Silver nanoparticles exhibit in vitro anthelmintic and antimicrobial activities against Dactylogyrus minutus (Kulwiec, 1927), and Aeromonas hydrophila in Cyprinus carpio Koi.

The study aimed to evaluate the in vitro anthelminthic and antimicrobial activity of silver nanoparticles (AgNPs) against Dactylogyrus minutus and Aeromonas hydrophila, pathogens of Cyprinus carpio Koi. Gill arches of the fish were removed and placed into six-well plates containing 10 mL of tank water with varying concentrations of AgNPs: 100, 400, 500, 600, and 800 mg/L, along with control groups using tank water and distilled water. Each group was tested in triplicate. Parasites were observed every 10 min for 300 min (5 h) using a stereomicroscope, and mortality rates were recorded. Anthelminthic efficacy was calculated at the end of the tests. For the in vitro antimicrobial test, the Minimum Inhibitory Concentration (MIC) of AgNPs was determined by adding 100 µL of Poor Broth (PB) culture medium to all 96 wells of a microplate. The first well was filled with 100 µL of AgNPs, followed by serial dilutions (1:2 ratio). Subsequently, 50 µL of A. hydrophila (1 × 107 CFU/mL) was added to all wells and incubated for 24 h at 28 °C. Results showed that 800 mg/L of AgNPs achieved 87% anthelminthic efficacy within 300 min, while 100 mg/L achieved 47% efficacy. The MIC showed bacterial growth inhibition at 125 mg/mL. Despite the 87% efficacy against parasites within 300 min, AgNPs did not reach 100% efficacy quickly, limiting their potential use in ornamental fish farming. Further studies are needed to assess the toxicity of AgNPs in fish.

Enriched Diet With Orange Essential Oil Citrus sinensis for Tambaqui Colossoma macropomum Promotes Growth Performance and Resistance Against Aeromonas hydrophila.

The objective of this study was to evaluate the effect of dietary supplementation with Citrus sinensis (L.) Osbeck essential oil on the growth, immune system, and resistance to Aeromonas hydrophila infection in Colossoma macropomum fingerlings. The experiment was conducted with five treatments (control diet, Tween80 diet, and diets supplemented with 200, 400, and 800 mg L-1 of C. sinensis essential oil) with four replicates. At the end of the experimental period, growth parameters were measured, and blood samples were collected for thrombogram, leukogram, and phagocytic activity analysis. A bacterial challenge with A. hydrophila was conducted for 96 h. C. macropomum fingerlings that were fed with 400 and 800 mg L-1 of C. sinensis essential oil had the highest growth parameters, with final weights of 533.18 ± 2.03 mg and 531.91 ± 2.67 mg, respectively, and an increase in the number of thrombocytes, lymphocytes, monocytes, and neutrophils, as well as higher phagocytosis rates compared to the control group. Regarding the challenge, fish in the 400 and 800 mg L-1 treatments also exhibited the lowest cumulative mortality rate (26.66% ± 3.33%). Therefore, supplementation with C. sinensis essential oil promotes growth, improves health, and enhances resistance to A. hydrophila in C. macropomum fingerlings.

The effect of Biokos™, a natural lipopeptide surfactant extracted from the bacterium Pseudomonas, on Epistylis infections in Carassius auratus.

In the aquaculture industry, biocides are routinely used to treat parasitosis in fish, and researchers are continually developing sustainable alternatives that can replace these harsh chemicals. In this context, the objective of this study was to evaluate the effectiveness of a new natural compound, BiokosTM, for the treatment against Epistylis sp. in Carassius auratus fish. The infestation was identified by the presence of whitish plaques on the integument of five animals, and the diagnosis was confirmed through skin scrapings. BiokosTM is a lipopeptide derived from the bacteria Pseudomonas that can destroy the functionality of the cell membrane of ciliated protozoa. The action of BiokosTM does not harm animals and the environment because the compound degrades into amino acids and fatty acids within days. A 0.15 m3 (150 L) aquarium was treated with an Ich-AwayTM water conditioner manufactured by the Danish company Sundew ApS, which has BiokosTM as the active ingredient. Six tablets were added to the water daily for two days, and new skin scrapings were performed. The fish were clinically well and no longer possessed lesions or parasites. The results obtained indicate that BiokosTM can be an innovative and more sustainable alternative for controlling epistyliasis in ornamental fish.

Exploring the antiparasitic and antimicrobial potential of Schinus terebinthifolius Raddi essential oil against fish and shrimp pathogens.

