Olfactory nerve response of masu salmon (Oncorhynchus masou Brevoort) and rainbow trout (O. mykiss Walbaum) to clove oil and MS-222

Two anaesthetics, clove oil and methane sulphonate (MS-222), were examined for their effects on the olfactory nerve response of masu salmon ( Oncorhynchus masou Brevoort) and rainbow trout ( O. mykiss Walbaum). Exposing both species to clove oil for 3 min at concentrations of 50 and 100 mg L⁻¹, or for 10 min at 50 mg L⁻¹, did not significantly reduce their olfactory response. Directly applying clove oil anaesthesia to the olfactory epithelium significantly reduced olfactory response though after 20 min, olfactory response recovered to 70% and 52% of pre-treatment levels in masu salmon and rainbow trout respectively. Compared with the post-anaesthetic recovery of responses after clove oil (50 mg L⁻¹), buffered MS-222 (100 mg L⁻¹) with NaHCO₃ (100 mg L⁻¹), and unbuffered MS-222 (100 mg L⁻¹) treatment for 3 min, the response after MS-222 treatment declined gradually and significantly, but not after clove oil and MS-222+NaHCO₃ treatments. Clove oil appears to be an effective and relatively safe anaesthetic for salmonids with little long-term impact on their olfactory response, which plays a crucial role in their life history.     

Anaesthetic efficacy and physiological responses to clove oil-anaesthetized kelp grouper Epinephelus bruneus

The efficacy of clove oil as an anaesthetic and at producing a physiological response (plasma cortisol and glucose) was evaluated in the kelp grouper, Epinephelus bruneus . To acquire complete anaesthesia in less than 3 min and recovery in <10 min, three doses of clove oil were tested at 18, 22 and 26 °C. Although higher anaesthetic doses resulted in shorter induction times and longer recovery times, and a lower temperature resulted in longer anaesthesia induction and slower recovery, we found the optimal dose and administering temperature of clove oil to be 250–300 mg L⁻¹ at water temperature of 18 °C, 150–200 mg L⁻¹ at water temperature of 22 °C and 50–100 mg L⁻¹ at water temperature of 26 °C respectively. Following the administration of 150 mg L⁻¹ of clove oil at 22 °C, the plasma cortisol level was highest (4.24 ± 1.571 μg dL⁻¹) after 12 h and the plasma glucose was highest (92.7 ± 9.61 mg dL⁻¹) after 2 h. These results should be useful to the aquaculture industry, where anaesthesia is necessary for a range of activities.     

Comparison of the effects of four anaesthetics on biochemical blood profiles of perch

This study investigated the feasibility of using clove oil, 2-phenoxyethanol, or Propiscin as an alternative to tricaine methane sulphonate (MS 222) as a fish anaesthetic, particularly in regard to reducing fish stress. The biochemical blood profiles of perch Perca fluviatilis L. anaesthetized with either MS 222 (100 mg L⁻¹), clove oil (33 mg L⁻¹), 2-phenoxyethanol (0.40 mL L⁻¹) or Propiscin (1.0 mL L⁻¹), and a non-anaesthetized control group were compared. Biochemical profiles were determined from blood samples collected before treatment in controls. For each anaesthetic tested, fish were divided into two groups, one sampled immediately after 10-min anaesthesia and a second, sampled 24 h after 10-min anaesthesia. The values determined in the present study suggested that internal organs and tissues of perch were slightly altered by MS 222, clove oil and 2-phenoxyethanol anaesthesia, but not by Propiscin anaesthesia.     

Effects of a peroxide-based commercial product on bacterial load of larval rearing water and on larval survival of two species of Sparidae under intensive culture: preliminary study

Larvae of two Mediterranean Sparidae species, Sparus aurata and Dentex dentex , were used to test the efficacy of a peroxide-based product (Ox-Aquaculture^©) on the reduction in bacterial load in larval rearing water and its effects on larval survival. Eleven-day-old S. aurata larvae and 15-day-old D. dentex larvae were exposed to different concentrations of Ox-Aquaculture^© (50, 100 and 200 mg L⁻¹, and 20 and 50 mg L⁻¹ respectively) for 1 h. Results indicated that 50 and 20 mg L⁻¹ were the most effective concentrations for the reduction in bacterial load (at least one order of magnitude) after 1 h treatment, without affecting larval survival and/or vitality in 11 dph S. aurata and 15 dph D. dentex larvae respectively. Ox-Aquaculture^© concentrations of 200 and 50 mg L⁻¹ during 1 h affected negatively final survival rate of the larvae of S. aurata and D. dentex respectively.     

