Phylogenomic analysis of the Ponticus clade: strains isolated from the spotted rose snapper (Lutjanus guttatus).

The family Vibrionaceae is classified into many clades based on their phylogenetic relationships. The Ponticus clade is one of its clades and consists of four species, Vibrio panuliri, V. ponticus, V. rhodolitus, and V. taketomensis. Two strains, CAIM 703 and CAIM 1902, were isolated from the diseased spotted rose snapper external lesion (Lutjanus guttatus), they were analyzed to determine their taxonomic position, a phylogenetic analysis was performed based on the 16S rRNA sequences proved that the two strains are members of the genus Vibrio and they belong to the Ponticus clade. Then, a phylogenomic analysis was performed with four type strains and four reference strains isolated from marine organisms and aquatic environments. Multilocus Sequence Analysis (MLSA) of 139 single-copy genes showed that CAIM 703 and CAIM 1902 belong to V. panuliri. The 16S rRNA sequence similarity value between CAIM 703 and CAIM 1902 was 99.61%. The Ponticus clade species showed Average Nucleotide Identity (ANI) values between 78 to 80% against the two strains for ANIb, except V. panuliri LBS2T (99% and 100% similarity). Finally, this analysis represents the first phylogenomic analysis of the Ponticus clade where V. panuliri strains are reported from Mexico.

Effects of feed restriction on growth performance, lipid mobilization, and gene expression in rose spotted snapper (Lutjanus guttatus).

The effects of feed deprivation were evaluated for 1 week and 2 weeks in Lutjanus guttatus juveniles. A significant reduction in body mass was observed in both feed deprivation schemes, as well as in hepatosomatic, viscerosomatic and mesenteric fat indexes. The composition of fasted fish was characterized by a decrease in lipid content; the liver displayed an intense reduction of lipid reserves in both fasted groups, and increased expression of the lysosomal acid lipase. 1 week after re-feeding, both experimental groups showed an increase in specific growth rate, feed intake and feed conversion ratio. A recovery in hepatic lipid reserves was observed, and the expression of the lysosomal acid lipase decreased, although lipid content in both groups was still significantly lower than in control groups. Hepatic expression of the growth hormone receptor decreased after fasting, and remained low even after the fish were fed, whereas the expression of insulin-like growth factor 1 and 2 increased after fasting and decreased in both groups when fish were fed again. Altogether, these results showed a partial compensatory growth response in L. guttatus juveniles after fasting, with enhanced growth rates and improved feed efficiency. Fish used stored lipid reserves as the main energy source, and the expression of growth-related and lipid mobilization marker genes in the liver showed similar patterns in both fasting schemes. Based on the results, we suggest as much as 1-week fasting intervals during grow-out programmes to reduce visceral fat and increase growth rate in this species.

Post-transcriptional silencing of myostatin-1 in the spotted rose snapper (Lutjanus guttatus) promotes muscle hypertrophy.

Muscle growth is regulated by several factors including the growth differentiation factor 8, known as myostatin, an inhibitor of myocyte differentiation and proliferation. Research on myostatin regulation was already conducted to improve growth rates in farmed animals, including aquatic species. To explore the effects of myostatin inactivation in a commercial marine fish (spotted rose snapper, Lutjanus guttatus) in vivo, we induced post-transcriptional silencing (knockdown) of myostatin-1 (mstn-1) by injecting dsiRNA directly into the muscle of juvenile fish (87 days post-hatch) using a commercial polymer as vehicle. Results show a significant decrease in mstn-1 expression starting at 2 days after injection and for up to 5 days. Knockdown of mstn-1 caused muscle fiber hypertrophy (but not hyperplasia); however, there were no significant changes in weight or length. Although still experimental, this study provides evidence that temporary knockdown of mstn-1 in a commercial marine fish in vivo promotes fiber hypertrophy and therefore could potentially help grow-out programmes in fish aquaculture.

