinely brought about by a combination of two related factors: (1) a rapid drop in water temperature which induces immunosuppression in catfish; and (2) maintenance of low water temperatures (∼10°C), which favour the proliferation of an ubiquitous fungus of the genus Saprolegnia to produce high levels of fungal zoospores (≥5 spores ml⁻¹). In this study, two commercial catfish ponds were monitored for over one year to determine if the above factors occurred in the field and could be correlated with outbreaks of winter saprolegniosis. It was noted that passages of severe cold weather fronts were able to drop pond water temperatures ≥10°C within 24h and that such decreases in water temperature were associated with immunosuppression of the catfish in the ponds. Furthermore, when Saprolegnia sp. zoospore levels were ≥5 spores ml^-1, the immunocompromised catfish exhibited overt signs of winter saprolegniosis. If one of the factors implicated in the induction of disease was missing, fish in the ponds remained healthy. In addition, the onset of disease in the ponds appeared independent of pH and oxygen, total ammonia nitrogen and un-ionized ammonia levels. Collectively, these field results confirm the laboratory-based hypothesis concerning the aetiology of winter saprolegniosis in channel catfish.     

Growth of juvenile tilapia, Oreochromis niloticus L. from Lakes Zwai, Langeno and Chamo (Ethiopian rift valley) based on otolith microincrement analysis

Abstract – Age and growth of juvenile tilapia, Oreochromis niloticus , from Lakes Zwai, Langeno and Chamo (Ethiopia) were studied from microincrements in otoliths. Growth in length was best described by the Gompertz model. Average growth rate of the fish was most rapid in Lake Chamo (0.39 mm  ·  day⁻¹, 1.14%  ·  day⁻¹), intermediate in Lake Zwai (0.20 mm  ·  day⁻¹, 0.72%  ·  day⁻¹) and slowest in Lake Langeno (0.16 mm  ·  day⁻¹, 0.62%  ·  day⁻¹). Similarly, back-calculation from otolith increment widths gave growth rates of 0.28 to 0.43 mm  ·  day⁻¹, 0.15 to 0.32 mm  ·  day⁻¹ and 0.11 to 0.28 mm  ·  day⁻¹ for Chamo, Zwai and Langeno fish, respectively. In addition, Fulton's condition factor was largest for Chamo tilapia and smallest for Langeno tilapia; the difference between fish from Langeno and Zwai was small. Rapid growth of juvenile O. niloticus in Lake Chamo was attributed to warm temperature and better food quality. ^Note     

Residues of [14C]-malachite green in eggs and fry of rainbow trout, Oncorhynchus mykiss (Walbaum), after treatment of eggs

Abstract. Rainbow trout, Oncorhynchus mykiss (Walbaum), eggs were exposed to [methane-¹⁴C] malachite green chloride on day 0 and on every third day thereafter through day 24, with a final treatment administered to fry on day 31. Eggs or fry were sampled before each treatment, and at selected times from day 31 to day 59. Malachite green equivalence in eggs and fry was determined by sample oxidation and liquid scintillation counting. Total malachite green residues increased throughout the exposure period to 0.271 ± 0.042 μg g⁻¹ (± SD) on day 31. Residues declined to 0.055 ± 0.011 μg g⁻¹ on day 59. The depuration phase declined monoexponentially with a half-life of 13.3 days for the absolute amount (μg sample⁻¹) and a half-life of 9.7 days for the concentration of malachite green residues (μg g⁻¹). Growth dilution accounted for the 25% increase in the elimination (9.7 days) of malachite green residues. Extracts from treated eggs and fry were analysed by reversed-phase liquid chromatography. Three peaks were resolved in treated eggs: chromatic malachite green, leuco malachite green and an unknown polar metabolite. Only two peaks were resolved in the fry: leuco malachite green and the unknown polar metabolite. The most prominent residue in all samples was leuco malachite green.     

