Effects of non-starch polysaccharides enzyme, phytase and citric acid on activities of endogenous digestive enzymes of tilapia (Oreochromis niloticus × Oreochromis aureus)

Non-starch polysaccharides (NSPs) enzyme, phytase and citric acid have been shown to increase digestibility in pigs, poultry and some fish. To examine their roles in digestion in tilapia ( Oreochromis niloticus  ×  Oreochromis aureus ), the present study was designed to evaluate their effects on activities of a number of endogenous digestive enzymes in tilapia. Neither NSPs enzyme (1 g kg⁻¹) nor phytase (1 g kg⁻¹) affected the activities of protease in digestive organs. In contrast, NSPs enzyme increased the activities of amylase in the hepatopancreas and intestine by 11.4% and 49.5%, respectively, while phytase (1 g kg⁻¹) increased the activities of amylase in the hepatopancreas and intestine by 14.0% and 24.1%, respectively. Citric acid (10 g kg⁻¹) increased the activities of protease in stomach by 29.6%, but reduced the activities of protease in the intestine by 35.1%. Citric acid increased the activities of amylase in the hepatopancreas and intestine by 30.7% and 29.4%. Lipase activities were not affected by NSPs enzyme, phytase or citric acid. Above results suggest that NSPs enzyme, phytase and citric acid primarily influence the activities of amylase in digestive tract of tilapia.     

Optimal dietary lipid level of silver barb, Puntius gonionotus fingerlings in relation to growth, nutrient retention and digestibility, muscle nucleic acid content and digestive enzyme activity

Five iso-nitrogenous (300 g kg⁻¹ diet) purified diets with graded level of lipid at 40 (D-1), 60 (D-2), 80 (D-3), 100 (D-4) and 120 (D-5) g kg⁻¹ diet were fed to Puntius gonionotus fingerlings for 90 days to determine their dietary lipid requirement. Two hundred and twenty-five fingerlings (average weight 2.34 ± 0.03 g) were equally distributed in five treatments in triplicate groups with 15 fish per replicate. Fifteen flow-through cement tanks of 100 L capacity with a flow rate of 0.5 L min⁻¹ were used for rearing the fish. Specific growth rate (SGR), feed conversion ratio (FCR), nutrient digestibility, retention, digestive enzyme activity, RNA : DNA ratio and whole-body composition were considered as the response parameters with respect to dietary lipid levels. Maximum SGR and minimum FCR with highest RNA : DNA ratio, whole-body protein content and digestive enzyme activity was found in D-3 group fed with 80 g kg⁻¹ diet lipid. Nutrient digestibility was similar in all the groups irrespective of the dietary lipid level. Maximum protein and energy retention was recorded at 80 g kg⁻¹ dietary lipid fed group. However, from the second-order polynomial regression analysis, the maximum growth of P. gonionotus fingerlings was found at 96.9 g lipid kg⁻¹ diet.     

Carbohydrate level in the diet of silver barb, Puntius gonionotus (Bleeker) fingerlings: effect on growth, nutrient utilization and whole body composition

Five iso-nitrogenous (300 g crude protein kg⁻¹ diet) semi-purified diets with graded levels of carbohydrate at 220 (D-1), 260 (D-2), 300 (D-3), 340 (D-4) and 380 (D-5) g kg⁻¹ diet were fed ad libitum to Puntius gonionotus fingerlings (average weight 0.59±0.01 g) in triplicate groups (20 fish replicate⁻¹) for a period of 90 days to determine the effect of the dietary carbohydrate level on the growth, nutrient utilization, digestibility, gut enzyme activity and whole-body composition of fish. Fifteen flow-through cement tanks of 100 L capacity with a flow rate of 0.5 L min⁻¹ were used for rearing the fish. The maximum weight gain, specific growth rate, protein efficiency ratio, RNA:DNA ratio, whole-body protein content, protease activity, protein and energy digestibility and minimum feed conversion ratio (FCR) were found in the D-2 group fed with 260 g carbohydrate kg⁻¹ diet. The highest protein and energy retention was also recorded in the same group. However, from the second-order polynomial regression analysis, the maximum growth and nutrient utilization of P. gonionotus fingerlings was 291.3–298.3 g carbohydrate kg⁻¹ diet at a dietary protein level of 300 g kg⁻¹ with a protein/energy (P/E) ratio of 20.58 −20.75 g protein MJ⁻¹.     

