Evaluation of the therapeutic effect of potassium permanganate at early stages of an experimental acute infection of Flavobacterium columnare in channel catfish,Ictalurus punctatus (Rafinesque)

The efficacy of potassium permanganate (KMnO₄) against the early stages of an experimental acute infection of Flavobacterium columnare in channel catfish, Ictalurus punctatus, was evaluated. Fish were experimentally challenged by waterborne exposure for 2 h to F. columnare after cutaneous abrasion, and treated with KMnO₄ at 2.0 mg L^?1 above the KMnO₄ demand at 0, 1, 2 or 4 h postchallenge for 24 h. Challenged non‐treated fish acted as a positive control and non‐challenged non‐treated fish acted as a negative control. Fish challenged and treated with KMnO₄ at 0, 1, 2 or 4 h postchallenge had mortalities of 26%, 63%, 64% and 83% respectively. The mortality of challenged fish treated with KMnO₄ at 0 h postchallenge (26%) was significantly less than the positive control (77%). The mortalities of challenged fish treated at 1, 2 or 4 h postchallenge were not significantly different from the positive control fish. The results suggest that KMnO₄ has a clear therapeutic value in early stages of columnaris infection but limited therapeutic value once the infection has progressed.     

Assessment of Aquaflor(®) , copper sulphate and potassium permanganate for control of Aeromonas hydrophila and Flavobacterium columnare infection in sunshine bass, Morone chrysops female × Morone saxatilis male

Two experiments were conducted to test the effectiveness of different therapeutants against a mixed infection of Aeromonas hydrophila and Flavobacterium columnare in sunshine bass. Experiment 1 evaluated copper sulphate, florfenicol‐medicated feed and potassium permanganate (KMnO₄) against a natural mixed infection. Experiment 2 further evaluated copper sulphate as a treatment to control an experimental mixed infection. In experiment 1, naturally infected untreated fish had the lowest final survival per cent, at 71%, while florfenicol‐medicated feed at 15 mg kg^?1 body weight for 10 days or copper sulphate at 2.1 mg L^?1 (1% of the total alkalinity) for 24 h produced the highest final survivals, at 90% and 88%, respectively. The final survival of the naturally infected fish administered florfenicol‐medicated feed was significantly different (P < 0.1) from the untreated fish. The survival curves for the florfenicol and the copper sulphate at 2.1 mg L^?1 were significantly improved from the untreated fish. In experiment 2, fish were challenged by waterborne exposure to A. hydrophila and F. columnare and either not treated or treated with copper sulphate at 2.1 mg L^?1. At the end of experiment 2, the per cent survival of the challenged fish treated with copper sulphate (99%) was significantly higher (P < 0.05) than the non‐treated (61%). The results illustrate clear benefit of florfenicol and copper sulphate against a mixed infection of A. hydrophila and F. columnare.     

Dietary copper effects survival of channel catfish challenged with Flavobacterium columnare

Columnaris disease is an important bacterial disease of commercially grown channel catfish, Ictalurus punctatus. Copper sulphate (CuSO₄) has been shown to be therapeutic and prophylactic as a water treatment for columnaris disease. Copper is an essential dietary component in animal feeds and CuSO₄ is typically included in base diets; a study was conducted to evaluate whether fish feed supplemented with additional CuSO₄ at 0, 40 and 80 mg kg^?1 of diet and fed at a daily rate of 3% body weight would affect survival to columnaris disease. Results indicate fish fed the copper‐supplemented diet for 2 weeks significantly increased survival following F. columnare challenge. This increase appeared to be dose‐dependent. The mean per cent survival (±SEM) for fish fed the base diet (unsupplemented) for 2 weeks and then challenged was 2.0% ± 1.1. Fish fed the base plus 40 mg CuSO₄ kg^?1 had a mean survival of 22.0% ± 11.0. Fish fed the base plus 80 mg CuSO₄ kg^?1 had a mean survival of 29.3% ± 13.4. The mean per cent survival for fish fed the base diet for 4 weeks and then challenged was 28.3% ± 9.0. Fish fed the base plus 40 mg CuSO₄ kg^?1 for 4 weeks had a mean survival of 12.5% ± 6.3. Fish fed the base plus 80 mg CuSO₄ kg^?1 for 4 weeks had a mean survival of 40.5% ± 8.1. There was a significant effect after 4 weeks with fish fed the base plus 80 mg CuSO₄ kg^?1 mg not with 40 mg kg^?1.     