Effect of population density on the growth, feed and protein conversion efficiency and biochemical composition of a tropical freshwater catfish, Mystus nemurus (Cuvier & Valenciennes)

Abstract. Six groups of tropical freshwater catfish, Mystus nemurus (Cuvier & Valenciennes)(mean weight. 20·45 ± l·5g), were reared in 0·34m³ fibreglass tanks at different stocking densities (105, 195, 285, 375, 465 and 555 specimens/m³ water) for 84 days. The objective of the study was to determine the effect of various stocking densities on the growth, nutrition, biochemical composition and survival of M. nemurus. The lowest growth rate appeared in fish at the highest density and the highest was observed in fish stocked in moderate density of 285 and 375 fish/m³ water. Fish production was also lower at relatively low stocking densities of 105 and 195 fish/m³. Food conversion ratio (FCR), protein efficiency ratio (PER) and biochemical composition of M. nemurus indicate that there exists an optimum stocking density which lies between 285 and 375 fish/m³.     

Growth and body composition of juveniles of Pterophyllum scalare (Lichtenstein) (Pisces; Cichlidae) at different densities and diets

Abstract. The effect on the growth of freshwater angelfish, Pterophyllum scalare (Lichtenstein) (Pisces; Cichlidae), of different densities of population and diet compositions was studied. The results show that increasing the population density from 50 fish/m² to 200/m² significantly decreased the rate of growth of fingerlings, and that this effect is correlated to the size of fish. No significant differences were observed in growth between diets containing 37%, 41% or 47% protein. However, the addition of live food (artemia) to the diet significantly raised the growth rate, especially in the higher protein diets.
Body composition shows no changes during the growth periods, with a mean of 64·68% body protein, 35·47% dry weight fat and no significant differences in the percentage of residual ash through the growth period.
The food conversion rate was very low, ranging from 1·15 to 1·31. Total production increased with density, from 4·0g/m²/day (50 fish) to 13·7g/m²/day (200 fish) over a period of 60 days.     

Histopathology, electron microscopy and isolation of channel catfish virus in experimentally infected European catfish, Silurus glanis L.

Abstract Two groups of European catfish, Silurus glanis L., fingerlings were infected with channel catfish virus (CCV) by either intraperitoneal injection with 10⁵ TCID₅₀ of CCV, or bathing in water containing 10⁵ TCID₅₀ of CCV per 1·0 ml. The virus was isolated from spleen, intestine and brain of CCV-injected fish at day 1 and the titres ranged from 10^2·1 to 10^3·3 TCID₅₀/g. However, the tissue distribution of CCV was irregular and no virus was isolated after day 3 post-exposure. In CCV-bathed fish, the virus was isolated only from the liver of one specimen at day 3 post-exposure. No clinical signs of CCV disease developed in any of the fish. Specimens in each regime from all sampling periods showed some minor histopathological changes, but there were no differences between treatments. Lesions included oedema and focal haemorrhage in the liver and the spleen was congested. Electron micrographs of tissue samples showed the presence of a few virus particles around the nuclei of kidney, spleen and intestinal cells, and in or around a myelinated nerve within the optic lobes of infected fish during the first 4 days of infection.     

Occurrence of viral haemorrhagic septicaemia in rainbow trout Salmo gairdneri Richardson reared in sea-water

Abstract. Viral haemorrhagic septicaemia (VHS) is reported for the first time in sea-water cultured rainbow trout. Heavy mortalities with typical signs and lesions A VHS virus (serotype 1) was isolated from the diseased fish. The mortalities were caused only by the VHS virus and 80 days post transfer of trout to sea-water the mortalities reached 85%, of the initial population.
The disease was experimentally transmitted to rainbow trout, both in sea-water 3·10⁴ pfu/ml of virus or by intramuscular injection of various doses of VHS₁ (7·10¹ 7·10⁴ or 7·10⁴ pfu per fish). Death occurred in all infected groups and started earlier in sea-water. Typical signs of VHS were observed in moribund fish. Viral multiplication was demonstrated to have occurred in fish organs.     

