Ocimum basilicum essential oil in pacu Piaractus mesopotamicus: anesthetic efficacy, distribution, and depletion in different tissues.

This study aimed to evaluate the anesthetic activity of Ocimum basilicum essential oil and the distribution and depletion of its major compounds in different tissues of the pacu, Piaractus mesopotamicus. Juveniles (319.08 ± 9.14 g) were individually anesthetized with six concentrations of essential oil from O. basilicum (150, 180, 210, 240, 270, and 300 mg L-1), while in a second experiment, fish (492.39 ± 51.51 g) were subjected to a 10 min immersion bath with essential oil from O. basilicum (300 mg L-1). After anesthetic recovery, blood and tissue samples of the brain, gills, liver, spleen, and white muscle were collected at 0, 0.5, 1.0, 3.0, 6.0, 12.0, and 24 h. A 300 mg L-1 concentration induced anesthesia in the shortest time (193.11 ± 9.31), while at 270 and 300 mg L-1 concentrations, the anesthetic recovery period was the longest (244.33 ± 12.44) Methyl chavicol and linalool were quantified in all tissue samples. The plasma concentrations of methyl chavicol differed (p < 0.05) at all evaluated times. Linalool decreased (p < 0.05) from 0 to 1 h and decreased again only after 12 h. Reduction percentages in 24 h were 92.9% for methyl chavicol, and 97.2% for linalool. Elimination of the compounds methyl chavicol and linalool is slower in the gills, where lower elimination constants (0.03 and 0.15 per h) and longer half-lives (25.84 and 4.53 h), respectively, are noted. In general, essential oil from O. basilicum compounds was readily eliminated, showing promising potential for use as an anesthetic in aquaculture.

Influence of Seasonality and Culture Stage of Farmed Nile Tilapia (Oreochromis niloticus) with Monogenean Parasitic Infection.

The aim of this study was to observe how abiotic and biotic factors in a tropical region influence the rate of monogenean parasitism in Nile tilapia (Oreochromis niloticus) that are farmed in net cages. A total of 240 sexually reversed fish were analyzed, and 20 from each culture stage were collected during each sampling month. Overall, 60 fish were sampled in April (autumn), 60 in August (winter), 60 in November (spring), and 60 in February (summer). Fish were collected from a commercial fish farm located in Capivara Reservoir in the lower Paranapanema River region of Paraná, Brazil. In total, 3290 monogenean parasites were collected from fish gills of the following species: Cichlidogyrushalli, C. thurstonae, Scutogyruslongicornis, C. cirratus, C. sclerosus, and C. tilapiae. Higher parasitological indices were observed in colder seasons with lower precipitation. Autumn had the highest parasitic infection values compared to the other seasons. The occurrence of monogenean parasites showed a negative correlation with season, in contrast to the culture stage, in which there was a positive correlation. These results may provide a means for establishing adequate fish farm management to predict periods of high monogenean infestation.

Ovicidal effect of essential oils of Lippia alba, Lippia sidoides and Lippia gracilis on the acanthocephalan Neoechinorhynchus buttnerae (Eoacanthocephala: Neoechinorhynchidae).

Acanthocephalosis in tambaqui Colossoma macropomum culture is a limiting factor for the production of the species, being the most recurrent, which makes it necessary to develop strategies to control parasitosis even in the early stages of parasite development. This study aimed to elucidate the morphological characteristics of eggs of the acanthocephalan Neoechinorhynchus buttnerae and the ovicidal effect of the essential oils of Lippia alba, Lippia sidoides, and Lippia gracilis. The parasites were collected from hosts naturally infected with N. buttnerae, and the eggs obtained from the female 'parasites' abdominal cavity were added to 12-well plates containing the essential oils at different dilutions. After the eggs (N = 300) were separated and measured (length and width) with the aid of an optical microscope coupled to a digital program, the viability of the treated eggs was monitored 24 h after treatments, L. gracilis essential oil was 100% effective at the highest concentration, whereas L. sidoides essential oil showed over 50% efficacy at 2 mg/ml, reaching 100% efficacy at all higher concentrations. Lippia. alba essential oil did not show satisfactory efficacy in preventing N. buttnerae egg hatching.