Purification and characterization of the amylase from a small abalone <Emphasis Type="Italic">Haliotis sieboldii</Emphasis>

ABSTRACT:   Amylase, with MW of 59 kDa, was purified from small abalone Haliotis sieboldii by ammonium sulfate fractionation, CM Sepharose Fast Flow and Sephacryl S-100 HR chromatographies. The optimal pH and temperature of purified amylase were 6.0 and 37°C, respectively. The purified enzyme was stable at pH 6.0–8.0 and low temperatures. It was activated by Ba²⁺, Mg²⁺, Ca²⁺, Co²⁺, Ni²⁺, Mn²⁺, K⁺, Ag⁺, Na⁺ and Li⁺, but completely or partially inhibited by Al³⁺, Cu²⁺, Cd²⁺, Hg²⁺ and Zn²⁺. EDTA could completely inhibit, while iodoacetamide, N-ethylmaleimide and urea partially inhibit the purified amylase. According to the digestion mode of various polysaccharides, the purified enzyme was considered to be an α-amylase.     

A dietary energy level of 14.6 MJ kg−1 and protein-to-energy ratio of 20.2 g MJ−1 results in best growth performance and nutrient accretion in silver barb Puntius gonionotus fingerlings

Five iso-nitrogenous (300 g protein kg⁻¹ diet) and iso-lipidic (80 g kg⁻¹ diet) semi-purified experimental diets with variable energy levels of 10.5 (D-1), 12.5 (D-2), 14.6 (D-3), 16.7 (D-4) and 18.8 (D-5) MJ kg⁻¹ diets were fed to Puntius gonionotus fingerlings (average weight 1.79 ± 0.02 g) in triplicate groups (15 healthy fishes per replicate) for a period of 90 days to assess the optimum dietary energy level and protein-to-energy ratio (P/E). Fifteen flow-through cement tanks of 100 L capacity with a flow rate of 0.5 L min⁻¹ were used for rearing the fish. Maximum specific growth rate, protein efficiency ratio, protein productive value, RNA : DNA ratio, whole body protein content, digestive enzyme activity and minimum feed conversion ratio was found in fish-fed diet D-3 with 14.6 MJ kg⁻¹ energy level. There were no improvements in all these parameters with the further rise in dietary energy level. Hence, it may be concluded that the optimum dietary gross energy level for maximum growth and nutrient utilization of silver barb is 14.6 MJ kg⁻¹ diet with a resultant P/E ratio of 20.2 g protein MJ⁻¹ diet, when the dietary protein and lipid are maintained at optimum requirement levels of 300 and 80 g kg⁻¹ diet, respectively, for this species.     

Evaluation of carbohydrate rich diets through common carp culture in manured tanks

Four diets (T₀–T₃) were formulated reducing the fishmeal (Indian) component by 100 g kg^–1 from 300 to 0 g kg^–1 and including proportionately increasing quantities of maize. Diets were fed for 120 days at 50 g kg^–1 body weight to triplicate groups of common carp (av. wt. 2.11–2.18 g) stocked at 1 m^–2 in mud bottomed cement tanks (18 m²), fertilized with poultry manure. Fish growth, SGR and FCR in the different treatments were statistically not significantly different ( P  > 0.05). PER was lowest for the 300 g fishmeal kg^–1 diet treatment (diet T₀), increasing with decrease in dietary fishmeal content (diets T₁–T₃). Fish survival ranged from 96.29 to 100%. Diets influenced carcass composition and digestive enzyme activity. A significant increase in lipid deposition was recorded with increasing dietary carbohydrate content. Amylase, protease and lipase activities were higher in fish fed with diets T₂ and T₃. The protein sparing effect of dietary carbohydrate and the economic implication of eliminating fishmeal from the diet are discussed.     

The synthetic antioxidant, ethoxyquin, adversely affects immunity in tilapia (Oreochromis niloticus)

Ethoxyquin (EQ) has been used as an antioxidant in livestock, aquaculture and pet foods. Animal food safety law has established the upper limit of EQ in animal feed at 150 mg kg⁻¹. However, the risk of EQ at the approved level for aquaculture feed (150 mg kg⁻¹) to fish health is unknown. Here, we examine the effect of EQ on the immunity of tilapia ( Oreochromis niloticus ). EQ concentration in the blood reached 0.16 mg L⁻¹ in fish fed EQ at the approved level. This level of EQ inhibited phagocytic activity of leucocytes in vitro and antibacterial activity of whole blood in vivo . Furthermore, pyknosis in the liver was observed throughout the duration of feeding. However, after 30 days of experimental challenge with feed containing 150 mg kg⁻¹ of EQ, no significant difference was observed in mortality. Although EQ at the approved level in feed causes immunosuppression in fish, the severity of immunosuppression does not lead to a lowering of disease resistance for short feeding periods.     