The susceptibility of salmonid fish to an atypical strain of Aeromonas salmonicida that infects goldfish, Carassius auratus (L.), in Australia

Abstract. An enzootic, Australian, atypical strain of Aeromonas salmonicida isolated from diseased goldfish, Carassius auratus (L.), was inoculated into Atlantic salmon, Salmo salar L., brown trout, S. trutta L., rainbow trout, S. gairdneri Richardson, and brook trout, Salvelinus fontinalis (Mitchill), fingerlings by intraperitoneal injection (i.p.) and by bath challenge, the latter with and without prior abrasion of skin. The 10-day LD₅₀ (i.p.) was estimated to be 7·4 × 10^-3 colony forming units (cfu) for Atlantic salmon, 3·0 × 10^-2 cfu for brown trout, 3·7 × 10² cfu for brook trout and 6·4 × 10³ cfu for rainbow trout. Brown, rainbow and brook trout succumbed to bath challenges with between 10⁵–10⁶ cfu/ml, developing ulcers of the skin and septicaemia. The organism was trasmitted from inoculated fish to five of 195 within-tank control fish via water and established a carrier state in one of 14 Atlantic salmon. It was concluded that the organism poses a significant threat to the salmonid farming industry and wild salmonid fisheries in Australia.     

Comparative sensitivity of carp, Cyprinus carpio L. and rainbow trout, Salmo gairdneri Richardson, to Renibacterium salmoninarum

Abstract. The pathogenicity of Renibacterium salmoninarum to carp, Cyprinus carpio L., and rainbow trout, Salmo gairdneri Richardson, was investigated. All carp injected with 4·8 × 10⁸ cells/fish, or 4·8 × 10⁷ cells/fish survived for 38 days. R. salmoninarum was isolated from all moribund fish, but not from the kidney of surviving fish, although R. salmoninarum antigen was detected in several of these fish by the dot blot assay. On the other hand, mortality in rainbow trout was 95% in the fish injected with 4·8 × 10⁸ cells/fish, and 15% in those which received 4·8 × 10⁷ cells/fish. R. salmoninarum antigen was detected by the dot blot assay in all surviving rainbow trout. The number of R. salmoninarum cells was immediately decreased by carp or rainbow trout serum, and the serum bactericidal activity of carp was higher than that of rainbow trout. Carp blood leucocytes had higher phagocytic activity than those of rainbow trout.     

Nutrient requirement for zoospore formation in two alariaceous plants Undaria pinnatifida (Harvey) Suringar and Alaria crassifolia Kjellman (Phaeophyceae: Laminariales)

ABSTRACT:   Sporophyll formation in two alariaceous plants, Undaria pinnatifida and Alaria crassifolia , was studied in relation to the nutrient requirements. The sporophylls of U. pinnatifida formed zoosporangia when they had N and P contents greater than 1.4 kgN/m³ and 0.74 kgP/m³. This indicates that these values are the critical nutrient levels for zoospore formation in U. pinnatifida . In the sporophytes of A. crassifolia , many sporophylls with zoosporangia showed nutrient contents higher than 6.25 kgN/m³ and 1.70 kgP/m³. These results suggest that the U. pinnatifida can form zoospores at lower levels of N and P contents than A. crassifolia . Both species often formed zoosporangial sori on the blades in the late period of each reproductive season. The fertile parts of the blade showed N and P contents higher than the critical levels. This phenomenon indicates that the blades have the capability to form zoosporangial sori if there is a sufficient accumulation of nutrients for zoospore formation. The zoospore formation on the blade seems to be accomplished by an overflow of excess nutrients from sporophylls into the blade, or by accumulating sufficient nutrients to form sori even if the sporophylls are not formed.     

