Methodologies for a sustainable development of estuarine ecosystems

As zonas costeiras, estuarinas e lagunares são consideradas áreas muito produtivas e dotadas de grande biodiversidade sendo, por isso, consideradas de elevado valor ecológico e económico. No entanto, nas últimas décadas tem vindo a verificar-se um aumento da contaminação destes ecossistemas como resultado de diversas actividades antrópicas. As abordagens actualmente disponíveis para avaliação do impacto da poluição em ecossistemas estuarinos e lagunares apresentam diversos tipos de lacunas, pelo que é importante desenvolver metodologias mais eficazes com organismos autóctones. Neste contexto, o objectivo central desta dissertação consistiu em desenvolver e validar métodos ecologicamente relevantes para avaliação da contaminação estuarina e dos seus efeitos, utilizando o góbio-comum (Pomatoschistus microps), quer como organismo-teste quer como espécie sentinela, devido à importante função que desempenha nas cadeias tróficas de diversos estuários da costa Portuguesa. A Ria de Aveiro foi seleccionada como área de estudo principalmente pelo facto de possuir zonas com diferentes tipos de contaminação predominante e de haver conhecimento científico de base abundante e de elevada qualidade sobre este ecosistema. Na primeira fase do estudo, foram investigados os efeitos agudos de dois hidrocarbonetos aromáticos policíclicos (HAPs) (benzo[a]pireno e antraceno), de um fuel-óleo e de dois metais (cobre e mercúrio) em P. microps, utilizando ensaios laboratoriais baseados em biomarcadores e em parâmetros comportamentais, os quais foram avaliados utilizando um dispositivo expressamente desenvolvido para o efeito, designado por speed performance device (SPEDE). Como biomarcadores foram utilizados parâmetros envolvidos em funções fisiológicas determinantes para a sobrevivência e desempenho dos animais (neurotransmissão, obtenção de energia, destoxificação e defesas anti-oxidantes), nomeadamente a actividade das enzimas acetilcolinesterase, lactato desidrogenase, CYP1A1, glutationa S-transferases, glutationa reductase, glutationa peroxidase, superóxido dismutase, catalase, tendo ainda sido determinados os níveis de peroxidação lipídica como indicador de danos oxidativos. De forma global, os resultados indicaram que os agentes e a mistura testados têm a capacidade de interferir com a função neurológica, de alterar as vias utilizadas para obtenção de energia celular, induzir as defesas antioxidantes e, no caso do cobre e do mercúrio, de causarem peroxidação lipídica. Foram ainda obtidas relações concentração-resposta a nível dos parâmetros comportamentais testados, nomeadamente a capacidade de nadar contra a corrente e a distância percorrida a nadar contra o fluxo de água, sugerindo que os agentes testados podem, por exemplo, diminuir a capacidade de fuga aos predadores, as probabilidades de captura de presas e o sucesso reprodutivo. Na segunda fase, tendo sido já adaptadas técnicas para determinação de vários biomarcadores em P. microps e estudada a sua resposta a dois grupos de poluentes particularmente relevantes em ecossistemas estuarinos e lagunares (metais e HAPs), foi efectuado um estudo de monitorização utilizando P. microps como bioindicador e que incluiu diversos parâmetros ecológicos e ecotoxicológicos, nomedamente: 20 parâmetros indicativos da qualidade da água e do sedimento, concentração de 9 metais em sedimentos e no corpo de P. microps, 8 biomarcadores e 2 índices de condição na espécie seleccionada. A amostragem foi efectuada em quatro locais da Ria de Aveiro, um considerado como referência (Barra) e três com diferentes tipos predominantes de contaminação (Vagueira, Porto de Aveiro e Cais do Bico), sazonalmente, durante um ano. Os resultados obtidos permitiram uma caracterização ecotoxicológica dos locais, incluindo informação sobre a qualidade da água, concentrações de contaminantes ambientais prioritários nos sedimentos e nos tecidos de P. microps, capacidade desta espécie para bioacumular metais, efeitos exercidos pelas complexas misturas de poluentes presentes em cada uma das zonas de amostragem nesta espécie e possíveis consequências para a população. A análise multivariada permitiu analisar de forma integrada todos os resultados, proporcionando informação que não poderia ser obtida analisando os dados de forma compartimentalizada. Em conclusão, os resultados obtidos no âmbito desta dissertação indicam que P. microps possui características adequadas para ser utilizado como organismoteste em ensaios laboratoriais (e.g. abundância, fácil manutenção, permite a determinação de diferentes tipos de critérios de efeito utilizando um número relativamente reduzido de animais, entre outras) e como organismo sentinela em estudos de monitorização da poluição e da qualidade ambiental, estando portanto de acordo com estudos de menor dimensão previamente efectuados. O trabalho desenvolvido permitiu ainda adaptar a P. microps diversas técnicas bioquímicas vulgarmente utilizadas como biomarcadores em Ecotoxicologia e validá-las quer no laboratório quer em cenários reais; desenvolver um novo bioensaio, utilizando um dispositivo de teste especialmente concebido para peixes epibentónicos baseado na performance natatória de uma espécie autóctone e em biomarcadores; relacionar os efeitos a nível bioquímico com parâmetros comportamentais que ao serem afectados podem reduzir de forma drástica e diversificada (e.g. aumento da mortalidade, diminuição do sucesso reprodutivo, redução do crescimento) a contribuição individual para a população. Finalmente, foi validada uma abordagem multidisciplinar, combinando metodologias ecológicas, ecotoxicológicas e químicas que, quando considerada de forma integrada utilizando análises de estatística multivariada, fornece informação científica da maior relevância susceptível de ser utilizada como suporte a medidas de conservação e gestão em estuários e sistemas lagunares.