Laboratory-controlled challenges of streptococcosis in Nile tilapia using the oral route (infected-feed) for infection.

The study aimed to evaluate the oral infected-feed, intragastric-gavage, and intraperitoneal routes of the Streptococcus agalactiae infection in Nile tilapia (Oreochromis niloticus). For this purpose, 270 juveniles of Nile tilapia, with an average weight of 2 g, were distributed in 18 experimental units of 90 L, acclimatized, and raised for 55 days, until reaching 50 g (median weight). The experimental design was entirely randomized in six treatments, three of which were composed by bacterial infection routes: intraperitoneal 100 µL fish-1 [108 CFU], intragastric 100 µL fish-1 [108 CFU], feed inoculum 100 µL g feed-1 [109 CFU], and three respective control groups. Clinical signs were observed, and mortalities monitored until reaching 50% in the infected groups. Then, tissue samples from the spleen, liver, intestine, brain, and blood were collected from 20 fish per treatment for histopathological and hemato-immunological analyses. In addition, a related mortality curve was established at the end of the experimental challenge. The intraperitoneal and intragastric routes were more aggressive than the oral inoculum, causing greater brain damage, acute hemato-immunological response, and early mortality. While the orally fed inoculum, fish presented brain lesions with less intensity, and a chronic haemato-immunological response, the mortalities occurred twice as long as the other routes. The present research demonstrated that the S. agalactiae oral (feed inoculum) administration can be an innovative methodology to future experimental challenges in aquaculture research.

Effects of sodium butyrate and Lippia origanoides essential oil blend on growth, intestinal microbiota, histology, and haemato-immunological response of Nile tilapia.

This study aimed to verify the effects of dietary supplementation with sodium butyrate and Lippia origanoides, combined and isolated, on the health and zootechnical performance of Nile tilapia juveniles Oreochromis niloticus. A total of 120 fish (5.38 ± 0.65 g) were randomly distributed in 12 experimental units and fed different experimental diets for 30 days, namely: commercial diet without supplementation (Unsupplemented); commercial diet supplemented with 0.5% sodium butyrate (Butyrate); commercial diet supplemented with 0.125% L. origanoides (Lippia) and commercial diet supplemented with a mixture of 0.5% sodium butyrate and 0.125% L. origanoides (Butyrate + Lippia). After preparing the experimental diets there was an increase in the pH of diet Butyrate when compared to the other diets. After 30 days the fish supplemented with Butyrate + Lippia showed reduction significate in the mean corpuscular haemoglobin, concentration of total heterotrophic bacteria in the intestine, and lymphocyte infiltrates in the liver. Besides that, the supplementation with Butyrate + Lippia promoted an increased number of intestinal villi compared to the fish Unsupplemented ones. Additionally, fish fed a diet containing only Lippia presented an increase in the villus perimeter in the posterior region of the gut and in the red blood cell number. Animals supplemented only with sodium butyrate demonstrated increased lactic acid bacterium in the gut and macrosteatosis in the liver, besides decreased melanomacrophages in the spleen. The use of sodium butyrate associated with essential oil had positive effects on the intestinal microbiota, intestinal structure, liver, and spleen integrity, suggesting a greater efficiency of the compounds when used together in the nutrition of Nile tilapia juveniles.

Autochthonous vs allochthonous probiotic strains to Rhamdia quelen.