This study aimed to perform in vitro antiparasitic and antimicrobial tests with the essential oil (EO) of Schinus terebinthifolius against of fish and shrimp. The chemical composition of the EO of S. terebinthifolius was determined by gas chromatography. For the antiparasitic test, the protozoan Epistylis sp. obtained from parasitized Oreochromis niloticus was used, and exposed to different concentrations of EO (2%, 1%, 0.5%, 0.25%), and control with 1% grain alcohol. The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) test with EO of S. terebinthifolius evaluated the antimicrobial potential, with serial dilutions starting at 2% and control with 1% grain alcohol, using the strains of Aeromonas hydrophila (2.2 × 108 CFU mL-1), Edwardsiella tarda, Vibrio parahaemolyticus, V. harveyi, and V. alginolyticus (2.0 × 108 CFU mL-1). Chemical analysis revealed that the major EO compounds of S. terebinthifolius were δ-3-Carene (56.00%) and α-Pinene (16.89%). In the antiparasitic test, the concentration of 2% EO showed 100% efficacy against Epistylis sp. within 5 min. In the antimicrobial tests, the concentration of 2% EO was effective against all bacteria tested. The EO of S. terebinthifolius demonstrated antiparasitic and antimicrobial activity at a concentration of 2%, standing out as an alternative to conventional antibiotics.

Antimicrobial and anthelmintic effects of copper nanoparticles against Koi carp parasites and their toxicity.

This study investigated the in vitro antimicrobial and anthelmintic effect of copper nanoparticles (CuNPs) against the bacterium Aeromonas hydrophila, the monogeneans Dactylogyrus minutus, Dactylogyrus extensus, Gyrodactylus cyprini, and the cestode Schyzocotyle acheilognathi, as well as their toxicity to Cyprinus carpio Koi. In the antimicrobial in vitro test, the inhibition zone method and minimum inhibitory concentration (MIC) were performed. In order to determine the time and efficacy of monogenean parasite mortality, the parasites were exposed to CuNP concentrations of 20, 50, 100, 150, 200, and 300 mg L-1, and a control group with tank water and one with copper sulphate pentahydrate (CuSO4.5H2O) at a concentration of 0.3 mg L-1, performed in triplicate. The parasites were observed every 10 min for 300 min, and mortality was recorded. For the cestodes, parasites were immersed in CuNP concentrations of 50, 100, 150, and 300 mg L-1. At the end of the in vitro tests, the anthelmintic efficacy of each treatment was calculated. To assess the tolerance and toxicity in fish, they were exposed to CuNP concentrations of 0.6, 1.25, 2.5, 5, 10, 20, and 50 mg L-1 for 12 h. The MIC demonstrated that CuNPs effectively inhibited the growth of A. hydrophila up to a dilution of 12,500 mg L-1 and showed an inhibition zone of 14.0 ± 1.6 mm for CuNPs. The results of anthelmintic activity showed a dose-dependent effect of concentration for both groups of parasites, with the most effective concentration being 300 mg L-1 in 120 min. In the toxicity test, the carps showed tolerance to lower concentrations. The study indicated that CuNPs were effective against the studied pathogens. However, it proved to be toxic to fish at high concentrations. The use of low concentrations is recommended still requires further investigation.

Hematoimmunological responses of juvenile Nile tilapia (Oreochromis niloticus) receiving the dietary supplementation of immunomodulators and different levels of vitamins after challenge with physical stress.

In this study, we analyzed the hematoimmunological effects of dietary supplementation with immunomodulators (ß-glucans + nucleotides) and different levels of vitamins on Nile tilapia (Oreochromis niloticus) after exposure to physical stress. The following four diet treatments were used: diets with indicated vitamin levels (Vitind), diets with Vitind + immunomodulator (Vitind + Immune), diets with high vitamin content (Vithigh), and those with Vithigh + immunomodulator (Vithigh + Immune). The experiment included 560 fish in 28 tanks (20 fish tank-1), with seven replicates per treatment. After 60 days of supplementation, the water temperature was set at 20 °C, and complete biometrics were performed. The animals were then subjected to physical stress with temperature oscillations of 20 ºC to 30 ºC/30 ºC to 20 ºC/20 ºC to 30 ºC. Hematoimmunological data from 140 animals were collected post-stress. Antimicrobial titer and total plasma protein levels were significantly higher in fish not receiving immunomodulator-supplemented diets (2.88 ± 0.43 log2 and 26.81 ± 4.01 mg∙mL-1, respectively) than in those that did. Conversely, the agglutination titer increased in fish fed with lower vitamin levels (3.33 ± 0.66 log2) compared to those with higher vitamin levels. Increased immunoglobulin levels were observed in fish fed diets co-supplemented with vitamins and immunomodulators, revealing an interaction between immunomodulators and dietary vitamin levels. In summary, the inclusion of immunomodulators in the diet enhanced the animals' resistance to physical stress and improved hematoimmunological parameters. Additionally, a high vitamin content in the diet did not modulate the immune responses in the animals.

