Molecular characterisation of Sardina pilchards larvae diet in Málaga Bay (SW Mediterranean Sea)

The study of predation in the early life stages of fish by direct observation of their stomach content is very difficult, mostly because prey lose all identifiable characteristics. We developed a multiplex-PCR based method, which detected within the gut of the larvae the presence/absence of the five main species of copepods that live in Malaga Bay: Clausocalanus parapergens, Oncaea waldemari, Paracalanus indicus, Temora stylifera and Acartia clausi. A set of five species-specific primers were designed and combined in a single multiplex PCR system, which allowed a time and cost effective screening of the samples. The method detected as little as 0.09 ng/μL of copepod DNA without cross-reactions with Sardina pilchardus DNA

The effect of nutritional condition on the growth to post-flexion of bluefin tuna larvae under cultured condition

These findings indicate that protein synthesis increase in proportion to the flexion stage which is strongly associated with the development of the digestive system and therefore the increase in the nutritional condition. Ontogenetic differences in the RNA and DNA content will be discussed in more detail. Also, the pattern obtained for Atlantic bluefin tuna larva will be compared with those of other species

Metabolic rates in Atlantic bluefin tuna larvae: first data and methodological challenges

Atlantic bluefin tuna is an emblematic scombrid species, but many physiological aspects during the larval stages are still unknown. The mechanisms of how fish larvae balance growth and activity are of great interest since metabolic costs are very high compared to juveniles and adults. However, there is a lack of information about metabolic costs in scombrid larvae. This lack of data is probably related to the challenges associated to larval handling before and during respirometry trials. In this study for the first time, we: i) estimate the relationship between routine metabolic rate and the larval dry weight (mass scaling exponent) at 26°C, ii) measure metabolism under light and darkness and iii) explore the influence of nutritional status (RNA:DNA ratio) on the inter-individual variability in metabolic rates. The relationship between metabolism and size (ranging from 0.6 to 23 mg) was near isometric (slope, b=0.99), in contrast to the allometric relationship observed in most species (b=0.87). Our results show no significant differences in oxygen consumption under light and darkness. A possible regulation (decrease) of their swimming activity with the consequent decrease in the oxygen consumption in light situations is discussed. Nutritional condition did not explain the inter-individual differences in oxygen consumption. This study first reports metabolic rates of Atlantic bluefin tuna larvae and discusses the challenges of performing bioenergetics studies with early life stages of Scombrids

Seasonal variability of oceanographic conditions off NW Galician waters influencing the early life trophic ecology of European hake Merluccius merluccius by stable isotope analysis

Stable isotopes of nitrogen (δ15N) and carbón (δ13C) have been used as tracers for analyzing the trophic ecology of European hake larvae collected during the winter and summer of 2012 off NW Galicia. The sampling times correspond to periods of high and low hake spawning activity, respectively. Trophic enrichment was analyzed using as baseline two different plankton size fractions, corresponding to micro- (55-200μm) and mesozooplankton (>200μm). During both seasons, the δ15N signatures of hake larvae showed higher values than the zooplankton fractions. The first survey carried out in late February was characterized by a preceding marked upwelling, low geostrophic velocities in the hydrographic circulation and an intense mixing of the water column. This environmental scenario is hypothesizing that originated a great nutrient availability for primary consumers showed its effect in the low δ15N values observed in plankton and hake larvae. During July, the environmental setting corresponded to period where the water column was stratified, an anticyclonic gyre was formed in the center of the study area that concentrated particles and weak upwelling conditions and strong geostrophic currents where found. This scenario was characterized hypothetically by low nutrient availability in which production is mainly based on regenerated nutrients which would explain the higher δ15N values found during this period. On the other hand, the greater δ13C values observed in summer in comparison to winter suggest differences in the energetic sources of carbon (Fig. 1). The discussion of results is centered in how oceanographic variability conditions signatures of stable isotope fractionation in the early life trophic food web influencing the variable trophic pathways of larval hake subject to contrasting environmental conditions, thereby affecting their larval nutritional condition as estimated by RNA/DNA ratios and the Fulton index. This study was financed by the projects: CRAMER-CTM2010-21856-CO3 y ECOPREGA-10MMA602021P

Shifts in the seasonal trophic ecology of larvae and juveniles of European hake (Merluccius merluccius): From the Galician upwelling system (NW Spain).

The trophic ecology of European hake (Merluccius merluccius) larvae and early-juveniles from the Iberian shelf off Galicia (NW Spain) are assessed by means of stable isotopes (SIA) and nutritional condition in conjunction with hydrobiological variations observed during the winter and summer season of 2012. Hake early-juveniles (18–47 mm SL) showed higher δ15N than larvae (3–11 mm SL) together with the microzooplankton (55–200 μm) and mesozooplankton (>200 μm) during both seasons. Low δ15N values and high variability in both zooplankton and hake larvae were found during winter coinciding with an unusually strong upwelling event. Inversely, high δ15N content in zooplankton in summer suggests strong nitrogen reutilization. However, hake larvae sampled in summer had a higher RNA:DNA ratio in line with higher Fulton condition factors than those sampled in winter. Higher δ13C values for microzooplankton and mesozooplankton and hake larvae in summer compared to late winter point to prey availability differences. The relationship between nitrogen and carbon stable isotopes versus size or weight suggests an ontogenetic shift in the diet of hake larvae. Early-juveniles had a lower isotopic niche width compared to larvae in both late winter and summer, indicating a trophic specialization related to changes following settlement process from planktonic life to demersal habitat. Higher trophic specialization was observed in summer, which recorded a narrower isotopic niche and higher trophic position estimations.En prensa2,04

