褐牙鲆和夏牙鲆杂交的遗传学研究

在进行褐牙鲆(Paralichthys olivaceus)和夏牙鲆(P. dentatus)的杂交及回交的基础上，利用染色体计数、AFLP、线粒体DNA和核基因部分序列等分析方法对褐牙鲆和夏牙鲆正反交和回交子代进行遗传学研究，探讨了褐牙鲆和夏牙鲆正反交不对称的遗传学基础及其生殖隔离现象，主要结果如下： 1. 褐牙鲆和夏牙鲆正反交的活力是不对称的，褐牙鲆♀×夏牙鲆♂的正交杂种活力正常，能够正常存活、生长和发育，而反交夏牙鲆♀×褐牙鲆♂的杂种体态畸形，孵出后不久死亡。染色体计数发现正交个体的染色体核型与父母本一致，均为48条端部着丝粒染色体；而反交杂种比亲本缺失了两条染色体，仅为46条端部着丝粒染色体，这表明反交杂种为非整倍体。进一步利用AFLP方法对遗传物质从亲本到子代的传递进行了分析，结果显示正反交遗传物质的传承方式存在很大差异。几乎所有亲本的AFLP位点(97.71%)均传递到正交子代。然而，仅有86.64%的AFLP位点从亲本传递到反交子代，反交子代中亲本位点的丢失比例显著高于正交子代和亲本种内交配子代的比例 ( P < 0.05)，这可能与反交杂种染色体丢失有关。进一步分析发现，杂交组中的偏分离标记高于对照组，尽管经2检验发现其差异并不显著 (P > 0.05)。 2. 对于可以成活的正交杂种进行培育达到性成熟后，利用褐牙鲆和夏牙鲆的精液分别与雌性杂交鲆的卵子进行母本回交实验。通过统计受精率、孵化率及杂交适合度值（CFM，受精率和孵化率相乘获得的结果）评估褐牙鲆和夏牙鲆的杂交可适度，结果表明正交及各回交组中的CFM值均显著低于褐牙鲆自交(P < 0.05)。同时，利用AFLP对回交子代基因组的变化进行了分析，发现回交中不仅存在亲本位点的丢失(褐牙鲆回交子代-回交1, 3.96%; 夏牙鲆回交子代-回交2, 6.03%)的现象，也存在非亲位点(回交1, 5.63%; 回交2, 3.28%)的现象。而且，两回交组合分别有27.40%和31.18%的AFLP标记偏离孟德尔遗传。 3. 利用线粒体DNA 16S rDNA、COⅠ基因及核基因rag1的部分序列对正反交及回交子代的线粒体及核DNA的传承进行分析，发现正反交子代的16S rDNA和线粒体DNA片段的同源性和母本一致，各回交组中16S rDNA和COⅠ基因片段与褐牙鲆的同源性较高 (98%)，这表明褐牙鲆和夏牙鲆杂交及回交遵循母性遗传规律。但在回交子代中发现16S rDNA和COⅠ基因具有多种单倍型。褐牙鲆和夏牙鲆的rag1基因具有高度的保守性，但在正交子代中发现rag1多种单倍型。 4. 进一步利用线粒体DNA的16S rDNA、COⅠ基因的部分序列对8种重要海水养殖鱼类的系统进化分析，计算了其种间的遗传距离。根据这几种鲆鲽鱼的杂交是否可行的试验结果，评价种间遗传距离与杂交可适度的关系，结果表明，这8种鲆鲽鱼类的种间遗传距离与杂交可适度呈显著的负相关 (r2 = 0.805，P < 0.01)，即种间遗传分化越大，杂交成功的可能性越小，这表明鲆鲽鱼类中可能存在物种进化的不亲和钟 (Incompatibility clock)。

Size hierarchies affecting the social interactions and growth of juvenile Japanese flounder, Paralichthys olivaceus

Japanese flounder Paralichthys olivaceus (T. & S.)juveniles were size-graded and divided into three groups (small, large, and mixture of small and large flounder), and their social interactions (feeding, aggressive attacking and activity) and growth were investigated. The growth of the small flounder was markedly suppressed by the presence of the large flounder. Large flounder did not significantly suppress the overall food intake of the small flounder but exhibited high aggressive attacking on them and consequently inhibited their activity. Size dominance showed little influence on the aggressive behavior, feeding, activity and growth of the large flounder. The large flounder could not effectively defend the food in excess during the experiments ruling out disproportional food acquisition as the primary mechanism responsible for the size hierarchy effect. Aggressive interaction of the large flounder on the small flounder might be an important cause for the growth retardation of the small flounder. In culture, size grading could markedly improve the growth and survival of the early juvenile flounder. (C) 2004 Elsevier B.V. All rights reserved.

