Monogeneans are major parasites of fish and cause large economic losses in aquaculture. Treatment for this parasitic infection is done with products that are mostly toxic to fish and the environment. Essential oils (EOs) of Melaleuca alternifolia and Mentha piperita and the oleoresin (OR) of Copaifera duckei were tested for their in vitro anthelmintic activity against the monogenean parasites (Anacanthorus penilabiatus and Mymarothecium viatorum) of pacu Piaractus mesopotamicus. Naturally infected gills were bathed with the herbal solutions (100, 200, 400, 800, and 1600 mg/L) and monitored every 15 min for 4 h. Because of its greater efficacy in vitro (p b 0.05) compared to the other herbal medicines, C. duckei OR was selected for in vivo testing. The in vivo treatment consisted of 10 and 50 mg/L baths of C. duckei OR for 10 min. Parasitological, hematological, and histological analyses were conducted post-bath and seven days after treatment. Parasite loads decreased by approximately 45% in fish treated with 50 mg/L of C. duckei OR. No hematological changes caused by treatment with C. duckei OR at 10 and 50 mg/L were observed. Histology revealed branchial and hepatic alterations in fish from all groups, whereas spleen and kidney tissues were not affected. Histopathological alterations observed in all fish were due to parasitism or nutritional/farming conditions. Hematological and histological results showed that short baths were safe for fish. Based on the strong anthelmintic activity observed, C. duckei OR offers a promising alternative treatment against monogenean parasites.
The effect of the essential oils (EOs) of peppermint, Mentha piperita L., and tea tree, Melaleuca alternifolia (Maiden & Betche) Cheel, on the haematological, biochemical, and immunological parameters and intestinal morphology of Nile tilapia, Oreochromis niloticus L., was evaluated. Fish (58.09 ± 5.87 g) were fed 100 mg/kg and 250 mg/kg of each EO and sampled on days 7, 14, 30 and 60 after starting supplementation. The haematological and biochemical parameters were not altered by the supplementation of EOs compared to the control (p > .05). With regard to the immunological parameters, the activation of the complement system of fish fed 250 mg/kg peppermint and 100 mg/kg and 250 mg/kg tea tree EOs were significantly higher compared to the control after 60 days of feeding (p < .05). The complement system plays an essential role in innate immunity and contributes significantly to the acquired immune response; thus, its activation through supplementation with EOs is promising for the formulation of nutritional additives in aquaculture. Regarding intestinal morphology, fish fed 250 mg/kg tea tree EO presented higher villus size compared to all other groups (p < .05), which represents a healthier gut. These fish present a larger intestinal surface, which can result in better absorption and utilization of the nutrients. Based on the responses found in this study, both EOs were considered promising for the formulation of feed additives for Nile tilapia.
Trypanosomes are flagellated parasite protozoans that prey especially on wild fish and have recently been described affecting fish in aquaculture. The present study was carried out during a mortality outbreak of Nile tilapia Oreochromis niloticus reared in net cages. Samples of 10 fish in the finishing phase showing unspecific signs, such as anorexia, skin darkening and gill paleness, were analyzed using hemogram, parasitology, microbiology, histopathology, electron microscopy and molecular identification. For microbiological analysis, the kidney was collected aseptically and cultured on tryptone soy agar for 48 h at 30°C. For parasitological analysis, skin, gills, intestinal contents and blood scrapings were examined. Photomicrographs of the parasite were measured. For molecular identification, blood samples were processed and sequenced for amplification of the 18S rDNA gene. Gills, liver, spleen, kidney, heart and intestine were sampled for histopathological processing. The microbiological results indicated that the fish were not infected with bacteria. Scrapings of the skin and gills revealed the massive presence of kinetoplastids, which were also observed in greater numbers than erythrocytes in the blood. Intestines were not affected by endoparasites. The morphometric characteristics indicated the presence of the Trypanosoma genus, which was confirmed in the sequenced samples, where 95% and 98% of the identity were Trypanosoma sp. In histopathology, all organs presented different levels of alteration, accompanied by large numbers of the parasite in small and large vessels. The main findings were the description of mast cell infiltrates in the gill and intestine, as well as multifocal aggregates of melanomacrophages in the liver, pancreas, spleen and kidney. Furthermore, the study addresses the newest features of clinical signs of infected fish and possible causes of infestations and compares the diagnosis of this hemoparasite with other hemoflagellates. To our knowledge, this study represents the first outbreak of Trypanosoma in Nile tilapia in South America. The authors warn of possible new cases of trypanosomiasis in aquaculture, recommending possible forms of containment and biosecurity measures.
In this study, the identification and characterization of potential probiotic bacteria present in natural microbiota of tambaqui Colossoma macromum were performed for the first time. Bacteria from the intestinal mucosa were isolated in agar Man, Rogosa and Sharpe (MRS). Only Gram‐positive and haemolysis‐negative isolates were selected (n = 152). The tests included morphological differentiation, gene sequencing, catalase, bile tolerance, pathogen antagonism, antimicrobial susceptibility and safety to the host. To differentiate the isolates, colonies were morphologically characterized. From 152 initial isolates, 6 strains with probiotic potential were selected and identified through sequencing of 16S rRNA gene of Enterococcus faecalis, Enterococcus hirae, Lactococcus lactis, Pediococcus pentosaceus, Staphylococcus hominis and Staphylococcus saprophyticus. The growth of the six strains was not affected by different levels of bile inclusion. All probiotic strains inhibited the growth of the Lactococcus garvieae pathogen in the antagonism test. When tested against Aeromonas hydrophila, L. lactis and E. hirae strains were prominent. All strains were resistant to some degree against the 11 antibiotics. With this study, it was possible to screen the following six strains with probiotic potential for inclusion in the diet of C. macropomum: S. hominis, S. saprophyticus, L. lactis, P. pentosaceus, E. hirae and E. faecalis.
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