Geographic variation in allometry in the guppy (Poecilia reticulata)

Variation in static allometry, the power relationship between character size and body size among individuals at similar developmental stages, remains poorly understood. We tested whether predation or other ecological factors could affect static allometry by comparing the allometry between the caudal fin length and the body length in adult male guppies (Poecilia reticulata) among populations from different geographical areas, exposed to different predation pressures. Neither the allometric slopes nor the allometric elevations (intercept at constant slope) changed with predation pressure. However, populations from the Northern Range in Trinidad showed allometry with similar slopes but lower intercepts than populations from the Caroni and the Oropouche drainages. Because most of these populations are exposed to predation by the prawn Macrobrachium crenulatum, we speculated that the specific selection pressures exerted by this predator generated this change in relative caudal fin size, although effects of other environmental factors could not be ruled out. This study further suggests that the allometric elevation is more variable than the allometric slope.

Blood lactate response and critical speed in swimmers aged 10-12 years of different standards

It has previously been shown that measurement of the critical speed is a non-invasive method of estimating the blood lactate response during exercise. However, its validity in children has yet to be demonstrated. The aims of this study were: (1) to verify if the critical speed determined in accordance with the protocol of Wakayoshi et al. is a non-invasive means of estimating the swimming speed equivalent to a blood lactate concentration of 4 mmol . l(-1) in children aged 10-12 years; and (2) to establish whether standard of performance has an effect on its determination. Sixteen swimmers were divided into two groups: beginners and trained. They initially completed a protocol for determination of speed equivalent to a blood lactate concentration of 4 mmol . l(-1). Later, during training sessions, maximum efforts were swum over distances of 50, 100 and 200 m for the calculation of the critical speed. The speeds equivalent to a blood lactate concentration of 4 mmol . l(-1) (beginners = 0.82 +/- 0.09 m . s(-1), trained = 1.19 +/- 0.11 m . s(-1); mean +/- s) were significantly faster than the critical speeds (beginners = 0.78 +/- 0.25 m . s(-1), trained = 1.08 +/- 0.04 m . s(-1)) in both groups. There was a high correlation between speed at a blood lactate concentration of 4 mmol . l(-1) and the critical speed for the beginners (r = 0.96, P < 0.001), but not for the trained group (r = 0.60, P > 0.05). The blood lactate concentration corresponding to the critical speed was 2.7 +/- 1.1 and 3.1 +/- 0.4 mmol . l(-1) for the beginners and trained group respectively. The percent difference between speed at a blood lactate concentration of 4 mmol . l(-1) and the critical speed was not significantly different between the two groups. At all distances studied, swimming performance was significantly faster in the trained group. Our results suggest that the critical speed underestimates swimming intensity corresponding to a blood lactate concentration of 4 mmol . l(-1) in children aged 10-12 years and that standard of performance does not affect the determination of the critical speed.