Effectiveness of copper sulphate,potassium permanganate and peracetic acid to reduce mortality and infestation of Ichthyobodo necator in channel catfish Ictalurus punctatus (Rafinesque 1818)

Ichthyobodo necator is a single‐celled biflagellate parasite, which in high density can cause significant mortality in young fish. Copper sulphate (CuSO₄), potassium permanganate (KMnO₄) and peracetic acid (PAA) were evaluated for effectiveness against ichthyobodosis. Treatments were: untreated control, 2.1 mg L^?1CuSO₄, 3.0 mg L^?1 KMnO₄, 1.5 mg L^?1 PAA and 3.0 mg L^?1 PAA, and were applied to flow‐through tanks on three consecutive days. The study was designed to simulate the flow‐through systems utilized in the commercial rearing of juvenile channel catfish (Ictalurus punctatus). Mortality was monitored daily to compare survival rate among treatments. Parasite intensity was assessed pre chemical exposure and 20–24 h after the third application to determine effectiveness of the treatment. An assessment was also done 7 days post application to investigate possible reoccurrence. Copper sulphate, KMnO₄ and PAA (3.0 mg L^?1) significantly reduced the infestation rate of I. necator. Copper sulphate significantly improved the survival of I. necator infested channel catfish after three flow‐through applications compared with the untreated control. The 3.0 mg L^?1 PAA resulted in significantly lower survival than the untreated control, the 1.5 mg L^?1 PAA and the KMnO₄ were not statistically different from the untreated control.     

In vitro comparisons of the inhibitory activity of florfenicol,copper sulphate and potassium permanganate towards Aeromonas hydrophila and Flavobacterium columnare

Aeromonas hydrophila and Flavobacterium columnare, the aetiological agents of motile aeromonas septicaemia (MAS) and columnaris disease respectively, have been recently causing crippling mortalities to the sunshine bass, Morone chrysops female ×Morone saxatilis male (Percichthyidae), industry in the United States. Isolates of A. hydrophila and F. columnare obtained from fish that died during farm outbreaks were subjected to in vitro evaluation of florfenicol (FFC), copper sulphate (CuSO₄) and potassium permanganate (KMnO₄). Florfenicol inhibited the growth of A. hydrophila and F. columnare more than CuSO₄ and KMnO₄. The minimum inhibition concentration (MIC) of FFC was 0.04 ± 0 and 0.2 ± 0.1 mg L^?1 for A. hydrophila and F. columnare respectively, while the 50% inhibition concentration (IC50) for A. hydrophila and F. columnare was 0.23 ± 0.01 and 0.4 ± 0.2 mg L^?1 respectively. Copper sulphate was more effective against A. hydrophila than KMnO₄; CuSO4 had a MIC of 83.2 ± 0 mg L^?1 compared to 158.0 ± 0 mg L^?1 for KMnO₄. Copper sulphate was also more effective against F. columnare than KMnO₄. The IC50 values of CuSO₄ and KMnO₄ towards F. columnare were 4.8 ± 0.3 and 8.7 ± 1.6 mg L^?1 respectively, and the minimum bactericidal concentration values of CuSO₄ and KMnO₄ towards F. columnare were 25.0 ± 0 and > 158.0 mg L^?1 respectively. In addition, F. columnare was more sensitive to CuSO₄ and KMnO₄ than A. hydrophila.     