Identification of factors affecting the growth and survival of the settling Japanese flounder larvae, Paralichthys olivaceus

A series of experiments were conducted to identify the factors that affected the growth and survival of the settling flounder larvae Paralichthys olivaceus. Settling larvae 24 days after hatching (DAH) were reared in 10-l experimental tanks up to 40 DAH, and two of the following factors were changed as controlled factors in each experiment: light regime (24L:0D or 12L:12D), prey density (1500, 3000, or 5000 Artemia l(-1)), shelter (sand or no sand) and stocking density (5, 10, or 15 fish l(-1)). Early settling larvae (24-35 DAH) experienced little mortality (less than 10% of the overall mortality) that was not significantly affected by above factors. In contrast, late settling larvae (36-40 DAH) suffered high cannibalistic mortality which was significantly influenced by each of the above factors. Larvae experienced significantly lower mortality at 10 fish l(-1) level than at other densities. Larvae at 15 fish l(-1) level had higher mortality than at 5 fish l(-1) when all other factors were identical. Larvae at 3000 and 5000 Artemia l(-1) treatments survived significantly better than at 1500 Artemia l(-1), but no significant differences in larval mortality were found between the two higher densities. Larvae suffered higher mortality at low prey density or at the absence of sand when they were exposed to longer photoperiod. Low stocking density significantly improved the growth of the settling larvae. The average daily instantaneous growth rate (G) at 5 and 15 fish l(-1) treatments were 0.050 and 0.034, with the coefficient of variation (CV) in final length at 16.4 and 23.5, respectively. Daily instantaneous growth rate increased significantly from 0.033 in the 1500 Artemia l(-1) to 0.041 and 0.045 in the 3000 and 5000 Artennia l(-1), respectively, but no significant difference in larval growth existed between the two higher prey densities. These findings suggested that the optimal prey density for growth and survival of the settling flounder larvae at a stocking density of 5 - 15 fish l(-1) was around 3000 Artemia l(-1) . Larvae that were exposed to 24L showed 20% increase in growth ( G = 0.046, CV = 18.7) than those exposed to 12L ( G = 0.037, CV = 20.5). Longer exposure to light significantly improved larval growth, provided sufficient food was available. Sand substrate did not show significant effects on larval growth, possibly because the larvae spent most of the time swimming or feeding in the water column during this stage. (C) 2003 Elsevier Science B.V. All rights reserved.

Cannibalism in Japanese flounder juveniles, Paralichthys olivaceus, reared under controlled conditions

Laboratory experiments were conducted to determine the effects of sand substratum, light, starvation, fish density and size variation on cannibalism among Japanese flounder juveniles, Paralichthys olivaceus. Cannibalistic behavior (biting and swallowing) increased significantly with starvation except in darkness, where juveniles could not find and attack their prey. Cannibalism occurred more frequently in heterogeneous than in the homogeneous size groups. Cannibalism was more frequent in light than in darkness at each starvation level. With sufficient food present, sand had no significant effect on cannibalism among homogeneous size juveniles, but played an important role in discouraging predation among members of the heterogeneous size groups. With sufficient food present, the rate of cannibalism was generally low and there were no significant differences in cannibalism among fish density levels of the homogeneous juveniles, With starvation, the cannibalism rate was significantly higher in high density groups than in the lower ones. Our results indicate that size variation, starvation and Light are the major factors affecting cannibalism in flounder juveniles, while sand and fish density only have a limited effect on cannibalism, depending on the nature of other factors. (C) 2000 Elsevier Science B.V. All rights reserved.