The aim of this study was to obtain an autochthonous probiotic candidate strain from the silver catfish (Rhamdia quelen) intestinal tract, comparing its in vivo performance with an allochthonous probiotic isolated from another fish, Nile tilapia (Oreochromis niloticus), in a growth performance assay. The study was divided in two parts: in vitro and in vivo assay followed by challenge with A. hydrophila. In the in vitro assay, the species-specific isolated strain Lactococcus lactis presented characteristics such as: absence of hemolysis, antagonism to bacterial pathogens isolated from freshwater fish, and considerable speed of duplication. In the in vivo trial, both fish supplemented with autochthonous or allochthonous strains presented an increase the final concentration of lactic acid bacteria in the intestinal tract of the fish after 60 days of dietary supplementation reaching concentrations of 1 × 107 CFU g-1 and 4 × 107 UFC.g-1, respectively. In addition, the autochthonous strain increased the mean corpuscular hemoglobin (MCH) of the treated animals, but no significant differences were observed in the other hemato-immunological and zootechnical parameters between treatments. After challenge with Aeromonas hydrophila, only animals that received autochthonous probiotic supplementation showed an increase in the serum total immunoglobulin concentration, but not enough to observe a significant difference in the survival rate between the treatments. Dietary supplementation of the probiotic allochthonous strain did not demonstrate any effects superior to those of the isolated autochthonous strain. Although the autochthonous strain did not present significant improvements in the other parameters evaluated in this study, it was able to inhibit bacterial pathogens in vitro, to increase the final concentration of LAB's and the amount of immunoglobulin after experimental challenge, demonstrating probiotic potential. This study demonstrated for the first time the isolation and in vivo use of an autochthonous probiotic strain isolated from silver catfish, as well as its comparative evaluation with the performance of allochthonous probiotic.

Toxicity of formalin for fingerlings of Cyprinus carpio var. koi and in vitro efficacy against Dactylogyrus minutus Kulwièc, 1927 (Monogenea: Dactylogyridae).

The toxicity of formalin on Cyprinus carpio var. koi and its anti-parasite effects against Dactylogyrus minutus (Monogenea) in in vitro tests is analyzed. Specimens of D. minutus were submitted to eight concentrations of formalin: 50, 75, 100, 125, 150, 175, 200, 250 mg L-1, in triplicate. Concentrations of formalin 100, 150 and 200 mg L-1 were then tested to determine the median lethal concentration of 50% of the fish per immersion bath. Fish behavior was also observed during the first 6 h of exposure. The 200 mg L-1 concentration was the most rapid efficacy for D. minutus, killing all parasites in 16 min. All parasites were killed in 47 min at concentration 100 mg L-1. Concentration 200 mg L-1 was the most lethal for fish in less than 24 h exposure, with 24 h LC50 at 135.44 (119.78-153.14) mg L-1. The therapeutic index was 2.05-30 min and 1.15-16 min. A short bath (1 h) is recommended in koi carp with a minimum concentration of 75 mg L-1 of formalin, not exceeding 100 mg L-1 for treatment against D. minutus.

Sylimarin as hepatic protector and immunomodulator in Nile tilapia during Streptococcus agalactiae infection.

This study investigated the use of silymarin, an extract obtained from the milk thistle (Silybum marianum) and its effects as a possible hepatoprotector in Nile tilapia (Oreochromis niloticus). Silymarin was used as feed additive to the diet at a concentration of 0.1% (1 kg per ton of dry ration) with the commercial product named Di-Heptarine S® (16% silymarin phosphatide). A total of 90 juvenile tilapia with approximately 45 days old and mean weight of 0.72 ±â€¯0.04 g were distributed in two groups, one fed with a diet with the hepatoprotector and the other without the additive. At the end of the assay (55 days after feeding), samples of blood were collected for hematological, immunological, histological (liver, spleen and intestine) and enzymatic analysis such as superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST). After 55 days all fish were challenged with Streptococcus agalactiae serotype Ib to verify the sylimarin effects on the immunological parameters and its protection effect while challenged. During the challenge period another biological material sample was collected for hematological, immunological and histopathological analysis (liver, spleen and intestine). Before the challenge, an increase on the count of thrombocyte was found in the supplemented fish. In the liver, dilation of the sinusoids was observed in unsupplemented fish while supplemented fish the alteration was less severe. No significant alteration was found in SOD, CAT and GST between the groups. Histological changes after the challenge were provoked by bacterial toxins as a result of inflammatory processes. Periacinar degeneration was less intense in unsupplemented fish when compared to supplemented fish. On the other hand, eosinophilic and lymphocytic infiltrate did occur in unsupplemented fish differently from supplemented fish which did not show the alteration. The survival was 28% higher in silymarin supplemented fish when compared to unsupplemented fish that presented no survival. Silymarin supplementation in the diet provided a hepatoprotective and immunomodulatory effect on Nile tilapia.

