Albendazole,levamisole and ivermectin are effective against monogeneans of Colossoma macropomum (Pisces: Serrasalmidae)

This study evaluated the efficacy of albendazole, ivermectin, levamisole, mebendazole and praziquantel on monogeneans of Colossoma macropomum, based on in vitro and in vivo assays. In vitro assays indicated that albendazole (500, 100, 1,500 and 2,000 mg/L), ivermectin (200, 250, 300 and 350 mg/L) and levamisole (50, 75, 100 and 125 mg/L) were 100% effective against Anacanthorus spatulatus, Notozothecium janauachensis, Mymarothecium boegeri and Linguadactyloides brinkmanni, while mebendazole (125, 150, 175 and 200 mg/L) and praziquantel (5, 10, 15 and 20 mg/L) were ineffective. Fish mortality in 24 hr therapeutic baths with 500 mg/L of albendazole was 6.6%, but the behaviour of the animals remained unchanged, while 200 mg/L of ivermectin caused lethargy, signs of hypoxia and 100% mortality within 2 hr, and 125 mg/L of levamisole caused no mortality. The efficacy of 500 mg/L of albendazole was 48.6% in the 24 hr baths, while that of 125 mg/L levamisole was 88.2%. Although ivermectin showed in vitro efficacy, the lowest concentration used in baths was highly toxic to fish. Therefore, we recommend the use of 125 mg/L of levamisole to control and treat monogenean infestations on C. macropomum in fish farming.     

Effects of a nanoemulsion with Copaifera officinalis oleoresin against monogenean parasites of Colossoma macropomum: A Neotropical Serrasalmidae

Monogeneans are ectoparasites that may cause losses in production and productivity in the aquaculture of Colossoma macropomum. Chemotherapeutics used in aquaculture usually have major adverse effects on fish; hence, the use of essential oils has been considered advantageous, but these are not soluble in water. Thus, the use of nanostructures to enhance water solubility of compounds and improve bioactivity may be very promising. This study investigated the antiparasitic activity of nanoemulsion prepared with Copaifera officinalis oleoresin (50, 100, 150, 200 and 300 mg/L), against monogenean parasites from the gills of C. macropomum. The particle size distribution and zeta potential suggested that a potentially kinetic stable system was generated. The nanoemulsion from C. officinalis oleoresin achieved high efficacy (100%) at low concentrations (200 and 300 mg/L) after 15 min of exposure. This was the first time that a nanoemulsion was generated from C. officinalis oleoresin using a solvent‐free, non‐heating and low‐energy method. Moreover, this was the first time that an antiparasitic against monogeneans on fish gills, based on nanoemulsion of C. officinalis oleoresin, was tested.     

Essential oil of Lippia grata (Verbenaceae) is effective in the control of monogenean infections in Colossoma macropomum gills,a large Serrasalmidae fish from Amazon

This study investigated the efficacy of Lippia grata essential oil (EO) against monogeneans of Colossoma macropomum and effects on the haematology after immersion baths. In the in vitro assays, the efficacy of 100, 250, 350 and 700 mg/L of L. grata EO and two controls were tested, of which one control was with the cultivation tank water and the other was with the tank water and 70% ethyl alcohol. Composition of majority bioactive compounds in the EO was carvacrol (48.12%), p‐cymene (24.39%) and γ‐terpinene (2.49%). Anacanthorus spathulatus, Notozothecium janauachensis, Mymarothecium boegeri and Linguadactyloides brinkmanni obtained of fish gills showed 100% immobilization in the in vitro assays when exposed to 700 and 350 mg/L after 30 min and 2 hr respectively. Fish showed tolerance to 700 mg/L of the EO, which was used in therapeutic in three consecutive baths, which caused lamellar hyperplasia in C. macropomum gills. The 700 mg/L of the EO in the therapeutic baths showed an efficacy of 95.1% against the monogeneans and caused decrease in haemoglobin, whereas control fish showed a reduction in mean corpuscular haemoglobin concentration. Results indicated that 700 mg/L of L. grata EO is a safe and effective concentration to treat C. macropomum infected with monogeneans.     

