ABSTRACT
Primordial germ cells transplantation is a unique approach for conservation and reconstitution of endangered fish species. This study aimed to establish techniques to culture dechorionated embryos in different incubation systems and also to determine anaesthetic concentration for fish recipients in the larval stage for subsequent primordial germ cell transplantation. Intact and dechorionated embryos were divided into three incubation systems: (1) a control group with manual replacement of the solution; (2) a closed environment with high oxygen with manual replacement of the solution; and (3) constant solution recirculation. This combination resulted in six treatments. For the evaluation of anaesthetics for larvae, the concentrations evaluated were 19.5 mM, 24.4 mM, 29.3 mM, and 34.2 mM of 2-phenoxyethanol. Anaesthesia concentration and recovery at different stages were evaluated. For transplantation, primordial germ cells of Astyanax altiparanae were transplanted into anaesthetised larvae (1 dph) of Prochilodus lineatus. Better results were obtained in the recirculation system for dechorionated embryos of P. lineatus for hatching (54.18%) and normal morphology (50.06%). The 2-phenoxyethanol anaesthetic with a dose of 29.3 mM resulted in shorter induction times, in addition to the recovery time between 5 and 10 min. By using this anaesthetic concentration at transplantation, GFP-positive cells were seen in two recipients, but the cells did not proliferate. This study established an effective incubation system for the development of the dechorionated embryo and determined an effective anaesthetic concentration for P. lineatus larvae. In addition, micromanipulation and transplantation of primordial germ cells in neotropical species were conducted for the first time.
Subject(s)
Anesthetics , Characiformes , Animals , Germ Cells , Embryo, Mammalian , Larva , Anesthetics/pharmacologyABSTRACT
Hyper motility is a negative factor in fish handling procedures due to the risks of damages to the animals. Chemicals are widely used to anesthetize fish during biometry and many other handling procedures. 2-Phenoxyethanol is largely employed, but many studies are necessary about tropical fish responses. The present research evaluated the anesthesia induction time for juvenile matrinxã submitted to eight different levels of 2-phenoxyethanol. The range of 2-phenoxyethanol concentration was 250-600mg liter-1. Induction time of anesthesia decreased as a function of 2-phenoxyethanol concentrations. Fish were safely anesthetized approximately after one minute of exposure to 2-phenoxyethanol batches in concentrations above 400mg/liter, and the recover period was about one minute for all anesthetic concentrations. 2-Phenoxyethanol is a safe anesthetic for juvenile matrinxã even in exposures up to 600mg liter-1 being recommended for many field procedures of fish handling.
Em muitos procedimentos de campo que englobam a manipulação de peixes vivos, os animais sofrem grandes riscos de se machucarem devido aos excessivos movimentos que realizam fora da água, sendo necessária a contenção dos animais através do uso de anestésicos. O 2-phenoxyethanol é um produto químico amplamente utilizado no país, sendo, entretanto ainda necessários mais estudos a respeito das respostas fisiológicas dos peixes tropicais às suas propriedades anestésicas. O presente trabalho avaliou o tempo de indução à anestesia de juvenis de matrinxã submetidos a oito diferentes concentrações de 2-phenoxyethanol. O tempo de indução à anestesia diminuiu em função das concentrações do anestésico testado no intervalo de 250-600mg litro-1. Os peixes foram anestesiados com segurança após um minuto, em banhos de 2-phenoxyethanol em concentração superior a 400mg litro-1. O período de recuperação dos animais foi de aproximadamente um minuto para todas as concentrações testadas. O 2-phenoxyethanol é um anestésico seguro para juvenis de matrinxã mesmo em elevadas concentrações até 600mg litro-1.
ABSTRACT
Hyper motility is a negative factor in fish handling procedures due to the risks of damages to the animals. Chemicals are widely used to anesthetize fish during biometry and many other handling procedures. 2-Phenoxyethanol is largely employed, but many studies are necessary about tropical fish responses. The present research evaluated the anesthesia induction time for juvenile matrinxã submitted to eight different levels of 2-phenoxyethanol. The range of 2-phenoxyethanol concentration was 250-600mg liter-1. Induction time of anesthesia decreased as a function of 2-phenoxyethanol concentrations. Fish were safely anesthetized approximately after one minute of exposure to 2-phenoxyethanol batches in concentrations above 400mg/liter, and the recover period was about one minute for all anesthetic concentrations. 2-Phenoxyethanol is a safe anesthetic for juvenile matrinxã even in exposures up to 600mg liter-1 being recommended for many field procedures of fish handling.
Em muitos procedimentos de campo que englobam a manipulação de peixes vivos, os animais sofrem grandes riscos de se machucarem devido aos excessivos movimentos que realizam fora da água, sendo necessária a contenção dos animais através do uso de anestésicos. O 2-phenoxyethanol é um produto químico amplamente utilizado no país, sendo, entretanto ainda necessários mais estudos a respeito das respostas fisiológicas dos peixes tropicais às suas propriedades anestésicas. O presente trabalho avaliou o tempo de indução à anestesia de juvenis de matrinxã submetidos a oito diferentes concentrações de 2-phenoxyethanol. O tempo de indução à anestesia diminuiu em função das concentrações do anestésico testado no intervalo de 250-600mg litro-1. Os peixes foram anestesiados com segurança após um minuto, em banhos de 2-phenoxyethanol em concentração superior a 400mg litro-1. O período de recuperação dos animais foi de aproximadamente um minuto para todas as concentrações testadas. O 2-phenoxyethanol é um anestésico seguro para juvenis de matrinxã mesmo em elevadas concentrações até 600mg litro-1.