Blood biochemical parameters and melanomacrophage centers in Nile tilapia fed essential oils of clove basil and ginger.

Nile tilapia (Oreochromis niloticus) is the most farmed freshwater fish in the world, however, disease outbreaks are the main cause of losses in production. Due to this, there is an increasing interest in natural products for enhancing disease resistance, without causing physiological impairment. The aim of this study was to verify the effect of the essential oils of clove basil (Ocimum gratissimum) and ginger (Zingiber officinale) on physiological and immunological parameters of nile tilapia. After 35 and 55 days of supplementation (0.5%, 1.0% and 1.5%), blood collection was performed for determination of metabolites (glucose, cholesterol, triglycerides, serum total protein and immunoglobulins) and lysozyme activity. Spleen tissue was also collected at 55 days for analyzing melanomacrophages centers. At 35 days, cholesterol showed significant reduction in the treatments 0.5% and 1.5 ginger and 1.5% clove basil. The serum triglycerides showed significant reduction in these same treatments, in addition to 1.0% clove basil. At 55 days all the supplemented groups showed significant reduction for both parameters. The lysozyme activity at 35 days was significantly higher in fish supplemented with 1.0% ginger and showed no difference among the treatments at 55 days. Regarding the melanomacrophage centers, at 35 days increased number in fish supplemented with 0.5% clove basil was observed but with no significant difference in the total percent area of spleen occupied by centers. The levels of glucose, total serum protein and immunoglobulins did not show significant differences in both sampling times. The addition of essential oils showed to be useful for improving physiological status of the fish, without excessive activation of defense mechanisms.

Probiotic supplementation in diet and vaccination of hybrid surubim (Pseudoplatystoma reticulatum♀ x P. corruscans♂) / Suplementação de probiótico na dieta e vacinação de surubim híbrido (Pseudoplatystoma reticulatum♀ x P. corruscans♂)

ABSTRACT: A supplementary diet with the probiotic bacteria Weissella cibaria on the efficacy of surubim hybrid immunization against a specific hemorrhagic septicemia caused by Aeromonas hydrophila was evaluated on the following treatments: fish fed a supplemented probiotic diet, vaccinated fish and vaccinated fish fed a supplemented probiotic diet, and untreated fish (control). Fish from the probiotic treatments were fed a diet containing W. cibaria for 41 days. On the 15th day of the experiment, fish from vaccine treatments were intraperitoneally vaccinated, with posterior oral booster for four days. One week after the oral booster, three fish from each experimental unit were sampled. The probiotic supplementation increased the number of thrombocytes and lysozyme concentration compared with surubim that did not receive W. cibaria in the diet. On the other hand, the vaccination increased agglutination titer, lysozyme concentration, and antimicrobial activity compared with surubim that were not vaccinated. However, there was no interaction between diet with probiotics and vaccination in the surubim hybrid in the analysed parameters.

RESUMO: A dieta suplementada com a bactéria probiótica Weissella cibaria sobre a eficácia da imunização de surubim híbrido contra a septicemia hemorrágica específica, causada por Aeromonas hydrophila, foi avaliada nos seguintes tratamentos: peixes alimentados com a dieta suplementada com probiótico, peixes vacinados, peixe vacinadas que receberam um probiótico na dieta e peixes não tratados (controle). Peixes dos tratamentos probióticos foram alimentados com uma dieta contendo W. cibaria por 41 dias. No 15o dia do experimento, os peixes dos tratamentos de vacinas foram vacinados intraperitonealmente, com posterior reforço oral por quatro dias. Uma semana após o reforço oral, três peixes de cada unidade experimental foram amostrados. A suplementação de probiótico aumentou o número de trombócitos e concentração lisozima em comparação com surubins que não receberam W. cibaria na dieta. Por outro lado, a vacinação influenciou no aumento do título de aglutinação, a concentração de lisozima e a atividade antimicrobiana, em comparação com os surubins que não foram vacinados. No entanto, não houve interação entre a dieta com probióticos e vacinação no híbrido surubim nos parâmetros analisados.