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.     

Nanoemulsion from essential oil of Pterodon emarginatus (Fabaceae) shows in vitro efficacy against monogeneans of Colossoma macropomum (Pisces: Serrasalmidae)

Outbreaks of diseases pose a major threat to sustainable aquaculture development worldwide. Application of herbal products to combat parasitic diseases provides an alternative approach for sustainable aquaculture. This study investigated the in vitro antiparasitic effects of an oil‐in‐water nanoemulsion prepared using the essential oil from Pterodon emarginatus, against monogeneans infesting Colossoma macropomum. Gill arches from C. macropomum (47.6 ± 14.5 g and 13.5 ± 1.4 cm) that were naturally parasitized by Anacanthorus spathulatus, Notozothecium janauachensis and Mymarothecium boegeri were immersed in different dispersions of the P. emarginatus nanoemulsions (0, 50, 100, 200, 400 and 600 mg/L). The major compounds presented in the essential oil of P. emarginatus were β‐elemene, β‐caryophyllene and α‐humulene. Characterization of these nanoemulsions showed that they have a small mean droplet size and low polydispersity index, which is concordant with stable systems. In this in vitro trial, the P. emarginatus nanoemulsion concentrations of 100, 200, 400 and 600 mg/L presented 100% helminthic efficacy against monogeneans of the gills of C. macropomum. The highest two concentrations used (400 and 600 mg/L) were seen to immobilize the parasites after only 15 min. Therefore, it would be worthwhile testing these concentrations in therapeutic baths against monogeneans of C. macropomum.     

Nanoemulsions with oleoresin of Copaifera reticulata (Leguminosae) improve anthelmintic efficacy in the control of monogenean parasites when compared to oleoresin without nanoformulation

This study compared the in vitro anthelmintic activity of Copaifera reticulata oleoresin (200, 400, 600, 800 and 1,000 mg/L) and of nanoemulsions prepared with this oleoresin (50, 100, 150, 200 and 250 mg/L) against monogeneans on the gills of Colossoma macropomum. The major compounds present in the oleoresin of C. reticulata were γ-macrocarpene (14.2%), α-bergamotene (13.6%), β-selinene (13.4%) and β-caryophyllene (11.7%). All concentrations of the nanoemulsion and the oleoresin without nanoformulation showed anthelmintic efficacy against monogeneans, and higher concentrations led to more rapid parasite mortality. Structural damages to the tegument of the parasites exposed to C. reticulata oleoresin were observed with scanning electron microscopy. At two hours of exposure, fish showed 100% tolerance to all nanoemulsion concentrations used in the in vitro assays, whereas 100% mortality was shown in the fish exposed to the oleoresin without nanoformulation after one hour. The results of this study suggest that nanoemulsions with oleoresin of C. reticulata have advantages in the control and treatment of monogenean infections in C. macropomum when compared to the oleoresin without nanoformulation. In addition, since nanoemulsions with the C. reticulata oleoresin are safe to control monogeneans, the efficacy of these nanoformulations may be assayed in therapeutic baths to treat C. macropomum infected by monogeneans.     

Trichlorfon is effective against Dawestrema cycloancistrium and does not alter the physiological parameters of arapaima (Arapaima gigas): A large Neotropical fish from the Amazon

This study investigated the in vitro and in vivo efficacy of trichlorfon against Dawestrema cycloancistrium, as well as its physiological effects on arapaima. Naturally parasitized arapaima gill arches were exposed in vitro to 100, 250, 500 and 750 mg/L of trichlorfon and a control group (only distilled water), in triplicate. Parasites were monitored, and mortality was used to determine the median effective concentration (EC₅₀). The 750 mg/L concentration demonstrated 100% in vitro efficacy against D. cycloancistrium after 60 min, while the intermediate (500 mg/L) and the lowest (100 and 250 mg/L) tested concentrations were completely efficient after 90 and 130 min, respectively. The EC_50-1h of trichlorfon for D. cycloancistrium was determined at 171.73 mg/L. Parasitized arapaima juveniles were exposed to a control group and 150 mg/L of trichlorfon in triplicate. Fish were exposed to two therapeutic baths for 60 min with 24-h intervals between treatments. Therapeutic baths with 150 mg/L of trichlorfon were 92.99% effective against D. cycloancistrium and did not bring about haematological alterations (erythrogram, white blood cell count, thrombogram, plasma glucose and total proteins). Therefore, 150 mg/L of trichlorfon can be used in therapeutic baths to control and treat D. cycloancistrium infestations with no physiological impairments for arapaima.     

