Spatial behavior of two coral reef fishes within a Caribbean Marine Protected Area

A better understanding of the key ecological processes of marine organisms is fundamental to improving design and effective implementation of marine protected areas (MPAs) and marine biodiversity. The movement behavior of coral reef fish is a complex mechanism that is highly linked to species life-history traits, predation risk and food resources. We used passive acoustic telemetry to study monthly, daily and hourly movement patterns and space use in two species, Schoolmaster snapper (Lutjanus apodus) and Stoplight parrotfish (Sparisoma viride). We investigated the spatial overlap between the two species and compared intra-specific spatial overlap between day and night. Presence-absence models showed different diel presence and habitat use patterns between the two species. We constructed a spatial network of the movement patterns, which showed that for both species when fish were detected by the array of receivers most movements were made around the coral reef habitat while occasionally moving to silt habitats. Our results show that most individuals made predictable daily crepuscular migrations between different locations and habitat types, although individual behavioral changes were observed for some individuals across time. Our study also highlights the necessity to consider multiple species during MPA implementation and to take into account the specific biological and ecological traits of each species. The low number of fish detected within the receiver array, as well as the intraspecific variability observed in this study, highlight the need to compare results across species and individuals to be used for MPA management.

Assessment of a data-limited, multi-species shark fishery in the Great Barrier Reef Marine Park and south-east Queensland

The status of five species of commercially exploited sharks within the Great Barrier Reef Marine Park (GBRMP) and south-east Queensland was assessed using a data-limited approach. Annual harvest rate, U, estimated empirically from tagging between 2011 and 2013, was compared with an analytically-derived proxy for optimal equilibrium harvest rate, UMSY Lim. Median estimates of U for three principal retained species, Australian blacktip shark, Carcharhinus tilstoni, spot-tail shark, Carcharhinus sorrah, and spinner shark, Carcharhinus brevipinna, were 0.10, 0.06 and 0.07 year-1, respectively. Median U for two retained, non-target species, pigeye shark, Carcharhinus amboinensis and Australian sharpnose shark, Rhizoprionodon taylori, were 0.27 and 0.01 year-1, respectively. For all species except the Australian blacktip the median ratio of U/UMSY Lim was <1. The high vulnerability of this species to fishing combined with life history characteristics meant UMSY Lim was low (0.04-0.07 year-1) and that U/UMSY Lim was likely to be > 1. Harvest of the Australian blacktip shark above UMSY could place this species at a greater risk of localised depletion in parts of the GBRMP. Results of the study indicated that much higher catches, and presumably higher U, during the early 2000s were likely unsustainable. The unexpectedly high level of U on the pigeye shark indicated that output-based management controls may not have been effective in reducing harvest levels on all species, particularly those caught incidentally by other fishing sectors including the recreational sector. © 2016 Elsevier B.V.

Detection of viral diseases in shrimp Litopenaeus vannamei in Iran with emphasis on white spot disease prevention by using marine alga extract

