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.     

Use of salt to control ichthyophthiriosis and prevent saprolegniosis in silver perch, Bidyanus bidyanus

The ciliate protozoan, Ichthyophthirius multifiliis , and the fungus, Saprolegnia parasitica , cause the diseases ichthyophthiriosis and saprolegniosis respectively. Both diseases are difficult to control and can cause high mortalities of freshwater fish, including the Australian silver perch, Bidyanus bidyanus (Mitchell). The efficacy of salt (NaCl) in controlling and preventing these diseases in silver perch was evaluated in aquaria and tanks. Low pH levels were also evaluated as a control for ichthyophthiriosis. Concentrations of 2 or 3 g L⁻¹ salt controlled infestations of I. multifiliis , and fish were free of both theronts and trophonts by day 8 at temperatures of 17.3–21.3 °C and by day 6 at 19.2–23.5 °C. Fish treated with 1 g L⁻¹ salt remained infested and all fish in a control treatment (0 g L⁻¹ salt) died. Although the mean survival rates of infested fish at pH levels of 5 or 6 were only 13.9% and 7.6%, respectively, there were no theronts or trophonts on surviving fish after 12 days. Silver perch harvested from a pond and treated with 2 or 3 g L⁻¹ salt did not become infected with S. parasitica and survival was 100%, whereas 16.6% of untreated (0 g L⁻¹ salt) fish became infected and survival was only 66.7%. A concentration of 2 g L⁻¹ NaCl is recommended for the control of ichthyophthiriosis and the prevention of saprolegniosis in silver perch held in tanks, aquaria and re-circulating aquaculture systems.     

Protective Immunity of Goldfish Against Ichthyophthirius multifiliis Infection Induced by Different Trophont Vaccine Preparations

Ichthyophthirius multifiliis, commonly called “Ich,” is a protozoan parasite that infects the epidermis and gills of freshwater fish. Here, we used goldfish (Carassius auratus) to determine whether the four vaccine preparations (live trophonts, formalin‐fixed trophonts, freeze‐thawed trophont lysates, and trophont cilial and cell cortical fractions) of I. multifiliis (Fouquet) can elicit resistance of immunized fish against subsequent theront challenge, and whether a relationship exists between in vitro immobilization of theronts in sera or mucus from immunized fish and host protective immunity against the parasite. Experimental goldfish were randomly divided into five groups with five parallel controls, with each group or subgroup consisting of 20 individuals. Each test goldfish was immunized with one of the four Ich vaccine preparations administered by one of the three routes: live trophonts by gavage (Group 1), formalin‐fixed trophonts by injection (Group 2) or by immersion (Group 3), freeze‐thawed trophont lysates by injection (Group 4), and trophont cilial and cell cortical fractions by injection (Group 5). The fish were then challenged by a standard Ich theront challenge on Day 21 postimmunization. For every 10 d following immunization, we tested the in vitro immobilization of Ich theronts by sera or mucus from immunized and control fish. We found that although all control (nonimmunized) and Group 3‐immunized goldfish died following theront challenge, more than half (55–65%) of the immunized fish from Groups 1, 2, 4, and 5 survived. Furthermore, except for the fish from Groups 3 and 4, the number of mean days to death in each test group was significantly higher than that in the respective control. These results indicate that goldfish immunized by injection or by gavage with any of the four vaccine preparations gained resistance to Ich infection. Further analysis indicated that this protective immunity was closely associated with the in vitro immobilization of Ich theronts by fish sera or mucus.     

Effects of formalin on total haemocytes count and histopathological changes in the shrimp Litopenaeus schmitti (Pérez‐Farfante & Kensley 1997)

Formalin is used in shrimp industry to eliminate ectoparasites and as a disinfectant. However, the effect of formalin on shrimp's defense mechanism is unknown. Several biomarkers were used to assess the immune response of juveniles of the shrimp Litopenaeus schmitti exposed to different concentrations of formalin (10, 25 and 50 mg L^?1) during 24 and 48 h. Formalin concentrations of 25 and 50 mg L^?1 produced a significant reduction in the total haemocyte count at both 24 and 48 h. Peroxidase and phenoloxidase activity did not show variations. The total protein concentration in haemolymph of shrimps was not significantly different between experimental groups. Histological analysis showed hyperplasia in the gills of animals exposed at 10 mg L^?1 for 24 h; however, the exposure at 25 and 50 mg L^?1 caused necrosis in gills, hepathopancreas and the antennal gland.     