Experimental pathogenicity in rainbow trout, Oncorhynchus mykiss (Walbaum), of two distinct morphotypes of long-spined Saprolegnia isolates obtained from wild brown trout, Salmo trutta L., and river water

Juvenile rainbow trout, Oncorhynchus mykiss (Walbaum), were experimentally infected to investigate the pathogenicity of 20 isolates of two morphotypes of long-haired Saprolegnia obtained from wild brown trout, Salmo trutta L., and river water in Spain. The trout were exposed to 2 × 10⁵ and 3 × 10⁵ L^–1 zoospores. Saprolegnia infection could not occur without 'ami-momi' treatment. Pathogenicity varied greatly among isolates as mortality ranged from 0 to 100% of the fish. There was a statistically significant difference ( P  < 0.001) between the mortality caused by morphotype I isolates and that produced by those of morphotype II. The most pathogenic isolates usually belonged to morphotype II, consisting of isolates which had secondary cysts with bundles of hooked hairs which were shorter and less numerous than those of morphotype I; the morphotype I isolates usually had low pathogenicity. Lesions were most frequently found on the fins. Cultures detected the presence of Saprolegnia in internal organs. Histopathology of the intestine suggests that Saprolegnia may reach this and other organs via the blood stream from surface lesions.     

Use of night-lights to attract food organisms into tropical marine fish cages

Abstract. A 91-day cage trial was conducted with juvenile seabass, Lates calcarifer (Bloch), and grouper, Epinephelus tauvina (Forskal), to ascertain the capacity of kerosene pressure lamps and fluorescent electric lamps as night-lights above the cages to attract pelagic food organisms into the cage and consequently sustain the survival and growth of the cultured fish. The experiment was conducted using 1×1×1·5m floating cages with three different net mesh sizes (1, 13, and 19mm) and four fish stocking densities (seabass—10, 20,30 and 40/m³; grouper —10/m³). A positive growth response and survival was observed with seabass and to a lesser extent with grouper with increasing net mesh size and decreasing fish stocking density. At the lowest tested density of 10 fish/m³ seabass survival increased from 5·0 to 95·0% and total cage fish biomass increased from −95·1% to +56·9% with an increase in net mesh size from 1 to 19mm over the 91-day culture trial, respectively. The results are discussed in relation to the current commercial marine finfish cage farming practices employed in Indonesia.     

Enzyme-linked immunosorbent assay (ELISA) for the detection of spring viraemia of carp virus (SVCV) in tissue homogenates of the carp, Cyprinus carpio L.

Abstract. An enzyme-linked immunosorbent assay (ELISA) for the demonstration of the virus of spring viraemia of carp (SVCV) in liver, kidney and spleen homogenates, and in infected cell cultures is described. The sensitivity of the method is 10^2·8–10^3·5 TCID₅₀ 0·lml⁻¹ of the examined fluid. The specificity has been confirmed by the ELISA inhibition test and by results of virological examinations. Contamination with bacteria or fungi of samples taken from dead fish had no effect on the results of ELISA. Specific anti-SVCV sera were used successfully for the production of conjugates for the direct immunoperoxidase and immunofluorescence detection of SVCV in infected cell cultures.     

Autecology of chub, Leudscus cephalus (L.), in a Greek stream, and the use of the pharyngeal bone in fish predator-prey studies

Abstract. The autccology of chub, Leudscus cephalus (L.), in the Renlina Stream, Greece, was carried out in an effort to obtain information on the age, growth, mortality and reproduction of this species. Fork length increment during the first year of life was 8·9cm, after which it decreased. Life span was short (maximum age 8 years) and the instantaneous mortality rates were higher for the older age-groups. Chub reached sexual maturity at the second year of life for both sexes and spawning occurred from the second fortnight of April into May, at a mean water temperature of 18°C. The relationship between fecundity ( F ) and fork length (1) was F = 0·0797 L^3·63 The average density and biomass were 1·06 fish.m⁻² and 11·38g.m⁻² respectively. The relationship between L and the pharyngeal bone size [gape ( G ), tip ( T ) and shank ( S )] in mm and caudal fin length ( C ) (cm) can be described by the equations: L = 0·8605 + 1·550 G , L = -0·0703 + 1·332 S, L = 1·092 + 1·615 T and L = 1·032 + 2·934 C .     