Nowadays, following was expanded shrimp breeding and culture; viral diseases have been main problem which threatened shrimp industry in the country. Therefore, shrimp samples were obtained from different stages of Litopenaeus vannmei life cycle (larval, post larval, juveniles, adults and broodstocks) based on clinical signs in the breeding center and shrimp farming from Bushehr, Khozestan and Sistan and Baluchestan provinces. Viral diseases were detected by PCR (Polymerase Chain Reaction), histopathology and transmission electron microscopy (TEM) methods. Results of the PCR were indicated present white spot virus (WSV) in juveniles, sub adults and adults shrimp with medium intensity from three provinces, but it was not showed in larval and post larval stages. Histopathological sections were indicated hypertrophy and basophilic Cowdry type A formation in nucleus cells of gill, haematopoietic, lymphoid and epithelial's cuticles and intestinal tissues which was associated with small vacuoles increased in B cells of hepatopancreas tissue of infection shrimps. Transmission electronic microscopic studies were demonstrated that the length and diameter virus was detected, respectively, 300 ± 20 nm and 75 ± 5 nm. Considerable, results of the PCR were only displayed IHHNV in juvenile, adult and broodstock shrimps from breeding and farming center of Bushehr province. The main lesion pathology was formed eosinophilic Cowdry type A in nucleus cells of gill, haematopoietic, lymphoid and epithelial's cuticles and intestinal tissues. Whereas penaeid shrimps are lack specific immune system, hence, in the present study was used of marine alga (Lurensia snideria) collected from along costal Persian Gulf of Bushehr province for viral diseases were prevented. Powder alga extract were added with a ratio of 1 % to shrimp diet. Total haemocyte count (THC) and total protein plasma (TPP) were increased after 5 days of oral administration diets. When shrimps were infected by with spot virus experimentally, THC and TPP gradually were increased in both two groups (shrimps fed with diet containing alga extract and without alga extract) after 48h. Nevertheless; THC, TPP and survival of shrimp fed with diet containing alga extract were more than shrimp control in 15 days. So, oral administration Lurensia snideria extract was capable prevention infected L. vannamei via stimulant specific immune system.

Assessment and Life-Cycle Analysis of Recycled Materials for Sustainable Highway

Recycled materials replacing part of virgin materials in highway applications has shown great benefits to the society and environment. Beneficial use of recycled materials can save landfill places, sparse natural resources, and energy consumed in milling and hauling virgin materials. Low price of recycled materials is favorable to cost-saving in pavement projects. Considering the availability of recycled materials in the State of Maryland (MD), four abundant recycled materials, recycled concrete aggregate (RCA), recycled asphalt pavement (RAP), foundry sand (FS), and dredged materials (DM), were studied. A survey was conducted to collect the information of current usage of the four recycled materials in States’ Department of Transportation (DOTs). Based on literature review, mechanical and environmental properties, recommendations, and suggested test standards were investigated separately for the four recycled materials in different applications. Constrains in using these materials were further studied in order to provide recommendations for the development of related MD specifications. To measure social and environmental benefits from using recycled materials, life-cycle assessment was carried out with life-cycle analysis (LCA) program, PaLATE, and green highway rating system, BEST-in-Highway. The survey results indicated the wide use of RAP and RCA in hot mix asphalt (HMA) and graded aggregate base (GAB) respectively, while FS and DM are less used in field. Environmental concerns are less, but the possibly low quality and some adverse mechanical characteristics may hinder the widely use of these recycled materials. Technical documents and current specifications provided by State DOTs are good references to the usage of these materials in MD. Literature review showed consistent results with the survey. Studies from experimental research or site tests showed satisfactory performance of these materials in highway applications, when the substitution rate, gradation, temperature, moisture, or usage of additives, etc. meet some requirements. The results from LCA revealed significant cost savings in using recycled materials. Energy and water consumption, gas emission, and hazardous waste generation generally showed reductions to some degree. Use of new recycled technologies will contribute to more sustainable highways.

Seaweed Aquaculture and Marine Biotechnology

Macroscopic marine algae, typically known as macroalgae or seaweeds, form an important living resource of the oceans, as primary producers. People have collected seaweeds for food, both for humans and animals for millennia. They also have been a source of nutrient rich fertilizers, as well as a source of gelling agents known as phycocolloids. More recently macroalgae are playing significant roles in medicine and biotechnology. Although Biotechnology and in particular marine biotechnology may have different meanings for different people, under the present context we will consider a broader definition. Marine biotechnology consists on the use of biological knowledge and/or the application of biological techniques on marine organisms, for the development of products in some way beneficial for humans. Seaweed aquaculture is, therefore a biotechnology activity. It is also one that can allow for further development of the industry. Today, seaweed cultivation techniques are standardized, routine and economical. Several factors, including understanding the environmental regulation of life histories and asexual propagation of thalli, are responsible for the success of large-scale seaweed cultivation. Presently, seaweed aquaculture represents approximately 23% of the world’s aquaculture production, including fish, crustaceans and other animals. A promising approach for the development of seaweed aquaculture, and aquaculture in general, is the integrated multi-trophic aquaculture (IMTA). In these systems, fed-aquaculture is combined with extractive organisms like bivalves and/or algae. The constraints and advantages of IMTA will be discussed. In particular, land based IMTA systems allow for much greater environmental and input controls. Traceability, security of supply, high-quality standards and safety should be the future of seaweed aquaculture and contribute for the development of marine biotechnology.