Effects of Water pH and Hardness on Infection of Silver Catfish,Rhamdia quelen,Fingerlings by Ichthyophthirius multifiliis

The aim of this study is to determine the effect of (1) different pH levels (5, 6, 7, 8, and 9) with water hardness 20 mg/L CaCO₃ and (2) pH 5 and 7 with varying water hardnesses (20, 60, and 120 mg/L CaCO₃) on the intensity of infection by Ichthyophthirius multifiliis in fingerlings of the silver catfish, Rhamdia quelen. In Experiment 1, the lowest and highest mortality rates were observed in fingerlings exposed to pH 5 and 9, respectively; in Experiment 2, fish kept at pH 5 and 7 with hardness 20 mg/L CaCO₃ showed significantly lowest and highest cumulative mortality, respectively, than those maintained in other conditions. The lowest intensity of trophonts/fingerling was observed at pH 5 and hardness 20 mg/L CaCO₃ in both experiments. We conclude that infection by I. multifiliis is less severe in silver catfish maintained at pH 5 and hardness 20 mg/L CaCO₃. High water hardness led to an increased intensity of trophonts and decreased survival in silver catfish kept at pH 5, but decreased intensity of trophonts and improved survival when the fish were kept at pH 7.     

Susceptibility of three strains of blue catfish,Ictalurus furcatus (Valenciennes), to Ichthyophthirius multifiliis

This study compared the susceptibility of three blue catfish strains (D&B, USDA 101 and USDA 102) to the parasite Ichthyophthirius multifiliis (Ich). In Trial I, a cohabitation study (all strains stocked communally) was conducted and fish were exposed to theronts at 0, 200, 1000, 5000 or 25 000 theronts fish^?1, respectively. All fish died when exposed to theronts at 5000 or 25 000 theronts fish^?1. When exposed to 1000 theronts fish^?1, USDA 102 strain of blue catfish showed significantly lower mortality (78.5%) compared to USDA 101 and D&B strains (92.7% and 100%). In Trial II, the same three strains of blue fish were evaluated for their susceptibility to Ich with strains challenged in separate tanks by adding Ich theronts at 0, 200 and 1000 theronts fish^?1, respectively. All D&B and USDA 101 blue catfish died; however, 42.3% of USDA 102 strain survived the infection when exposed to 1000 theronts per fish. The results indicate that there are differences among strains of blue catfish for susceptibility to Ich, and these differences will be useful in the development of improved catfish germplasm for commercial aquaculture.     

Efficacy of formalin,iodine and sodium chloride in improvement of egg hatching rate and fry survival prior to the onset of exogenous feeding in yellow perch

Egg disinfection is considered the most important routine work in hatcheries to avoid fungal and/or bacterial infection of fish eggs. The aim of this study was to determine the effectiveness of three disinfectants: formalin, iodine and sodium chloride on the hatching success of yellow perch eggs. The disinfectants were tested in triplicate at different concentrations for 15 and 30 min bath treatments. Two experiments were conducted; formalin at five concentrations (25, 50, 100, 150 and 200 mg L^?1) and 25 mg L^?1 iodine were tested in the first experiment. The second experiment involved formalin at three concentrations (250, 500 and 1000 mg L^?1), iodine at three concentrations (50, 100 and 250 mg L^?1) and sodium chloride at three concentrations (500, 1000 and 3000 mg L^?1) were used. Iodine and sodium chloride‐treated eggs hatched earlier than formalin‐treated eggs. The highest mean percentage of eyed stage, hatching rate and survival to first feeding fry was observed at 200 mg L^?1 formalin for 30 min, 50 mg L^?1 iodine for 15 min and 500 mg L^?1 sodium chloride for 30 min. High concentrations of formalin (1000 mg L^?1), iodine (250 mg L^?1) and sodium chloride (1000 and 3000 mg L^?1) showed toxicity to yellow perch eggs, resulting in low hatching rate and survival to first feeding fry. We recommended formalin at a concentration of 150–200 mg L^?1 for 30 min as an effective, easily available and low‐cost disinfectant for routine use to improve yellow perch hatchability.     