Potência atada na máxima fase estável de lactato e índices do desempenho aeróbio de nado

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effetti idrodinamici prodotti dal costume tecnico sul nuotatore

L’introduzione dei costumi tecnici nel nuoto ha portato miglioramenti senza precedenti sulla prestazione. I miglioramenti nella velocità di nuoto sono stati attribuiti dalla letteratura a riduzioni nelle resistenze idrodinamiche sul nuotatore. Tuttavia, gli effetti specifici dovuti all’utilizzo di questo tipo di costume non sono ancora completamente chiariti. Questa tesi aveva l’obiettivo di indagare gli effetti del costume tecnico sul galleggiamento statico, sulla posizione del corpo e sulla resistenza idrodinamica in avanzamento passivo. Nello studio preliminare sono stati misurati la spinta idrostatica, i volumi polmonari dinamici e la circonferenza toracica di 9 nuotatori che indossavano un costume tradizionale o un costume tecnico in gomma sintetica. Indossare il costume tecnico ha determinato una riduzione significativa del galleggiamento statico, e la compressione toracica causata da questo tipo di costume potrebbe avere una relazione con la significativa riduzione dei volumi polmonari misurati quando il nuotatore indossa questo tipo di costume. Un successiva analisi prevedeva il traino passivo di 14 nuotatori che mantenevano la miglior posizione idrodinamica di scivolamento indossando un costume tradizionale, tecnico in tessuto e tecnico in gomma. La posizione del corpo in avanzamento è stata misurata con un’analisi cinematica. La resistenza passiva indossando i costumi tecnici è risultata significativamente minore per entrambi i costumi tecnici rispetto alla prova con costume tradizionale. L’analisi condotta attraverso modelli di regressione lineari ha mostrato che una parte della riduzione della resistenza passiva era legata a proprietà intrinseche dei costumi tecnici. Tuttavia, anche l’area di impatto frontale determinata dall’inclinazione del tronco del soggetto in scivolamento e l’inclinazione degli arti inferiori hanno mostrato una marcata influenza sulla resistenza idrodinamica passiva. Pertanto, la riduzione di resistenza idrodinamica durante lo scivolamento passivo effettuato con costume tecnico da nuoto è attribuibile, oltre all’effetto del materiale di composizione del costume, ad una variazione della posizione del corpo del nuotatore.

Effect of thermal acclimation on female resistance to forced matings in the eastern mosquitofish

All copulations in the eastern mosquitofish, Gambusia holbrooki, are coercive-and-achieved by force. Female G. holbrooki never appear to cooperate with males, but vigorously resist matings at all times. We examined the role of females within a sexually coercive mating system by investigating the ability of female G. holbrooki to resist forced copulations after acclimation to 16 degrees C and 32 degrees C for 4-5 weeks. We also examined burst swimming performance of female G. holbrooki after acclimation, as this trait is likely to underlie a female's ability to resist forced matings. We predicted that if female G. holbrooki indiscriminately resist matings from all males, acclimation would enhance female resistance at their acclimation temperature. However, we found that it did not. We also predicted that if females are able to influence the outcome of mating interactions, acclimation to an optimal thermal environment may induce females to reduce resistance. In support of this prediction, females acclimated at 32 degrees C were able to modify their resistance behaviour between exposure to 16 degrees C and 32 degrees C. The rate of copulations experienced by 32 inverted perpendicular C-acclimated females was 2.5 times greater at 32 degrees C than at 16 degrees C. In addition, acclimation at 32 degrees C significantly enhanced burst swimming performance at 32 degrees C but no effect of acclimation was observed at 16 degrees C. Our results suggest that female G. holbrooki are able to play a greater role in determining the outcome of sexual coercive mating interactions than previously thought. (c) 2006 The Association for the Shidy of Animal Behavioor. Published by Elsevier Ltd. All rights reserved.