Ontogeny changes and weaning effects in gene expression patterns of digestive enzymes and regulatory digestive factors in spotted rose snapper (Lutjanus guttatus) larvae.

The study of digestive physiology is an important issue in species that have been introduced in aquaculture like the spotted rose snapper (Lutjanus guttatus). The aims of this study were to describe the expression of digestive enzymes (trypsinogen, chymotrypsinogen, α-amylase, lipoprotein lipase, phospholipase A and pepsinogen) and their relation with orexigenic (neuropeptide Y, NPY) and anorexigenic (cholecystokinin, CCK) factors during the larval development and to evaluate the effect of weaning in their expression. The results showed that the transcripts of all the assayed digestive enzymes, with the exception of pepsinogen, and NPY and CCK were already present in L. guttatus from the hatching stage. The expression of all the enzymes was low during the yolk-sac stage (0-2 days after hatching, DAH), whereas after the onset of exogenous feeding at 2 DAH, their expression increased and fluctuated throughout larval development, which followed a similar pattern as in other marine fish species and reflected changes in different types of food items and the progressive maturation of the digestive system. On the other hand, weaning of L. guttatus larvae from live prey onto a microdiet between 25 and 35 DAH significantly affected the relative expression of most pancreatic digestive enzymes during the first weaning days, whereas chymotrypsinogen 2 and lipoprotein lipase remained stable during this period. At the end of co-feeding, larvae showed similar levels of gene expression regardless of the diet (live prey vs. microdiet), which indicated that larvae of L. guttatus were able to adapt their digestive capacities to the microdiet. In contrast, feeding L. guttatus larvae with live feed or microdiet did not affect the expression of CCK and NPY. The relevance of these findings with regard to current larval rearing procedures of L. guttatus is discussed.

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.

Biología reproductiva de Lutjanus guttatus (Perciformes: Lutjanidae) en el Parque Nacional Natural Utría, Pacífico colombiano / Reproductive biology of Lutjanus guttatus (Perciformes: Lutjanidae) in Utría National Park, Colombian Pacific

Protected areas are important for natural population conservation since they work as refuge, feeding and breeding areas, where specimens should be exempt from human pressure. The generation of better criteria for management decisions and conservation of fishery resources, is based on the reproductive aspects of species that support fishing activities, since this information is related to the abundance, size and frequency of capture. With this aim, the reproductive biology of the spotted snapper Lutjanus guttatus was studied from April 2008 and February 2009 at the Utría National Park, Colombian Pacific. For this, we analyzed the volume of catch, size structure, sex ratio, fecundity, maturity size, breeding areas and seasons (n=278), of daily landings of 21 units of artisanal fisheries in ten fishing grounds in the Park. Form all landings, we evaluated a total of 4 319 individuals belonging to 84 species. Based on the number of individuals, Lutjanus guttatus ranked third in catches representing 6.4% (278 individuals), and 16th with 1.8% (95.79kg), based on catch biomass. The average weight was 0.34kg ±0.25kg, while 29cm±6.4cm for total length. The total length-weight relationship had the best fit (Kruskal-Wallis, p<0.05, n=272) with the equation PT=0.00000885* L T3.09. The occurrence of mature fish and high condition factors suggested a spawning season in June, September and October in sandy and rocky shores. This species showed an asynchronical gonadal development, with a mean sexual maturity size estimated in 23.5cm total length, and an absolute fecundity of 156 253.11 oocytes (mode of 4µm diameter). We concluded that L. guttatus medium sizes observed indicated a fishing pressure on small size specimens (the minimum size being 18cm); thus, we recommend the implementation of minimum catch sizes based on the criterion of size at maturity LT100 (25.5cm) and to apply seasonal fishing closures during the highest reproductive activity (June- October). However, it is necessary to obtain additional biological information with multi-year monitoring to improve fisheries management criteria in the area.