Antiparasitic activity of the antimicrobial peptide HbβP‐1, a member of the β‐haemoglobin peptide family

A family of antimicrobial peptides (AMPs) derived from the β‐subunit of haemoglobin was recently isolated from channel catfish, Ictalurus punctatus, infected with Ichthyophthirius multifiliis (ich), an important freshwater fish parasite that causes ichthyophthiriosis. We previously discovered that one of these AMPs, HbβP‐1, had strong cidal activity against ich as well as another ectoparasite, Tetrahymena pyriformis. HbβP‐1 toxicity was specific, primarily affecting the trophozoite (trophont) stage of ich. Here, we show that HbβP‐1 acts more rapidly to kill smaller (presumably less mature) trophonts of ich, taking almost twice as long to kill larger trophonts (P < 0.0001). It acts more rapidly than an unrelated AMP, piscidin 1, which is haemolytic and also lethal to ich trophonts. HbβP‐1 is potently and selectively lethal to the trophont stage of the dinoflagellate ectoparasite, Amyloodinium ocellatum, one of the most important pathogens of warmwater marine fish. HbβP‐1 has no effect on the fish gill cell line feeder layer (G1B cells) used to propagate Amyloodinium, further suggesting a highly selective action. These findings suggest that HbβP‐1 or related AMPs might function in protecting marine as well as freshwater fish and that HbβP‐1 has highly selective activity against specific life stages of important fish ectoparasites.     

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.     

Development of a feeding strategy for silver perch, Bidyanus bidyanus (Mitchell), based on restricted rations

To develop a feeding strategy for the Australian freshwater fish silver perch (Bidyanus bidyanus Mitchell), a series of eight experiments was done in 1 m³ cages in an aerated, earthen pond to determine the effects of feeding rate (% body weight) and feeding frequency (no. of feeds day^?1) on the growth and feed conversion ratio (FCR) of fingerlings and larger fish under ambient water temperatures over the range 13.8–30.6°C. Fish were fed extruded pellets of a silver perch diet containing 34% digestible protein and 14 MJ kg^?1 digestible energy. Commercial silver perch farmers were consulted about feeding practices for large fish (>500 g) and at water temperatures below 12°C, and winter feeding practices for other warmwater species were used to complete the strategy. In the feeding experiments, growth and FCR increased with increasing feeding rates to a level above which only FCR increased. Optimal feeding rates and frequencies were those which resulted in maximal growth, while minimizing effort (feeding frequency) and FCR. The highest feeding frequency required for maximal growth, including that of small fingerlings was twice (2 ×) daily, and the optimal feeding rates varied with water temperature and size of fish. The optimal daily regimes were: small fingerlings (initial mean weight, 2.0 g) 7.5% 2 × at a mean temperature of 23.3°C; fingerlings (14.9–27.7 g) 7.5% 2 × at 27.1°C, 5.0% 2 × at 23.7°C and 2.0% 1 × at 16.8°C; and large silver perch (162.5–510.6 g) 0.5% 1 × daily or 1.0% on alternate days at 15.6°C, 1.0% 1 × at 17.3°C, 3.0% 2 × at 24.1°C and 2.0% 2 × at 27.9°C. It is suggested that regimes of 0.5% 1 × daily for fingerlings (<50 g) and 0.5% 1 × on alternate days for larger fish are used at temperatures of 9–12°C, and 0.5% 3 days week^?1 and 0.5% 1 day week^?1 for fingerlings and larger fish, respectively, at 6–9°C. Feed inputs should not exceed 150 kg ha^?1 day^?1 in ponds less than 0.3 ha and 100 kg ha^?1 day^?1 in larger ponds. Our research has established a feeding strategy for silver perch based on restricted rations.     

Survival, growth and biochemical parameters of silver catfish, Rhamdia quelen (Quoy & Gaimard, 1824), juveniles exposed to different dissolved oxygen levels

Silver catfish, Rhamdia quelen (Quoy & Gaimard, 1824), is an endemic species from Latin America that is raised in cultivation ponds, and consequently may be exposed to low oxygen levels. Therefore, the objective of this study was to verify the lethal concentration (CL_{50?96 h}) of dissolved oxygen levels for silver catfish juveniles. In addition, the effects of different dissolved oxygen levels (1.96±0.08, 3.10±0.10, 4.14±0.09, 5.20±0.07 and 6.16±0.03 mg L^?1) on growth and metabolic parameters (glycogen, glucose, protein, lactate levels and catalase activity) were also investigated. CL_{50?96 h} was 0.52 mg L^?1 (CI 0.42–0.61 mg L^?1) or 6.7% oxygen saturation. After exposure of silver catfish to hypoxia for 30 days, there were no changes in biochemical parameters indicating the use of an anaerobic pathway by the fish. However, the dissolved oxygen levels influenced silver catfish juvenile behaviour, survival and growth, and under the experimental conditions 5.2 mg L^?1 (or 65.6% oxygen saturation) is the minimum oxygen level recommended for the growth of this species.     

