Connectivity patterns inferred from the genetic structure of white seabream (Diplodus sargus L.)

Themarine environment seems, at first sight, to be a homogeneousmediumlacking barriers to species dispersal. Nevertheless, populations of marine species show varying levels of gene flow and population differentiation, so barriers to gene flow can often be detected. Weaimto elucidate the role of oceanographical factors ingenerating connectivity among populations shaping the phylogeographical patterns in the marine realm, which is not only a topic of considerable interest for understanding the evolution ofmarine biodiversity but also formanagement and conservation of marine life. For this proposal,we investigate the genetic structure and connectivity between continental and insular populations ofwhite seabreamin North East Atlantic (NEA) and Mediterranean Sea (MS) aswell as the influence of historical and contemporary factors in this scenario using mitochondrial (cytochrome b) and nuclear (a set of 9 microsatellite) molecular markers. Azores population appeared genetically differentiated in a single cluster using Structure analysis. This result was corroborated by Principal Component Analysis (PCA) and Monmonier algorithm which suggested a boundary to gene flow, isolating this locality. Azorean population also shows the highest significant values of FST and genetic distances for both molecular markers (microsatellites and mtDNA). We suggest that the breakdown of effective genetic exchange between Azores and the others' samples could be explained simultaneously by hydrographic (deep water) and hydrodynamic (isolating current regimes) factors acting as barriers to the free dispersal of white seabream(adults and larvae) and by historical factors which could be favoured for the survival of Azorean white seabream population at the last glaciation. Mediterranean islands show similar genetic diversity to the neighbouring continental samples and nonsignificant genetic differences. Proximity to continental coasts and the current system could promote an optimal larval dispersion among Mediterranean islands (Mallorca and Castellamare) and coasts with high gene flow.

Ultramorphology and histochemistry of fat body cells from last Instar larval of the Pachycondyla (=Neoponera) villosa (Fabricius) (Formicidae: Ponerinae)

As células do corpo gorduroso de Pachycondyla (=Neoponera) villosa distribuem-se como uma única camada entre a cutícula e o trato digestivo, formando um conjunto de células agrupadas e recobertas por uma fina membrana. Não foram identificados tipos celulares distintos por meio da ultramorfologia, porém a histologia revelou três tipos celulares distintos: os trofócitos, mais abundantes, as células de urato, distribuídas por entre os trofócitos, e os enócitos, menos abundantes que os demais. Os enócitos são comumente observados próximos da cutícula. Histoquimicamente, os trofócitos apresentaram reação positiva para proteínas básicas no núcleo e no citoplasma e reação fortemente positiva nos grânulos citoplasmáticos. O teste para carboidratos foi fortemente positivo em todo o citoplasma, enquanto para os lipídeos observou-se reação positiva nas vesículas citoplasmáticas. em relação às células de urato, estas apresentaram reação positiva para proteínas básicas no núcleo e citoplasma, por entre as vesículas. Essas células não apresentaram reação para o teste de PAS e Sudan Black B. Quanto aos enócitos, estes apresentaram citoplasma fracamente positivo ao PAS e fortemente positivo ao Sudan Black B e para o azul de bromofenol.

Food intake of Macrobrachium rosenbergii during larval development

This work investigates the acceptance of different food types and sizes by Macrobrachium rosenbergii during each larval stage. Food intake of dry and wet formulated diets of four different size classes (250-425, 425-710, 710-1000 and 1000-1190 mum), as well as Artemia nauplii, was determined. Larvae of each zoeal stage were stocked in beakers and fed ad libitum. After 30-45 min, the digestive tract of each larva was observed under a stereomicroscope. Acceptance was evaluated by food intake frequency (FFI). There was no significant interaction (P<0.05) between inert diet size and FFI for each larval stage. Therefore, food intake during larval development is independent of food particle size. The ingestion of Artemia nauplii, was significantly higher by larvae between stages II and VI. Between stages VII and XI, FFI for Artemia nauplii and wet diet was similar, while the FFI of the dry diet was similar to live food between stages IX and XI. The wet diet was ingested by more than 50% of the larvae only from stage VII onwards, while the dry diet from stage VIII onwards. These results indicate that larvae could be fed Artemia nauplii only until stage VI. Diet supplementation should start from stage VII onwards, using food particles varying from 250 to 1190 mum. (C) 2003 Elsevier B.V. B.V. All rights reserved.