Substantial changes in the genetic basis of tadpole morphology of Rana lessonae in the presence of predators

Predator-induced morphological plasticity is a model system for investigating phenotypic plasticity in an ecological context. We investigated the genetic basis of the predator-induced plasticity in Rana lessonae by determining the pattern of genetic covariation of three morphological traits that were found to be induced in a predatory environment. Body size decreased and tail dimensions increased when reared in the presence of preying dragonfly larvae. Genetic variance in body size increased by almost an order of magnitude in the predator environment, and the first genetic principal component was found to be highly significantly different between the two environments. The across environment genetic correlation for body size was significantly below 1 indicating that different genes contributed to this trait in the two environments. Body size may therefore be able to respond to selection independently in the two environments to some extent.

Investigations on the Chilean kelp crab Taliepus dentatus

Physiological responses of larval stages can differ from those of the adults, affecting key ecological processes. Therefore, developing a mechanistic understanding of larval responses to environmental conditions is essential vis-à-vis climate change. We studied the thermal tolerance windows, defined by lower and upper pejus (Tp) and critical temperatures (Tc), of zoea I, II, and megalopa stages of the Chilean kelp crab Taliepus dentatus. Tp limits determine the temperature range where aerobic scope is maximal and functioning of the organism is unrestrained and were estimated from direct observations of larval activity. Tc limits define the transition from aerobic to anaerobic metabolism, and were estimated from the relationship between standard metabolic rate and temperature. Zoea I showed the broadest, Zoea II an intermediate, and megalopae the narrowest tolerance window (Tp). Optimum performance in megalopae was limited to Tp between 11 and 15°C, while their Tc ranged between 7 and 19°C. Although Tc may be seldom encountered by larvae, the narrower Tp temperatures can frequently expose larvae to unfavorable conditions that can drastically constrain their performance. Temperatures beyond the Tp range of megalopae have been observed in most spring and summer months in central Chile, and can have important consequences for larval swimming performance and impair their ability to avoid predators or settle successfully. Besides the well-documented effects of temperature on development time, variability in field temperatures beyond Tp can affect performance of particular larval stages, which could drive large-scale variability in recruitment and population dynamics of T. dentatus and possibly other invertebrate species.

Les impacts de mitochondries étrangères sur le phénotype : une étude holistique chez le ventre rouge du nord Chrosomus eos

Les organismes cybrides souffrent généralement d'une altération de la spécificité des interactions mito-nucléaires, résultant en une détérioration du phénotype. Toutefois, diverses études démontrent que le transfert de mitochondries peut occasionnellement être positif. À l'heure actuelle, de nombreuses questions demeurent quant au degré d'influence de ces transferts sur les différents niveaux d'organisation du phénotype. Afin de répondre à ces questions, les formes sauvages et cybrides du poisson Chrosomus eos sont étudiées. Ainsi, le premier volet de ce projet de recherche démontre un impact des mitochondries Chrosomus neogaeus à différents niveaux d'organisation du phénotype des poissons C. eos, lorsque les formes sauvages et cybrides sont retrouvées en allopatrie. Le deuxième volet de cette thèse révèle, quant à lui, que ces modifications phénotypiques ne sont pas suffisantes pour induire un évènement de spéciation entre les deux biotypes, lorsqu'en sympatrie. De plus, cette étude suggère que la coévolution mito-nucléaire peut ne pas être une condition sine qua non à la perpétuation des individus en milieu naturel. Finalement, l'approche holistique considérée dans le troisième volet de cette recherche atteste de l'influence des mitochondries C. neogaeus à différents niveaux d'organisation du phénotype de C. eos, lorsque les formes sauvages et cybrides sont sympatriques. Cette influence est moins prononcée que celle observée à partir de biotypes allopatriques. Combinés, ces chapitres contribuent à une meilleure compréhension des liens existant entre les mitochondries et le phénotype d'un individu.