Las áreas protegidas son importantes para la conservación de las poblaciones naturales ya que funcionan como zonas de refugio, alimentación y reproducción en la que los organismos deberían estar eximidos de la presión antrópica, pero en aquellas áreas donde se permite la pesca, es elemental conocer los aspectos reproductivos de las especies que soportan la presión pesquera, ya que ésta información relacionada con las abundancias, tallas y frecuencias de captura pueden ofrecer mejores criterios para tomar decisiones de manejo y conservación de los recursos pequeros. La biología reproductiva del pargo lunarejo Lutjanus guttatus se estudió entre abril 2008 y febrero 2009 en el Parque Nacional Utría, Pacífico colombiano. Se analizaron los volúmenes de captura, estructura por tallas, proporción de sexos, fecundidad, talla media de madurez, áreas y épocas de reproducción (n=278). La longitud total osciló entre 18 y 56cm con una media y desviación estándar de 29±6.4cm. La talla media de madurez fue estimada en 23.5cm de longitud total, la fecundidad absoluta fue 156 253.11 ovocitos, con una moda de 4μm de diámetro y un desarrollo gonadal asincrónico. La presencia de individuos maduros y de un alto factor de condición sugieren épocas de desove en junio, septiembre y octubre en litorales rocosos y arenosos.

Population characteristics of spotted rose snapper lutjanus guttatus caught as shrimp bycatch in the gulf of California / Características poblacionales del pargo lunarejo lutjanus guttatus capturado con la fauna de acompañamiento del camarón en el golfo de california / CARACTERÍSTICAS POPULACIONAIS DO "PARGO LUNAREJO" Lutjanus guttatus CAPTURADO COM A FAUNA DE ACOMPANHAMENTO DO CAMARÃO NO GOLFO DE CALIFÔRNIA

There are limited population biology studies of the spotted rose snapper Lutjanus guttatus. Adults of this highly valued commercial species are fished with gillnets and hook-and-line, while juveniles are caught as shrimp bycatch and usually discarded. The effects of this practice have not been studied. As a first step, this study assessed some population parameters of juvenile snapper caught by the Gulf of California shrimp fishery. We looked for early growth stages and determined by the least squares method the weight to standard length relationship as W= 0.000092, SL3.0509. Length frequency distributions were constructed; using the ELEFAN I method, von Bertalanffy growth parameters were found to be L¥= 515mm (standard length) and K= 0.13. Natural mortality (M= 0.35) was estimated from Pauly’s empirical and Ralston equations; and total mortality was calculated by the catch curve equation. The recruitment pattern extended throughout the year, with spring and summer peaks. The sex ratio was 1:1 and the length at capture was 80mm (standard length). With an apparently high fishing mortality levels, it is recommended that abundance and distribution studies be performed to determine the impact of shrimp fishing on this population.

Son escasos los estudios sobre la biología poblacional del pargo lunarejo Lutjanus guttatus. Los adultos de esta especie, de alto valor comercial, son capturados con redes agalleras y anzuelos mientras que los juveniles son capturados como fauna de acompañamiento de la pesca de camarón y usualmente descartados. Los efectos de esta práctica no han sido evaluados. Como un primer paso, en este estudio se estiman algunos parámetros poblacionales de juveniles del pargo lunarejo capturados en la pesquería de camarón del Golfo de California. Se indagaron estadios de desarrollo y se determinó mediante mínimos cuadrados la relación longitud estándar-peso como W= 0,000092; SL3,0509. Se construyeron distribuciones de frecuencia de talla, y usando el método ELEFAN I se encontraron los parámetros de crecimiento de von Bertalanffy L¥= 515mm (longitud estándar) y K= 0,3. La mortalidad natural (M= 0,35) fue estimada por la ecuación empírica de Pauly y la ecuación de Ralston, y la mortalidad total se calculó mediante la ecuación de la curva de captura. El patrón de reclutamiento se extendió a lo largo del año, con máximos en primavera y verano. La proporción sexual fue 1:1 y la talla media de captura fue 80mm (longitud estándar). Con una tasa de mortalidad pesquera aparentemente alta, se recomienda evaluaciones de la abundancia y distribución de la especie para determinar el impacto de la pesquería del camarón sobre esta población.