Ingestion rates of Artemia nauplii for different larval stages of Macrobrachium rosenbergii

The functional response between ingestion rate and food concentration was determined for each larval stage of Macrobrachium rosenbergii. Artemia franciscana nauplii were supplied at 2,4, 6, 8, 10 and 12 per milliliter. The nauplii were counted by sight using a Pasteur pipette and transferred to Petri dishes containing 40 ml of brackish water (12 parts per thousand) lying on the top of black plastic. One larva at each stage was individually placed into each Petri dish containing different food density. After 24 h, each larva was removed from the Petri dish and the leftover nauplii were counted. The amount consumed was determined by the difference between the initial and final number of nauplii. Ingestion rate (I) increased as food density (P) increased and was defined by the model I=I-m(1-e(-kP)). The results suggest four levels of ingestion during larval development. The first level includes stages II, III and IV, with average maximum consumption of about 40 nauplii/day; the second level includes stages V and VI, with consumption of approximately 55 nauplii/day; the third level includes stages VII and VIII, with consumption of 80-100 nauplii/day. The fourth level includes stages IX, X and XI, in which the high values for maximum ingestion (Im) exceed the load capacity of the medium. The low values for constant k (that may correspond to the adaptability of the food to prey characteristics, such as, size, mobility, etc.) obtained for stages IX, X and XI indicated that Artemia is not an adequate prey and there is necessity of a supplementary diet. The best relationship between predator and prey seemed to occur during stage IV Results obtained in the present work may subsidize future researches and serve as a guideline for practical considerations of feeding rates. (C) 2003 Elsevier B.V. B.V. All rights reserved.

Comportamento alimentar do camarão de agua doce, Macrobrachium rosenbergii (De Man) (Crustacea, Palaemonidae) durante a fase larval: análise qualitativa

Feeding behavior of the larvae's prawn, Macrobrachium rosenbergii de Man, 1879, in each stage of development was studied face alArtemia Linnaeus, 1758 nauplii, dry and wet diets. Groups of ten larvae in each stage were placed in 500 ml beakers at 29ºC in filtered brackish water. After the fasting period of acclimatization, the three types of food were offered separately to the larvae and the feeding behavior was observed during 30 min. Feeding perception, capture, grasp and ingestion of food were used to determine feeding pattern. Stage I lawac do not eat. The feeding behavior of larvae was similar on the stages II - XI. The perception in distance does not occur in most individuais; the feeding process generally started with the physrcal contact of food with the animal. Food is captured by thoracic appendages (maxillipeds until stage III, and maxillipeds + pereopods in stages IV - XI) and particles are grasped and handled by maxillipeds. Inopposition at the capture, the ingestion is a seletive process. Selective responses decrease in later stages and larvae become omnivorous in greater degree.

Influence of food restriction on the reproduction and larval performance of matrinxã, Brycon amazonicus (Spix and Agassiz, 1829)

Este estudo avaliou o efeito da restrição alimentar e realimentação na reprodução de fêmeas e no crescimento inicial e sobrevivência de larvas de matrinxã, Brycon amazonicus. Matrizes distribuídas em 8 viveiros (15 peixes/tanque) foram alimentadas diariamente (em 4 tanques - G1) e alimentados em ciclos de 3 dias de alimentação seguidos de 2 dias de restrição (em 4 tanques - G2) por 6 meses antes da desova. Na indução à desova, 57% das fêmeas no G1 e 45% no G2 desovaram. Os pesos médios dos oócitos foram 208,1 g (G1) e 131,6 g (G2), sendo os oócitos G2 menores (1,017 ± 0,003 mm) que os oócitos de G1 (1,048 ± 0,002 mm). As taxas de fertilização (71,9 ± 12,6% e 61,2 ± 13,7%) e de eclosão (61,3 ± 33,9% e 67,5 ± 23,4%) entre os G1 e G2 não diferiram. Larvas foram coletadas na eclosão e às 24, 48 e 72 horas de incubação para medida do crescimento e as restantes transferidas para aquários e amostradas 1, 5, 9 e 15 dias depois. Na transferência, as larvas G1 e G2 tinham pesos similares (1,5 ± 0,15 e 1,46 ± 0,07 mg), mas o comprimento das larvas G2 era maior (6,2 ± 0,13 e 6,7 ± 0,14 mm). Ao 9° dia, quando é recomendada a transferência dos juvenis para tanques externos, os juvenis G2 tinham peso (13,6 ± 0,26 e 18,9 ± 0,07 mg) e comprimento (11,8 ± 0,09 e 14,5 ± 0,04 mm) maiores, mas no 15º dia os juvenis G1 eram maiores em peso (90,2 ± 1,19 e 68,6 ± 0,77 mg) e comprimento (18,8 ± 0,16 e 18,5 ± 0,04 mm). Aos 15 dias, a prole das fêmeas submetidas à restrição alimentar apresentou sobrevivência mais alta que a prole das fêmeas alimentadas diariamente (24,7 ± 2,07% e 19,2 ± 1,91%). A restrição alimentar imposta às fêmeas de matrinxã, apesar de reduzir o número de fêmeas que desovaram e a quantidade de oócitos extrusados, não afetou a fertilização e eclosão das larvas e melhorou a sobrevivência final das larvas.