Context dependency of trait repeatability and its relevance for management and conservation of fish populations

Repeatability of behavioural and physiological traits is increasingly a focus for animal researchers, for which fish have become important models. Almost all of this work has been done in the context of evolutionary ecology, with few explicit attempts to apply repeatability and context dependency of trait variation toward understanding conservation-related issues. Here, we review work examining the degree to which repeatability of traits (such as boldness, swimming performance, metabolic rate and stress responsiveness) is context dependent. We review methods for quantifying repeatability (distinguishing between within-context and across-context repeatability) and confounding factors that may be especially problematic when attempting to measure repeatability in wild fish. Environmental factors such temperature, food availability, oxygen availability, hypercapnia, flow regime and pollutants all appear to alter trait repeatability in fishes. This suggests that anthropogenic environmental change could alter evolutionary trajectories by changing which individuals achieve the greatest fitness in a given set of conditions. Gaining a greater understanding of these effects will be crucial for our ability to forecast the effects of gradual environmental change, such as climate change and ocean acidification, the study of which is currently limited by our ability to examine trait changes over relatively short time scales. Also discussed are situations in which recent advances in technologies associated with electronic tags (biotelemetry and biologging) and respirometry will help to facilitate increased quantification of repeatability for physiological and integrative traits, which so far lag behind measures of repeatability of behavioural traits.

Intraspecific individual variation of temperature tolerance associated with oxygen demand in the European sea bass (Dicentrarchus labrax)

The European sea bass (Dicentrarchus labrax) is an economically important fish native to the Mediterranean and Northern Atlantic. Its complex life cycle involves many migrations through temperature gradients that affect the energetic demands of swimming. Previous studies have shown large intraspecific variation in swimming performance and temperature tolerance, which could include deleterious and advantageous traits under the evolutionary pressure of climate change. However, little is known of the underlying determinants of this individual variation. We investigated individual variation in temperature tolerance in 30 sea bass by exposing them to a warm temperature challenge test. The eight most temperature-tolerant and eight most temperature-sensitive fish were then studied further to determine maximal swimming speed (U-CAT), aerobic scope and post-exercise oxygen consumption. Finally, ventricular contractility in each group was determined using isometric muscle preparations. The temperature-tolerant fish showed lower resting oxygen consumption rates, possessed larger hearts and initially recovered from exhaustive exercise faster than the temperature-sensitive fish. Thus, whole-animal temperature tolerance was associated with important performance traits. However, the temperature-tolerant fish also demonstrated poorer maximal swimming capacity (i.e. lower UCAT) than their temperature-sensitive counterparts, which may indicate a trade-off between temperature tolerance and swimming performance. Interestingly, the larger relative ventricular mass of the temperature-tolerant fish did not equate to greater ventricular contractility, suggesting that larger stroke volumes, rather than greater contractile strength, may be associated with thermal tolerance in this species.

Les impacts de mitochondries étrangères sur le phénotype : une étude holistique chez le ventre rouge du nord Chrosomus eos

