Refrigeração do sêmen da garoupa-verdadeira Epinephelus marginatus / Refrigerated storage of dusky grouper Epinephelus marginatus sperm

Foram estudados diferentes diluentes no processo de refrigeração do sêmen da garoupa-verdadeira. Inicialmente, a taxa e a duração da motilidade e a concentração espermática foram avaliadas, caracterizando a qualidade do sêmen fresco. Para os testes de refrigeração a 4ºC, diferentes diluentes foram testados em atmosfera normal e modificada (100% oxigênio). O sêmen fresco apresentou concentração espermática de 3,1±0,2 x 109 células mL-1, motilidade média de 90%, permanecendo móvel, em média, por 3.060 segundos. No experimento de refrigeração, a taxa e a duração média da motilidade foram mantidas adequadas durante 144 horas para o diluente A (70%; 3100 segundos) em atmosfera normal. Na atmosfera modificada, a qualidade do sêmen caiu drasticamente durante as primeiras 24 horas, independentemente do diluente empregado, não propiciando vantagem em sua utilização. A refrigeração do sêmen da garoupa-verdadeira permite manter por até 144 horas uma apropriada qualidade espermática.(AU)

This study aimed to evaluate different extenders in refrigerated storage of dusky grouper sperm. Initially, motility rate, motility time, and sperm density were analyzed to characterize the fresh sperm quality. Different diluents were used in the refrigerated storage sperm at 4ºC in normal atmosphere and modified atmosphere (100% oxygen). The fresh sperm has a spermatozoa density of 3.1±0.2 x 109 spermatozoa mL-1, motility rate 90% and motility time of 3,060 seconds. In the experiment of refrigerated storage, the motility rate and the motility time were maintained appropriate for 144 hours for the extender A (70%; 3,100 seconds) in normal atmosphere. In the modified atmosphere, the sperm quality decreased drastically during the first 24 hours, independently of the diluent used, not propitiating advantages. The refrigerated sperm of dusky grouper is a viable alternative for the short-term storage, being possible, to maintain for up to 144 hours an appropriate sperm viability.(AU)

Desarrollo osteológico de la columna vertebral y del complejo caudal de larvas de Lutjanus guttatus (Perciformes: Lutjanidae) en condiciones de cultivo / Osteological development of the vertebral column and caudal complex of Lutjanus guttatus (Perciformes: Lutjanidae) larvae under rearing conditions

The spotted rose snapper (Lutjanus guttatus) is an important commercial species in Mexico with good culture potential. The osteological study at early stages in this species is an important tool to confirm normal bone structure and for the detection of malformations that may occur during early development. This study was carried out in order to evaluate and describe the normal osteological development of the vertebral column and caudal complex of this species grown under controlled conditions. For this, a total of 540 larvae of L. guttatus, between 2.1 and 17.5mm of total length (TL), were cultured during 36 days; culture conditions were 28ºC, 5.74mg/L oxygen and 32.2ups salinity with standard feeding rates. To detect growth changes, a sample of 15 organisms was daily taken from day one until day 36 of post-hatch (DPH). Samples were processed following standard techniques of clearing, and cartilage (alcian blue) and bone staining (alizarin red). Results showed that the vertebral column is composed of ten vertebrae in the abdominal region, and 14 vertebrae including the urostyle in the caudal region. The development of the axial skeleton starts with the neural arches and haemal arches at 3.8mm TL. Caudal elements such as the hypurals and parahypural began to develop at 4.1mm TL. Pre-flexion and flexion of the notochord and the formation of all hypurals were observed between 5.3 and 5.8mm TL. Ossification of the vertebrae in the abdominal region and in some neural arches initiated at 9.5mm TL. In the caudal region, all the neural and haemal arches ossified at 10.2mm TL. All the abdominal vertebrae and their respective neural arches and parapophyses ossified at 11.2mm TL, while the elements of the caudal complex that ossified were the hypurals, parahypurals and modified haemal spines. All caudal fin rays, 12 neural spines and 3 haemal arches were ossified by 15.5mm. The complete ossification process of this specie under laboratory culture conditions was observed when larvae reached 17.3mm TL on 36 DPH. Detailed analysis of the osteological structures will allow a reference description to evaluate and detect malformations that may occur during the larval culture of the spotted rose snapper.