Effects of fish size and water temperature on the acute toxicity of boron to Japanese flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis> and red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis>

ABSTRACT:     The acute toxicities of boron were examined for Japanese flounder Paralichthys olivaceus and red sea bream Pagrus major in terms of fish size and water temperature. Japanese flounder of 0.1–70.0 g and red sea bream of 0.6–20.3 g were exposed to different concentrations of boron for 96 h at 20°C under semi-static conditions. In both fish species, the median lethal concentration (LC₅₀) for 96 h of boron increased linearly with increasing fish size, ranging from 108 to 252 mg B/L for the flounder, and from 97 to 172 mg B/L for the sea bream. The effect of water temperature on acute toxicity was examined for Japanese flounder of 0.6 and 1.5 g at 10, 15, 20 and 25°C, and for red sea bream of 0.6 and 2.4 g at 12, 15, 20 and 25°C. The toxicity of boron for the flounder increased linearly with increasing water temperature. The 96 h LC₅₀ values ranged from 299 to 108 mg B/L for the 0.6 g flounder and from 350 to 113 mg B/L for the 1.5 g flounder. A similar trend was shown for the 2.4 g red sea bream; however, the relationship for the 0.6 g red sea bream was not significant.     

Effect of combined photoperiod, water calcium concentration and pH on survival, growth, and moulting of juvenile crayfish (Procambarus clarkii ) cultured under laboratory conditions

The red swamp crayfish, Procambarus clarkii (Girard), is one of the most commonly farmed freshwater species in inland China due to its high market value and consumer demand. The aim of this study was to determine the optimum combinations of photoperiod, water calcium concentration and pH for juvenile survival, growth and moulting. In our orthogonal experiment, the three environmental factors were varied at three levels (photoperiod: 16L:8D, 12L:12D and 8L:16D; calcium concentration: 45.5, 65.5 and 85.5 mg L⁻¹; and pH: 6.8, 7.8 and 8.8). Range analysis showed that the maximum survival of juvenile crayfish occurred at photoperiods of 16L:8D or 8L:16D, water calcium concentration of 45.5 mg L⁻¹ and pH of 7.8; maximum weight gain at photoperiod 16L:8D, water calcium concentration 65.5 mg L⁻¹ and pH 7.8; maximum length increase at photoperiod 16L:8D, water calcium concentration 65.5 mg L⁻¹ and pH 7.8; and the highest moult frequency at photoperiod 12L:12D, water calcium concentration 65.5 mg L⁻¹ and pH 7.8. Analysis of variance indicated that photoperiod, water calcium concentration and pH significantly influenced only the weight gain of juvenile crayfish ( P <0.05). Taking growth into consideration, we suggest that a photoperiod of 16L:8D, calcium concentration of 65.5 mg L⁻¹ and pH 7.8 might be optimal conditions for rearing juvenile P. clarkii .     

Potential benefit of clove oil sedation on animal welfare during salmon smolt, Salmo salar L. transport and transfer to sea

The purpose of this study was to determine the effect of clove oil (4.0 mg L⁻¹) sedation, compared with non-sedation, on the primary (plasma cortisol), secondary (osmoregulation) and tertiary (mortality) stress responses in Atlantic salmon smolts during transport and transfer to sea. Clove oil sedation during on- and off-loading sufficiently reduced the primary stress response to lower mortality (2.1%) during transfer to sea compared with unsedated fish, which experienced a mortality rate above 12.2%. The unsedated fish experienced an acute mortality that stabilized only 6 days after the transport. None of the secondary stress responses measured in this experiment could contribute towards explaining this phenomenon, with the possible exception of plasma magnesium (Mg²⁺). Plasma Mg²⁺ differed between the groups; while plasma Mg²⁺ in the clove oil sedated group returned to pre-stress levels 72 h after transport, the unsedated group showed no such recovery even 1 week after transport, which may indicate a disturbance in the hydromineral balance, and provides a plausible explanation for the delayed mortality in this group. Eugenol-based anaesthetics appear to be promising as a stress-reducing sedative for Atlantic salmon smolts, and, if used properly, this chemical could improve animal welfare and survivability during and after common aquaculture-related incidents.     