Les organismes cybrides souffrent généralement d'une altération de la spécificité des interactions mito-nucléaires, résultant en une détérioration du phénotype. Toutefois, diverses études démontrent que le transfert de mitochondries peut occasionnellement être positif. À l'heure actuelle, de nombreuses questions demeurent quant au degré d'influence de ces transferts sur les différents niveaux d'organisation du phénotype. Afin de répondre à ces questions, les formes sauvages et cybrides du poisson Chrosomus eos sont étudiées. Ainsi, le premier volet de ce projet de recherche démontre un impact des mitochondries Chrosomus neogaeus à différents niveaux d'organisation du phénotype des poissons C. eos, lorsque les formes sauvages et cybrides sont retrouvées en allopatrie. Le deuxième volet de cette thèse révèle, quant à lui, que ces modifications phénotypiques ne sont pas suffisantes pour induire un évènement de spéciation entre les deux biotypes, lorsqu'en sympatrie. De plus, cette étude suggère que la coévolution mito-nucléaire peut ne pas être une condition sine qua non à la perpétuation des individus en milieu naturel. Finalement, l'approche holistique considérée dans le troisième volet de cette recherche atteste de l'influence des mitochondries C. neogaeus à différents niveaux d'organisation du phénotype de C. eos, lorsque les formes sauvages et cybrides sont sympatriques. Cette influence est moins prononcée que celle observée à partir de biotypes allopatriques. Combinés, ces chapitres contribuent à une meilleure compréhension des liens existant entre les mitochondries et le phénotype d'un individu.

Effects of dry-land strength and conditioning programs in age group swimmers

Context: Even though dry-land S&C training is a common practice in swimming, there are countless uncertainties over it effects in performance of age group swimmers. Objective: To investigate the effects of dry-land S&C programs in swimming performance of age group swimmers. Participants: A total of 21 male competitive swimmers (12.7±0.7 years) were randomly assigned to the Control Group (n=7) and experimental GR1 and GR2 (n=7 for each group). Intervention: Control group performed a 10-week training period of swim training alone, GR1 followed a 6-week dry-land S&C program based on sets/repetitions plus a 4-week swim training program alone and GR2 followed a 6-week dry-land S&C program focused on explosiveness, plus a 4-week program of swim training alone. Results: For the dry-land tests a time effect was observed between week 0 and week 6 for vertical jump (p<0.01) in both experimental groups, and for the GR2 ball throwing (p<0.01), with moderate-strong effect sizes. The time*group analyses showed that for performance in 50 m, differences were significant, with the GR2 presenting higher improvements than their counterparts (F=4.156; ƿ=0.007; η2=0.316) at week 10. Conclusions: The results suggest that 6 weeks of a complementary dry-land S&C training may lead to improvements in dry-land strength. Furthermore, a 4-week adaptation period was mandatory to achieve beneficial transfer for aquatic performance. Additional benefits may occur if coaches plan the dry-land S&C training focusing on explosiveness.

Effects of taper on swimming force and swimmer performance after an experimental ten-week training program

Papoti, M., L.E.B. Martins, S.A. Cunha, A.M. Zagatto, and C.A. Gobatto. Effects of taper on swimming force and swimmer performance after an experimental ten-week training program. J. Strength Cond. Res. 21(2):538-542. 2007.- The purpose of this research was to examine how an 11-day taper after an 8.5-week experimental training cycle affected lactate levels during maximal exercise, mean force, and performance in training swimmers, independent of shaving, psychological changes, and postcompetition effects. Fourteen competition swimmers with shaved legs and torsos were recruited from the São Paulo Aquatic Federation. The training cycle consisted of a basic training period (endurance and quality phases) of 8.5 weeks, with 5,800 m·d -1 mean training volume and 6 d·wk -1 frequency; and a taper period (TP) of 1.5 weeks' duration that incorporated a 48% reduction in weekly volume without altering intensity. Attained swimming force (SF) and maximal performance over 200m maximal swim (Pmax) before and after taper were measured. After taper, SF and Pmax improved 3.6 and 1.6%, respectively (p < 0.05). There were positive correlations (p < 0.05) between SF and Pmax before (r = 0.86) and after (r = 0.83) the taper phase. Peak lactate concentrations after SF were unaltered before (6.79 ± 1.2 mM) and after (7.15 ± 1.8 mM) TP. Results showed that TP improved mean swimming velocity, but not in the same proportion as force after taper, suggesting that there are other factors influencing performance in faster swimming. © 2007 National Strength & Conditioning Association.