Identification and partial characterization of selected proteolytic enzymes in the digestive system of giant freshwater Prawn Macrobrachium rosenbergii (De Man) Postlarvae

Biochemical assays and substrate SDS-PAGE were conducted to partially characterize and identify various types of proteases present in the digestive tract of PL₁₅ giant freshwater prawn ( Macrobrachium rosenbergii ). Casein hydrolytic assay of the enzyme extracts showed major proteolytic activities at pH 3.0, 6.0 and 9.0, while assay of preincubated enzyme extracts with phenylmethylsulphonyl fluoride (PMSF), a serine protease inhibitor produced a 33.17% reduction in alkaline protease activity. When specific inhibitors tosyl-lysine chloromethyl ketone and tosyl-phenylalanine chloromethyl ketone were used, they resulted in a reduction in activity of proteases in the enzyme extracts by 82.41% and 55.03%, respectively, confirming the presence of trypsin and chymotrypsin, while ethylenediamine tetraacetic acid produced protease activity reduction in 33.92% showing the presence of metalloproteases in the digestive tract of the prawn. Further characterization of the alkaline proteases using SDS-PAGE technique, after incubating the extract in the presence or absence of specific inhibitors, produced six bands corresponding to molecular masses of between 13.48 and 136.1 kDa; two trypsin bands of 13.48 and 36.4 kDa, three chymotrypsin bands in the range of 23.0–73.4 kDa and one for metalloprotease of 136.1 kDa, all of which were identified from a zymogram. This study suggests that protein digestion in M. rosenbergii is initiated by an acid protease followed by a combination of action of alkaline proteases: trypsin, chymotrypsin and metalloproteases.     

Distribution and biomass of an underfished vendace, Coregonus albula, population in a mesotrophic German reservoir

Abstract  The distribution and overall biomass of an underfished vendace, Coregonus albula L., population in the mesotrophic Henne Reservoir (Germany) was studied using hydroacoustics and gill nets. Additionally, midwater trawling was carried out. Overall fish biomass, based on five hydroacoustic surveys (June to September 2002), ranged from 188 kg ha⁻¹ in early August to 302 kg ha⁻¹ in September 2002. The overall mean fish biomass was 256 kg ha⁻¹ (±48 kg ha⁻¹ SD). Biomass of fish smaller than 25 cm total length (mostly vendace) varied from 56 kg ha⁻¹ in August to 99 kg ha⁻¹ in September, with an overall mean fish biomass of 74 kg ha⁻¹ (±17 kg ha⁻¹ SD). The echograms showed temporal variation in fish distribution in Henne Reservoir. In June, fish were fairly evenly distributed over the whole reservoir but in September a dense aggregation of fish (mostly vendace) was found in the deeper water layers near the dam. The distribution of vendace stock, its impact on water quality and fisheries management, biomanipulation and effort for mass removal are discussed.     

Continuous exposure to infectious pancreatic necrosis virus during early life stages of rainbow trout, Oncorhynchus mykiss (Walbaum)

Rainbow trout, Oncorhynchus mykiss (Walbaum), were exposed continuously to infectious pancreatic necrosis virus (IPNV) at 0, 10¹, 10³ or 10⁵ plaque forming units (pfu) L⁻¹ of water to estimate the effects of chronic IPNV exposure on early life stages. Fish density averaged 35 fish L⁻¹ (low density) or 140 fish L⁻¹ (high density), and the tank flow rate was 250 mL⁻¹ min. Virus exposure began at 6 days before hatch and continued until fish were 44 days old. Cumulative per cent mortality, analysis of survival and hazard functions, and discrete-time event analysis were used to explore the patterns of survival and mortality. In eggs and fish exposed to IPNV, mortality significantly greater than in the 0 pfu L⁻¹ exposure did not occur until IPNV concentration was 10⁵ pfu L⁻¹ at low fish density and 10³ pfu IPNV L⁻¹ at high fish density. These results suggest that in the natural aquatic environment, where rainbow trout densities are likely to be considerably lower than in this study, mortality resulting from infection with IPNV will very likely not occur when ambient concentrations of virus are ≤10³ pfu IPNV L⁻¹. In aquaculture rearing units, trout density is likely to be as high or higher than the densities used in this study. Therefore, continuous inputs of virus at concentrations greater than 10¹ pfu L⁻¹ may result in IPN epidemics in aquaculture facilities.     