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.     

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.     

Evaluation of sodium percarbonate as a bath treatment for amoebic gill disease in Atlantic salmon

Amoebic gill disease (AGD), caused by Neoparamoeba perurans, is a major health challenge for Atlantic salmon aquaculture globally. While freshwater bathing for 2 hr is effective in reducing infection severity, there is need for more rapid and lower cost alternatives. To this end, a combination of sodium percarbonate (SPC) in freshwater was examined for its treatment efficacy. Initial in vitro studies showed a reduction in amoeba viability when exposed for 30 min to freshwater containing >500 mg/L SPC. Subsequently, AGD‐affected salmon were bathed for 30 min in 16°C freshwater containing 100, 500 or 1,000 mg/L SPC, or for 2 hr in 16°C freshwater to mimic industry practice. Treatment at the highest SPC concentration caused extensive gill damage and substantial mortality. Neither occurred to a significant extent at lower SPC concentrations. Gill pathology of surviving fish 10 days post‐treatment (dpt) was comparable to or more severe than pre‐treatment, and significantly (p < .001) more severe than in 2 hr freshwater bathed fish. N. perurans DNA was confirmed by qPCR in all treatment groups at 10 dpt. The data indicate that a 30‐min exposure to SPC in freshwater is not a suitable alternative to existing freshwater treatment of AGD.     

Effects of increasing concentrations of fine suspended sediment on the survival,growth, condition factor and gill condition of artificially reared juveniles of Oncorhynchus masou,Salvelinus leucomaenis and Plecoglossus altivelis

Little is known about the effect of chronic turbidity created by fine suspended sediment on the health and growth of artificially reared Oncorhynchus masou, Salvelinus leucomaenis and Plecoglossus altivelis—three important aquaculture species in Japan. This study compared survival rate, daily growth rate, condition factor (estimated from the ratio of standard length to weight) and gill condition among like‐sized juveniles exposed to fine suspended sediment (<2‐µm fraction) at concentrations of 0, 50, 100 and 500 mg/L, after 21 days. Even at 500 mg/L, there was almost no damage to the gill condition in all species. In O. masou, the daily growth rate was unexpectedly high in the 500‐mg/L treatment group, and condition factor was higher at sediment concentrations of 100 and 500 mg/L than at 0 mg/L. In S. leucomaenis, daily growth rate was significantly lower in the 500‐mg/L treatment group as compared with all other groups, but condition factor did not significantly differ among the treatments. The survival rates of O. masou and S. leucomaenis did not significantly differ among the treatment groups. In P. altivelis, neither survival rate, daily growth rate nor condition factor significantly differed among the treatments. The concentration of fine suspended sediment that affects the growth and condition factor of each of these species is likely related to their natural habitat.     

Efficacy and toxicity of praziquantel in helminth‐infected barbel (Barbus barbus L.)

This study evaluated efficacy and toxicity of the pyrazinoisoquinoline anthelmintic praziquantel (PZQ) in barbel infected with metacercariae of Diplostomum spathaceum and adult Pomphorhynchus laevis, and assessed antioxidant biomarkers and the lipid peroxidation response in juvenile barbel post‐treatment. The estimated 96‐hr LC50 of PZQ was 28.6 mg/L. For evaluation of efficacy, barbel naturally infected with D. spathaceum were exposed to a 10 and 20 mg/L PZQ 4‐day bath treatment. Both concentrations were 100% effective against D. spathaceum and significantly (p < .01) affected the activity of catalase, superoxide dismutase, glutathione reductase and glutathione‐S‐transferase as well as levels of reduced glutathione in liver and muscle. The efficacy of orally administered PZQ was assessed in adult barbel naturally infected with P. laevis. Fish were administered 10, 30 and 50 mg/kg of body weight and examined via gut dissection after 6 days. The 50 mg/kg dose significantly decreased the intensity of infection. Praziquantel is a feasible bath treatment for barbel infected with D. spathaceum and has potential for oral treatment of broodfish infected with P. laevis.     