肉桂活性成分肉桂醛杀灭离体多子小瓜虫效果

研究从肉桂中分离纯化出具有抑杀多子小瓜虫活性的化合物。以乙醇为提取剂,用索氏超声提取法从肉桂中提取浸膏,再用石油醚、乙酸乙酯、甲醇为萃取剂,萃取不同有效组分,以不同浓度分别进行杀灭离体多子小瓜虫实验,发现石油醚萃取物的杀虫效果最优。然后对石油醚萃取物采用硅胶层析柱和制备型高效液相色谱进行分离纯化,利用质谱和核磁波谱分析,最终鉴定其杀虫活性成分为肉桂醛;将肉桂醛溶于二甲基亚砜并用二倍梯度稀释法配成不同浓度的药液测试其对离体小瓜虫的杀灭活性。结果表明,100%杀灭滋养体和感染性幼虫的剂量分别为50和8 mg/L,半数有效浓度分别为13.9和1.8 mg/L;使用剂量在50 mg/L可完全抑制小瓜虫包囊孵化。     

No reason for keeping 0+ perch (Perca fluviatilis L.) with the prey fish

Growth and survival of 0+ perch were studied in 4 ponds with the topmouth gudgeon (Pseudorasbora parva) and without it. In the end of April, all ponds were stocked by free-swimming perch larvae (120,000 ind ha^?1). In June, topmouth gudgeon was introduced as forage fish (40 kg ha^?1) into two of the experimental ponds. Topmouth gudgeon significantly influenced neither the total abundance of zooplankton nor the abundance of its groups (Rotifera, Cladocera, Copepoda). The most important food item for perch (TL > 29 mm) was macroinvertebrates (especially Chironomidae). In the ponds with topmouth gudgeon, copepods and cladocerans were more important than in ponds without it. Specific growth rate of perch was 0.01 mm day^?1 in all ponds. Final mean total length (TL ± SD) of perch was 73 ± 13 and 70 ± 6 mm in the ponds with topmouth gudgeon and without it, respectively. Only 1 % of the perch reached higher TL than that recorded in the ponds without the topmouth gudgeon. Survival rate of perch varied from 12 to 36 % depending on ponds. Piscivory was recorded in all ponds from the age of 57 days (post-stocking); however, perch siblings were preferred to topmouth gudgeon. The highest asymptotic growth (L∞ = 88 mm) was calculated in the pond stocked with topmouth gudgeon. This corresponded with the highest cannibalism and lowest survival rate (12 %). Perch growth rate increased till 42–53 days of perch age and then started to decrease. There was no significant influence of potential prey fish (topmouth gudgeon) on the growth of 0+ perch; however, two size cohorts were found in the ponds with the topmouth gudgeon. Presence or absence of littoral macrophytes seems to be more substantial for rearing perch in ponds than stocking with the prey fish.     

Effects of diets with different plant proteins on the performance of silver perch (Bidyanus bidyanus Mitchell) and on water quality in earthen ponds

As part of a project to develop least‐cost diets with low levels of fish meal, silver perch (Bidyanus bidyanus Mitchell) fingerlings (mean weight, 11.8 g) were stocked at a density of 7500 fish ha⁻¹ into 0.1‐ha earthen ponds and fed one of two diets containing 33% digestible protein, 13 MJ kg⁻¹ digestible energy, similar nutrient specifications, but with different levels of fish meal and plant proteins. The reference diet SP35 had 27% fish meal, 28% wheat, 20% soybean and 11% sorghum, while the diet silver perch least‐cost (SPLC) had 10% fish meal, 20% peanut meal, 19% wheat, 17% lupins, 16% canola, 8% soybean and 5% blood meal; there were three replicate ponds for each diet. Fish were fed a restricted ration up to 5% body weight day⁻¹ and cultured for 10 months. Survival ranged from 85.9% to 94.3% and was not affected by diet. The mean weight (550 g), specific growth rate (SGR; 1.28% day⁻¹), absolute growth rate (AGR; 1.9 g fish⁻¹ day⁻¹) and production rate (4.5 tonnes ha⁻¹ year⁻¹) were significantly higher (P<0.05) and feed conversion ratio (FCR=1.8) was significantly lower for fish fed SP35 compared with fish fed SPLC (413 g, 1.18% day⁻¹, 1.4 g fish⁻¹ day⁻¹, 3.3 tonnes ha⁻¹ year⁻¹, 2.4). From October (spring) to March (autumn), turbidity was significantly lower (P<0.05) in SPLC ponds than in SP35 ponds, and fish were observed avoiding or ingesting and then expelling SPLC pellets. In February and March, infestations of the ectoparasitic copepod Ergasilus sp. were found on silver perch fed SPLC, and there was 5% post‐harvest mortality of these fish. The high inclusion levels of plant proteins, particularly peanut meal and canola in SPLC, may have provided anti‐nutritional factors and/or reduced the palatability and intake of the diet, adversely affecting the performance and health of silver perch, and water quality in the ponds. Our study demonstrates the value of evaluating new aquaculture diets under practical conditions over a complete growing period.     