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 varying salinities on the toxicity of potassium permanganate to larval and juvenile striped bass, Morone saxatilis (Walbaum)

Abstract. Eight-day-old striped bass, Morone saxatilus (Walbaum) larvae, and one-month-old juveniles were held in varying salinities and exposed to a range of concentrations of potassium permanganate. Lethal concentrations of potassium permanganate for 50% of the larval fish exposed for96h (96h LC₅₀) were 1middot;02, 2 middot;11, 1 middot;41 and 1 middot;73mg/1 for salinities of 0, 3, 6and 9 g/l, respectively; The 96h LC₅₀S for juveniles were 0 middot;96, 3 middot;26, 1 middot;63 and 1 middot;48 mg/l for salinities of 0, 5, 10 and 15g/l, respectively. There were significant differences in 96h LC₅₀ values at different salinities for both life stages. The 96 h LC₅₀ values derived for both life stages revealed a decrease in toxicity as salinity increased from 0 to 3–5g/l. Results indicated potassium permanganate to be toxic in fresh water to striped bass larvae and fingerlings at recommended treatment levels (3 middot;0mg/l).     

Comparative growth performances of diploid and triploid Chinese shrimp Fenneropenaeus chinensis (Osbeck, 1765) under different salinities

This study aimed at evaluating the ploidy effects on growth performances of Chinese shrimp ( Fenneropenaeus chinensis Osbeck, 1765) reared in different salinities under laboratory conditions. In the acute salinity experiment, there was no difference ( P >0.05) in tolerance observed in triploid and diploid shrimp due to abrupt salinity changes. The lethal salinity for 50% of the individuals in 96 h at 23–25 °C was about 2 g L⁻¹ in both triploids and diploids. While for the chronic salinity experiment, statistical analyses confirmed that the differences in growth performances including the specific growth rate (SGR), the feeding rate (FR), feed conversion efficiency (FCE) and intermoult period (IP) between triploid and diploid were related to salinity. Diploid shrimp reared in 20 g L⁻¹ exhibited highest SGR ( P <0.05), while triploids performed well in 20 and 30 g L⁻¹ salinities ( P <0.05). Based on the survival and growth data, the optimal salinity for the culture of diploid F. chinensis should be 20 g L⁻¹ and for triploids it should be between 20 and 30 g L⁻¹.     

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.     

Further development of bithionol therapy as a treatment for amoebic gill disease in Atlantic salmon, Salmo salar L.

This study examined the efficacy of bithionol as a prophylactic or therapeutic oral treatment for Atlantic salmon (AS), Salmo salar , affected by amoebic gill disease (AGD). Furthermore, it explored the interaction of bithionol oral therapy with the current standard treatment (a freshwater bath for at least 3 h). The efficacy of three medicated feeds was determined in the trial by feeding AGD-affected AS at 1% body weight (BW) day⁻¹ either oil coated commercial feed (control) or prophylactic and therapeutic bithionol at 25 mg kg⁻¹ feed. Feeding commenced 2 weeks prior to exposure to Neoparamoeba spp. at 300 cells L⁻¹ and continued for 49 days post-exposure (PE). Bithionol when fed as a 2-week prophylactic or therapeutic treatment at 25 mg kg⁻¹ feed delayed the onset of AGD pathology and reduced the percentage of gill filaments with lesions. Administration of a 3-h freshwater bath at 28 days PE significantly reduced amoeba numbers to a similar level across all treatments; in contrast, gross gill score and percent lesioned filaments were reduced to different extents, the control having a significantly higher score than both bithionol treatments. Following the freshwater bath, clinical signs of AGD increased at a similar level across all treatments, albeit controls were significantly higher than the bithionol treatments immediately following freshwater treatment. This study demonstrated that bithionol at 25 mg kg⁻¹ feed, when fed as a 2-week prophylactic or a therapeutic treatment, delayed and reduced the intensity of AGD pathology and warrants further investigation as a treatment for AGD-affected AS.     