Use of duckweed (Lemna sp.) grown in sugarmill effluent for milkfish, Chanos chanos Forskal, production

Abstract. Oxidation treatment of sugarmill waste using duckweed ( Lemna sp.) as part of the system, to reduce effluent nutrient concentrations and biological oxygen demand (BOD), was evaluated during a 6 month milling season in Negros Oriental, Philippines. Mean ammonia concentration in effluent water was reduced from 0·87 to 0·31 mg/l NH₃-N and orthophosphate from 0·93 to 0·51 mg/l P₂O₅, while mean BOD was reduced from 611 to 143mg/t BOD₅, after treatment. Seasonal mass fish kills in the adjoining bay no longer occurred during the 3 years following introduction of this treatment. The mean duckweed production (dry weight) was 8–8g/m²/day. Duckweed was harvested from a 1·9ha area of the system and transferred to an adjacent milkfish, Chanos chanos Forskal, farm. Its fertilization effect, in terms of lablab production ( lablab is the biological complex of bluegreen algae, diatoms, bacteria and various animals which forms a mat at the bottom or floats in patches), was evaluated in the milkfish ponds. This was compared with ponds fertilized with either inorganic fertilizers or cow manure, in the traditional way. Lablab growth was significantly increased using duckweed, with ash-free dry weight production averaging 32g/m²/day following fertilization with duckweed compared with 4g/m²/day using inorganic fertilizers. Milkfish net production averaged 320 kg/ha/90-day crop in inorganically fertilized ponds. 545kg/ha/90-day crop for cow manure and 820kg/ha/90-day crop in duckweed-fertilized ponds. The system is described and the benefits of this integrated waste treatment-fish production facility are discussed.     

Vaccination of channel catfish, Ictalurus punctatus (Rafinesque), by immersion and oral booster against Edwardsiella ictaluri

Abstract. A commercially prepared vaccine against Edwardsiella ictaluri was used to vaccinate 12-day-old channel catfish fry by immersion, or by immersion plus an oral booster 2 months later. One month after the fish were fed the booster vaccine, they were challenged by waterborne exposure to 2·1 × 10⁶ cells ml⁻¹ of E. ictaluri. Immersion only vaccinated fish suffered 6·7% mortality and immersion plus oral-boosted fish had a 3·3% mortality. Mortality among non-vaccinated controls was 96·7% and was significantly ( P < 0·01) above the vaccinated mortality. The relative per cent survival for the immersion-only fish was 93·1, while it was 96·6 for the immersion plus oral-boosted fish. Agglutinating antibody titres of the vaccinated fish were significantly ( P < 0·05) higher than the control fish. When the ponds were drained 6 months after stocking, 42·7% of non-vaccinated, 56·3% of immersion-only and 70·8% of immersion plus oral-boosted fish were harvested. Survival of immersion plus orally-boosted fish was significantly ( P < 0·05) higher than the controls of immersion-only fish. Duplicate populations of immersion plus oral-booster-vaccinated fish grew 34% and 56% faster, respectively, on an average daily gain than the control fish, while immersion-only fish in one pond grew 20% less per day and fish in the second pond grew 48% faster.     

Differences in performance, flesh composition and water output quality in relation to density and feeding levels in rainbow trout, Oncorhynchus mykiss (Walbaum), farming

Abstract. The aim of this research was lo evaluate the effect of density and feeding level (initially 8 or 16kg/m³ and 1·3% or 2% of present biomass respectively) on productive traits, meat composition and water quality output in farmed rainbow trout, Oncorhynchus mykiss (Walbaum). Individual fish growth shows significant differences in relation to density and feeding level; feed conversion rate is positively Influenced by the lower feeding level and low rearing density. At low density, specific growth rate, protein efficiency ratio and coefficient of variation of individual final weights are more favourable than at high density. Dry matter and lipid composition of meat are positively affected by feeding level; whereas no differences are recorded on the amino acid and fatty acid profiles. Water quality shows differences due to density (NO₃ and NH₄⁺) and feeding level (NO₂, NH₄⁺ and P).     