Performance and sensory evaluation of silver perch (Bidyanus bidyanus Mitchell) fed soybean or meat meal-based diets in earthen ponds

The performance of silver perch fed a commercially available diet based on meat meal (38%), grain legumes (18%), oilseeds (10%), wheat millrun (20%), fishmeal (5%) and fish oil (3%) was compared with experimental diets based on alternative protein sources in two experiments. In Experiment 1, two experimental diets contained similar contents of fishmeal and fish oil as the commercially available reference diet, but soybean (25%) and wheat millrun (>31%) were used to reduce animal protein meals by approximately 50%. The digestible protein and digestible energy of the two experimental diets was either slightly lower (31.5% and 12.8 MJ kg^?1) or slightly higher (34.9% and 14.3 MJ kg^?1) than the reference diet (32.1% and 13.2 MJ kg^?1). In Experiment 2, the two experimental diets contained no fishmeal but included higher amounts of rendered animal meals (41–48%). One of the diets had similar digestible protein to the reference diet (32%) while the other had only 25% digestible protein. Silver perch (38 g for Experiment 1 and 59 g for Experiment 2) were stocked into each of nine 0.1 ha earthen ponds with fish in three ponds fed each diet for 191 days (Experiment 1) or 187 days (Experiment 2). Survival was >94% in all ponds in both experiments. In Experiment 1, growth rates and feed conversion ratios (FCRs) ranged from 2.1 to 2.4 g fish^?1 day^?1 and 1.7 to 1.9 respectively. Growth rates were significantly (P<0.05) lower for fish fed the experimental diet with the lowest digestible energy content. Growth rates for fish fed the other experimental diet and the reference diet were similar (P>0.05). In Experiment 2, growth rates and FCRs ranged from 2.3 to 2.4 g fish^?1 day^?1 and 1.6 to 1.7. There were no significant differences in fish performance indices for any of the three diets although experimental power was low (power=0.31). A blind consumer sensory evaluation (taste panel) of fish fed the three diets in Experiment 2 rated fish as ‘highly acceptable’. The diet with the lowest digestible protein content produced the best fish in terms of ‘smell liking’, ‘flavour liking’, ‘muddy flavour strength’ and ‘fresh flavour strength’. These results confirm that soybean meal and/or rendered animal protein ingredients including meat meal and poultry offal meal, and wheat can form the basis for high‐performance, low‐cost diets for intensive pond culture of silver perch.     

Effectiveness of eugenol sedation to reduce the metabolic rates of cool and warm water fish at high loading densities

Effects of eugenol (AQUI‐S^®20E, 10% active eugenol) sedation on cool water, yellow perch Perca flavescens (Mitchill), and warm water, Nile tilapia Oreochromis niloticus L. fish metabolic rates were assessed. Both species were exposed to 0, 10, 20 and 30 mg L^?1 eugenol using static respirometry. In 17°C water and loading densities of 60, 120 and 240 g L^?1, yellow perch controls (0 mg L^?1 eugenol) had metabolic rates of 329.6–400.0 mg O₂ kg^?1 h^?1, while yellow perch exposed to 20 and 30 mg L^?1 eugenol had significantly reduced metabolic rates of 258.4–325.6 and 189.1–271.0 mg O₂ kg^?1 h^?1 respectively. Nile tilapia exposed to 30 mg L^?1 eugenol had a significantly reduced metabolic rate (424.5 ± 42.3 mg O₂ kg^?1 h^?1) relative to the 0 mg L^?1 eugenol control (546.6 ± 53.5 mg O₂ kg^?1 h^?1) at a loading density of 120 g L^?1 in 22°C water. No significant differences in metabolic rates for Nile tilapia were found at 240 or 360 g L^?1 loading densities when exposed to eugenol. Results suggest that eugenol sedation may benefit yellow perch welfare at high densities (e.g. live transport) due to a reduction in metabolic rates, while further research is needed to assess the benefits of eugenol sedation on Nile tilapia at high loading densities.     