Short-term baths with essential oils of Lippia sidoides,Ocimum gratissimum and Zingiber officinale influence blood parameters and survival of tambaqui (Colossoma macropomum) after infection with Aeromonas hydrophila

This study evaluated the effect of short-term baths with the essential oils (EOs) of Lippia sidoides, Ocimum gratissimum and Zingiber officinale on the survival, haematological, biochemical and histopathological parameters of native fish tambaqui (Colossoma macropomum), challenged with Aeromonas hydrophila. After challenge, the fish infected with A. hydrophila received therapeutic baths of 60 min, for five consecutive days. Treatments were as follows: challenged and not treated; challenged and treated with 10 mg L^?1 of gentamicin; challenged and treated with EO of L. sidoides (2.5 and 5.0 mg L^?1); challenged and treated with EO of O. gratissimum (5.0 and 10.0 mg L^?1); challenged and treated with Z. officinale EO (5.0 and 10.0 mg L^?1). The highest survival rates were 89.5% (fish treated with 5.0 mg L^?1 of O. gratissimum EO) and 75.0% (fish treated with 5.0 mg L^?1 of L. sidoides EO). Reductions in the haematocrit percentage, haemoglobin and erythrocyte number were observed in fish treated with L. sidoides and Z. officinale EOs compared with control fish. Mild to moderate damage to the liver tissue of fish after challenge and EO treatments were observed. It is suggested that baths containing O. gratissimum and L. sidoides EOs consist in a viable treatment to increase survival rates of C. macropomum infected with A. hydrophila.     

Acute and chronic effects of aqueous ammonia on marbled spinefoot rabbitfish,Siganus rivulatus (Forsskål 1775)

Ammonia is a metabolite of aquatic organisms which might reach deleterious levels in intensive fish farms. The aim of the present study was to determine median lethal concentrations (96‐h LC₅₀) of total ammonia nitrogen (TA‐N) on marbled spinefoot rabbitfish (Siganus rivulatus) and chronic effects of TA‐N on survival, growth and behaviour of juvenile rabbitfish over a 50 day period. In the first experiment, fish were exposed to 0, 2, 4, 6, 8, 10, 12, 14, 16, 18 and 20 mg L^?1 TA‐N for 96 h and survival evaluated. In the second experiment, 12 fish were stocked per 50‐L tank and treated with one of 0, 2, 4, 6, 8, 10 and 12 mg L^?1 TA‐N with three replicate tanks per treatment. Survival and growth were determined and histopathological alterations of gills due to chronic ammonia exposure were studied by light and electron microscopy. The 96‐h LC₅₀ values were 16–18 mg L^?1 TA‐N. In the chronic exposure experiment, fish reared in water with 0 mg L^?1 TA‐N had 100% survival and had 50% weight increase in 50 days. Fish at 2 and 4 mg L^?1 TA‐N all died whilst fish in 6, 8, 10 and 12 mg L^?1 TA‐N survived and grew albeit less than in treatment 0 mg L^?1. Gills from ammonia treated fish displayed severe histological and ultrastructural alterations including hyperplasia, hypertrophy and fusion of secondary lamellae, aneurysms and presence of pleomorphic altered cells. Chronic exposure to ammonia is deleterious to marbled spinefoot rabbitfish and low concentrations of ammonia appear to kill the fish in <50 days whilst fish can survive for more than 50 days at concentrations between 6 and 12 mg L^?1 TA‐N.     

Effects of dietary copper sources on growth,tissue copper accumulation and physiological responses of Japanese sea bass (Lateolabrax japonicus) (Cuvier, 1828) fed semipurified or practical diets

An 8‐week trial was conducted to compare the efficacy of a chelated copper (Cu) with the inorganic Cu (CuSO₄), in the diet of Japanese sea bass (Lateolabrax japonicus). Six diets were prepared based on two basal diets (semi‐purified or practical) at two Cu inclusion levels (0 and 4 mg kg^?1 from or chelated‐Cu), accordingly named as S0, SI4, SM4, P0, PI4 and PM4 respectively. Each diet was fed to quadruplicate groups of Japanese sea bass (initial weight: 65.0 ± 0.03 g) during the trial. Fish fed practical diets had higher feed intake, specific growth rate, protein efficiency rate (PER), liver Cu accumulation, liver Na⁺K⁺‐ATPase and plasma alkaline phosphatase, but lower hepatosomatic index than the groups fed semi‐purified diets (P < 0.05). Higher PER was exhibited by fish fed chelated‐Cu diets than that of fish fed CuSO₄ diets (P < 0.05). Although liver Cu accumulation was significantly higher in fish fed practical diets,, liver Cu content increased significantly with increasing dietary copper levels only in fish fed semipurified diets. Chelated‐Cu has higher efficiency than CuSO₄ when fed in practical diets. Poor growth performance of fish fed semipurified diets might induce underestimating the nutrient requirement of Japanese sea bass.     