High interannual variability in connectivity and genetic pool of a temperate clingfish matches oceanographic transport predictions

Adults of most marine benthic and demersal fish are site-attached, with the dispersal of their larval stages ensuring connectivity among populations. In this study we aimed to infer spatial and temporal variation in population connectivity and dispersal of a marine fish species, using genetic tools and comparing these with oceanographic transport. We focused on an intertidal rocky reef fish species, the shore clingfish Lepadogaster lepadogaster, along the southwest Iberian Peninsula, in 2011 and 2012. We predicted high levels of self-recruitment and distinct populations, due to short pelagic larval duration and because all its developmental stages have previously been found near adult habitats. Genetic analyses based on microsatellites countered our prediction and a biophysical dispersal model showed that oceanographic transport was a good explanation for the patterns observed. Adult sub-populations separated by up to 300 km of coastline displayed no genetic differentiation, revealing a single connected population with larvae potentially dispersing long distances over hundreds of km. Despite this, parentage analysis performed on recruits from one focal site within the Marine Park of Arrábida (Portugal), revealed self-recruitment levels of 2.5% and 7.7% in 2011 and 2012, respectively, suggesting that both long- and short-distance dispersal play an important role in the replenishment of these populations. Population differentiation and patterns of dispersal, which were highly variable between years, could be linked to the variability inherent in local oceanographic processes. Overall, our measures of connectivity based on genetic and oceanographic data highlight the relevance of long-distance dispersal in determining the degree of connectivity, even in species with short pelagic larval durations.

“Step by step”: high frequency short-distance epizoochorous dispersal of aquatic macrophytes

Aquatic macrophytes can successfully colonise and re-colonise areas separated by space and time. The mechanisms underlying such “mobility” are not well understood, but it has often been hypothesised that epizoochory (external dispersal) plays an important role. Yet, there is only limited, and mostly anecdotal, evidence concerning successful epizoochorous dispersal of aquatic macrophytes, particularly in the case of short-distance dispersal. Here we examine in situ and ex situ dispersal of aquatic macrophytes, including three invasive alien species. A high frequency of Lemna minor Linnaeus dispersal was observed in situ, and this was linked to bird-mediated epizoochory. We concluded that wind had no effect on dispersal. Similarly, in an ex situ examination Lemna minuta Kunth and Azolla filiculoides Lamarck, were found to be dispersed with a high frequency by mallard ducks (Anas platyrhynchos). No dispersal was measured for Elodea nuttalli (Planchon) H. St. John. It is concluded that short-distance or “stepping-stone” dispersal via bird-mediated epizoochory can occur with high frequencies, and therefore can play an important role in facilitating colonisation, range expansion and biological invasion of macrophytes.

Determinación de temperatura y densidad óptima en cultivo larval de lenguado Paralichthys adspersus (Steinda Chner, 1867) en condiciones de laboratorio

La presente tesis tiene como objetivo determinar la temperatura y densidad óptimas del cultivo larval de lenguado Paralichthys adspersus bajo condiciones controladas de laboratorio. Para el experimento de temperatura se usaron larvas de 2 días después de eclosión (DDE) hasta el día 46 DDE a una densidad de 10 larvas /litro, donde las temperaturas evaluadas fueron 16°, 18°, 20° y 22° con tres repeticiones por tratamientos. Para el experimento de densidad se usaron larvas de 5 DDE hasta el día 35 DDE, con una temperatura promedio de 19°, donde las densidades evaluadas fueron de 10larvas/litro, 20larvas/litro y 30 larvas/litro con tres repeticiones por tratamiento. Para ambos ensayos se realizó la evaluación de la longitud total (mm) y sobrevivencia; para el ensayo de temperatura de larvas de P. adspersus se obtuvo la más alta sobrevivencia a 18° C con 17 ±0,05% y una longitud de 12,25 ±1,06 mm. Para el ensayo de densidad de larvas de P. adspersus, se obtuvo la más alta sobrevivencia en el tratamiento de 10 larvas /litro con 10 ±3,72% y una longitud de 9,35 ±0,48 mm.