Seeds of Cucurbita maxima and Carica papaya are effective for controlling monogeneans in the gills of Leporinus macrocephalus.

This study was carried out to evaluate the anthelminthic efficacy of seeds of Cucurbita maxima and Carica papaya for controlling monogeneans in the gills of Leporinus macrocephalus, besides hepatosomatic and splenosomatic index and condition factor of host. The fish were fed with seeds of C. maxima or C. papaya for seven days, and these treatments did not cause any mortality among them. Jainus leporini, Urocleidoides paradoxus, Urocleidoides eremitus and Tereancistrum parvus were the monogeneans found, and their prevalence in fish fed with seeds of C. papaya was 100%, while in fish fed with C. maxima the prevalence was 42.8%. Fish fed with seeds of C. papaya showed decreased in intensity and abundance of monogeneans, while fish fed with seeds of C. maxima presented decreased in abundance. Feeding of L. macrocephalus with seeds of C. maxima or C. papaya had efficacy of 69.6 and 67.8%, respectively. The hepatosomatic index of fish fed with seeds of C. maxima or C. papaya was not affected by the treatments. However, the splenosomatic index and condition factor of fish fed with C. maxima seeds decreased. Seeds of C. maxima and C. papaya may be used for controlling monogeneans of L. macrocephalus in fish farming.

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

This study investigated the acute toxicity (LC50-24 hr ) effects of the essential oil of Cymbopogon citratus for adult Argulus sp. and Dolops discoidalis, before and during oviposition. In vitro acute toxicity (LC50-24 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 LC50-24 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 LC50-24 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.

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%), ß-selinene (13.4%) and ß-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.

Toxicological, hematological and immunological effects of levamisole and ivermectin diet supplementation on Colossoma macropomum (Serrasalmidae).

This study investigated the toxicological, hematological and immunological effects of diets supplemented with levamisole and ivermectin on Colossoma macropomum. Fish were fed for 24, 96 and 240 h with diets containing 300, 600, 900 and 1200 mg of levamisole kg-1 of feed. None of the levamisole concentrations caused either mortality or behavioral alterations among the fish over 10 d of feeding. In comparison, a single feeding of ivermectin at 4500, 9000, 13500 or 18000 mg kg-1 caused 100% mortality among the fish within 10 h. There were no alterations in erythrocyte parameters or albumin in any treatments with levamisole after 24, 96 and 240 h of feeding. At concentrations of 900 and 1200 mg kg-1, levamisole caused decreases in the albumin-to-globulin ratio compared to the control group and the 300 and 600 mg kg-1 diet treatment groups. Levels of glucose and total plasma protein were higher in the fish fed with 600, 900 and 1200 mg kg-1 than in the controls and the 300 mg kg-1 diet treatment group. After 10 d of receiving feed supplemented with levamisole, globulin levels were higher in the 600 mg kg-1 group than in the controls. Respiratory burst activity of leukocytes also increased in the fish supplemented with the 900 and 1200 mg kg-1 diets compared to the controls and other treatments. This is the first study to investigate how diets supplemented with these drugs affect C. macropomum. Our research indicates that all the levamisole concentrations tested can be used in the diet of C. macropomum for antiparasitic treatments against helminth species, and that dietary treatments with levamisole can stimulate components of the innate immune system.

Anthelminthic potential of the Ficus insipida latex on monogeneans of Colossoma macropomum (Serrasalmidae), a medicinal plant from the Amazon.

INTRODUCTION: Herbal therapy is a potentially beneficial alternative for fish aquaculture, since it may be cheaper and more effective than chemotherapy. The aim of this study was to investigate the in vitro efficacy of Ficus insipida latex on monogeneans of Colossoma macropomum gills. MATERIALS AND METHODS: To evaluate the anthelmintic activity, four concentrations of F. insipida latex (250, 500, 750 and 1000 µL/L) and exposure time to cause immobilization on monogeneans were used. In addition, two control groups, being one with water from the breeding tank and the other with breeding water from the tank + 70% ethyl alcohol were used. RESULTS: At the concentration of 250 µL/L of F. insipida latex, the immobilization of the monogeneans occurred after 4 h of exposure, while at the concentration of 500 µL/L, it occurred after 2 h. At the concentrations of 750 and 1000 µL/L of F. insipida latex, the immobilization of the monogeneans occurred after 1 h and 30 min, respectively. After exposure to 250 µL/L of F. insipida latex, 100% of immobilization of monogeneans was observed within 4 h, to 500 and 750 µL/L, 100% immobilization occurred within 4 h and to 1000 µL/L, 100% mortality occurred after 2 h. CONCLUSIONS: Thus, we recommended the use of 1000 µL/L of F. insipida latex for therapeutic baths in of C. macropomum against monogeneans, after previous test of toxicity.

