ABSTRACT. The present study describes the methodology used at IMARPE for the capture, acclimation and management of P. adspersus broodstock using recirculating aquaculture systems (RAS). RAS improved the water quality and maintained the environmental parameters during the acclimation period, temperature (17.2±1ºC), oxygen (8.1±0.7 mg L -1 ), pH (7.3±0.2), ammonia (0.004±0.003 mg L -1 ), nitrite (0.52±0.2 mg L -1 ) and nitrate (3.45±2.6 mg L -1 ). Fish began to be fed normally from day 15 post-capture, once or twice a day using live fish (Odonthestes regia regia, Mugil cephalus), crustacean (Emerita analoga), fresh food (Engraulis ringens and Dosidicus gigas) and artificial feed. A significant loss in the weight of the fish was registered during the first days of captivity, followed by a continuous increase in both sexes. The specific growth rate was positive from the third month of captivity, being the relative growth rate 24.5% and 16.2% in August 2010 in males and females, respectively. Different internal and external parasites were detected in the fish, being Entobdella sp. and Philometra sp. the prevailing parasites observed during samplings. Keywords: Paralichthys adspersus, reproduction, recirculating aquaculture system, broodstock management, culture, Peru. Manejo de reproductores del lenguado Paralichthys adspersus (Steindachner, 1867)usando sistemas de recirculación en acuicultura RESUMEN. El presente trabajo, describe la metodología desarrollada en IMARPE para la captura, aclimatación y acondicionamiento de ejemplares adultos de P. adspersus en sistemas de recirculación (SRA), con la finalidad de formar un stock de reproductores. El SRA permitió manejar parámetros medioambientales estables durante el periodo de acondicionamiento, como: temperatura del agua (17,2±1ºC), oxígeno disuelto (8,1±0,7 mg L -1 ), pH (7,3±0,2), amonio (0,004±0,003 mg L -1 ), nitrito (0,52±0,2 mg L -1 ) y nitrato (3,45±2,6 mg L -1 ). Se dio inicio a la alimentación el día 15 post-captura, utilizando alimento vivo (Odonthestes regia regia, Mugil cephalus), crustáceos (Emerita analoga), alimento fresco (Engraulis ringens y Dosidicus gigas) y artificial semihúmedo. Durante los primeros días de acondicionamiento los peces mostraron una disminución en el peso, hasta su adaptación a las condiciones de cultivo, luego de lo cual se produjo un incremento continuo en ambos sexos. La tasa específica de crecimiento fue positiva a partir del tercer mes y la tasa de crecimiento relativo mostró que en agosto 2010, el peso promedio se incrementó 24,5% en machos y 16,2% en hembras. Se realizó un análisis patológico a los ejemplares capturados y se observó la presencia de diferentes parásitos internos y externos, entre ellos predominaron Entobdella sp. y Philometra sp. Latin American Journal of Aquatic ResearchPalabras clave: Paralichthys adspersus, reproducción, sistema de recirculación en acuicultura, manejo de reproductores, cultivo, Perú.
RESUMEN. Para optimizar las técnicas de reproducción en cautiverio de lenguado Paralichthys adspersus; se elaboró una metodología para la criopreservación de sus espermatozoides. Para ello, se evaluó el efecto del dimetil sulfóxido (DMSO) como agente crioprotector en tres concentraciones diferentes (1,0; 1,5 y 2,0 M) sobre la motilidad espermática y su posterior congelación con cinco diferentes tasas: -7,5; -10; -12,5; -20 y -30°C min -1 , utilizando un congelador automático programable. Los mayores porcentajes de motilidad espermática post congelamiento-descongelamiento (40,5 ± 13%) fueron obtenidos al utilizar una tasa de -10°C min -1 y DMSO sin encontrar diferencias significativas entre las tres diferentes concentraciones de DMSO (P < 0,05). Posteriormente, se evaluó el efecto de un crioaditivo no permeable (vitelo de huevo de gallina o VHG), para determinar si era posible incrementar los porcentajes de motilidad espermática. Se utilizó una solución crioprotectora compuesta por DMSO, en cada una de las tres concentraciones diferentes, adicionando VHG al 10% v/v. Los mayores porcentajes de motilidad espermática (71,71 ± 13%) también se obtuvieron a la tasa de congelamiento de -10°C min -1 , sin encontrar diferencias significativas entre las concentraciones de la solución crioprotectora en la cual fue incubada la muestra de espermatozoides (P < 0,05). Al comparar los porcentajes de motilidad espermática post-descongelación utilizando el crioprotectante DMSO, se obtuvieron diferencias significativas cuando se agregó VHG. Palabras clave: crioaditivos, crioprotectantes, motilidad espermática, tasa de congelamiento, lenguado, Chile. Cryopreservation of flounder Paralichthys adspersus spermatozoaABSTRACT. To optimize the techniques of captive breeding of flounder Paralichthys adspersus, a methodology was developed for the cryopreservation of spermatozoa of this species. The effect on sperm motility post-thawing, using three different concentrations (1.0; 1.5 and 2.0 M) of DMSO as cryoprotective agent and five different freezing rates -7.5; -10; -12.5; -20 and -30°C min -1 , with an automatic programmable freezer was evaluated. The highest percentages of post-thawing sperm motility were obtained by freezing sperm at -10°C min -1 , no significant differences (P < 0.05) were founded between the three different concentrations of DMSO used. Subsequently, we evaluated the effect of a non-permeable cryo-additive (chicken egg yolk, VHG), in order to obtain an increasing of the percentage of sperm motility. We used a cryoprotectant solution including DMSO at three different concentrations adding 10% VHG v/v. The highest percentages of sperm motility (71.71 ± 13%) were obtained at the freezing rate of -10°C min -1 without significant differences between the three concentrations of the cryoprotectant solution in which the sample was incubated sperm (P < 0.05). A high significant difference between the sperm motility percentages postthawing using DMSO and DMSO with VHG, was observed.
The present work shows a practical and of cheap methodology for Engraulis ringens sperm cryopreservation, with the purpose to optimize its reproduction techniques in captivity. The investigation was divided in three stages. The first one consisted to evaluate toxicity of four cryoprotectants; dimethylsulfoxide (ME 2 SO), ethanol (ET), propylene glycol (PG), glycerol (GL) to three molars concentrations (0.5 M; 1.0 M and 1.5 M) on spermatic motility. Motility percentages of sperms incubated in ME 2 SO were significantly bigger than that observed in others three cryoprotectants. In the second stage, the optimal freezing rates were determinate, using ME 2 SO only. In this case, the biggest motility percentages were obtained with 1.5 M ME 2 SO with freezing rates of-20 to-40 ºC min-1 (61.3 ± 7.6% and 53.8 ± 4.9% respectively). Finally, the effect of addition of not permeable cryoprotectant in cryoprotective solution (egg hen yolk, VHG) was evaluated to optimize the post-thawing spermatic motility; nevertheless, the results did not change significantly respect to control without VHG.
Inducción del desove y espermiación de anchoveta peruana Engraulis ringens(Jenyns, 1842) en cautiverio mediante la inyección de un análogo de GnRH
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