An experimental investigation on aspects of infectious salmon anaemia virus (ISAV) infection dynamics in seawater Atlantic salmon, Salmo salar L.

This study investigated infection dynamics of infectious salmon anaemia virus (ISAV) by conducting two experiments to examine minimum infective dose and viral shedding of ISAV. In terms of minimum infective dose, the high variability between replicate tanks and the relatively slow spread of infection through the population at 1 × 10¹ TCID₅₀ mL⁻¹ indicated this dose is approaching the minimum infective dose for ISAV in seawater salmon populations. A novel qPCR assay incorporating an influenza virus control standard with each seawater sample was developed that enabled the quantity of ISAV shed from infected populations to be estimated in values equivalent to viral titres. Viral shedding was first detected at 7 days post-challenge (5.8 × 10⁻² TCID₅₀ mL⁻¹ kg⁻¹) and rose to levels above the minimum infective dose (4.2 × 10¹ TCID₅₀ mL⁻¹ kg⁻¹) on day 11 post-challenge, 2 days before mortalities in ISAV inoculated fish started. These results clearly demonstrate that a large viral shedding event occurs before death. Viral titres peaked at 7.0 × 10¹ TCID₅₀ mL⁻¹ kg⁻¹ 15 days post-infection. These data provide important information relevant to the management of ISA.     

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     

Evaluation of lipid spray beads for the delivery of water-soluble materials to a marine suspension-feeder, the Manila clam Tapes philippinarum (Deshayes 1853)

We describe the development and evaluation of a new microparticle for delivering low-molecular weight, water-soluble materials to suspension feeders. Spray beads successfully incorporated materials dissolved in an aqueous phase or as dry particulate, within a triacylglyceride bead composed of tripalmitin, 600 mg g⁻¹ tripalmitin/400 mg g⁻¹ triolein, or 600 mg g⁻¹ tripalmitin/400 mg g⁻¹ fish oil.
Riboflavin was successfully incorporated (up to 44 mg g⁻¹ lipid) and retained (up to 98% over 24 h in seawater) as dry particles in all three mixtures of lipid. Aqueous oxytetracycline hydrochloride or polymeric dye were incorporated (45.6 mg g⁻¹ lipid and 18.1 mg g⁻¹ lipid, respectively) and retained best (99% and 94%, respectively) in spray beads composed of tripalmitin. The addition of triolein or fish oil to the lipid bead reduced incorporation and retention efficiencies for aqueous core materials by up to 75%.
Manila clam seed readily ingested and digested lipid microparticles, spray beads and lipid-walled microcapsules. Microparticles composed of tripalmitin were excreted with their payloads intact. Intact microparticles composed of 600 mg g⁻¹ tripalmitin/400 mg g⁻¹ fish oil were largely absent in faecal strands suggesting successful release and delivery of microparticle contents to clams.
Spray beads composed of tripalmitin softened with 400 mg g⁻¹ fish oil represent an effective microparticle type for delivering low-molecular weight, water-soluble materials to aquatic suspension feeders.     

Effects of dietary protein to metabolizable energy ratios and total protein concentrations on the performance of yellow perch Perca flavescens

Juvenile yellow perch Perca flavescens were fed semipurified diets with varying protein to metabolizable energy ratios (PME, g protein MJ⁻¹ metabolizable energy) and nutrient densities in three experiments to determine recommended dietary protein and energy concentrations. Experiment 1 fish (18.6 g) were fed diets containing 450 g crude protein kg⁻¹ dry diet and 14.5–18.8 MJ ME kg⁻¹ dry diet for 10 weeks. No differences were found in the growth of experiment 1 fish fed the different diets. Experiment 2 fish (21.9 g) were fed diets containing 15.7 MJ ME kg⁻¹ dry diet and 210–420 g crude protein kg⁻¹ dry diet for 8 weeks. Fish fed the diet containing 340 g kg⁻¹ protein (diet PME = 22) exhibited the greatest weight gain. Experiment 3 fish (27.1 g) were fed diets with a PME of 22 and varying nutrient density (yielding 205–380 g crude protein kg⁻¹ dry diet) for 8 weeks. No differences were found in the growth of experiment 3 fish. Yellow perch fed the semipurified diets exhibited increased liver fat content, liver size and degree of liver discoloration compared with fish fed a commercial fish meal-based diet. Liver changes may have resulted from high dietary carbohydrate levels. We conclude that a protein level of 210–270 g kg⁻¹ dry diet is suitable for juvenile yellow perch provided that the dietary amino acid profile and carbohydrate content are appropriate for yellow perch.     