Growth, physiological and immune responses of Arapaima gigas (Arapaimidae) to Aeromonas hydrophila challenge and handling stress following feeding with immunostimulant supplemented diets.

The current study tested the efficacy of a dietary immunostimulant additive (Aquate Fish™®) on the growth performance, and on the physiological and immune responses of Arapaima gigas. Two trials were carried out: a feeding trial for 30 days with the experimental diets and a challenge trial for 7 days, in which fish were bacterial challenge (Aeromonas hydrophila) following by 60 s handling stress. During the feeding trial, fingerlings were fed diets supplemented with 0 (control), 6, 9 and 12 g Aquate Fish™®/kg diet. Dietary supplementation did not influence feed intake, feed conversion and condition factor, but increased the final biomass, number of erythrocytes, thrombocytes, leukocytes, lymphocytes, monocytes, hemoglobin, glucose, globulins and plasma triglycerides in fish fed at a concentration of 12 g/kg diet. After bacterial infection, mortality occurred only in fish fed control treatment, whereas respiratory burst of leukocytes, number of leukocytes and lymphocytes increased in fish that received 12 g of dietary supplementation. The results indicated that dietary supplementation with 12 g of Aquate Fish™® improved biomass and immunity performance of A. gigas fingerlings, without negatively affecting blood biochemical parameters.

Antiparasitic activity, histopathology and physiology of Colossoma macropomum (tambaqui) exposed to the essential oil of Lippia sidoides (Verbenaceae).

In vivo and in vitro antiparasitic activity of the essential oil of Lippia sidoides and blood and histological alterations were assessed in Colossoma macropomum (tambaqui). Essential oil concentrations of 10, 20, 40, 80, 160 and 320mg/L were assayed in vitro against monogenoideans Anacanthorus spathulatus, Notozothecium janauachensis and Mymarothecium boegeri from fish gills. Lippia sidoides essential oil concentrations of 320 and 160mg/L were 100% effective against monogenoideans in 10min and 1h of exposure, respectively. However, the effectiveness of 100% concentrations of 80mg/L and 40mg/L occurred in 3 and 6h, respectively. In the in vivo tests, juvenile fish were submitted to 60min of baths with 10mg/L and 15min of baths with 20mg/L of the essential oil of L. sidoides. These therapeutic baths were not efficient against Ichthyophthirius multifiliis, and monogenoideans present in the gills of C. macropomum. In addition, 10 and 20mg/L of the essential oil of L. sidoides caused an anesthetic effect on the fish and did not influence total glucose and protein plasma levels; however, it decreased the number of total erythrocytes in fish exposed to the higher concentration of this essential oil. Severe alterations and irreversible damage were observed in the fish gills just after L. sidoides essential oil baths and after 24h of recovery. The most recurrent lesions found were hyperplasia and fusion of the lamellar epithelium, vasodilation, detachment of the gill epithelium and lamellar aneurism, epithelial breakdown with hemorrhage, congestion, edema and necrosis, proliferation of the mucous cells and chloride cells and lamellar hypertrophy. Therefore, since the essential oil of L. sidoides has in vitro antiparasitic activity and low concentrations of it have shown toxic effects, the bioactive potential of its main chemical components should be investigated, as well as more efficient forms of its administration in therapeutic baths in order to eliminate fish parasites.

Effect of high levels of dietary vitamin C on the blood responses of matrinxã (Brycon amazonicus).