El pargo flamenco (Lutjanus guttatus) es una especie de importancia comercial en México con un gran potencial para su cultivo. El estudio osteológico en estadios tempranos de esta especie bajo condiciones controladas, es una herramienta importante para el conocimiento de su estructura ósea normal y poder detectar las malformaciones que se puedan presentar. El objetivo del presente trabajo se realizó para conocer y describir el desarrollo osteológico normal de la columna vertebral y el complejo caudal de 540 larvas de 2.1 a 17.5mm de longitud total (LT) bajo condiciones de cultivo a 28°C, 5.74mg/L de oxígeno y 32.2UPS de salinidad. Diariamente se tomó una muestra de 15 organismos desde el día uno hasta el 36 después de la eclosión (DDE) y se procesaron con las técnicas de clareado y tinción de cartílago (azul aciano) y hueso (rojo alizarina) para llevar a cabo la descripción de las estructuras. La columna vertebral se divide en región abdominal con diez vértebras y región caudal compuesta por 14 vértebras incluido el urostilo. El desarrollo del esqueleto axial inicia con la formación de los arcos neurales y hemales a los 3.8mm de LT. A los 4.1mm de LT empieza la formación de los hipurales y parahipural que son elementos caudales. Entre los 5.3 y 5.8mm de LT se observó en pre-flexión y flexión del notocordio y la formación de todos los hipurales. La osificación de las vértebras en la región abdominal y en algunos arcos neurales inició a los 9.5mm de LT. A los 10.2mm de LT se osificó la región caudal y todos los arcos neurales y hemales. A los 11.2mm LT se osificaron todas las vértebras abdominales con sus respectivos arcos neurales y los parapófisis, mientras que los elementos del esqueleto caudal que se osificaron fueron los hipurales, parahipurales y las espinas hemales modificadas. A los 15.5mm de LT se osificaron los radios de la aleta caudal y 12 espinas neurales y 3 hemales. El proceso de osificación de las larvas de esta especie en condiciones de cultivo se completó a los 17.3mm LT o 36 DDE. El análisis detallado de las estructuras osteológicas, permitirá una descripción de referencia para evaluar y detectar las malformaciones que se puedan presentar durante el cultivo larvario.

Caracterización del ciclo de desarrollo embrionario y larval de sarda chiliensis chiliensis (Alva, 1987) en un sistema acuícola de recirculación (SAR) / Characterization of the stages in the cycle of embryo development sarda chiliensis chiliensis (Alva, 1987) in an aquatic re-circulation system (SAR)

Los sistemas acuícolas de recirculación (SAR) son utilizados cada vez más en la acuicultura, especialmente en el norte de Chile en donde las condiciones ambientales son propicias para el cultivo de especies nativas como el bonito (Sarda chiliensis chiliensis), del cual se desconoce su desarrollo embrionario y larval, lo que complica evaluar la factibilidad técnica y económica de desarrollar su cultivo en SAR. Por ello se determinó las bases para su cultivo a partir de enero de 2013, durante 45 días. Los periodos embrionarios caracterizados fueron 31 en 71,83 horas, definidos en 5 estadios: Mórula, Blástula, Gástrula, Neurulación y Metamería. Del desarrollo larval se observaron 35 periodos en 519 horas, un estadio prejuvenil observado a las 591 horas y uno juvenil. Los huevos fueron translúcidos de tipo telolecítico de crecimiento lento en relación a Sarda sarda y Sarda ch. lineolata (especies con las que fue comparada), ya que le tomó casi 3 días eclosionar. Las tasas de sobrevivencia y eclosión fueron altas (86,316% ±9,808 y 89,596% ±11,683 respectivamente), por esto las condiciones serían propicias para su desarrollo. Las larvas en tanto, presentaron más semejanzas con Sarda sarda, en los primeros periodos de desarrollo. El periodo crucial fue entre las 81 a 89 horas de vida, cuando se reabsorbió la gota oleosa; ésta además fue un carácter diferenciador con las otras 2 especies, ya que fue la única que presentó 1 gota oleosa en el periodo larval. Se infiere la factibilidad de un hatchery de Sarda ch. chilensis, debido a las semejanzas con Sarda sarda, y dado el éxito obtenido de ésta en cultivo. De los caracteres morfométricos se sugiere monitorear en el futuro, los que representan las mayores variabilidades dentro de los estadios de desarrollo, que fueron longitud y alto de cabeza y longitud y alto de ojo.