Early life history of larvae and early juvenile atlantic horse mackerel trachurus trachurus off the portuguese west coast

Early life history traits (ELHTs) are key to understand recruitment patterns in marine animals. However, for reef fishes, studies on ELHTs are mainly focused on tropical systems and little is known for temperate reefs. In this study we used SMURFs (Standard Monitoring Units for the Recruitment of Reef Fishes) to collect fish in a temperate rocky reef system (Arrábida Marine Park, Portugal) on a weekly basis for three months during the recruitment period. Six sub-surface SMURFs sampled 2490 Atlantic horse mackerel (Trachurus trachurus) postlarvae and juveniles. Sagittal and lapilli otoliths were extracted from a subsample of 296 fish and ELHTs, such as size and age at settlement, growth rate and age at first secondary growth formation were examined. Additionally, we tested three growth curves and selected the best suited to back-calculate the hatching pattern based on the lengths of all sampled fish. Standard length ranged from 6.13 mm to 48.56 mm and subsampled fish were aged between 19 days to 44 days. Age and size at settlement were estimated between 19 days and 36 days for individuals of 6.13 mm and 24.95 mm, respectively. Otolith shape changed clearly with increasing age and, on average, secondary growth started to form on day 33 (±3 days). Age/length relationship was well described by a Gompertz growth model which was used to back-calculate hatching dates. Four distinct hatching cohorts were identified with fish of the earliest cohort showing a faster body and otolith growth. This study indicates that the nearshore environment might have an important role in the early growth, development and hence recruitment of Atlantic horse mackerel. Information on the early life history of Atlantic horse mackerel is key to understand recruitment processes for this economically and biologically important species.

Multibiomarker assessment of cadmium-based quantum dots effects in the marine mussel Mytilus galloprovincialis

Tese de Doutoramento, Ciências do Mar, da Terra e do Ambiente, Ramo: Ciências e Tecnologias do Ambiente, Especialização em Ecotoxicologia, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2016

Environmental variables, habitat discontinuity and life history shaping the genetic structure of Pomatoschistus marmoratus

Coastal lagoons are semi-isolated ecosystems exposed to wide fluctuations of environmental conditions and showing habitat fragmentation. These features may play an important role in separating species into different populations, even at small spatial scales. In this study, we evaluate the concordance between mitochondrial (previous published data) and nuclear data analyzing the genetic variability of Pomatoschistus marmoratus in five localities, inside and outside the Mar Menor coastal lagoon (SE Spain) using eight microsatellites. High genetic diversity and similar levels of allele richness were observed across all loci and localities, although significant genic and genotypic differentiation was found between populations inside and outside the lagoon. In contrast to the FST values obtained from previous mitochondrial DNA analyses (control region), the microsatellite data exhibited significant differentiation among samples inside the Mar Menor and between lagoonal and marine samples. This pattern was corroborated using Cavalli-Sforza genetic distances. The habitat fragmentation inside the coastal lagoon and among lagoon and marine localities could be acting as a barrier to gene flow and contributing to the observed genetic structure. Our results from generalized additive models point a significant link between extreme lagoonal environmental conditions (mainly maximum salinity) and P. marmoratus genetic composition. Thereby, these environmental features could be also acting on genetic structure of coastal lagoon populations of P. marmoratus favoring their genetic divergence. The mating strategy of P. marmoratus could be also influencing our results obtained from mitochondrial and nuclear DNA. Therefore, a special consideration must be done in the selection of the DNA markers depending on the reproductive strategy of the species.