Application of the rpoS gene for the detection of Vibrio anguillarum in flounder and prawn by polymerase chain reaction

Vibrio anguillarum , an opportunistic fish pathogen, is the main species responsible for vibriosis, a disease that affects feral and farmed fish and shellfish, and causes considerable economic losses in marine aquaculture. In this study, we used polymerase chain reaction (PCR) to detect V. anguillarum . PCR specificity was evaluated by amplifying the rpoS gene, a general stress regulator, in six strains of V. anguillarum and 36 other bacterial species. PCR amplified a species-specific fragment (689 bp) from V. anguillarum . Furthermore, the PCR assay was sensitive enough to detect rpoS expression from 3 pg of genomic DNA , or from six colony-forming units (CFU) mL⁻¹ of cultured V. anguillarum . However, the assay was less sensitive when genomic DNA from the infected flounder and prawn was used (limit of detection, 50 ng and 10 ng g⁻¹ tissue, respectively). These data demonstrate that PCR amplification of the rpoS gene is a sensitive and species-specific method to detect V. anguillarum in practical situations.     

Effects of dietary phosphorus and lipid levels on utilization and excretion of phosphorus and nitrogen by rainbow trout (Oncorhynchus mykiss). 2. Production-scale study

In a 8-week production-scale experiment at a commercial trout farm, the effects of dietary lipid level and phosphorus level on phosphorus (P) and nitrogen (N) utilization of rainbow trout (initial mean weight 99 g) were assessed. A low-phosphorus, high-lipid experimental diet (457 g protein, 315 g lipid, 9.1 g P  kg^–1 dry diet) was compared with a commonly used commercial diet (484 g protein, 173 g lipid, 13.6 g P  kg^–1 dry diet). P and N budgets were constructed using data from the production-scale experiment and digestibility data for the two diets. In addition, orthophosphate and ammonia-N waste were measured in effluent over one 24-h period. Relative to the commercial diet, the experimental diet resulted in significantly increased feed efficiency ratio, N retention and P retention, and substantially reduced dissolved, solid and total P waste (g kg^–1 dry feed). Although N retention resulting from the experimental diet was higher, this was attributable to higher N (protein) digestibility of the experimental diet. Solid N waste (g kg^–1 dry feed) resulting from the experimental diet was substantially lower, but dissolved N waste (g kg^–1 dry feed) was not significantly different relative to the commercial diet. Mean effluent orthophosphate production (mg day^–1 kg^–1 fish) of fish fed the experimental diet was substantially lower than that of fish fed the commercial diet ( P  < 0.05), but effluent ammonia-N production (mg day^–1 kg^–1 fish) was not significantly affected by dietary treatment.     

Effect of dietary lipid level on muscle composition in Atlantic salmon Salmo salar

This study was conducted to determine the effects of feeding increasing lipid concentrations (310, 380 and 470 g kg^–1 lipid on dry weight) in diets based mainly on herring byproducts to Atlantic salmon Salmo salar . The diets were isonitrogenous, varying in dietary lipid content at the expense dietary starch. Average fish weight increased from 1.2 kg in April to 2.2–2.7 kg at the end of the feeding trial in September. Significantly greater growth was found in fish fed either the 380 g kg⁻¹ or the 470 g kg⁻¹ lipid diets compared with the 310 g kg⁻¹ lipid diet. Muscle lipid content increased in all dietary groups on a wet weight basis from 7.7 ± 1.4% to 12 ± 3% in salmon fed the 310 g kg⁻¹ lipid diet, and to 16 ± 2% in salmon fed the 380 g kg⁻¹ and 470 g kg⁻¹ lipid diets. In fish of similar weight there was a positive correlation between dietary lipid and muscle lipid concentrations. Low concentrations of muscle glycogen were detected in fish fed each of the diets, while muscle vitamin E concentrations slowly decreased as muscle lipid increased. Muscle fatty acid composition reflected dietary fatty acid profiles, containing similar percentages of total saturated, monoenic and n-3 fatty acids (20:5n-3 and 22:6n-3) in fish from all dietary treatment groups. However, a higher ratio of n-3/n-6 was found in muscle from fish fed the 470 g kg⁻¹ lipid diet compared with the other two groups. Blood chemistry values varied somewhat, but all values were within normal ranges for Atlantic salmon of these sizes.     