Does Ralstonia eutropha,rich in poly‐β hydroxybutyrate (PHB), protect blue mussel larvae against pathogenic vibrios?

The natural amorphous polymer poly‐β‐hydroxybutyrate (PHB‐A: lyophilized Ralstonia eutropha containing 75% PHB) was used as a biological agent to control bacterial pathogens of blue mussel (Mytilus edulis) larvae. The larvae were supplied with PHB‐A at a concentration of 1 or 10 mg/L for 6 or 24 hr, followed by exposure to either the rifampicin‐resistant pathogen Vibrio splendidus or Vibrio coralliilyticus at a concentration of 10⁵ CFU/ml. Larvae pretreated 6 hr with PHB‐A (1 mg/L) survived a Vibrio challenge better relative to 24 hr pretreatment. After 96 hr of pathogen exposure, the survival of PHB‐A‐treated mussel larvae was 1.41‐ and 1.76‐fold higher than the non‐treated larvae when challenged with V. splendidus and V. coralliilyticus, respectively. Growth inhibition of the two pathogens at four concentrations of the monomer β‐HB (1, 5, 25 and 125 mM) was tested in vitro in LB₃₅ medium, buffered at two different pH values (pH 7 and pH 8). The highest concentration of 125 mM significantly inhibited the pathogen growth in comparison to the lower levels. The effect of β‐HB on the production of virulence factors in the tested pathogenic Vibrios revealed a variable pattern of responses.     

Evaluation of levamisole as a feed additive for growth and health management of hybrid striped bass (Morone chrysops × Morone saxatilis)

Three separate experiments were conducted with juvenile hybrid striped bass to determine the effects of levamisole as a feed supplement on growth performance, innate immunity and specific antibody responses, as well as resistance to Streptococcus iniae and Aeromonas hydrophila infection. A basal diet was formulated to contain 40% protein, 10% lipid and an estimated digestible energy level of 3.5 kcal/g. Four levels (100, 250, 500 and 1000 mg/kg) of levamisole were added to the basal diet in place of cellulose. After the 3-week feeding period in feeding trial 1, enhanced weight gain was generally observed in fish fed the diets supplemented with a low level (< 500 mg/kg) of levamisole. Dietary supplementation of levamisole at 100 mg/kg significantly (P < 0.05) enhanced growth and feed efficiency, compared to fish fed the basal diet. Fish fed diet supplemented with 1000 mg levamisole/kg diet showed chronic toxicity signs of inferior growth, reduced feed intake and feed efficiency. Although the extracellular superoxide anion production of head kidney cells, hematocrit, serum lysozyme and peroxidase were not significantly affected by dietary treatments, intracellular superoxide anion production of head kidney macrophages of fish fed 250 mg levamisole/kg diet was significantly (P < 0.05) higher than that of fish fed the other four diets. The hypothesized benefits of dietary supplementation of levamisole in reducing mortality from infection of pathogenic bacteria including S. iniae and A. hydrophila were not proven after two separate 3-week feeding trials and disease challenges in the present study. Agglutinating antibody titers of individual fish were highly varied and not significantly influenced by dietary levamisole at the tested levels.     