Lethal concentration of Cymbopogon citratus (Poaceae) essential oil for Dolops discoidalis and Argulus sp. (Crustacea: Argulidae)

This study investigated the acute toxicity (LC_50-24 hr) effects of the essential oil of Cymbopogon citratus for adult Argulus sp. and Dolops discoidalis, before and during oviposition. In vitro acute toxicity (LC_50-24 hr) was tested using 20, 40, 60, 80, 100, 120, 140 and 160 μg/L of C. citratus essential oil, and two control groups (one with cultivation tank water and one with cultivation tank water + alcohol) were used. Specimens of Argulus sp. and D. discoidalis submitted to acute toxicity were evaluated using histological procedures. The major chemical compounds of C. citratus essential oil were geranial (47.5%), neral (35.6%) and myrcene (6.7%). The LC_50-24 hr for Argulus sp. adults was 67.97 μg/L, while for D. discoidalis it was 59.55 µg/L. In the oviposition of both species of argulids, maximum mortality began with treatments of 140 μg/L, while the LC_50-24 hr for Argulus sp. and D. discoidalis was 83.98 μg/L and 82.48 μg/L, respectively. In both argulid species exposed to C. citratus essential oil, morphological alterations were observed only in the eyes, and they occurred in the ommatidium and rhabdomeres and were dependent on the concentration of C. citratus essential oil and the parasite species.     

Gill amoebae from freshwater rainbow trout (Oncorhynchus mykiss): In vitro evaluation of antiparasitic compounds against Vannella sp.

Gill parasitic infections challenge farming of rainbow trout (Oncorhynchus mykiss, Walbaum) in freshwater facilities. Apart from flagellates (Ichthyobodo, (Pinto) and ciliates (Ichthyophthirius (Fouquet), Ambiphrya (Raabe), Apiosoma (Blanchard), Trichodinella (Sramek-Husek) and Trichodina (Ehrenberg)), we have shown that amoebae are prevalent in Danish trout farms. Gills were isolated from farmed rainbow trout in six fish farms (conventional and organic earth pond and recirculated systems) and placed on non-nutrient agar (NNA) moistened with modified Neff's amoeba saline (AS) (15°C). Gill amoebae from all examined fish colonized the agar and were identified based on morphological criteria showing species within the genera Trinema (Dujardin) (family Trinematidae), Vannella (Bovee) (family Vannellidae). In addition, hartmannellid amoebae were recorded. We established a monoculture of Vannella sp., confirmed the genus identity by PCR and sequencing and performed an in vitro determination of antiparasitic effects (dose–response studies) of various compounds including sodium chloride (NaCl), hydrogen peroxide, peracetic acid, formalin, aqueous garlic and oregano extracts and a Pseudomonas H6 surfactant. All amoebae were killed in concentrations of 16.90 mg/ml (garlic), 17.90 mg/ml (oregano), NaCl (7.5 mg/ml), hydrogen peroxide (100 µg/ml), peracetic acid (0.03 µg/ml), formaldehyde (25 µg/ml) and the Pseudomonas H6 surfactant (250 µg/ml).     

Effect of Dimethyl sulfoxide (DMSO) and egg yolk on sperm motility,fertility and hatching rates of depik Rasbora tawarensis (Pisces: Cyprinidae) eggs after short‐term cryopreservation

Cryoprotectant is the crucial factor in the cryopreservation process. In general, there are two types of cryoprotectant, permeating and non‐permeating cryoprotectants. Dimethyl sulfoxide (DMSO) and egg yolk are common permeating and non‐permeating cryoprotectants respectively. Hence, the objective of the present study was to determine the best proportion of DMSO and egg yolk for the cryopreservation of Rasbora tawarensis sperm. A completely randomized experimental design was used in this study which involves two types of cryoprotectant and their combination at different concentrations, namely 5% DMSO, 5% egg yolk, 5% DMSO + 5% egg yolk and 2.5% DMSO + 2.5% egg yolk. Every treatment was conducted in three replicates. Combination of 5% DMSO + 5% egg yolk gave the best results cryoprotectant treatment had significant effects on sperm motility, fertilization and hatching rate of the R. tawarensis eggs (p < .05). It is concluded that the best proportion of cryoprotectants for sperm cryopreservation in this species is 5% DMSO + 5% egg yolk.     