Neste estudo analisamos os efeitos hematoimunológicos da suplementação dietética com imunomoduladores (ß-glucanos+nucleotídeos) e diferentes níveis de vitaminas na tilápia do Nilo (Oreochromis niloticus) após exposição ao estresse físico. Foram utilizados quatro tratamentos: dietas com níveis indicados de vitaminas (Vitind), dietas com Vitind + imunomodulador (Vitind+Immune), dietas com alto teor de vitaminas (Vithigh) e dietas com Vithigh + imunomodulador (Vithigh+Immune). O experimento incluiu 560 peixes em 28 tanques (20 peixes tanques-1), com sete repetições por tratamento. Após 60 dias de suplementação, a temperatura da água foi fixada em 20 °C e realizada biometria completa. Os animais foram submetidos a estresse físico com oscilações de temperatura de 20 ºC a 30 ºC/30 ºC a 20 ºC/20 ºC a 30 ºC. Dados hematoimunológicos de 140 animais foram coletados pós-estresse. O título antimicrobiano e os níveis de proteína plasmática total foram significativamente maiores em peixes que não receberam dietas com imunomodulador (2,88±0,43 log2 e 26,81±4,01 mg∙mL−1) do que naqueles que receberam. Por outro lado, o título de aglutinação aumentou em peixes alimentados com níveis mais baixos de vitaminas (3,33±0,66 log2) comparado àqueles com níveis mais elevados. Níveis aumentados de imunoglobulinas foram observados em peixes alimentados com dietas co-suplementadas com vitaminas e imunomoduladores, revelando interação entre imunomoduladores e níveis de vitaminas na dieta. Em resumo, a inclusão de imunomoduladores na dieta aumentou a resistência dos animais ao estresse físico e melhorou os parâmetros hematoimunológicos. Além disso, o alto teor de vitaminas na dieta não modulou as respostas imunológicas dos animais.

Effects of a multi-strain Bacillus probiotic on the intestinal microbiome, haemato-immunology, and growth performance of Nile tilapia.

The study evaluated dietary supplementation with a feed additive composed of multi-strain Bacillus for Nile tilapia Oreochromis niloticus. In vitro and in vivo assays employing culture-based microbiological methods and metagenomics were performed. Additionally, the study assessed the haemato-immunology, intestinal microbiome, and growth performance of the animals. For this, 30 juvenile Nile tilapia were used in the in vitro assay and 180 (60 + 120) in the in vivo assays. In the in vitro assay, we found evidence of adhesion of the probiotic bacteria to the intestinal mucus of fish, corroborated in the 15-day in vivo assay, in which the count of B. licheniformis was significantly higher in fish fed with probiotic when compared to fish of the control group. Furthermore, in the 50-day in vivo trial, a metagenomic analysis provided evidence for the modulation of the intestine microbiome of Nile tilapia by dietary supplementation of the probiotic. In addition, there was an increase in species richness, higher abundance of potentially probiotic autochthonous species and a lower abundance of Aeromonas sp. when the animals were fed the supplemented diet. Finally, no significant differences were observed in growth performance and haemato-immunological analyses, suggesting no harm to fish health when the product was supplemented for 15 and 50 days. The in vitro results indicate that the multi-strain probiotics were able to adhere to the intestinal mucus of Nile tilapia. Additionally, a modulation of the intestinal microbiome was evidenced in the in vivo assay.

Effects of supplementation with different zinc-based products on the growth and health of Nile tilapia.