Effects of digestible protein levels in isonitrogenous diets on growth performance and tissue composition of juvenile Atractoscion nobilis

A study was undertaken to estimate the effects of isonitrogenous diets (ca. 604 g kg⁻¹ crude protein) containing formaldehyde-treated (FT) fish meal and graded levels of digestible protein (DP) (541, 491, 372, 347 and 247 g kg⁻¹) on growth performance and tissue composition of juveniles white seabass. Five diets were formulated to contain increasing levels of FT fish meal (from 0 to 384 g kg⁻¹) and decreasing levels of non-treated fish meal (from 480 to 96 g kg⁻¹). Each dietary treatment was fed in triplicate to apparent satiation to groups of 25 fish for 50 days. Significantly higher growth performance and feed conversion ratio were obtained in fish-fed diets containing 491 or 541 g kg⁻¹ DP, compared with all other treatments. Apparent digestibility coefficient of protein in the diets was not significantly affected by the inclusion of treated fish meal in the diets. Estimation of protein requirements using a broken-line regression analysis indicated that maximum weight gain would be obtained with a diet containing 503 ± 23 g kg⁻¹ DP. The results from this study suggest that a single-diet formulation using protein treated with formaldehyde as filler might be useful to estimate the requirement of DP for fish.     

Acute LD50 and kidney histopathology following injection of erythromycin (Erythro-200) and its carrier in spring chinook salmon, Oncorhynchus tshawytscha (Walbaum)

We evaluated the toxicity of commercial injectable erythromycin (Erythro-200) and the injectable drug carrier alone (PEG-400, ethyl alcohol and ethyl acetate) in sub-adult spring chinook salmon, Oncorhynchus tshawytscha (Walbaum). Both compounds were toxic to salmon when administered in high dosages. The 50% lethal dose (LD₅₀) of the erythromycin within its carrier (Erythro-200) at 12 °C was estimated at 429 mg erythromycin kg^–1 or 2.145 mL kg^–1 when measured by volume. The LD₅₀ of the drug carrier alone was 2.95 mL kg^–1 by volume or equivalent to 3.21 g kg^–1 of the carrier. We evaluated and scored the histopathology in kidney sections removed from fish 96 h after injection with Erythro-200, the drug carrier alone, or sterile phosphate buffered saline (PBS) that served as a reference control. Kidneys from fish injected with saline had lowest average pathology scores; those injected with the carrier had moderate scores; and the pathology score was highest in tissues from erythromycin-injected fish. In tests at 12 °C, vacuoles were more prominent in proximal and distal tubules of fish injected with higher dosages of 403 and 545 mg erythromycin kg^–1 than in tissues from fish injected with 299 mg kg^–1.     

Herpesviral haematopoietic necrosis virus (CyHV-2) infection: case studies from commercial goldfish farms

Herpesviral haematopoietic necrosis is a disease of goldfish, Carassius auratus , caused by Cyprinid herpesvirus-2 (CyHV-2) infection. Quantitative PCR was carried out on tissue homogenates from healthy goldfish fingerlings, broodfish, eggs and fry directly sampled from commercial farms, from moribund fish submitted to our laboratory for disease diagnosis, and on naturally-infected CyHV-2 carriers subjected to experimental stress treatments. Healthy fish from 14 of 18 farms were positive with copy numbers ranging from tens to 10⁷ copies μg⁻¹ DNA extracted from infected fish. Of 118 pools of broodfish tested, 42 were positive. The CyHV-2 was detected in one lot of fry produced from disinfected eggs. Testing of moribund goldfish, in which we could not detect any other pathogens, produced 12 of 30 cases with 10⁶–10⁸ copies of CyHV-2 μg⁻¹ DNA extracted. Subjecting healthy CyHV-2 carriers to cold shock (22–10 °C) but not heat, ammonia or high pH, increased viral copy numbers from mean copy number (±SE) of 7.3 ± 11 to 394 ± 55 μg⁻¹ DNA extracted after 24 h. CyHV-2 is widespread on commercial goldfish farms and outbreaks apparently occur when healthy carriers are subjected to a sharp temperature drop followed by holding at the permissive temperature for the disease.