Aquatic re-circulation systems (SAR) are being used more and more in agriculture, especially in the north of Chile where the environment conditions are suitable for breeding in SAR (aquatic re-circulation system) of species such as Bonito (Sarda chiliensis chiliensis), whose embryo or larvae origins are unknown, which complicates the evaluation of a technical and economical possibility of developing its breeding in SAR. Due to this, bases for its breeding were determined in January 2013, for 45 days. The characterized embryo periods were 31 in 71, 83 hours, defined in 5 stages: Morula, Blastula, Gastrula, Neurulation and Metamery. From the larvae development 35 periods in 519 hours were observed, a pre-juvenile stage observed at 591 hours and a juvenile one. The eggs were translucent of a telolecitic type of a slow growing in relation to Sarda sarda and Sarda ch. lineolata, since it took them almost three days to hatch. The surviving and hatching rates were high (86,316±9,808% and 89,596±11,683% respectively), so the conditions for their development would be appropriate. Meanwhile, the larvae presented more similarities with Sarda sarda in the first periods of development. The crucial period was between the 81 and 89 hours of life, when the oily drop was re-absorbed; besides, this was a distinguishing feature with the other two species because it was the only one that presented just one oily drop in the larvae period. The possibility of a Sarda ch. chiliensis hatchery is inferred, due to the similarities with Sarda sarda, and because of the success obtained in this breeding. From the morphometric characters, future monitoring is suggested, the ones that represent the most variables in the stages of development such as length and height of the head, and length and height of the eye.

Desarrollo embrionario del pargo colorado Lutjanus colorado (Jordan & Gilbert, 1882) / Embryonic development of pargo colorado Lutjanus colorado (Jordan & Gilbert, 1882)

El pargo colorado (Lutjanus colorado) es una especie con un alto valor comercial en el mercado mexicano, con potencial para su cultivo. Hasta la fecha no existen estudios sobre su reproducción, cultivo larvario y engorda en cautiverio. El presente trabajo es el primer reporte sobre la descripción a detalle del desarrollo embrionario de la especie bajo condiciones de cultivo. Los huevos fertilizados viables del pargo colorado son pelágicos, esféricos, transparentes y con una sola gota de aceite. Midieron 0,77±0,09 mm de diámetro y la gota de aceite 0,14±0,01 mm. La primera división ocurrió a las 0,05 horas post fertilización (HPF). La eclosión se llevó a cabo a las 17,22 HPF bajo las condiciones del presente estudio. Las larvas recién eclosionadas midieron 1,8±0,1 mm de longitud total (LT). El desarrollo embrionario de esta especie fue similar a la descrita para especies de la misma familia. Los resultados del presente estudio aportan información básica para iniciar el desarrollo de la biotecnología para la producción de semilla de esta especie a escala comercial.

The Colorado snapper (Lutjanus colorado) is one of the most commercially important fish species in México and it is considered a suitable candidate for culture. Until now, no research has been carried out on its reproduction, larviculture and fattening in captivity. This study is the first description of embryonic development of this species under controlled conditions. Fertilized eggs of Colorado snapper are pelagic, spherical and transparent and contain one drop of oil. Eggs measured 0.77±0.09 mm and the drop of oil 0.14±0.01 mm. First cell division occurred at 0.05 h post-fertilization (HPF), hatching at 17.22 HPF under the above described conditions. Larvae total length (LT) was 1.8±0.1 mm. Embryonic development of this species was similar to other lutjanidae species. These results provide basic information for developing the necessary biotechnology for commercial seed production of the Colorado snapper.

Growth differentiation factor 9 and bone morphogenetic protein 15 expression in previtellogenic oocytes and during early embryonic development of Yellow-tail Kingfish Seriola lalandi