The in vitro efficacy of oxytetracycline against re‐isolated pathogenic Aeromonas hydrophila carrying the cytolytic enterotoxin gene through hybrid catfish,Clarias macrocephalus (Günther, 1864) × Clarias gariepinus (Burchell, 1822) in Thailand

Aeromonas hydrophila is a pathogen infecting farmed hybrid catfish, Clarias macrocephalus (Günther, 1864) × Clarias gariepinus (Burchell, 1822) which incurs substantial economic losses in Thailand. The study aimed at a genetic tracking of A. hydrophila infection and the in vitro assessment of the efficacy of antibiotics against its virulent strains. Five clinical strains from catfishes and Nile tilapia were employed. They were 3‐passage re‐isolated through healthy hybrid catfish and the cytolytic enterotoxin gene (AHCYTOEN) of individuals was traced. Each of the re‐isolates at a dose of ~6.67 × 10⁵ CFU/g was intraperitoneally injected into ~15 g‐healthy hybrid catfish and their pathogenicity were observed for 7 days. It was found that AHCYTOEN was carried over whereas typical signs of motile aeromonas septicaemia were found in the specimens. The bacterial strains of Nile tilapia origin did not induce mortality but those of catfish origins (80%–100% rate of mortality). The strains were susceptible to the tetracycline antibiotics, and oxytetracycline produced MIC₅₀ and MBC as low as 0.007–0.031 μg/ml and 1–8 μg/ml respectively. As oxytetracycline specifically inhibited pathogenic A. hydrophila in vitro, it is recommended that an appropriate dosage regimen of the drug should be established.     

Bioassay‐guided isolation and identification of active compounds from Macleaya microcarpa (Maxim) Fedde against fish pathogenic bacteria

Four alkaloids (Sanguinarine, 6‐Methoxyl‐dihydro‐chelerythrine, Cryptopine and β‐Allocryptopine) were isolated from aerial parts of Macleaya microcarpa (Maxim) Fedde using bioassay‐guided isolation method, and the inhibitory activity of ethanolic extract, various fractions and these four alkaloids against four fish pathogenic bacteria (Aeromonas hydrophila, Aeromonas salmonicida, Vibrio anguillarum and Vibrio harveyi) was assessed in vitro using the agar dilution method and the microdilution assay method respectively. A. hydrophila was the most sensitive strain to all the tested compounds. Minimum inhibitory concentration (MIC) values were lower for sanguinarine against all tested Gram‐negative strains than other three alkaloids, with MIC values of 12.5 mg L^?1 for A. hydrophila and 50 mg L^?1 to other pathogenic bacteria. Followed by 6‐methoxyl‐dihydro‐chelerythrine, which showed considerable antibacterial activity with MIC values of 80 mg L^?1 for A. hydrophila, 100 mg L^?1 for V. harveyi, and 125 mg L^?1 for both V. anguillarum and A. salmonicida. Cryptopine and β‐allocryptopine revealed similar inhibitory activity with MIC values of 100 mg L^?1 for A. hydrophila and 200 mg L^?1 for other three bacterial species. These finding provided evidence that extract, as well as isolated compounds from M. microcarpa might be potential sources novel antibacterial agents for the treatment of fish infectious diseases.     

Effect of combined fish meal : soybean meal ratio,vitamin C and fish oil supplementations in diet on the growth and reproduction of red swamp crayfish,Procambarus clarkii (Crustacea: Decapoda)

Fish meal (FM), vitamin C (VC) and fish oil (FO) play important roles in the growth and reproduction in aquatic animals. To determine the optimum dietary combinations of FM:SM (soybean meal) ratio, VC and FO supplementations for the growth and reproduction of female crayfish Procambarus clarkii, we used the L9 (3³) orthogonal array, which arranged three factors varied at three levels (FM:SM at 1:3, 1:2 and 1:1.5; VC supplementation at 0.02%, 0.04% and 0.06%; FO inclusion at 1.90%, 2.85% and 3.80%). Range analysis showed that the highest weight gain (WG) of the crayfish occurred at FM:SM of 1:1.5, VC supplementation of 0.04% and FO inclusion of 1.90%, and the maximum gonadosomatic index (GSI), egg laying amount (ELA) and spawning rate (SR) were obtained at FM:SM 1:1.5, VC supplementation 0.06% and FO inclusion 3.80%. Analysis of variance indicated that all the three factors had no significant effects on WG. Fish oil supplementation significantly affected GSI, ELA and SR (P<0.05), while FM:SM did not significantly influence any of these indicators. Taking the cost into consideration, we suggest that a dietary FM:SM of 1:3, VC supplementation 0.06% and FO inclusion 3.80% might be the optimal combinations for the reproduction of P. clarkii.     