Zinc is one of the essential microelements for the metabolism of animals. Zinc nanoparticles may have higher bioavailability due to their low specific surface area, facilitating absorption by fish. The present study aimed to evaluate the effects of supplementation with different zinc-based products on the growth and health of Nile tilapia Oreochromis niloticus. Zinc, in different sizes (nanoparticles or bulk) and forms (inorganic or organic), were used as a supplement in the tilapia diet at a dose of 15 mg kg feed-1 for 60 days. At the end of the feeding trial, production performance, hemato-immunological parameters, activity of antioxidant system enzymes, exposure to Streptococcus agalactiae and zinc concentration in the muscle were examined. After the bacterial challenge, the mean corpuscular hemoglobin concentration (MCHC) significantly increased in the fish treated with organic zinc, inorganic nano zinc, and organic nano zinc, while in the control group (inorganic zinc), MCHC remained unchanged. Regarding defense cells, dietary inorganic nano zinc increased the number of basophils (1.50 ± 1.10) compared to organic zinc (0.80 ± 0.90). Lymphocyte count increased after the challenge only in the organic zinc treatments (bulk and nanoparticles). Neutrophils decreased in the control (inorganic zinc) (2.20 ± 1.70) and inorganic nano zinc (2.60 ± 2.70) treatments after the challenge. When compared before and after the bacterial challenge, the plasma antimicrobial titer significantly increased after the bacterial challenge in all treatments. No significant differences were observed for total proteins, enzymes (SOD and CAT), cumulative survival and zinc deposition on fillet. In conclusion, organic zinc in nanoparticles or bulk size increased Nile tilapia innate defense during bacterial infection. However, the other parameters evaluated were not affected by zinc particle size or form (organic or inorganic), indicating that further evaluations should be conducted with organic zinc in nanoparticles or bulk size in the tilapia diet.

Lippia sidoides essential oil at concentration of 0.25% provided improvements in microbiota and intestine integrity of Danio rerio.

The study evaluated the effects of dietary supplementation with Lippia sidoides essential oil on the microbiota and intestinal morphology of Danio rerio. For this, 448 fish were randomly distributed in 28 tanks divided into a control group fed a commercial diet without supplementation, a group fed a commercial diet containing grain alcohol and five groups fed a commercial diet containing essential oil of L. sidoides (LSEO) at concentrations of 0.25%, 0.50%, 0.75%, 1.00% and 1.25%. After the period of dietary supplementation, biological materials were collected for microbiological and histological analyses. There were no significant differences regarding the microbiological count between the groups. Diversity of the microbiome was higher in 0.25% group than in control group. LSEO inhibited the growth of potentially pathogenic bacteria. Fish fed LSEO0.25% showed greater intestinal histomorphometric indices. The inclusion of LSEO at 0.25% in the diet of D. rerio provided improvements in fish microbiota and intestine integrity.

O estudo avaliou os efeitos da suplementação dietária com óleo essencial de Lippia sidoides sobre a microbiota e a morfologia intestinal de Danio rerio. Para isso, 448 peixes foram distribuídos aleatoriamente em 28 tanques divididos em grupo controle alimentado com dieta comercial sem suplementação, grupo alimentado com dieta comercial contendo álcool de cereais e cinco grupos alimentados com dieta comercial contendo óleo essencial de L. sidoides (LSEO) em concentrações de 0,25%, 0,50%, 0,75%, 1,00% e 1,25%. Após o período de suplementação alimentar, foram coletadas amostras biológicas para análises microbiológicas e histológicas. Não houve diferenças significativas quanto à contagem microbiológica entre os grupos. A diversidade do microbioma foi maior no grupo de 0,25% do que no grupo controle. LSEO inibiu o crescimento de bactérias potencialmente patogênicas. Peixes alimentados com LSEO 0,25% apresentaram maiores índices histomorfométricos intestinais. A inclusão de LSEO a 0,25% na dieta de D. rerio proporcionou melhorias na microbiota e na integridade intestinal dos peixes.

Trace metal biomonitoring in the farming of tambaqui (Colossoma macropomum), an Amazonian neotropical fish.