In two waters: contemporary evolution of lagoonal and marine white seabream (Diplodus sargus) populations

Brackish water ecosystems are often exposed to wide variations in environmental variables, including temperature and salinity, which may cause strong selective pressures on organisms modifying the genetic patterns of species. The aim of this work was to test whether there is a ‘divergence-with-gene flow’ in coastal lagoon populations of white seabream (Diplodus sargus) (Ria Formosa, S Portugal and Mar Menor, SE Spain) respect to four marine populations, by using partial sequences of cyt b mitochondrial gene and information from nine microsatellite loci. Genetic diversity was highest in both coastal lagoons (Mar Menor and Ria Formosa) considering mitochondrial and nuclear markers. Although some of FST population pairwise comparisons were not significant, analyses of molecular variance (AMOVAs) detected differences between groups (coastal lagoon and marine) close to significance. Also, only two haplotypes (Cytb-17 and Cytb-18) were detected in both coastal lagoon sampling sites and these localities (Mar Menor and Ria Formosa) showed the highest number of singletons, some of them with a high number of mutations, as has been already described for other Mar Menor populations (Pomatochistus marmoratus and Holothuria polii). Also, several tests detected significant positive and balancing selection considering mtDNA and microsatellite data. These data support the hypothesis of selection as one of the drivers of the genetic differences found between coastal lagoon and marine populations. The life strategy adopted by Diplodus sargus in coastal lagoons allows it to decrease its mortality rate and improve the heritability of its genes. Also, the increase time spent in coastal lagoons with different temperatures and salinities favours the fitness selection and the maintenance of exclusive haplotypes and genotypes in coastal lagoon inhabitants favouring the ‘divergence-with-gene-flow’.

Feeding ecology and life-history strategy of nesting males in a fish with long parental care, Lusitanian toadfish (Halobatrachus didactylus, Batrachoididae)

The Lusitanian toadfish, Halobatrachus didactylus, like other batrachoidids, is a benthic fish species with nesting behaviour during the breeding season. During this prolonged period it engages in mating activities and remains in the nest providing parental care. It is not known whether males feed while providing parental care but it is likely that their limited mobility may restrict their diet and influence their fitness. As a consequence, egg cannibalism could occur as a life-history strategy. The aim of the present study is to ascertain the feeding behaviour of nesting males, in comparison to mature non-nesting males, and to identify potential life-history traits related to egg cannibalism. Nest-holders were sampled from artificial nests placed in an intertidal area of the Tagus estuary, only exposed during spring low tides. The diet of nest-holders was compared with that of non-nesting mature males from the same area, captured by otter trawl. The present study demonstrates that despite their constrained mobility nest-holders feed during the breeding season, although in a more opportunistic fashion than non-nesting males. Nest-holders showed a generalist feeding behaviour, with a more heterogeneous diet. Egg cannibalism was not related to male condition, paternity or brood size but showed a higher incidence early in the season when water temperatures were lower. The results suggest a possible seasonal trade-off strategy between care and energy recovery, triggered by environmental factors, where under unfavourable conditions to sustain viable eggs the male may recover energy by eating eggs, thus benefiting future reproductive success, later in the season.

System innovation and life cycle thinking in packaging value chain: the circularity of plastics.