Juvenile matrinxã (Brycon amazonicus) were fed with different dietary vitamin C (ascorbic acid or AA) concentrations for 2 months in order to evaluate its effect on the physiological condition of this species. Five treatments were tested: Reference diet-RD (350 mg AA kg(-1) and 45% crude protein commercial feed); C400, C500, C600 and C800 with supplementation of 400, 500, 600 and 800 mg AA kg(-1). Fish fed 800 mg AA kg(-1) supplemented diets presented greater weight gain and survival. Hematocrit, hemoglobin concentrations and erythrocytes showed significant increase (p<0.05) in C600 and C800 relatively to the others. Total leukocyte values were significantly higher in fish fed vitamin C supplemented diets relatively to those fed RD and leukocytes count of C600 and C800 were the highest among the treatments. Monocytes were the only cells of fish defense mechanism that presented variation, and C600 and C800 presented the lowest values (P<0.05). Glucose level was significantly different relatively to RD only in C400 and the plasmatic total protein values for fish fed vitamin C supplemented diets were higher than for fish fed the RD. Fish plasmatic ions (Na(+) and Cl(-)) were significantly lower for C500, C600 and C800 relatively to C400 and RD. The physiological profile and the growth performance of matrinxã suggest that diet supplemented with 800 mg AA kg(-1) can improve its performance and increase the survival rate.

Influence of diets supplemented with vitamins C and E on pirarucu (Arapaima gigas) blood parameters.

This study evaluated the influence of diets supplemented with 500, 800, 1200 mg kg-1 of vitamin C (ascorbic acid or AA) and vitamin E (alpha-tocopherol or alpha-T) on the physiological responses of pirarucu fed for 2 months. Weight and mortality were not affected by dietary vitamin type or their concentrations. Significant increase (p<0.05) on the red blood cells count was obtained on treatments with 800 and 1200 mg AA kg-1 and on the hemoglobin concentration on treatment with 500 mg alpha-T kg-1 relatively to control. Mean corpuscular volume presented a significant decrease (p<0.05) on treatment with 800 and 1200 mg AA kg-1 when compared to control. Mean corpuscular hemoglobin concentration was significantly high (p<0.05) on treatment with 500 mg alpha-T kg-1. Only in vitamin C treatments, we noticed a significant increase (p<0.05) in the number of leucocytes relative to control. All fish in the vitamin-supplemented treatments, except 500 mg AA kg-1, had high total protein values compared to control. Fish treated with 800 or 1200 mg alpha-T kg-1 also showed increases in plasma glucose concentrations. Our results suggest that 800 and 1200 mg AA kg-1 are probably the most suitable concentrations for pirarucu diets, although high vitamin E diets are not necessary for quantitative leucocyte increases for this species.

The influence of dietary vitamin C and E supplementation on the physiological response of pirarucu, Arapaima gigas, in net culture.

This study evaluated the efficacy of dietary vitamin C (ascorbic acid or AA), vitamin E (alpha-tocopherol or alpha-T), and C+E supplementation on the blood parameters of Arapaima gigas grown in net cages for 45 days. Four treatments were tested: control (commercial feed); C800; E500 and C+E (800+500) with supplementation of 800 mg AA kg(-1), 500 mg alpha-T kg(-1) and 800+500 mg AA+alpha-T kg(-1), respectively. Hematocrit (Ht), red blood cells (RBC), and hemoglobin concentration (Hb) (oxidative status indicators), thrombocytes and leukocytes (immunological indicators), plasma protein and glucose were evaluated. Fish fed vitamin C and C+E supplemented diets showed greater weight gain and survival. Dietary vitamin C and C+E diet supplementation resulted in increased Ht, Hb, RBC, MCHC, total leukocytes, total proteins, thrombocytes and eosinophils compared to the control and alpha-T. The alpha-tocopherol-supplemented diet reduced the number of total thrombocytes, lymphocytes and neutrophils and increased glucose and eosinophils relatively to the control. In general, leukocytes and thrombocytes were good indicators of the efficiency of vitamin on the defense mechanism of the A. gigas reared in cages. Results indicate that high alpha-T diet supplementation provides no benefit for the maintenance of the oxidative or the immunological status of A. gigas. However, it was demonstrated that high dietary AA improves A. gigas immunological status. Red blood cell indices and immune system indicators showed no synergistic effect between the vitamins after supplementing the A. gigas diet with alpha-T+AA.