Biología y pesquería del lenguado paralichthys adspersus, con especial referencia al área norte del litorial peruano, departamento de Lambayeque

El lenguado común, Paralichthys adspersus (Steindachner), es una especie que se captura frecuentemente en la pesca artesanal costera y se distribuye latitudinalmente a lo largo del litoral peruano; longitudinalmente puede alejarse hasta la isóbata de 200 m, por influencia del calentamiento de las aguas, sobre todo durante los eventos El Niño. Dado el interés comercial que este lenguado representa se consideró conveniente efectuar un estudio biológico-pesquero, el mismo que se realizó en el Laboratorio IMARPE de San José, durante los años de 1991 a 1997, a base de muestreos realizados en el área de pesca de Lambayeque (6° a 7°20' 5). Se le determinó como pez predador de las especies de peces que comparten su habitat. En las muestras capturadas no se registró el estadía gonadal inmaduro (I). La mayor frecuencia del estadío desovante (VI) ocurre entre los meses de octubre y febrero (primavera-verano), considerada como la época de reproducción. siendo la talla media de desove de 60,4 cm para las hembras y 43,1 cm para los machos. En las muestras no se hallaron peces con edades de 1 y 2 años, posiblemente debido a la selectividad de las redes de pesca. En los machos e registraron tallas menores, con edades hasta de 5 años; y las hembras tuvieron tallas mayores y edades hasta de 9 años. Los parámetros de crecimiento fueron calculados en L = 87,8 cm; P = 9,118 g; K= 0,20 y to = 0,46. Los índices de abundancia relativa, estimados como captura/viaje, por caletas o lugar de desembarque, fueron calculados entre 1 78,1 kg en Puerto Pizarro y O, 9 kg en Parachique y con promedios generales por caleta de 21,8 kg/viaje de pesca, 14,5 kg/ día de pesca y 314 kg/mes-caleta. Para el área de Lambayeque los índices variaron de 70,1 a 31,5 kg/lancha-mes y de 30,8 a 11,8 kg/viaje-mes, para los años 1991-1997, siendo los años de 1996 y 1997 los que presentaron los más bajos índices.

Notas sobre cultivo de los lenguados Paralichthys adspersus y Etropus ectenes en ambiente controlado

Determina la factibilidad del cultivo del recurso lenguado en ambiente controlado, dando como resultado que es posible el cultivo de esta especie, en especial de P. adspersus, que se adapta fácilmente al cautiverio y a las condiciones abióticas del medio, en comparación al recurso E. ectenes.

Compendio metodológico para la reproducción del "Lenguado" Paralichthys adspersus en cautiverio

La acuicultura se está convirtiendo en una actividad de desarrollo importante para Perú, teniendo un crecimiento a una tasa de 20% anual. Existe una gran variedad de especies con potencial acuícola, como es el lenguado (P. adspersus). En tal sentido, en el presente compendio se describen los procedimientos para la reproducción del lenguado en condiciones de cautiverio, desde la captura hasta métodos de evaluación de la calidad de puestas, a fin de contribuir al desarrollo de la tecnología de cultivo de esta especie. Este documento es resultado de las experiencias realizadas en el proyecto: “Producción de semilla del lenguado Paralichthys adspersus en cautiverio: I Mejoramiento de la calidad y cantidad de desoves”- Contrato Nº051-FINCyT-PIBAP-2009”, ejecutado en el Instituto del Mar del Perú (IMARPE).

Seguimiento en cautiverio de la maduración gonadal y desove del lenguado Paralichthys adspersus (Steindachner)

Se describe el método de observaciones in vivo de la madurez gonadal de Paralichthys adspersus. Las observaciones microscópicas de las gónadas se realizaron con muestras provenientes de las canulaciones intra-ováricas e intra-testiculares, se analizó y describió el desarrollo ovocitario y espermatogénico en individuos mantenidos en condiciones de laboratorio para determinar el estado de madurez gonadal y el momento adecuado para la inducción hormonal. Se interpreta el significado de la atresia ovocitaria en las gónadas de lenguado.

Escala de madurez gonadal del lenguado Paralichthys adspersus (Steindachner, 1867)

Se presenta la escala de madurez gonadal del lenguado Paralichthys adspersus, elaborada en base al análisis y procesamiento histológico de 96 ovarios y 66 testículos de ejemplares capturados en la costa central del Perú y mantenidos en cautiverio. Los ovarios fueron clasificados micro y macroscópicamente, teniendo en cuenta el desarrollo de gametos y gonadas en: virginal, reposo, en maduración, maduro, desovante y recuperación; y a los testículos en virginal, reposo, en maduración, maduro, expulsante y post expulsante. La comparación de las características macro y microscópicas de las gónadas de peces en cautiverio no mostraron ninguna diferencia con respecto a las gónadas de los peces en su ambiente natural.