BACKGROUND: During fish oocyte maturation, specific molecules are expressed and accumulated within oocyte until fertilization and embryo development. Special attention have been paid in members of the transforming growth factor (TGF-ß) superfamily; growth differentiation factor 9 (GDF9/gdf9) and bone morphogenetic protein 15 (BMP15/bmp15), which exert regulatory functions during oocyte maturation and follicle development. However, little attention has been paid to the involvement of these molecules during embryogenesis considering its importance for the formation of a good quality egg and subsequent embryo survival. The purpose of this study was to analyze the expression of gdf9 andbmp15 in previtellogenic oocytes and during early embryonic development in Seriola lalandi, a pelagic fish with increasing prospect for its aquaculture development, which however, show high mortality at embryo and larval stages. RESULTS: Through RT-qPCR it was found that gdf9 expression was higher in previtellogenic oocytes decreasing after ovulation. This expression profile agrees with its participation in early stages of the follicular development. The transcripts for bmp15 also showed the highest levels in previtellogenic oocytes, however this expression was lower than obtained with gdf9. Conversely, in recently spawned oocytes mRNA bmp15 levels were highest than observed to gdf9. This, is consequent with the main role proposed for this growth factor at the final fish oocyte maturation: avoid the ovulation of an immature oocyte. During embryo development, low levels of mRNA were detected to gdf9, with an increase in 48 H post-fertilization embryos. The bmp15 expression did not change throughout development and was higher than gdf9 at 16 cells, blastula and appearance embryos stages. CONCLUSIONS: Both (gdf9 and bmp15) expression profiles in previtellogenic oocytes and newly spawned eggs are consistent with the described functions for these growth factors in vertebrate ovarian physiology in early and late stages of the follicular development. So, these genes could be considered as quality biomarkers at these stages. However, further studies of these proteins throughout folliculogenesis, are necessaries to fully understand their functions during the oocyte formation. In addition, the persistent expression of these growth factors during development, allows us to speculate possible roles in embryonic processes, which must also be addressed.

Embryonic development of aplodinotus grunniens (perciforme: sciaenidae) in Tenosique, Tabasco, Mexico / Desarrollo embrionario de Aplodinotus grunniens (perciforme: sciaenidae) en Tenosique, Tabasco, México

The description of the embryonic and larval development was based on 30 eggs and 30 larvae. Breeders were collected on the Usumacinta River, Tabasco, Mexico and were subjected to manual fertilization in May 2011. Fertilized eggs were spherical, pelagic, translucent and non-adhesive and had an average diameter of 1.07 mm (SD = 0.10), showing a concentric lipid droplet of 0.85 mm (SD = 0.02). The embryo occupied 100 percent of the perivitelline space at hour 14:08 post-fertilization and the egg measured 1.18 mm (SD = 0.14) at this stage. Eclosion started 17 hours post-fertilization when larvae had a length of 2.54 mm (SD = 0.66). Eight preanal and 13 postanal myomeres were observed exhibiting a light brown color. The morphologic characters recorded in this study are similar to the anatomy of embryos and larvae of the Sciaenidae family. The diameter of the eggs and the length of the larvae for A. grunniens are slightly larger than other Sciaenidae species: A. regius, C. striatus, B. chrysora, C. regalis, L. xanthurus, P. cromis, G. lineatus and S. ocellatus. This is the first study of its kind and its results contribute to the knowledge of the species reproductive biology.

La descripción del desarrollo embrionario y larval se basó en 30 huevos y larvas 30. Los reproductores se recolectaron en el río Usumacinta, Tabasco, México, y se sometieron a la fertilización manual en mayo de 2011. Los huevos fertilizados fueron esféricos, pelágicos, translúcido y no adhesivos presentaron un diámetro promedio de 1.07mm (SD = 0,10), mostrando una gota lipidica concéntrica de 0,85 mm (SD = 0,02). El embrión ocupo el 100 por ciento del espacio perivitelino a las 14:08 horas después de la fertilización y el huevo midió 1,18 mm (SD = 0,14) en esta etapa. La eclosión comenzó a las 17 horas después de la postfertilización, las larvas presentaron una longitud promedio de 2,54 mm (SD = 0,66). Se observaron ocho miómeros preanal y 13 postanal exhibieron una coloración marrón. Los caracteres morfológicos registrados en este estudio son similares a la anatomía de los embriones y larvas de la familia Sciaenidae. El diámetro de los huevos y la longitud de las larvas de A. grunniens son ligeramente más grandes que otras especies de Sciaenidae: A. regius, C. striatus, B. chrysora, C. regalis, L. xanthurus, P. cromis, G. lineatus y S. ocellatus. Este es el primer estudio de su tipo y sus resultados contribuyen al conocimiento de la biología reproductiva de la especie.