Effects of flow velocity on growth,metabolism and nitrogen and phosphorus budget of Haliotis discus hannai Ino in a recirculating aquaculture system

To study the effects of different flow velocities on growth, metabolism and nitrogen (N) and phosphorus (P) budget of Haliotis discus hannai Ino in a recirculating aquaculture system, abalones (shell length: 45.71 ± 3.09 mm, body weight: 8.62 ± 0.75 g) were cultured at three different flow velocities with four replicates each: high (400 L/hr), medium (300 L/hr) and low (200 L/hr). At the end of the 70 days experiment, the survival rate, food intake and P utilization rate of abalones in the 200 L/hr group were significantly lower than those in any other group (p < .05), whereas the total ammonia nitrogen and nitrite nitrogen concentrations in the water were significantly higher than those in any other group (p < .05). The body weight specific growth rate, food conversion efficiency, abalone‐harvested N and P outputs and N utilization rate were ranked as follows: 300 > 400 > 200 L/hr (p < .05). In the 300 L/hr group, the activities of hexokinase, pyruvate kinase and lactate dehydrogenase were significantly lower than those in the 400 L/hr group, whereas the activity of succinate dehydrogenase was significantly higher than that in the other two groups (p < .05). Aerobic metabolism was the main source of energy acquisition for abalones in the 300 L/hr group. In the 400 L/hr group, the expression levels of manganese superoxide dismutase and thioredoxin peroxidase significantly increased from Day 35 to Day 70, but they still were significantly lower than the values in the 200 L/hr group (p < .05). In the 200 L/hr group, the expression levels of catalase and heat shock protein 70 were significantly higher than those in any other group, and they significantly increased from Day 35 to Day 70 (p < .05). These results indicate that maintaining flow velocity at 300 L/hr will minimize oxidative damage due to deterioration of water quality, reduce the energy consumption required for organisms to resist the impact of water flow, improve the N and P utilization rates of the recirculating aquaculture system, and reduce the cost of culturing abalones.     

Assessment of induction and recovery times of anaesthesia in Astyanax bimaculatus using 2‐phenoxyethanol and the essential oils of Melaleuca alternifolia and Ocimum gratissimum

This study aimed to determine the lowest effective concentration (LEC) of 2‐phenoxyethanol (2‐Phe) and essential oils (EOs) of Melaleuca alternifolia and Ocimum gratissimum able to induce anaesthesia in Astyanax bimaculatus. LEC was assumed to be the required concentration to carry out anaesthesia in less than 3 min with recovery in <5 min and sufficient to carry out routine aquaculture procedures. The following concentrations were tested: 2‐Phe (100, 200, 300, 400, 500 and 600 µl/L); M. alternifolia (200, 300, 400, 500 and 600 µl/L); and O. gratissimum (10, 20, 30, 40, 50, 60, 70, 100, 150, 250 and 300 µl/L). Results show that all three agents evaluated have effective anaesthetic properties for use in Astyanax bimaculatus with LEC for 2‐Phe, M. alternifolia and O. gratissimum of 200, 300 and 60 µl/L respectively.     

Finishing plant diet supplemented with microalgae meal increases the docosahexaenoic acid content in Colossoma macropomum flesh

The need for a sustainable aquaculture is increasing the use of plant ingredients in replacement to fishmeal and fish oil in diets for tambaqui Colossoma macropomum, which is leading to not detectable levels of docosahexaenoic acid (DHA) in its flesh. We evaluated the effect of a finishing plant diet supplemented with 5% of microalgae meal from Schizochytrium sp. (MD) on tambaqui growth, on proximal composition and fatty acid content of its flesh, comparing it to a non‐supplemented diet. One hundred and sixty‐two fish (489.67 g) were distributed into six tanks (2,000 L) and fed the experimental diets for a 90‐day period. Three fish per tank were euthanized for analyses every 15 days. The MD did not affect the growth and proximal composition of fish flesh. The MD increased the DHA content (from 14.81 to 38.60 mg/g of lipids) and the n‐3:n‐6 ratio (from 0.16 to 0.51) in the flesh of fish, beginning on the 15th day and reaching the highest DHA content on the 71st day (39.81 mg/g of lipids). We recommend C. macropomum to be fed with a finishing diet supplemented with microalgae meal for 71 days before slaughter to improve the DHA content and n‐3:n‐6 ratio in the flesh.     