POTENCIAL PROBIÓTICO DE CEPAS DE BACILLUS CON PROPIEDADES ANTIMICROBIANAS FRENTE A PATÓGENOS DE PECES

El objetivo de este trabajo ha sido aislar, identificar y caracterizar cepas bacterianas procedentes de ambiente marino, fitoplancton, zooplancton, huevos, larvas de lenguado y diferentes juveniles (branquias, intestino, piel), que pudieran tener actividad probiótica y actividad antibacteriana frente a distintos patógenos de peces. Se aislaron un total de 217 cepas bacterianas. Tras un primer cribado mediante ensayos de inhibición en placa frente a patógenos, se seleccionaron 9 cepas y se identificaron utilizando la secuenciación de su rRNA 16S. Los resultados mostraron que las mejores candidatas eran cepas del género Bacillus. Varios de estas posibles cepas candidatas se caracterizaron in vitro para determinar su potencial probiótico, mediante ensayos de inhibición en co-cultivo con V. anguillarum, resistencia a bilis de pez, y adherencia a mucus intestinal de pez. Nuestros resultados mostraron que algunas de las cepas seleccionadas pertenecientes al género Bacillus, podrían ser consideradas como potenciales probióticos frente a patógenos de peces, por lo que el beneficio de la administración de estas cepas a cultivos de peces será posteriormente estudiada

Actividad bacteriocinogénica de bacterias aisladas de digestivos de peces marinos salvajes

El crecimiento exponencial que ha sufrido la acuicultura en las últimas décadas ha dado lugar a un incremento en número y virulencia de las enfermedades bacterianas que afectan a los peces cultivados. Hasta la fecha, el control de estas enfermedades se ha realizado a través del uso de vacunas y antibióticos, así como de la desinfección del agua y el control biológico, el uso de inmunoestimulantes no-específicos y suplementos dietéticos, entre otros. Pero una mala gestión de estas enfermedades, asociada sobre todo al abuso de antibióticos, ha sido asociada a la aparición de reservorios de bacterias resistentes a antibióticos, tanto en animales como en el ambiente acuático, con el consiguiente peligro para la salud humana (1, 2). El empleo de bacteriocinas como alternativa a los antibióticos parece ser una alternativa plausible para el control y tratamiento de estas enfermedades de una manera inocua y sostenible (3, 4). Pese a que en los últimos años se ha intensificado la búsqueda de cepas productoras de bacteriocinas frente a bacterias patógenas marinas, desafortunadamente todavía son pocas las cepas totalmente caracterizadas (5). El presente estudio tiene por objetivo caracterizar cepas bacterianas con actividad bacteriocinogénica aisladas de intestinos de peces salvajes del Mar Cantábrico. Para ello se procesaron un total de 19 intestinos pertenecientes a 19 ejemplares, cada uno de una especie diferente capturados en el Mar Cantábrico (Cantabria). De estas muestras se aislaron un total de 198 colonias en medio de cultivo TSA, en base a su morfología, color y brillo, que fueron seleccionadas para comprobar su actividad antagonista frente a patógenos de peces de acuicultura. Como cepas indicadoras patógenas de peces se emplearon: Vibrio splendidus y Vibrio anguillarum, y cepas patógenas aisladas de cultivos de lenguado senegalés (Solea senegalensis). Se detectaron 15 cepas de bacterias marinas que presentaban actividad antagonista frente al menos una de las cepas patógenas, de las cuales 4 mostraron actividad inhibitoria frente a más de una especie patógena. La identificación de las cepas procedentes del medio marino seleccionadas se realizó mediante la secuenciación del gen 16S ARN, presentando una homología >99% a Myroides sp. y Alcaligenes sp. Finalmente se estudió el efecto de los parámetros físico-químicos en la actividad bactericida (pH, temperatura y proteasas)

Molecular characterization of the diet of the planktonic community in Málaga Bay (NW Alboran Sea)

The seasonal changes in structure and functioning of the pelagic trophic web in Málaga Bay (NW Alboran Sea) are related to the annual hydrological cycle. However, time series analyses have shown that the relationship between interannual hydrological variability and the plankton community composition is weak. This might be due to different human-induced pressures (nutrient pollution, coastal fisheries) acting on different compartments of the trophic web. The net effect of all these factors would depend on how the ecosystem channels changes in the composition and abundance of each trophic level. Interactions of phytoplankton-ciliates-zooplankton might have a central role in the regulation of the trophic web in Málaga Bay, although the trophic relations of the dominant groups remain still undefined. In order to identify the dominant trophic relationships we aimed to characterise the diet of key ichthyo- and mesozooplankton species in the field. Given that gut content preys (phyto- and microplankton) are fragile and not easy to identify visually, we developed species-specific molecular markers to detect their presence/absence within the predators gut