Compared to other, plastic materials have registered a strong acceleration in production and consumption during the last years. Despite the existence of waste management systems, plastic_based materials are still a pervasive presence in the environment, with negative consequences on marine ecosystem and human health. The recycling is still challenging due to the growing complexity of product design, the so-called overpackaging, the insufficient and inadequate recycling infrastructure, the weak market of recycled plastics and the high cost of waste treatment and disposal. The Circular economy package, the European Strategy for plastics in a circular economy and the recent European Green Deal include very ambitious programmes to rethink the entire plastic value chain. As regards packaging, all plastic packaging will have to be 100% recyclable (or reusable) and 55% recycled by 2030. Regions are consequently called upon to set up a robust plan able to fit the European objectives. It takes on greater importance in Emilia Romagna where the Packaging valley is located. This thesis supports the definition of a strategy aimed to establish an after-use plastics economy in the region. The PhD work has set the basis and the instruments to establish the so-called Circularity Strategy with the aim to turn about 92.000t of plastic waste into profitable secondary resources. System innovation, life cycle thinking and participative backcasting method have allowed to deeply analyse the current system, orientate the problem and explore sustainable solutions through a broad stakeholder participation. A material flow analysis, accompanied by a barrier analysis, has supported the identification of the gaps between the present situation and the 2030 scenario. Eco-design for and from recycling (and a mass _based recycling rate (based on the effective amount of plastic wastes turned into secondary plastics), valorized by a value_based indicator, are the key-points of the action plan.

Molecular mechanisms controlling physiological plasticity in marine mussels under the influence of natural and anthropogenic stress factors

Marine mussels are exceptionally well-adapted to live in transitional habitats where they are exposed to fluctuating environmental parameters and elevated levels of natural and anthropogenic stressors throughout their lifecycle. However, there is a dearth of information about the molecular mechanisms that assist in dealing with environmental changes. This project aims to investigate the molecular mechanisms governing acclimatory and stress responses of the Mediterranean mussel (Mytilus galloprovincialis) by addressing relevant life stages and environmental stressors of emerging concern. The experimental approach consisted of two phases to explore (i) the physiological processes at early life history and the consequences of plastic pollution and (ii) the adult physiology processes under natural habitats. As the first phase, I employed a plastic leachate (styrene monomer), and polystyrene microplastics to understand the modulation of cytoprotective mechanisms during the early embryo stages. Results revealed the onset of transcriptional impairments of genes involved in MXR-related transporters and other physiological processes induced by styrene and PS-MPs. In the second phase, as a preliminary analysis, microbiota profile of adult mussels at the tissue scale and its surrounding water was explored to understand microbiota structures that may reflect peculiar adaptations to the respective tissue functions. The broader experiment has been implemented to understand the variability of transcriptional profiles in the mussel digestive glands in the natural setting. All the genes employed in this study have shown possibilities to use as molecular biomarker responses throughout the year for monitoring the physiology of mussels living in a particular environment and, in turn, more properly detecting changes in the environment. As a whole, my studies provide insights into the interactions between environmental parameters, and intrinsic characters, and physiology of marine bivalves, and it could help to interpretation of responses correctly under stress conditions and climate change scenarios.

Impact of anthropogenic stressors on the microbial communities in marine holobionts and ecosystems

Free-living or host-associated marine microbiomes play a determinant role in supporting the functioning and biodiversity of marine ecosystems, providing essential ecological services, and promoting the health of the entire biosphere. Currently, the fast and restless increase of World’s human population strongly impacts life on Earth in the forms of ocean pollution, coastal zone destruction, overexploitation of marine resources, and climate change. Thanks to their phylogenetic, metabolic, and functional diversity, marine microbiomes represent the Earth’s biggest reservoir of solutions against the major threats that are now impacting marine ecosystems, possibly providing valuable insights for biotechnological applications to preserve the health of the ocean ecosystems. Microbial-based mitigation strategies heavily rely on the available knowledge on the specific role and composition of holobionts associated microbial communities, thus highlighting the importance of pioneer studies on microbial-mediated adaptive mechanisms in the marine habitats. In this context, we propose different models representing ecologically important, widely distributed, and habitat-forming organisms, to further investigate the ability of marine holobionts to dynamically adapt to natural environmental variations, as well as to anthropogenic stress factors. In this PhD thesis, we were able to supply the characterization of the microbial community associated with the model anthozoan cnidaria Corynactis viridis throughout a seasonal gradient, to provide critical insights into microbiome-host interactions in a biomonitoring perspective. We also dissected in details the microbial-derived mitigation strategies implemented by the benthonic anthozoan Anemonia viridis and the gastropod Patella caerulea as models of adaptation to anthropogenic stressors, in the context of bioremediation of human-impacted habitats and for the monitoring and preservation of coastal marine ecosystems, respectively. Finally, we provided a functional model of adaptation to future ocean acidification conditions by characterizing the microbial community associated with the temperate coral Balanophyllia europaea naturally living at low pH conditions, to implement microbial based actions to mitigate climate change.