In Brazil, studies evaluating the concentration of trace metals in fish farms are scarce. Therefore, studies investigating the presence and levels of these metals in aquatic biota, particularly in fish tissues, are crucial for developing appropriate strategies to mitigate the impact of possible toxic metals. Herein, we investigated the levels of trace metals (Cr, Mn, Co, Ni, Cu, Zn, Al, Ag, Cd, Pb, Fe, Na, Mg, Ca, K, and Ba) in water, feed, and sediment, as well as the bioconcentration and bioaccumulation factors in tambaqui muscles (Colossoma macropomum). For this purpose, eight commercial fish farms, which are also engaged in other agricultural activities, were selected. Fe, Zn, Mg, and Cr concentration in tambaqui muscles exceeded the limits set by the Brazilian regulations for daily consumption by adults. Mn, Zn, Al, Pb, Fe, Na, Cu, Co, Ag, Cd, and Ba levels were substantially higher in the liver tissue of tambaquis than those in the muscle tissue. The most prevalent metals found in the feed were Ca, K, Na, Mg, and Fe. However, the levels of Cr and Cd in the sediment, as well as Pb, Mn, Cu, and Fe in the water, exceeded the maximum limits allowed by the Brazilian legislation. The highest bioconcentration factors were observed for the metals, Na, Zn, and K, with concentrations up to 4.74, 12.61, and 72.08 times, respectively, higher in tambaqui muscle compared to those in water. The bioaccumulation factors for Ca, Zn, Mg, Na, and K were 2.90, 6.96, 21.21, 212.33, and 492.02 times, respectively, higher in the muscle tissue than those in the sediment values in fishponds across all fish farms. Therefore, our findings suggest that tambaquis have a remarkable ability to bioaccumulate trace metals, particularly the essential ones, and can be categorized as a bioindicator species for environmental quality. Furthermore, we observed that, although water exhibits the highest prevalence and diversity of elements above the values recommended by the Brazilian legislation, sediment is the primary source of trace metal contamination for tambaquis.

Dietary supplementation with Lippia sidoides essential oil improves organ integrity but the specific activity of antioxidant enzymes is dose-dependent in Danio rerio.

The nutritional quality of food can affect the health of animals. This study examined the effects of dietary supplementation with Lippia sidoides essential oil (LSEO) on the physiology of Danio rerio. Four hundred fourty-eight fish were divided into 28 tanks and subjected to different dietary treatments: a control group with no supplementation, a group with grain alcohol supplementation and five groups with LSEO at concentrations of 0.25%, 0.50%, 0.75%, 1.00% and 1.25%. After 15 days, histological and enzymatic analyses were conducted. The 0.25% LSEO group exhibited lower glutathione peroxidase and catalase activity compared to the 1.00% group. Additionally, fish in the 0.25% LSEO group showed improved liver, kidney and splenic integrity indices. These findings support the inclusion of 0.25% LSEO in the diet of D. rerio, suggesting potential benefits for fish physiology and encouraging further research on phytotherapeutics in fish diets.

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.

Bioaccumulation of trace metals in Neoechinorhynchus buttnerae and in its fish host tambaqui (Colossoma macropomum) from fish farms.

Fish parasites are excellent bioindicators of environmental contamination because they respond quickly to water pollutant chemicals, and they can accumulate high concentrations of trace metals compared to their hosts. Here, we investigated the bioaccumulation pattern of the following: Cd, Ca, K, Na, Mg, Fe, Al, Zn, Ba, Mn, Cu, Pb, Cr, Ni, and Co. We investigated the presence of trace metals in the acanthocephalan parasite Neoechinorhynchus buttnerae, and the bioaccumulation factors (BAFs) of metals were tested in the food, muscle, and liver of its host fish Colossoma macropomum (tambaqui). We used samples from four commercial fish farms that also conduct other agribusiness activities. Tissues of the fish along with their parasites were subjected a trace metal concentration analysis by inductively coupled plasma optical emission spectrometry. Most of metals showed significantly higher presence in N. buttnerae than in tambaqui (p < 0.05), with increased level of Na, Pb, Ca, Mn, Zn, Al, and Fe in fish muscle and that of Cr, Ni, Zn, Al, Ca, and Ba in fish liver. Considering all the fish farms, the highest values of BAF were observed for Fe, Al, Zn, and Mn with concentrations up to 35.63, 26.88, 14.12, and 6.66 times higher in acanthocephalan tissues than in the fish muscle, respectively. Moreover, Ba, Ca, and Al showed concentrations up to 18.11, 12.18, and 11.77 times higher in acanthocephalan than in the liver of tambaqui. Our results indicate that the higher the levels of these metals in the parasite, the lower their concentrations in the muscular and hepatic tissues of the fish. Therefore, we suggest that N. buttnerae can directly influence the concentrations of trace metals in the fish tissues and accumulate both essential trace (Fe, Zn, Mn, and Ca) and toxic elements (Al, Pb, and Ba) in the host.

Efficacy of chlorine, sodium chloride and trichlorfon baths against monogenean Dawestrema cycloancistrium parasite of pirarucu Arapaima gigas.