The olive oil by‐product in ‘rainbow trout Onchorynchus mykyss (Walbaum)’ farming: productive results and quality of the product

The aim of this work is to investigate olive oil by‐product [vegetation water (VW)] inclusion in rainbow trout diet and its effect on the productive traits and the quality of the product. Two levels of VW inclusion were used and one control group was included. Fish diets were isonitrogeonous (crude protein 40%) and isoenergetic (18 MJ kg^?1 DM). Two thousand and four hundred rainbow trout were used. An in vivo digestibility experiment was performed in order to determine diets’ digestibility. All the fish diets and fillets were analysed to determine the proximate and fatty acid composition. On final fish fillet, lipid oxidation was determined at 0, 24, 48, 72, 96 and 192 h of storage using the 2‐thiobarbituric acid method (thiobarbituric acid reactive substances). Aroma analyses on the final cooked and raw fillet were performed using an electronic nose. The VW inclusion partially reduced protein digestibility. The fish growth varied between 1.08% and 1.1% day. The supplemental level of VW led to a better antioxidant status of fish fillet, in particular, in the fillet sample after 72 h of fillet conservation. Principal component analysis in raw and cooked fish fillet indicates that the VW inclusion in the fish diet led to aroma modification on fish fillet.     

Early development of the silver catfish Rhamdia quelen (Quoy & Gaimard, 1824) (Pisces:Heptapteridae) from the São Francisco River Basin, Brazil

The silver catfish, Rhamdia quelen , is endemic to North, Central and South America with high aquaculture potential and wide acceptance in the market. Breeder fish were subjected to induced reproduction through hypophysation using a crude common carp pituitary extract. Egg characteristics, oocyte surface ultrastructure and histology of larval ontogenesis until whole yolk resorption were described for the first time for this species. Oocytes and semen were obtained by manual extrusion, and fertilization was conducted using the dry method. After fertilization, eggs were kept in incubators at 24 °C. The embryonic development was monitored using a stereomicroscope every 10 min until hatching. To analyse the larval development, larvae samples were collected from incubators daily until the fifth day, fixed in Bouin's fluid and subjected to routine histological techniques. The oocyte extrusion occurred 8 h after the second hormone dose at 26 °C. The oocytes were spherical, non-adhesive and yellow, with a diameter of 1471.75±47.63 μm. Scanning electron microscopy revealed a thin jelly coat covering the zona radiata in the animal pole around the micropyle. The blastopore closure occurred within 8 h after fertilization, and the fertilization rate was 79.9±5.2% at 24 °C. Embryonic development was completed within 25 h 30 min after fertilization. The complete resorption of the yolk and the formation of the digestive system organs and the mouth opening occurred on the fifth day, indicating a need for exogenous feeding. The results of this study provide information important for improvement in R. quelen culture and management.     

In vitro and in vivo antifungal activity of Satureja cuneifolia ten essential oil on Saprolegnia parasitica strains isolated from rainbow trout (Oncorhynchus mykiss,Walbaum) eggs