Monitoring of Mediterranean marine benthic biodiversity through novel, standardized, and integrated approaches

Marine healthy ecosystems support life on Earth and human well-being thanks to their biodiversity, which is proven to decline mainly due to anthropogenic stressors. Monitoring how marine biodiversity changes trough space and time is needed to properly define and enroll effective actions towards habitat conservation and preservation. This is particularly needed in those areas that are very rich in species compared to their low surface extension and are characterized by strong anthropic pressures, such as the Mediterranean Sea. Subtidal rocky benthic Mediterranean habitats have a complex structural architecture, hosting a panoply of tiny organisms (cryptofauna) that inhabit crevices and caves, but that are still unknown. Different artificial standardized sampling structures (SSS) and methods have been developed and employed to characterize the cryptofauna, allowing for data replicability and comparability across regions. Organisms growing on these artificial structures can be identified coupling morphological taxonomy and DNA barcoding and metabarcoding. The metabarcoding allows for the identification of organisms in a bulk sample without morphological analysis, and it is based on comparing the genetic similarities of the assessed organisms with barcoding sequences present in online barcoding repositories. Nevertheless, barcoded species nowadays represent only a small portion of known species, and barcoding reference databases are not always curated and updated on a regular basis. In this Thesis I used an integrative approach to characterize benthic marine biodiversity, specifically coupling morphological and molecular techniques with the employment of SSS. Moreover, I upgraded the actual status of COI (cytochrome c oxidase subunit I) barcoding of marine metazoans, and I built a customized COI barcoding reference database for metabarcoding studies on temperate biogenic reefs. This work implemented the knowledge about diversity of Mediterranean marine communities, laying the groundworks for monitoring marine and environmental changes that will occur in the next future as consequences of anthropic and climate threats.

Relationship between phenotype and environment in marine calcifying organisms: exploring growth and shell properties of different mollusks species in past and modern scenario

Coastal ecosystems represent an inestimable source of biodiversity, being among the most productive areas on the planet. Despite the great ecological and economic value of those environments, many threats endanger the species living in this ecosystem, like the rapid warming and the sea acidification, among many other. Benthic calcifying organisms (e.g. mollusks, corals and echinoderms) in particular, are among the most exposed to those hazards. These organisms use calcium carbonate as a structural and protective material through the biomineralization process, biologically controlled by the organism, but nevertheless, strongly influenced by the environmental surroundings. Evaluating how a changing environment can influence the process of biomineralization is critical to understand how those species of great ecological and economic importance will face the ongoing climate change. This thesis investigates the mechanism of biomineralization in different mollusks’ species of the Adriatic Sea, providing detailed descriptions of shells skeletal, biometric and growth parameters. Applying a multidisciplinary and multi-scale research approach, the influence of external environmental factors on the process of shell formation has been investigated. To achieve this purpose analysis were conducted both on current populations and on fossil remain, which allows to investigate ecological responses to past climate transitions. Mollusks’ shells in fact are one of the best tools to understand climate change in the past, present and future, since they record the environmental conditions prevailed during their life, reflected on the geochemical properties, microstructure and growth of the shell. This approach allowed to overcome the time scale limit imposed by field and laboratory survey, and better understand species long term adaptive response to changing environment, a crucial issue to define proper conservation and management strategies. Furthermore, the investigation of fossil record of mollusks assemblages offered the opportunity to evaluate the long-term biotic response to anthropogenic stressors in the north Adriatic Sea.