Evaluation of potassium permanganate against an experimental subacute infection of Flavobacterium columnare in channel catfish, Ictalurus punctatus (Rafinesque)

An experiment was performed to evaluate the efficacy of potassium permanganate (KMnO₄) as a prophylactic and therapeutic treatment of an experimental subacute infection of Flavobacterium columnare in channel catfish, Ictalurus punctatus . Fish were cutaneously abraded and divided into five treatment groups: (i) challenged by waterborne exposure to F. columnare and not treated with KMnO₄ (positive control), (ii) challenged and simultaneously treated with KMnO_4, (iii) challenged and treated with KMnO₄ at 1, 6 and 9 days post-challenge, (iv) not challenged and treated with KMnO₄ at 1, 6 and 9 days post-challenge (first negative control) and (v) not challenged and not treated (second negative control). The dosing of KMnO₄ was 2.0 mg L⁻¹ above the potassium permanganate demand for 2 h duration. The survival of the group challenged and simultaneously treated with KMnO₄ (99%) was significantly higher than the positive control (78%) and was not significantly different from the negative control groups. The challenged fish treated with KMnO₄ post-challenge had 7% higher survival than the positive control (85% compared with 78%), but that difference was not statistically significant. The results demonstrate that KMnO₄ has a clear prophylactic value but probably a marginal therapeutic value once the infection has established.     

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.     

Effects of oestradiol-17β or 17α-methyltestosterone administration on gonadal differentiation of largemouth bass Micropterus salmoides (Lacepède)

Monosex populations can be a valuable management tool in culture of larger size largemouth bass (>400 g). In this study, we investigated the effective mode and duration of oestrogen and androgen administrations to produce monosex largemouth bass populations. The experiment consisted of nine treatments. In oral administration groups, we fed 40-day-old fry either 200 mg of an oestradiol-17β (E₂) kg⁻¹ diet or 60 mg of a 17α-methyltestosterone (MT) kg⁻¹ diet for 30, 45 or 60 days. In bath treatments, we immersed fry in a 1 mg MT L⁻¹ solution for 5 h a day on three or six occasions. For control treatment, we fed fry an ethanol-treated diet for 45 days. The frequency of females in the control group was 53.1%. Oral administration of E₂ at all durations resulted in slight increases in the frequency of females (59.8–70.5%). Both modes of androgen administration at all durations were ineffective in altering phenotypic sex. The experimental results of our study indicated that male differentiation passed the point of being completely and functionally influenced by exogenous oestrogens, while female differentiation had already taken place and was no longer responsive to exogenous androgens in 40-day-old (33.5 mm) largemouth bass fry.     

Total calcium and inorganic phosphate in the blood plasma of farmed rainbow trout, Oncorhynchus mykiss

Reference (physiological) value intervals determined by the lower 2.5% and upper 97.5% quantiles were calculated for total calcium (Ca _t ) and inorganic phosphate (P) in farmed rainbow trout, Oncorhynchus mykiss , at an age of 10–12 months in raceway culture. Total calcium levels were significantly ( P ≤0.0009) greater in males (2.63–4 mmol L⁻¹, n =47, mean weight 359±142 g) than in immature females (2.48–3.97 mmol L⁻¹, n =410, mean weight 400±145 g). The reference range for P in immature females and males was 3.09–9.32 mmol L⁻¹ ( n =467, mean weight 401±144 g). The distribution and density of the quantiles in the tested reference group were made possible by the use of histograms, which showed normal distributions for Ca _t in males but not for Ca _t and P in immature females and males, in which a sinistral asymmetry was found.     

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.     

Pharmacokinetics of oxolinic acid in gilthead sea bream, Sparus aurata L.