Dietary Levamisole as Immunostimulant for Striped Surubim,Pseudoplatystoma reticulatum

Immunostimulants are better suited for disease control than using drugs and chemicals as remedies in animal production. This study evaluates effects of dietary levamisole on growth and immunological parameters response of Pseudoplatystoma reticulatum. A trial was set up in a completely randomized design with six treatments – 0.0, 50, 100, 200, 300, and 500 mg levamisole per kg of feed (n = 3) – fed until apparent satiation, twice a day, for 60 d to striped surubim (77.68 ± 2.5 g) randomly stocked in 18 plastic tanks (300 L; 10 fish per tank). Dietary levamisole affected lysozyme activity and concentration, the best effects were recorded for fish fed 247 mg levamisole per kg of feed, that is, levamisole positively influenced innate immune system of striped surubim. However, dietary levamisole did not affect serum protein, albumin and globulins, as well as leukocyte respiratory burst activity and leukocyte proliferation. No differences in growth parameters were recorded. Dietary levamisole improved immunological responses of striped surubim.     

Hatching rate of Chinese sucker (Myxocyprinus asiaticus Bleeker) eggs exposed to total dissolved gas (TDG) supersaturation and the tolerance of juveniles to the interaction of TDG supersaturation and suspended sediment

Flood discharge containing excessive suspended sediments (SS) can result in total dissolved gas (TDG) supersaturation downstream of dams. It has been confirmed that high TDG levels and excessive SS cause serious threats to fish survival. To explore the effects of TDG supersaturation on the hatchability of Chinese sucker (Myxocyprinus asiaticus Bleeker) eggs, eggs were exposed to 100%, 125%, 130% and 135% TDG supersaturated water. The results showed that the hatching rates were 78.25%, 75.35%, 71.30% and 67.50% at the 100%, 125%, 130% and 135% TDG respectively. The hatching rate decreased with increasing TDG levels. Furthermore, acute lethality experiments were carried out to investigate the interaction of TDG supersaturation and SS on Chinese sucker. Half‐year‐old juveniles were exposed to TDG supersaturated water with SS concentrations of 0, 200, 600 and 1,000 mg/L, and the TDG levels were 125%, 130%, 135% and 140% respectively. The fish exhibited abnormal behaviours and signs of gas bubble disease. Mortality increased with increasing TDG levels and SS concentrations. The Chinese suckers only survived for 2.5 hr at 140% TDG and 1,000 mg/L SS. Moreover, an increase in SS in TDG supersaturated water could cause a decrease in the median lethal time (LT₅₀). When the TDG level reached 140%, the LT₅₀ declines significantly at 600 and 1,000 mg/L SS. The interaction of TDG supersaturation and SS was significant at LT₅₀ (F = 22.81, df = 9, p < 0.001). The results indicate that the combined effect of TDG and SS could be great on Chinese sucker.     

Effects of linalool on physiological responses of Cyprinus carpio (Linnaeus, 1758) and water physico‐chemical parameters during transportation

This study investigated the potential of linalool as an anaesthetic during transportation of common carp, Cyprinus carpio. The fish were transported at a loading density of ~103 g/L for 3 hr in 12 plastic bags (3 L water and 6 L pure oxygen) divided into four triplicated treatments: control (without linalool), L50 (50 mg linalool/L), L100 (100 mg linalool/L) and L200 (200 mg linalool/L). After 3‐hr transportation, serum physiological responses and water physico‐chemical parameters were compared among the treatments. Results showed that water total and un‐ionized ammonia increased and dissolved oxygen decreased in all treatments after transportation. Water total ammonia and dissolved oxygen levels in the linalool‐treated bags were significantly lower than the control. After transportation, the control fish showed significant elevation in serum cortisol and glucose levels; however, the levels were significantly lower than the linalool‐treated fish. All fish showed similar serum lactate levels, significantly lower than the value obtained before transportation. The control fish showed serum urea levels similar to the level obtained before transportation; however, the linalool‐treated fish had significantly higher serum urea levels compared to the control and before transportation levels. After transportation, all fish had decreased serum chloride and sodium levels compared to the before transport level. Overall, the present results showed that linalool is not beneficial for carp transportation in plastic bags, because it reacts with water oxygen, increases stress in fish, interferes with ammonia excretion and has no benefits in preventing ion loss.     