In the present study, the chemical composition and the antifungal properties against Saprolegnia parasitica (in vitro and in vivo) of the essential oils of thyme (Satureja cuneifolia) from Mediterranean region of Turkey were evaluated for the first time. The composition of oils was analysed using gas chromatography/mass spectrometry (GC/MS). The major constituents of oil of S. cuneifolia were cavracrol (46,84%) and cymene (16.90%). Antifungal effects of S. cuneifolia essential oil against S. parasitica strains (A1 and E1) were detected by disc diffusion and tube dilution assays. The antifungal effect of S. cuneifolia was determined to be stronger against S. parasitica E1 isolate (MIC 50 μL mL^?1, MLC 250 μL mL^?1) compared with S. parasitica A1 isolate (MIC 50 μL mL^?1, MLC 500 μL mL^?1). Following in vitro assays, effective doses of S. cuneifolia for disease control in rainbow trout eggs experimentally infected with S. parasitica were investigated. For this aim, infected eggs were treated with the essential oil (0, 5, 10, 20 and 50 ppm) during incubation period (21 days) after fertilization. Formalin (5 mL L^?1) was used as positive control. Hatching rate of eggs at the end of incubation period were calculated. The highest hatching rates were recorded in S. parasitica E1 strain at 5 and 10 ppm concentrations of S. cuneifolia and in S. parasitica A1 strain at 10 and 20 ppm (P < 0.05).     

A survey of the distribution of the PKD‐parasite Tetracapsuloides bryosalmonae (Cnidaria: Myxozoa: Malacosporea) in salmonids in Norwegian rivers – additional information gleaned from formerly collected fish

The malacosporean Tetracapsuloides bryosalmonae was detected in kidneys from Atlantic salmon parr in 64 of 91 sampled Norwegian rivers. Using real‐time PCR, this parasite was found to be present in Atlantic salmon parr in rivers along the whole coast, from the northernmost and southernmost areas of the country. In addition, T. bryosalmonae was found in kidneys from brown trout parr in 17 of 19 sampled rivers in south‐east Norway, and in Arctic charr sampled in the River Risfjordelva, located at the northernmost edge of the European mainland. In conclusion, T. bryosalmonae has a widespread distribution in salmonids in Norwegian watercourses. Proliferative kidney disease (PKD) caused by T. bryosalmonae and PKD‐induced mortality has been observed in salmonids in several Norwegian rivers and it can be speculated that more PKD outbreaks will occur as a result of climate change.     

Silver catfish Rhamdia quelen immersion anaesthesia with essential oil of Aloysia triphylla (L'Hérit) Britton or tricaine methanesulfonate: effect on stress response and antioxidant status

Responses to anaesthesia with essential oil (EO) of Aloysia triphylla (135 and 180 mg L^?1) and tricaine methanesulfonate (MS222) (150 and 300 mg L^?1) were assessed in silver catfish. Exposure to the anaesthetics elicited a stress response in the species. In the case of MS222, it was displayed as a release of cortisol into bloodstream, elevation in hematocrit and plasma ion loss. The EO presented cortisol‐blocking properties, but increased haematocrit and disturbances of hydromineral balance were observed. Liver antioxidant/oxidant status of EO and MS222‐anaesthetized silver catfish was also estimated. The synthetic anaesthetic induced lipoperoxidation, notwithstanding increased catalase contents, whereas the naturally occurring product was capable of preventing the formation of lipid peroxides, possibly due to combined actions of catalase and glutathione‐S‐transferase. Anaesthetic efficacy was also tested via induction and recovery times. Overall, the promising results obtained for the physiological parameters of the EO‐treated fish counterbalanced the slight prolonged induction time observed for 180 mg L^?1. As for 135 mg L^?1, both induction and recovery times were lengthy; despite that, the EO was able to promote oxidative protection and mitigate stress. None of the MS222 concentrations prompted such responses concomitantly.     

Chemical,bioactive properties and in vitro digestibility of spray‐dried fish silages: Comparison of two discard fish (Equulites klunzingeri and Carassius gibelio) silages

The fermented fish silages produced with Streptococcus spp., Lactobacillus brevis, Lactobacillus plantarum, Pediococcus acidilactici and Enterococcus gallinarum, and formic acid silages were compared for production of two discard fish silages (Equulites klunzingeri and Carassius gibelio). The E/NE ratio of spray‐dried fish silages was determined in range of 0.80–1.10 for E. klunzingeri and 0.80–0.90 for C. gibelio silages. Pediococcus acidilactici and En. gallinarum groups had greater antioxidant activity than other silage groups. The DPPH radical scavenging ability was found as 6.14%–14.71% and 6.99%–13.36% for E. klunzingeri and C. gibelio silages, respectively. The OMD, ME and NE_L values were determined in range of 69.74%–80.08%, 6.38–8.65 MJ/kg DM and 6.45–7.49 MJ/kg DM, respectively for spray‐dried E. klunzingeri silages and 81.18%–86.62%, 8.97–9.61 MJ/kg DM and 7.61–8.08 MJ/kg DM, respectively, for spray‐dried C. gibelio silages. According to the nutritional and chemical evaluation, spray‐dried fish silages have great potential as a feed components because of high rate of digestibility and nutritious components.     