Anaesthetic efficacy of eugenol on iridescent shark,Pangasius hypophthalmus (Sauvage, 1878) in different size classes

Anaesthetic efficacy of eugenol was investigated on iridescent shark, Pangasius hypophthalmus. Fish (2, 5, 10 and 20 g) subjected to 20–200 mg L^?1 eugenol and behavioural response as well as induction and recovery times were recorded. Induction and recovery times were significantly affected by eugenol concentration as well as fish weight (P < 0.05). Generally, 27–300 s after exposure to 20–200 mg L^?1 eugenol, iridescent sharks reached stage 3 anaesthesia (suitable for general handling). Fish entered stage 4 anaesthesia (suitable for surgery and blood sampling) over 54–710 s exposure to such concentrations. Recovery time was 109–600 s in all weight classes as well as eugenol concentrations. Mortality (44–100%) was only observed in 2 g fish when subjected to 110–170 mg L^?1 eugenol. This study, for the first time, showed behavioural response of iridescent shark to anaesthesia as well as effectiveness of eugenol as anaesthetic in this important aquaculture‐ornamental species. According to the models obtained in this study, minimum eugenol concentrations to induce anaesthesia over less than 3 min were 53.8–81.5 mg L^?1 in 2–20 g fish. Likewise, maximum eugenol concentrations in which fish recovered over less than 5 min were 65.9–105.8 mg L^?1 in 2–20 g fish.     

Vaccination strategies with oral booster for surubim hybrid (Pseudoplatystoma corruscans × P. reticulatum) against haemorrhagic septicaemia

This study evaluated the efficiency of differently prepared vaccines against Aeromonas hydrophila in the hybrid surubim (Pseudoplatystoma corruscans × P. reticulatum). Survival and haemato‐immunological parameters were compared between the treatments: non‐vaccinated fish (C); bacterin‐vaccinated fish (B); bacterin plus oral booster vaccinated fish (B+O); bacterin and toxoid‐vaccinated fish (B+T) and bacterin, toxoid and oral booster‐vaccinated fish (B+T+O). Fourteen‐days vaccinated fish from B+O and B+T+O were fed with an oral booster for 4 days. After 1 week, the fish were intraperitoneally challenged with 2 × 10⁸ CFU mL^?1 of A. hydrophila. Fish from the treatment B+T+O showed the lowest cumulative mortality (11.36%) 96 h after challenge, compared with other treatments (22.72–44.04%), and a relative survival of 74%. Serum immunoglobulin in B+T+O fish was higher than in other treatments. All vaccinated fish showed an increased agglutination titre when compared with non‐vaccinated fish, both before and after challenge. Fish fed with oral booster showed an increase in phagocytic percentage before and after challenge. It can be inferred that the oral booster vaccination was efficient in reducing mortality in hybrid surubim by enhancing the response against haemorrhagic septicaemia due to A. hydrophila infection.     

Hatch rate of channel catfish Ictalurus punctatus (Rafinesque 1818) eggs treated with 100 mg L−1 copper sulphate pentahydrate

Catfish hatcheries use copper sulphate pentahydrate (CuSO₄·5H₂O) as an economical control for saprolegniasis on eggs. This study determines hatch rate of channel catfish, Ictalurus punctatus (Rafinesque 1818), eggs in hatching troughs containing 23.8 °C flow‐through well water when treated with 100 mg L^?1 CuSO₄·5H₂O (10 times the proposed therapeutic dose). Eggs were treated daily until the embryos reached the eyed stage. Fry survival in the control and 100 mg L^?1 CuSO₄·5H₂O treatments was significantly different (15% and 71% respectively). This study demonstrates that there is a considerable margin of safety in using CuSO₄·5H₂O as a catfish egg treatment to control saprolegniasis.     

