This study investigated the in vivo and in vitro antiparasitic effects of the essential oil of Lippia alba and the bloodrelated and histopathological alterations that it causes in Colossoma macropomum. In the in vitro trial, the anthelminthic effects of 160, 320, 640, 1280 and 2560 mg/L of the essential oil were tested against monogenoideans (Anacanthorus spathulatus, Notozothecium janauachensis and Mymarothecium boegeri) of the gills of this fish, which are its natural parasites. The concentrations of 1280 mg/L and 2560 mg/L showed 100% efficacy after 20 min of exposure to the essential oil, while at lower concentrations this efficacy against the gill monogenoideans only occurred after 2-3 h of in vitro exposure. However, in the controls, mortality of all of these monogenoideans only occurred after 9 h. A total of 240 fry were distributed into four treatments (20 fish per repetition) and three repetitions were used in the in vivo trial for baths with 100 and 150 mg/L of the essential oil of L. alba, for 30 min. The efficacy in this trial against Ichthyophthirius multifiliis in fish exposed to 100 and 150 mg/L of the essential oil was 40.7% and 50.3%, respectively. However, for monogenoideans, there was efficacy of 14.0% only in the fish exposed to 100 mg/L of the essential oil used. Moreover, the fish exposed to these concentrations of the essential oil presented increased plasma glucose levels, thus denoting signs of stress. Severe lesions such as hyperplasia, fusion of the lamellar epithelium, capillary dilatation, epithelial detachment, lamellar aneurysm, epithelial rupture with hemorrhage, congestion, edema, necrosis, mucous cell proliferation, chloride cells and lamellar hypertrophy were observed in the gills of the fish exposed to 100 and 150 mg/L of the essential oil of L. alba. Alterations to total protein levels, hemoglobin, hematocrit, red blood cell number, thrombocytes number, white blood cell number, lymphocytes, eosinophils and blood neutrophil number was also observed in these fish. The essential oil of L. alba showed great potential for antiparasitic treatment, given that it had high in vitro efficacy against monogenoideans and in vivo efficacy against the protozoon I. multifiliis. Because of the low concentrations of the essential oil (100 and 150 mg/L) that were tolerated by the fish and thus could be used in the therapeutic baths, the efficacy against monogenoideans was low. This indicates that there is a need for additional strategies for using this essential oil in antiparasitic treatments, since the concentrations that eliminate these ectoparasites are toxic for the hosts. Lastly, this was the first study on the antiparasitic activity of L. alba. Statement of Relevance The manuscript entitled "Antiparasitic activity of the essential oil of Lippia alba on ectoparasites of Colossoma macropomum (tambaqui) and its physiological and histopathological effects", represents original article on use of the essential oil of Lippia alba on ectoparasites of tambaqui, an important finfish of Amazon regio...
As espécies de plantas do gênero Lippia Linn., pertencentes à família Verbenaceae, possuem grande distribuição geográfica e são facilmente encontradas em países tropicais. No Brasil, ocorrem principalmente a Lippia alba Mill.) N. E. Brown, Lippia gracilis Schauer, Lippia grandis Schau, Lippia origanoides Kunth, Lippia sidoides Cham. e Lippia triplinervis Gardner. Há muito tempo, diversas espécies de Lippia são usadas na medicina popular por suas atividades biológicas e terapêuticas. Por isso, diferentes espécies dessas plantas vêm sendo testadas, devido ao potencial bioativo, para o tratamento de diferentes doenças do homem e animais. O objetivo deste estudo foi concatenar e discutir dados da literatura sobre o uso de Lippia spp. na medicina veterinária e aquicultura. Os constituintes majoritários dessas plantas são o timol, carvacrol, geranial, linalol, pcimeno, carvona, neral, limoneno, β-cariofileno, óxido cariofileno, mirceno e γ-terpineno. Porém, as concentrações de tais constituintes podem variar em função de diversos fatores aqui discutidos. Estudos in vivo e in vitro usando principalmente L. alba, L. sidoides, L. gracilis, L. origanoides e L. gracilis comprovaram atividade antimicrobiana, antiparasitária, antinflamatória, analgésica, anestésica e antitumoral em animais, indicando grande potencial dessas espécies para uso na medicina veterinária. Algumas espécies de Lippia estão sendo utilizadas também na aquicultura. Assim, produtos obtidos dessas plantas são recursos promissores, necessitando de estudos para desenvolvimento de tecnologias que possibilitem seu uso na sanidade e produção animal.
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.
This study examined the in vitro and in vivo, histopathological, anti-parasitic and hematopathological effects of the essential oil of Lippia origanoides on Colossoma macropomum. Essential oil concentrations of 10, 20, 40, 80, 160 and 320 mg•L −1 were tested in vitro against monogenoideans (Anacanthorus spathulatus, Notozothecium janauachensis and Mymarothecium boegeri) from the gills of C. macropomum. Concentrations of 320 and 160 mg•L −1 were 100% effective against these parasites within 20 and 60 min of exposure, respectively. The 80 mg•L −1 concentration was approximately 80% effective with 3 h of exposure, reaching 100% with 6 h of exposure. The 40 mg•L −1 concentration was also 100% effective with 6 h of exposure. The other concentrations were only weakly effective in vitro. Parasite mortality in controls (water or water + alcohol) began after 3 h, with 100% mortality after 8 h. In vivo tests, in which fry of C. macropomum were placed in baths with 20 mg•L −1 of the essential oil for 60 min, and 40 mg•L −1 for 30 min, did not lead to reductions in parasite abundances. In addition, the essential oil had an anaesthetic effect on fish, increased total protein levels, increased monocyte and neutrophil numbers, and reduced haematocrit. Slight to moderate and severe damage was observed in the gills of C. macropomum fingerling immediately after exposure to the essential oil, and 24 h after the treatments were applied, with no difference between treatments. Histological changes observed in the gills after exposure to concentrations of 20 and 40 mg•L −1 of L. origanoides essential oil were: hyperplasia and fusion of the lamellar epithelium, capillary dilation, displacement of the lamellar epithelium, and lamellar aneurism and epithelial rupturing with haemorrhaging. Oedema, mucous and chloride cell proliferation, lamellar hypertrophy, congestion and necrosis were less frequently observed. It can be concluded that the essential oil of L. origanoides was dose-dependent in vitro effect against monogenoidean parasites of C. macropomum. Unfortunately, the low concentrations tolerated by the fish in the vivo assay (20 and 40 mg•L −1) was not effective. Statement of relevance: The manuscript represents original research on use of the essential oil of Lippia origanoides against ectoparasites of Colossoma macropomum, an important finfish of Amazon region. This manuscript includes treatment in vitro against monogenoideans, and in vivo against protozoans and monogenoideans. Besides, histopathological and hematological analysis of the fish exposed to different concentrations of L. origanoides, a medical plant from North, Central and South America, were performed.
O estado do Amapá é o único estado brasileiro cuja bubalinocultura supera quantitativamente a bovinocultura. Os dados referentes ao perfil do gado abatido nos estados são restritos aos órgãos de defesa e inspeção oficiais, portanto informações sobre a origem, a espécie e o quantitativo abatido tornam-se muitas vezes indisponíveis nos meios de consulta. O objetivo deste
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