Effects of dietary levels of essential oil (EO) extract from bitter lemon (Citrus limon) fruit peels on growth,biochemical, haemato‐immunological parameters and disease resistance in Juvenile Labeo victorianus fingerlings challenged with Aeromonas hydrophila

Essential oils (EOs) are used in the food industry because of their biological activity. We evaluated the effects of administration of essential oil (EO) extracted from bitter lemon (Citrus limon) fruit peels on the growth performance, biochemical, haemato‐immunological parameters and possible disease resistance in fingerlings (4 weeks old) Labeo victorianus challenged with Aeromonas hydrophila. Fish were divided into five groups and fed diets supplemented with C. limon fruit peels EO extract at 1%, 2%, 5% and 8% [as fed basis] and treatment compared with control group fed diet without C. limon fruit peels EO extract. The experiment was executed in triplicate. Concentration of plasma cortisol, glucose, triglyceride and cholesterol decreased while that of total protein and albumin increased as dietary inclusion of the EO extract of C. limon fruit peels was increased from 2% to 5%. Meanwhile haemato‐immunological parameters including red blood cell (RBC), white blood cell (WBC) counts, haematocrit (Htc), mean cell haemoglobin (MCH), mean cell haemoglobin concentration (MCHC) and neutrophiles increased with increasing dietary inclusion from 1% to 5% inclusion of C. limon fruit peels EO extract. Serum immunoglobulins, lysozyme activity and respiratory burst increased with increasing dietary levels up to 5% inclusion of EO extract of C. limon fruit peels. We demonstrate that formulation of feeds by incorporating upto 5% the EO extract from C. limon fruit peels significantly improved biochemical, haematological and immunological response in juvenile fish resulting to lower mortality than the untreated groups and appear to be effective antibacterial against A. hydrophila.     

Replacement of fishmeal by soya protein concentrate in the diets of Totoaba macdonaldi (Gilbert, 1890) juveniles: effect on the growth performance,in vitro digestibility,digestive enzymes and the haematological and biochemistry parameters

The potential of soya protein concentrate (SPC) as an alternative protein source in diets for Totoaba macdonaldi juveniles was evaluated. Seven isonitrogenous and isolipidic diets were formulated containing 15–100% SPC (SPC15, SPC30, SPC45, SPC60, SPC75, SPC90 and SPC100) to replace fishmeal (FM‐protein), and a FM‐protein‐based diet without SPC was used as a reference diet (RD). Each diet was randomly assigned to triplicate groups of 20 totoaba (50 ± 1.0 g) and was fed twice daily to apparent satiation. After 60 days of experiment, effects on totoaba were evaluated. Growth performance in fish fed RD to SPC45 was similar. The maximum FM‐protein replacement for weight gain (g kg ABW^?1 day^?1) was estimated to be 34.17% using a broken‐line model. In vitro digestibility of fish fed RD, SPC15 and SPC30 was similar. Trypsin activity was higher in fish fed the RD (9.38 ± 0.52 mU × 10^?3 mg protein^?1. Chymotrypsin activity was similar in fish fed RD, SPC15 and SPC30. Activity of alkaline protease and phosphatase was similar in fish fed RD, SPC15 and SPC30. Red blood cells (RBC) were the highest in fish fed RD to SPC60. Fish fed the RD, SPC15 and SPC30 present the highest haematocrit (HT) and haemoglobin (HB) content. Diets SPC90 and SPC100 presented similar MCV and MCHC. Total protein (TP) profile overall decreased in fish fed SPC90 and SPC100, pointing to nutritional hypoproteinemia due to deficient digestion and absorption. These results indicate that SPC‐based diets could be used efficiently by totoaba with FM‐protein replacement of less than 45%.