The effects of dietary vitamin C on growth performance,serum enzymes activities and resistance to Vibrio alginolyticus challenge of yellow drum Nibea albiflora

A 9‐week feeding trial was conducted to determine the optimal dietary vitamin C requirement and its effects on serum enzymes activities and bacterial resistance in the juvenile yellow drum Nibea albiflora (initial weight 33.2 ± 0.10 g). Six practical diets were formulated containing vitamin C 2.1, 45.3, 89.6, 132.4, 178.6 and 547.1 mg kg^?1 diet supplied as l ‐ascorbyl‐2‐monophosphate. The fish fed 547.1 mg kg^?1 diet showed a significantly higher survival than that fed 2.1 mg kg^?1 diet. The weight gains and specific growth rate of the fish fed 2.1 mg kg^?1 diet were significantly lower than those of the fish fed 89.6–547.1 mg kg^?1 diets. The liver vitamin C concentration firstly increased with increasing dietary vitamin C supply from 2.1 to 178.6 mg kg^?1 diet and then stabilized. The serum superoxide dismutase activities of the fish fed 547.1 mg kg^?1 diet were significantly lower than those of the fish fed 2.1–89.6 mg kg^?1 diet. The fish fed 2.1 mg kg^?1 diet had a significantly higher alkaline phosphatase activity than those in the other groups except the 45.3 mg kg^?1 group. Fish that received diets containing vitamin C at 547.1 mg kg^?1 had significantly higher nitro blue tetrazolium and lysozyme activity, and fish that received diets containing vitamin C at 45.3–547.1 mg kg^?1 exhibited resistance against Vibrio alginolyticus infection. The dietary vitamin C requirement of the juvenile yellow drum was established based on broken‐line model of weight gain to be 142.2 mg l ‐ascorbyl‐2‐monophosphate kg^?1 diet.     

Comparison of Copper Sulfate Concentrations to Control Ichthyophthiriasis in Fingerling Channel Catfish

ABSTRACT

Fingerling channel catfish Ictalurus punctatus were exposed to Ichthyophthirius multifiliis-infested fish until immature trophonts developed. The fish were transferred to individual static fiberglass tanks filled with 600 L of pond water (total alkalinity and total hardness was 220 mg/L and 101 mg/L, respectively) and were treated with 0, 1.1, 2.2, 3.3, or 4.4 mg/L copper sulfate (CuSO₄ · 5H₂O) every other day for four treatments to evaluate its effectiveness to control mortality associated with ichthyophthiriasis. Water temperature was maintained at 18 ± 1°C. Fish were observed for ten days post-treatment and mortalities were recorded. Results indicate that half of the recommended dose (1.1 mg/L CuSO₄) is needed to effectively control an occurrence of ichthyophthiriasis under the conditions of this study. However, fish culturists should be aware that effective CuSO₄ treatment of ichthyophthiriasis on channel catfish raised in ponds may be influenced by water chemistry characteristics and suspended materials such as pond sediments.     

Gill damage to juvenile orange‐spotted grouper Epinephelus coioides (Hamilton, 1822) following exposure to suspended sediments

Juvenile Epinephelus coioides were exposed to three nominal concentrations (8, 32, 128 mg wet sediments L^?1) of suspended sediments (SS) from Port Shelter (PS), Mirs Bay (MB) and Victoria Harbour (VH) for 10 and 30 days using semi‐static system. Sediments from VH contained higher concentrations of Cu and polycyclic aromatic hydrocarbons (PAHs) than sediments from PS and MB. Gill damages including lamellar blood sinus dilation and vascular congestion were prevalent after just 10 days of exposure to SS. Fish exposed to SS at the highest concentration of 128 mg L^?1 showed higher incidence of lamellar aneurism. Hyperplasia in the base of lamellae was recorded in fish that had been exposed to contaminated SS from VH. Significant increases in the density of chloride cells and mucous cells were found on the gills of fish that had been exposed to 128 mg L^?1 of SS from PS. Clogging of gills by SS produced hypoxic‐like responses in fish. Polluted sediments from VH produced addictive or synergistic effects between SS and chemical contaminants on fish.     