A study on Flexibacter columnaris infection in loach, Misgurnus anguillicaudatus (Bleeker, Günther)

Abstract. Infectivity of Flexibator columnaris to loach, Misgurnus anguillicaudatus (Blecker, Günther), was studied with variation conditions and the presence of other bacteria in the water medium. Invasion of fish by the poathogen varied with different water conditions. The highest prevalence of infection was observed in an experimentally formulated water containing 0·03% NaCl 0·01% KCl, 0·002% CaCl₂2H₂O and 0·004% MgCI₂ 6H₂O. The percentage of infection in tap water was similar to that of the formulated water. The infeetion rate was low in the individual salt solutions, and no infection occurred in distilled water. Even in the formulated water, the pathogen failed to infect the fish when Citrobacter freundii, Aeromonas hydrophila or Pseudomonas fluorescens were present at densities 100 times that of F. columnaris. However, in such cases, the numbers of F. columnaris in the mucus of the fish body surface did not increase much throughout the experimental period. In contrast, F. columnaris suceessfully invaded the fish in the presence of either Streptococcus sp., Flavobacterium flavescens or Flavobacterium fuscum , even when the numbers of these bacteria were 1000 times that of F. columnaris. In this ease, F. columnaris gradually increased its numbers on the fish body surface.     

Influence of photoperiod, frequency of stripping and presence of females on sperm output in turbot, Scophthalmus maximus (L.)

Abstract. The effect of four environmental conditions was investigated upon sperm output in turbot, Scophthalmus maximus (L.), submitted to three different rhythms of stripping. Males kept under a natural light cycle and under a 6-month contracted light programme released a similar sperm output in terms of total volume of semen produced per fish during the experimental period (4·9 ± 0·9ml), mean sperm concentration (29·4 ± 2·8 × 10⁹ spermatozoa/ml) and total sperm number (163·2 ± 40·5 × 10⁹ spermatozoa). Attempts to stimulate spermiation for a second time just after the end of the natural reproduction period resulted in the release of low sperm output (total volume of semen: 1·6 ± 0·4 ml; mean sperm motility: 2 min 36s ± 0 min 47s; mean sperm concentration: 47·6 ± 10·2 × 10⁹ spermatozoa/ml; total sperm number: 84·5 ± 25·3 × 10⁹ spermatozoa). Stripping frequency had no effect on total volume of semen, mean sperm motility and total sperm number. Monthly collection did not modify sperm samples in relation to stripping rank. However, decreasing volume, motility and sperm concentration were observed when males were stripped fortnightly and weekly. During the natural spawning period, the presence of females in the tank enhanced mean sperm motility (from 3 min 27s + 0 min 52s to 6 min 38s ± 1 min 38s).     

Serratia marcescens: a potential pathogen for fish

Abstract. Characterization of a red pigmented enterobacterium isolated from natural population of white perch, Morone americanus (Gmelin), during the course of a bacteriological survey in the Back River (Baltimore, Maryland, USA) indicated that the bacterium belonged to the species Serratia marcescents. The virulence properties of this isolate (RB 469), studied in comparison with the reference strain of S. marcescens ATCC 1800 and S. plymuthica K1R, revealed that all the strains were highly pathogenic for fish with LD₅₀s raging from 5 × 10³ to 1 × 10⁵. Similarly, te extracellular products (ECP) from the three isolates were lethal for fish (LD₅₀ ranging from 0·22 to 4·8 μg protein g^-1 fish). However, only ECP from strains with strong proteolytic activity (the white perch isolate and S. plymuthica ) were cytotoxic for both in fish and homoiothermic cell cultures and both activities were completely destroyed by heating at 100°C for 10min. In contrast, only the two S. marcescens strains which possessed phospholipase active were pathogenic for mice and produced enterotoxins. None of the Serratia strains displayed dermonecrotic factor in rabbits. All these lindings indicate that a direct relationship between eytotoxicity and virulence cannot be established.