This is the first study on the pharmacokinetic parameters of oxolinic acid (OA) in gilthead sea bream, Sparus aurata L. The kinetic profile of OA was studied after a single intravascular injection (20 mg kg⁻¹) in 100 g fish at 20 °C. The distribution half-life ( t _1/2α) and the elimination half-life ( t _1/2β) of the drug were found to be short (0.51 and 12.60 h, respectively). The drug penetration from the plasma to the tissues was adequate as the apparent volume of distribution of the drug at steady-state ( V _d(ss)) was found to be 2.11 L kg⁻¹. The mean residence time ( MRT ) of OA was short (14.25 h) and the total clearance rate ( Cl _T) of the drug was low (0.15 L kg⁻¹ h⁻¹). The bioavailability ( F %) of OA following oral administration (30 mg kg⁻¹) was also low (14%). Maximum values were observed for muscle at 0.5 h after injection, with levels declining as with subsequent sampling. At the first two time points (0.5 and 1 h) plasma levels of OA were higher than muscle, however, the reverse was evident for subsequent samples. Following oral administration, highest muscle levels were found at 16 h and, with the exception of the 24-h sampling, muscle OA concentrations were higher than plasma at all time points. The fast elimination of OA suggests short withdrawal times with reference to human consumption of treated fish.     

Comparison of lipid-walled microcapsules and lipid spray beads for the delivery of water-soluble, low-molecular-weight materials to aquatic animals

Microparticles (< 40 μm diameter) composed of 600 mg g⁻¹ tripalmitin/400 mg g⁻¹ fish oil were used to encapsulate the low-molecular-weight (mol. wt 460) antibiotic oxytetracycline in the form of either oxytetracycline hydrochloride (OTC.HCl) or oxytetracycline hemicalcium salt (OTC.HEM). Dry, finely ground particles of core material were encapsulated in spray beads. Dissolved core material was encapsulated in lipid-walled microcapsules.
Oxytetracycline (OTC) was most efficiently delivered (≈ 46.5 mg g⁻¹ lipid after 24 h suspension in seawater) as a hemicalcium salt in spray beads. Lipid-walled microcapsules were most efficient for delivering OTC (≈ 8.7 mg g⁻¹ lipid) as OTC.HCl dissolved in 0.2  M HCl at a concentration of 300 mg mL⁻¹.
Spray beads containing OTC.HEM were very stable over 1 month in storage. Lipid-walled microcapsules containing aqueous OTC.HCl lost ≈ 30% of their core material during storage. Freeze-drying of both microparticle types did not improve storage of spray beads, but showed promise for reducing leakage from lipid-walled microcapsules during storage and delivery to suspension feeders.     

Evaluation of diquat against an acute experimental infection of Flavobacterium columnare in channel catfish, Ictalurus punctatus (Rafinesque)

A trial was performed to evaluate the efficacy of diquat (6,7-dihydrodipyrido[1,2-a:2',1'-c]pyrazinediium dibromide) against an acute experimental infection of Flavobacterium columnare in channel catfish, Ictalurus punctatus . Diquat is an Environmental Protection Agency-approved herbicide and has the potential to be legally and practically used against columnaris. Channel catfish were challenged, by cutaneous abrasion, and waterborne exposure to F. columnare and treated once at 22-h post-challenge with 2.5, 5.0, 10.0 and 15 mg L⁻¹ of diquat active ingredient for 6 h. At the conclusion of the trial, 21-day post-challenge, diquat at 5.0, 10.0 and 15 mg L⁻¹ significantly ( P  < 0.05) reduced the mortality of infected fish from 95% in the challenged non-treated fish to 68%, 59% and 49%, respectively. In vitro , the minimum inhibitory concentration (MIC) of 23 isolates of F. columnare was assayed. The majority of the isolates had an MIC value of 5 μg mL⁻¹ (15 of the 23 isolates). Infected fish exhibited acute clinical signs similar to a natural infection. The skin had severe ulcerative necrotizing dermatitis and the muscles had severe necrotizing myositis. The gills had severe multifocal necrotizing branchitis. The results demonstrate that diquat would reduce mortalities caused by an acute columnaris infection.