Dietary myo‐inositol requirement for juvenile gibel carp (Carassius auratus gibelio)

An 11‐week growth trial was conducted to determine dietary myo‐inositol (MI) requirement for juvenile gibel carp (Carassius auratus gibelio). Myo‐inositol was supplemented to the basal diet to formulate six purified diets containing 1, 56, 107, 146, 194 and 247 mg MI kg^?1 diet, respectively. Each diet was fed to triplicate groups of juvenile gibel carp (initial body weight 3.38 ± 0.27 g, mean ± SD) in a flow‐through system. The diets were randomly assigned to different fish tanks. Fish fed ≥ 107 mg MI kg^?1 diet had significantly higher weight gain (WG), feed efficiency (FE) and protein efficiency ratio than those fed 1 mg MI kg^?1 diet. Fish fed ≥ 56 mg MI kg^?1 diet had higher feeding rate and survival compared with fish fed 1 mg MI kg^?1 diet. Dietary supplemental inositol did not affect fish liver inositol concentration. Fish fed ≥ 56 mg MI kg^?1 diet had higher body dry matter, crude protein and gross energy and lower hepatosomatic index than fish fed 1 mg MI kg^?1 diet. Dietary inositol supplementation decreased fish body ash. Quadratic regression of weight gain indicated that the myo‐inositol requirement to maximum growth for juvenile gibel carp was 165.3 mg MI kg^?1 diet.     

Effects of formalin on water quality and parasitic monogeneans on silver perch (Bidyanus bidyanus Mitchell) in earthen ponds

Infestations of parasitic monogenean trematodes (Lepidotrema bidyana and Gyrodactylus sp.) on freshwater silver perch (Bidyanus bidyanus Mitchell) in earthen ponds were treated with formalin (37% formaldehyde). Concentrations of 30 and 40 mg L^?1 formalin were effective, but fish in ponds treated with 20 or 25 mg L^?1 remained infested. At temperatures of 24.1–26.9°C, concentrations of 30 or 40 mg L^?1 formalin caused dissolved oxygen (DO) to decline from 10.1–11.9 to 3.0–3.3 and 1.2–1.7 mg L^?1, respectively, within 36–42 h of treatment. In addition, pH declined from 7.2–8.4 to 6.3–6.7, within 36 h and turbidity decreased over 48 h. In the ponds where DO was 1.2–1.7 mg L^?1, silver perch showed signs of severe stress, but continuous aeration (10 hp ha^?1) for 3 days and inflow of well‐oxygenated water for 6–8 h prevented mortalities. At temperatures of 13.2–15.7°C, concentrations of 30 or 40 mg L^?1 formalin caused DO to decline from 9.0–10.0 to 6.0–8.1 mg L^?1 and pH from 7.0–7.3 to 5.9–6.6 within 72 h. Total ammonia‐nitrogen increased over 72 h in ponds treated with 30 or 40 mg L^?1 formalin. Fish became re‐infested with L. bidyana in all ponds within 30 days of treatment. A concentration of 30 mg L^?1 formalin is recommended as a treatment for monogeneans on silver perch in ponds, but aeration is necessary to maintain adequate water quality at higher temperatures.     

Comparative effects of dietary l‐carnitine supplementation on diploid and triploid rainbow trout (Oncorhynchus mykiss)

This study compared the effects of dietary l ‐carnitine and ploidy on growth performances and fatty acid content in rainbow trout (Oncorhynchus mykiss). Fish [initial body weight (BW) = 30 g] reared under high density (50 kg biomass m^?3) were hand‐fed in triplicate (3 tanks treatment^?1, 34 fish tank^?1) twice a day, with three nutritionally identical diets containing 15, 200 or 530 mg l ‐carnitine kg^?1 of diet. No significant growth differences were observed over a 56‐day grow‐out period, during which BWs increased threefold. Growth performances and survival were not significantly affected by either ploidy or dietary l ‐carnitine content, although daily growth index showed an increasing trend (2.52–2.65% day^?1) with increasing dietary l ‐carnitine. Body l ‐carnitine content increased significantly with dietary l ‐carnitine content. Diploid fish had higher plasma ammonia (716–725 μmol L^?1) and osmolality (297–303 mOsm) levels than triploid trout (523–649 μmol L^?1 and 285–291 mOsm, respectively). l ‐carnitine, ploidy and their interaction showed to affect significantly the concentration of several fatty acids. Palmitoleic (16:1), oleic (18:1) and erucic (22:1) acids showed lower concentrations, while the eicosadienoic (20:2n‐6) and arachidonic (20:4n‐6) acids were elevated in liver of triploid fish. Eicosapentaenoic acid (22:5n‐3) was significantly higher in fish fed 200 mg than in fish fed 15 mg l ‐carnitine.     