Embryonic and larval development of Eugerres mexicanus (Perciformes: Gerreidae) in Tenosique: Tabasco, Mexico

Most studies on Eugerres mexicanus mainly consider biogeographic and systematic aspects and rarely address reproductive characteristics, which are useful for fishery population management plans. This study aimed at evaluating the ontogeny of E. mexicanus, based on 30 embryos and 30 larvae sampled by induced spawning of breeders, taken in February 2009 from the Usumacinta River in Tenosique, Tabasco, Mexico. All descriptions of the embryonic development were based on morphometric and meristic data and followed standard methods. Eggs, recovered at the gastrula stage, had an average diameter of 1.17mm (SD=0.08). The bud stage appeared during the first three hours of development, in which the posterior side was adhered to the vitellus; Kupffer´s vesicle was visible. Yolk-sac larvae hatched 18 hours after fertilization, exhibiting a light brown color and an average total length of 2.94mm (SD=0.70); the preflexion stage was reached eight days after hatching, with a total average length of 4.67mm (SD=0.50) and a total notochord length of 4.45mm (SD=0.50). The flexion stage was reached on the 16th day, with an average total length of 6.66mm (SD=1.53), while postflexion was reached on the 24th day, with 10.33mm (SD=1.45). The pre-juvenile stage was reached on the 33rd day, with a total length of 14.30mm (SD=0.93), showing IX spines and 10 rays and III spines and eight rays in the dorsal and anal fins, respectively. The juvenile stage was reached by the 45th day, with an average length of 28.16mm (SD=1.93) and average weight of 4.75g (SD=1.49). Prejuveniles showed an initial pigmentation with dark colored dots in the superior and inferior jaw and dispersed on the head, while juveniles presented the same pigmentation pattern, decreasing towards the margin of the caudal peduncle. In conclusion, the embryonic developmental stages of E. mexicanus were typical for the Gerreidae group. However, their morphometric characters were slightly different since the diameter and size of the drop of oil were bigger than those reported for marine species. In addition, regarding pigmentation, the yolk-sac larvae of E. mexicanus were olive and yellow on the margin of the notochord, which differs from those reported for other species. This is the first recorded report on the reproductive biology and early life development of this species.

La ontogenia se basó en 30 embriones y 30 larvas, obtenidos mediante la inducción del desove de reproductores provenientes de la ribera del río Usumacinta en Tenosique, Tabasco, México, recolectados en febrero de 2009. La descripción se fundamentó en el registro morfométrico y merístico. Los huevos fueron recuperados en estado de gástrula y presentaron un diámetro de promedio de 1.17mm (SD=0.08). Durante las primeras tres horas de desarrollo embrionario, se presentó la etapa de capullo, en la que se observó la región caudal adherida al vitelo, apreciándose la vesícula de Kupffer. Las larvas con saco eclosionaron a las 18 horas pos-fertilización, fueron de color marrón claro con un promedio de 2.94mm; (SD=0.70) de longitud total y alcanzaron la preflexión a los ocho días post-eclosión con una longitud total promedio de 4.67mm; (SD=0.50) y una longitud total del notocordio de 4.45; (SD=0.50). A los 16 días de la eclosión alcanzaron la flexión, con un promedio de 6.66mm; (SD=1.53) de longitud total. La postflexión se presentó a los 24 días con 10.33mm; (SD=1.45). Al llegar a los 33 días, se presentó la fase prejuvenil y llegaron a medir 14.30; (SD=0.93) de longitud total, presentando IX espinas y 10 radios en la aleta dorsal y III y ocho en la aleta anal. Los juveniles midieron 28.16; (SD=1.93) de longitud a los 45 días, con un peso promedio de 4.75g; (SD=1.49). Los prejuveniles presentaron una pigmentación inicial en la mandíbula superior e inferior con tintes oscuros en forma de puntos y de manera dispersa sobre la cabeza. En los juveniles se observó el mismo patrón de pigmentación, disminuyendo hacia el margen del pedúnculo caudal. Las características descriptivas de la etapa de desarrollo embrionario de E. mexicanus son típicas del período de desarrollo de los peces de la familia Gerreidae, en particular en el caso de las especies E. brasilianus y E. lineatus que habitan en ambientes marinos. Sin embargo, sus caracteres morfométricos son diferentes con respecto al diámetro y el tamaño de los huevos y de la gota de aceite, ya que en E. mexicanus son más grandes que los de las especies marinas y son similares a los de los peces de agua dulce. Con respecto a la pigmentación, la larva con saco de E. mexicanus presenta un olor olivo y amarillo sobre el margen del notocordio, lo cual difiere a lo reportado para E. lineatus,ya que en esta muestra un grupo de melanóforos entre los miomeros nueve al 13 como principal característica, y para Diapturus peruvianus por presentar tres manchas en el margen dorsal de los intestinos desde la inserción de la aleta pectoral hasta el ano. Los resultados de este estudio son los primeros registrados para esta especie y han generado información sobre aspectos de su biología reproductiva y el desarrollo de la vida temprana.