São escassos os estudos sobre a biologia populacional do "pargo lunarejo" Lutjanus guttatus. Os adultos desta espécie, de alto valor comercial, são capturados com redes de emalhar e anzóis enquanto que os juvenis são capturados como fauna de acompanhamento da pesca de camarão e usualmente descartados. Os efeitos desta prática não tem sido avaliados. Como um primeiro passo, neste estudo se estimam alguns parâmetros populacionais de juvenis do "pargo lunarejo" capturados na pescaria de camarão do Golfo da Califôrnia. Indagaram-se estágios de desenvolvimento e se determinou mediante mínimos quadrados a relação longitude estandar-peso como W= 0,000092; SL3,0509. Construiram-se distribuições de frequência de tamanho, e usando o método ELEFAN I se encontraram os parâmetros de crescimento de von Bertalanffy L¥= 515mm (longitude estandar) e K= 0,3. A mortalidade natural (M= 0,35) foi estimada pela equação empírica de Pauly e a equação de Ralston, e a mortalidade total se calculou mediante a equação da curva de captura. O padrão de recrutamento se extendeu ao longo do ano, com máximos em primavera e verão. A proporção sexual foi 1:1 e o tamanho médio de captura foi 80mm (longitude estandar). Com uma taxa de mortalidade pesqueira aparentemente alta, se recomenda avaliações da abundância e distribuição da espécie para determinar o impacto da pescaria do camarão sobre esta população.

Reproducción y crecimiento del pargo mancha Lutjanus guttatus (Pisces: Lutjanidae) en el Golfo de Nicoya, Costa Rica

Reproduction and growth of the fish Lutjanus guttatus (Pisces: Lutjanidae) in the Gulf of Nicoya, Costa Rica. The fish Lutjanus guttatus is important in the fisheries of Golfo de Nicoya, Costa Rica where they are captured with varied gear mainly by the artisanal fleet. We measured specimens from the commercial catch of 2002-2006. Gonadal state was determined macroscopically and age with otoliths The total length-total weight relationship was y = 0.0236x2.8153 and total length-eviscerated weight y = 0.0216x2.8129. Simplified relationships were y = 0.0173x3 and y = 0.0162x3. There is year-round reproduction with peaks in March (dry season) and September (rainy season). Male-female sexual ratio was 1:1. The age-length key for the gulf shows availability until the 6 years of age. The von Bertalanffy growth curve is L (t) = 65,9 (1-e -0,13(t+2,66)). Rev. Biol. Trop. 57 (1-2): 125-131. Epub 2009 June 30.

Una de las pesquerías más importantes en el Golfo de Nicoya es la del pargo mancha (Lutjanus guttatus); su captura se realiza con diversas artes y principalmente por la flota artesanal. Estudiamos especímenes de la captura comercial artesanal entre el 2002 y 2006. Se analizaron las relaciones morfométricas para aplicar la ecuación P = aLb; el análisis del estado de madurez gonadal se hizo macroscópicamente; se determinó la edad a través de la lectura de anillos anuales de los otolitos sagitales y los parámetros de crecimiento fueron aplicados en la ecuación de von Bertalanffy: L (t) = * [1-exp (-K*(t-t0))]. La relación longitud total-peso total es y = 0.0236x 2.8153 y longitud total-peso eviscerado y = 0.0216x2.8129; las relaciones simplificadas correspondientes fueron y = 0.0173x3 y y = 0.0162x3. La reproducción es todo el año con dos picos en marzo y setiembre (época seca y lluviosa, respectivamente). La proporción macho-hembra es 1:1. Se generó para la población del Golfo de Nicoya una clave edad-talla donde su utilidad se presenta como adecuada hasta los 6 años de edad. La curva de crecimiento von Bertalanffy es L (t)=65,9(1-e -0,13(t+2,66)).