The effect of β‐1,3‐glucan derived from Euglena gracilis (Algamune™) on the innate immunological responses of Nile tilapia (Oreochromis niloticus L.)

Algamune^? is a commercial additive produced from Euglena gracilis, providing a rich source of the β‐1,3‐glucan paramylon. Isolated kidney phagocytes of Nile tilapia were incubated with graded doses (0, 8, 16, 32, 64 and 128 μg/ml) of Algamune^? and purified paramylon to gauge their ability to elicit the production of reactive oxygen species. A linear response was observed for extracellular superoxide anion for both sources but only Algamune^? for intracellular superoxide anion. After corroborating the immunostimulant properties ex vivo, a feeding trial was conducted to evaluate the dietary supplementation of Algamune^? (0, 100, 200, 400 and 800 mg/kg of diet) for Nile tilapia. Fish were fed for 3 weeks, after which, fish were sampled for blood and head kidney phagocytes. The remaining fish were challenged with Streptococcus iniae. Macrophage extracellular superoxide anion production was significantly elevated in fish fed diets with 200 mg of Algamune^? kg^?1 when compared to fish fed the basal diet. Even though the disease challenge did not show statistical differences, it is worth mentioning that fish fed intermediate doses of Algamune^? had lowest numerical mortality values. Therefore, Algamune^? was demonstrated to enhance some immunological responses of tilapia both in ex vivo and in vivo.     

Virulence assay of rhizoid and non‐rhizoid morphotypes of Flavobacterium columnare in red tilapia,Oreochromis sp., fry

Numerous isolates of Flavobacterium columnare were previously recovered from red tilapia, Oreochromis sp., exhibiting columnaris‐like disease in Thai farms, and the phenotypic and genetic characteristics were described. The objective of this study was to determine the virulence of two morphotypes (rhizoid and non‐rhizoid colonies) of F. columnare and to determine their ability to adhere to and persist in red tilapia fry. The results showed that the typical rhizoid isolate (CUVET1214) was a highly virulent isolate and caused 100% mortality within 24 h following bath challenge of red tilapia with three different doses. The non‐rhizoid isolate (CUVET1201) was avirulent to red tilapia fry. Both morphotypes adhered to and persisted in tilapia similarly at 0.5 and 6 h post‐challenge as determined by whole fish bacterial loads. At 24 and 48 h post‐challenge, fry challenged with the rhizoid morphotype exhibited significantly higher bacterial loads than the non‐rhizoid morphotype. The results suggested that an inability of the non‐rhizoid morphotype to persist in tilapia fry may explain lack of virulence.     

Acute toxicity of ammonia in Meagre (Argyrosomus regius Asso, 1801) at different temperatures

Argyrosomus regius (3.0 ± 0.9 g) were exposed to different concentrations of ammonia in a series of acute toxicity tests by the static renewal method at three temperature levels (18, 22 and 26°C) at a pH of 8.2. Low temperature clearly increased the tolerance of the fish to total ammonia nitrogen (TAN) and unionized ammonia (NH₃) (P < 0.05). While the 96‐h LC₅₀ values of TAN were 19.79, 10.39 and 5.06 mg L^?1, the 96‐h LC₅₀ of NH₃ were 1.00, 0.70 and 0.44 mg L^?1 at 18, 22 and 26°C respectively. The safe levels of NH₃ for A. regius was estimated to be 0.10, 0.07 and 0.04 mg L^?1 at 18, 22 and 26°C respectively (P < 0.05). This study clearly indicates that A. regius is more sensitive to ammonia than other marine fish species cultured on the Mediterranean and Eastern Atlantic coasts.