This study investigated the efficacy of sodium chloride (SC) and trichlorfon (T) against Dawestrema cycloancistrium and its physiological effects on Arapaima gigas. The efficacy of chlorine (C) as a prophylactic disinfectant was also evaluated. In vitro test with 15 treatments were: SC 4, 8, 10, 12, 14 g/L, T0.1, 0.4, 0.8, 1.6, 3.2 mg/L, and C500, 1000, 3000, 6000 mg/L. Scanning electron microscopy was performed to evaluate parasite damage. The in vivo test was as follows: control, 4 h short baths, once a day, for four consecutive days (SC12 g/L, T5 mg/L); 24 h long baths, for 2 days in 24 h intervals (SC10 g/L, T5 mg/L). In vitro exposure to SC12 and 14 g/L caused 100% mortality of monogeneans at 45 and 60 min, while at T3.2 and 1.6 mg/L 100% of monogeneans died at 30 and 60 min, respectively. In vitro exposure to C resulted in complete mortality after 2-5 min exposure. The SC and T LD50-96 h were 9.9 g/L and 9.73 mg/L, respectively. All in vivo treatments presented efficacy above or close to 90%, with low survival in the long baths. C, starting at 500 mg/L for 5 min, can be used as a disinfectant. Short baths with SC12 g/L and T5 mg/L are recommended for D. cycloancistrium infestations in Arapaima.

Dermocystidium sp. infection in farmed striped catfish Pangasianodon hypophthalmus farmed in Ceará state, Northeastern Brazil.

The genus Dermocystidium infects a wide range of animals. The host infection often occurs through the ingestion of endospores. The diagnosis depends on wet mounts and histopathological analysis of the affected tissue. The aim of this study was to investigate the incidence of Dermocystidium sp. infection on the skin of farmed striped catfish (Pangasianodon hypophthalmus) from a fish farm located in Fortaleza, Ceará state, northeastern Brazil. From these observations, we determined that 100% of the analyzed animals were infected with Dermocystidium sp. The wet mount and histopathology of the fish lesions revealed spore-filled cysts between the dermis and epidermis, encapsulated by connective tissue. Owing to a lack of research on the parasite and its prevalence among different fish species in Brazil and the rest of the world, additional studies are required to understand their endemicity in fish farms of Brazil, and consequently develop better disease prevention methods and increase the overall productivity.

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.

Antiparasitic potential of alternative treatments against larval stages of Lernaea cyprinacea.

This study evaluated the potential of alternative treatments against larval stages of Lernaea cyprinacea. For in vitro test, the nanoemulsified oils of Pinus sp. acicule and oleoresin were evaluated, along with Biogermex® (commercial product based on citrus biomass). For this, the motility of five larvae of the same stage (nauplii or copepodite) were evaluated in a 96-well microplate. Using the best results, on the in vivo test, fries of Rhamdia quelen were submitted to a long-term immersion bath (96 h) containing different concentrations of the product diluted directly in the water. It was possible to notice the antiparasitic potential of the oleoresin and the acicule of Pinus sp., as well as the citrus biomass extract against the parasites. The nanoemulsified oils successfully inhibited the development of nauplii (10 mg L-1 in 24 h) and the fries showed to be tolerant to the presence of the compound (LC50 96 h-16.74 mg L-1). The concentration of 30.5 mg L-1 of Biogermex® eliminated the copepodites within 24 h, being more efficient than Pinus sp. when tested at the same stage, at the times analyzed. The results obtained indicate a potential use of these compounds as prophylactic agents against L. cyprinacea.

Distribution of cysts holding Ascocotyle (Phagicola) longa metacercariae in tissues and organs of mugilid.

Ascocotyle (Phagicola) longa is an etiological agent of human phagicolosis. Mugilids are the second intermediate host, the first being Heleobia australis, and mugilids predatory birds and mammals are its definitive hosts. The occurrence of cysts holding A. longa metacercariae is described in mugilids with a prevalence of up to 100%. The wide geographical distribution of A. longa and its intermediate hosts coupled with the rise in the consumption of raw or poorly cooked fish may elevate the risk of human infection. Therefore, in this study, we aimed to verify the distribution pattern of cysts holding A. longa in mugilids. The tissue and organ samples of these fish were processed in a domestic blender and examined under a stereoscopic microscope to identify the cysts holding the digenetic metacercariae. Of the 24 (100%) fish samples that were analyzed, 12 of Mugil curema and 12 of Mugil liza possessed cysts holding A. longa metacercariae. Digenetic cysts were identified to be present in the gills, heart, stomach, liver, intestines, mesentery, and muscular tissues collected from M. curema and M. liza. Conclusively, in M. curema, the cysts holding A. longa metacercariae were found to be distributed randomly throughout the fish body in almost every tissue and organ that was examined.

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.