Further evaluation of the efficacy of emamectin benzoate for treating Pseudocapillaria tomentosa (Dujardin 1843) in zebrafish Danio rerio (Hamilton 1822)

Pseudocapillaria tomentosa is a pathogenic nematode parasite, causing emaciation and severe inflammatory lesions in the intestines in zebrafish Danio rerio (Hamilton 1822). Emamectin benzoate is commercially available analogue of ivermectin used for treating salmon for sea lice, under the brand name SLICE^®, and we have used this for treating zebrafish with the P. tomentosa. Here, SLICE^®, 0.2 per cent active emamectin benzoate, was used for oral treatments at 0.35 mg emamectin benzoate/kg fish/day for 14 days starting at 7 days post‐exposure (dpe). Another experiment entailed initiating treatment during clinical disease (starting at 28 dpe). Early treatment was very effective, but delaying treatment was less so, presumably due to inappetence in clinically affected fish. We evaluated emamectin benzoate delivered in water, using Lice‐Solve? (mectinsol; 1.4% active emamectin benzoate) in two experiments. Application of four 24‐hr treatments, space over 7 days was initiated at 28 dpe at either 0.168 or 0.56 mg emamectin benzoate/L/bath, and both treatments completely eradicated infections. This was 3 or 10 times manufacture's recommended dose, but was not associated with clinical or histological side effects.     

Disinfection of almaco jack (Seriola rivoliana Valenciennes) eggs: Evaluation of three chemicals

Almaco jack (Seriola rivoliana Valenciennes) is an excellent candidate for aquaculture due to its fast growth rate and high market value. While S. rivoliana have adapted well to captivity, survival at early life stages can be improved to increase profitability during production. A wide range of variables cause larval mortalities but high bacterial loads in rearing tanks are often correlated with these losses. The aim of this study was to investigate the effect of egg disinfection on bacterial load and hatch rate of S. rivoliana. Disinfectants tested included formalin (F100 and F200; 100 and 200 mg/L, respectively, for 60 min), hydrogen peroxide (HPO; 300 mg/L for 10 min) and peracetic acid/hydrogen peroxide (PAA/HPO; 15.7 mg/L/39.6 mg/L for 1 min). Concentrations and contact times were determined based on current use in marine aquaculture and preliminary trials. Eggs treated with HPO and F100 had significantly higher hatch rates than the untreated control group. All treatments significantly decreased total Vibrio counts compared to untreated eggs; however, total bacterial counts were only decreased following treatments with PAA/HPO and F200. To prevent egg mortality due to bacterial overgrowth, consideration should be given to the use of surface disinfection using HPO or F100. Future studies should investigate the use of peracetic‐based products at lower doses.     

A fructose‐based extender protects Colossoma macropomum spermatozoa against chilling injuries

Our study assessed the efficiency of a formulated new extender in maintaining viability and morphological integrity of Colossoma macropomum spermatozoa under chilling storage. Semen was diluted in the test extender and BTS? (Beltsville Thawing Solution) and exposed to a short‐term storage at 4.6 ± 0.6°C for 96 hr. Both extenders were able to maintain 17% ± 8% motile spermatozoa by the end of experiment. Sperm dilution in test extender did not affect the morphologically normal cells (61% ± 6%) up to 48 hr of chilling, being higher than in BTS? (50% ± 6%) (p < 0.05). After 96 hr, samples kept in the test extender had 50% of normal spermatozoa, whereas those kept in BTS? presented only 38% of normal cells. Chilling storage increased the incidence of cells with strongly coiled flagella in BTS^?. Our study is the first to evaluate in detail the spermatozoa morphology as indicative of C. macropomum semen viability. The new extender was able to protect the spermatozoa against increase in coiled flagellum injuries.