Metazoarios parásitos del lenguado sureño Paralichthys lethostigmaJordan y Gilbert, 1884 (Pisces: paralichtydae) de la Laguna Madre San Fernando, Tamaulipas, México

Tesis (Maestría en Ciencias con Especialidad en Parasitología) UANL

Margolisianum bulbosum n. gen., n. sp. (Nematoda: Philometridae) from the Southern Flounder, Paralichthys lethostigma (Pisces: Bothidae), in Mississippi Sound, USA

A new species of a philometrid nematode, Margolisianum bulbosum, is described from the subcutaneous tissue in the mouth (larvigerous females), head (males, ovigerous, and larvigerous females), and eye (preovigerous females) of the southern flounder, Paralichthys lethostigma, from Mississippi Sound. It is placed in a new genus diagnosed by the combination of 8 large, paired but separate cephalic papillae; no inner cephalic papillae; an esophagus with a separate, muscular anterior bulb; a prominent mononuclear esophageal gland; and variable, irregularly distributed cuticular bosses in the females, as well as a vestigial rectum, particularly in larvigerous females. Some female specimens exhibit rows of lateral grooves and longitudinal ridges near the posterior end. Males have two small slightly subequal spicules, a barbed gubernaculum, 4 pairs of small cephalic papillae, and a bipartite hypodermal extension within a membranous cuticle on the posterior end. Males, ovigerous females, and larvigerous females appear to be present year round in this sporadic infection in Mississippi.

Lectotypification of the name Helianthus dentatus Cav., basionym of Viguiera dentata (Cav.) Spreng. (Asteraceae: Heliantheae)

A recent molecular phylogenetic analysis (Schilling & Panero 2011) proposed to narrow the concept of the paraphyletic genus Viguiera Kunth (1818: 176; Asteraceae: Heliantheae) to embrace only the type, V. dentata (Cavanilles 1794: 10) Sprengel (1826: 615). Robinson (1977) emphasized the peculiarities of Viguiera dentata as the "distinctive and unique" presence of hairs on the filaments of the anther, the disk corollas throat less than twice as long as the lobes and densely scabrous below, a combination of features that differs from most member of the genus. During taxonomic studies of Viguiera and related genera, it was discovered that V. dentata has never been typified. The basionym, Helianthus dentatus, was described by Antonio Jose Cavanilles in Icones et Descriptiones Plantarum based on plants grown in the Real Jardin Botanico from seed sent by Martin Sesse from Mexico (Blanco 2000). Previous workers including Blake (1918) and Robinson (1977) have accepted the plate in Cavanilles as the type for Helianthus dentatus Cav.

Effects of tributyltin on the immune system of Japanese flounder (Paralichthys olivaceus)

Effects of tributyltin (TBT) which has been used for antifouling paint of ship's hulls and fishing nets on the immune system in Japanese flounder (Paralichthys olivaceus) were investigated. After short-term exposure to a high level of TBT, leucocytes in the head kidney from 1-year-old flounder were examined for the proportion of neutrophils in total leucocytes. Also examined were their respiratory burst activities using flow cytometry, the reduction of nitroblue tetrazolium (NBT) and lysozyme activities. Furthermore, long-term exposures to a relatively low level of TBT using young flounder were also carried out. The proportion of neutrophils in total leucocytes prepared from head kidney in each fish exposed to TBT at 20 microg/L for 5 days and the reduction of NBT by leucocytes prepared from the same experimental conditions increase compared to the control group. The contents were 42.0+/-6.8 and 52.5+/-6.3%, respectively. Significant differences of the NBT reduction were observed between 0 and 20 microg/L TBT exposure groups. On the other hand, the respiratory burst activity of cells in the exposure group clearly showed a tendency to decrease compared to the control group. Furthermore, high level of TBT also inhibited lysozyme activity which plays an important role for the bacteriocidal procedures. However, similar results were not obtained in the exposure group with a relatively low level of TBT. To determine the immunotoxic effects of TBT, infection experiments using pathogens which are naturally occurring should be further investigated.