Determination of histological and genotoxic effects of formalin on Nile tilapia (Oreochromis niloticus L.)

Formalin (37–40% formaldehyde) is one of the most effective, widely used chemical in pisciculture for its antiparasitic, antifungal and prophylactic activities. It is used in paints, cleaning products and textile industry, as well. Genotoxic and histological effects of sublethal formalin exposure on Nile tilapia (Oreochromis niloticus) are investigated in this study. The semi‐static acute test was employed. One‐tenth, one‐fifth and half of the 96‐h LC₅₀ value, 148 mg L^?1, were used as sublethal exposure concentrations: 15, 30 and 75 mg L^?1 for 24, 48, 96 and 168 h respectively. The micronucleus test was applied to investigate the genotoxic effects on fish erythrocytes at the end of predetermined exposure periods and all tissues of the fish were sampled for histological examination. The micronucleus frequencies increased significantly in all exposure groups when compared with their control groups in a dose and time dependent manner (P < 0.05). Results of histological examination showed no histopathological findings in the gonads, heart, skeletal muscle, spleen and intestine tissues after sublethal formalin exposure. Passive hyperaemia of liver tissue in varying proportions, hydropic degeneration, fatty degeneration in high concentrations, branchitis on gill tissues in varying proportions, epithelial lifting, telangiectasia, bleeding in kidney and hyperaemia were found in Nile tilapia exposed to formalin at different concentrations and durations. As a result, it is thought that formalin leads to tissue damage and shows genotoxic effects even at 15 mg L^?1 concentration in O. niloticus.     

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.     

Salt in the Food and Water as a Supportive Therapy for Ichthyophthirius multifiliis Infestation on Silver Catfish, Rhamdia quelen, Fingerlings

This study verified the efficacy of common salt (NaCl) in the food and water as a supportive therapy for Ichthyophthirius multifiliis and growth of silver catfish, Rhamdia quelen, fingerlings. After appearance of the first white spots following experimental exposure, indicating infestation by this parasite, fish were transferred to eighteen 40‐L polypropylene tanks and kept for 30 d. Experimental diets were prepared with five different levels of salt supplementation: 0.0, 1.2, 2.5, 5.0, and 6.0% for 30 d. In addition, another group of fish were fed a diet without salt supplementation but exposed to 4 g of salt/L of water. Fingerlings were collected at different times for analyses of whole‐body Na⁺, K⁺, and Cl^?. Dietary salt supplementation provoked a transitory increase in whole‐body Na⁺, K⁺, and Cl^? in infested catfish fingerlings, but the best stabilization of these ion levels was observed in specimens exposed to salt in the water. Silver catfish fed a diet containing 1.2% salt showed higher weight and biomass after 30 d compared to those fed with 6% salt. The use of 4 g of salt/L of water improved survival of infested silver catfish, but dietary salt supplementation was ineffective as a therapy for ichthyophthiriasis.     

Essential oils to control ichthyophthiriasis in pacu,Piaractus mesopotamicus (Holmberg): special emphasis on treatment with Melaleuca alternifolia

In vitro effect of the Melaleuca alternifolia, Lavandula angustifolia and Mentha piperita essential oils (EOs) against Ichthyophthirius multifiliis and in vivo effect of M. alternifolia for treating ichthyophthiriasis in one of the most important South American fish, Piaractus mesopotamicus (Holmberg), were evaluated. The in vitro test consisted of three EOs, each at concentrations of 57 μL L^?1, 114 μL L ^?1, 227 μL L^?1 and 455 μL L ^?1, which were assessed once an hour for 4 h in microtitre plates (96 wells). The in vitro results demonstrated that all tested EOs showed a cytotoxic effect against I. multifiliis compared to control groups (P < 0.05). The in vivo treatment for white spot disease was performed in a bath for 2 h day^?1 for 5 days using the M. alternifolia EO (50 μL L ^?1). In this study, 53.33% of the fish severely infected by I. multifiliis survived after the treatment with M. alternifolia (50 μL L ^?1) and the parasitological analysis has shown an efficacy of nearly 100% in the skin and gills, while all the fish in the control group died. Furthermore, the potential positive effect of M. alternifolia EO against two emergent opportunistic bacteria in South America Edwardsiella tarda and Citrobacter freundii was discussed.