From lab to application: Cultivating limnetic microalgae in seawater coupled with wastewater for biodiesel production on a pilot scale.

The technology of cultivating salt-tolerant limnetic microalgae in seawater reduces the freshwater demand and costs of biodiesel production. However, all current trials still occur on the bench scale, and efforts for pilot-scale operation are urgently needed. This study firstly optimised the diameter of the photobioreactors (PBRs) to 0.2 m, as the single module to produce lipid-rich Golenkinia sp. SDEC-16 because of the better algal growth and light attenuation in the PBRs, and then established a 1000 L algal cultivation system. In a medium of seawater supplemented with monosodium glutamate wastewater at a ratio of 1:1000 (S-MSGW), the biomass productivity was 0.26 g/L/d, which was approaching the 0.30 g/L/d obtained in BG11, and the lipid productivity (98.99 mg/L/d) was doubled in comparison to growth in BG11. C16-C18 accounted for more than 98% of the total fatty acid in S-MSGW, and the biodiesel properties also met the biodiesel standards. The input cost of the biodiesel in this pilot-scale system was estimated to be 2.2 $/kg. When considering the carbon reduction and diversified application of the biomass, Golenkinia sp. would annually capture 186.77 kg/m3 PBR of CO2, and yield an output-to-input ratio (OIR) of 3.4 in S-MSGW, higher than the 2.8 in BG11.

The physiology of rapid ecological specialization: A look at the Midas cichlids.

Understanding the process of speciation is a primary goal of evolutionary biology, yet the question of whether speciation can reach completion in the presence of gene flow remains controversial. For more than 50 years, the cichlids of Africa, and more recently those in South and Central America, have served as model systems for the study of speciation in nature. Cichlids are distinguished by their enormous species richness, their diversity of behavioural and trophic adaptations, and their rapid rate of divergence. In both Africa and South and Central America, the repeated interaction of geology, new founder events and adaptive evolution has created a series of natural experiments with speciation occurring both within and between waterbodies of differing ages. In the "From the Cover" paper in this issue of the Journal of Molecular Ecology, Raffini, Schneider, Franchini, Kautt and Meyer move beyond the question of which mechanisms drive speciation, and instead show that divergent morphologies and physiologies translate into adaptive traits. They investigate differences in physiology and gene expression profiles in a benthic/limnetic species pair of Midas cichlidsin a 24,000-year-old Nicaraguan crater lake. While recently diverged, these two species demonstrate significant ecological, but limited genetic differentiation. The authors find that the distinct morphotypes translate into relevant differences in swimming performance and metabolic rates that correspond to differential gene expression profiles. Hence, the authors take an integrative approach examining the impacts of morphological differences on performance and niche partitioning: an approach that can advance our understanding of the drivers of morphological and physiological divergence during speciation.

Divergent Allometric Trajectories in Gene Expression and Coexpression Produce Species Differences in Sympatrically Speciating Midas Cichlid Fish.

The mechanisms of speciation without geographic isolation (i.e., sympatric speciation) remain debated. This is due in part to the fact that the genomic landscape that could promote or hinder species divergence in the presence of gene flow is still largely unknown. However, intensive research is now centered on understanding the genetic architecture of adaptive traits associated with this process as well as how gene expression might affect these traits. Here, using RNA-Seq data, we investigated gene expression of sympatrically speciating benthic and limnetic Neotropical cichlid fishes at two developmental stages. First, we identified groups of coexpressed genes (modules) at each stage. Although there are a few large and well-preserved modules, most of the other modules are not preserved across life stages. Second, we show that later in development more and larger coexpression modules are associated with divergence between benthic and limnetic fish compared with the earlier life stage. This divergence between benthic and limnetic fish in coexpression mirrors divergence in overall expression between benthic and limnetic fish, which is more pronounced later in life. Our results reveal that already at 1-day posthatch benthic and limnetic fish diverge in (co)expression, and that this divergence becomes more substantial when fish are free-swimming but still unlikely to have divergent swimming and feeding habits. More importantly, our study describes how the coexpression of several genes through development, as opposed to individual genes, is associated with benthic-limnetic species differences, and how two morphogenetic trajectories diverge as fish grow older.

Lipid productivity in limnetic Chlorella is doubled by seawater added with anaerobically digested effluent from kitchen waste.

BACKGROUND: An economical strategy for producing microalgae as biofuel feedstock is driven by the freshwater and nutrients input. In this study, seawater was applied to limnetic algal cultivation and the behavior of algae in seawater media was observed including growth, lipid synthesis, and ultrastructure. To make seawater cater algae, a kind of wastewater, anaerobically digested effluent from kitchen waste (ADE-KW), was used as nutrient sources. RESULTS: Pure seawater cannot support the growth demand of freshwater microalga, due to high salinity and lack of nutrients. However, it is the conditions triggered the algae to synthesize lipids of 60%, double of lipid content in standard medium BG11. Introducing 3 or 5% ADE-KW (volume percentage) into seawater made algal growth reach the level attained in BG11, while lipid content compared favourably with the level (60%) in pure seawater. This method achieved the goal of fast growth and lipid accumulation simultaneously with the highest lipid productivity (19 mg/L â€Šday) at the exponential stage, while BG11 obtained 10.55 mg/L â€Šday at the stationary stage as the highest lipid productivity, almost half of that in seawater media. Moreover, the condition for highest lipid productivity enlarged algal cells compared to BG11. Under the condition for highest lipid productivity, Chlorella sorokiniana SDEC-18 had enlarged cells and increased settling efficiency compared to BG11, which facilitated harvest in an energy saving way. CONCLUSIONS: The results suggested that combining seawater with ADE-KW to cultivate microalgae had a double function: nutrients and water for algal growth, and high salinity for stimulating lipid accumulation. If this technology was operated in practice, freshwater and non-waste nutrient consumption would be completely obviated.

Ecosystem size predicts eco-morphological variability in a postglacial diversification.

Identifying the processes by which new phenotypes and species emerge has been a long-standing effort in evolutionary biology. Young adaptive radiations provide a model to study patterns of morphological and ecological diversification in environmental context. Here, we use the recent radiation (ca. 12k years old) of the freshwater fish Arctic charr (Salvelinus alpinus) to identify abiotic and biotic environmental factors associated with adaptive morphological variation. Arctic charr are exceptionally diverse, and in postglacial lakes there is strong evidence of repeated parallel evolution of similar morphologies associated with foraging. We measured head depth (a trait reflecting general eco-morphology and foraging ecology) of 1,091 individuals across 30 lake populations to test whether fish morphological variation was associated with lake bathymetry and/or ecological parameters. Across populations, we found a significant relationship between the variation in head depth of the charr and abiotic environmental characteristics: positively with ecosystem size (i.e., lake volume, surface area, depth) and negatively with the amount of littoral zone. In addition, extremely robust-headed phenotypes tended to be associated with larger and deeper lakes. We identified no influence of co-existing biotic community on Arctic charr trophic morphology. This study evidences the role of the extrinsic environment as a facilitator of rapid eco-morphological diversification.

The temporal window of ecological adaptation in postglacial lakes: a comparison of head morphology, trophic position and habitat use in Norwegian threespine stickleback populations.

BACKGROUND: Studying how trophic traits and niche use are related in natural populations is important in order to understand adaptation and specialization. Here, we describe trophic trait diversity in twenty-five Norwegian freshwater threespine stickleback populations and their putative marine ancestor, and relate trait differences to postglacial lake age. By studying lakes of different ages, depths and distance to the sea we examine key environmental variables that may predict adaptation in trophic position and habitat use. We measured trophic traits including geometric landmarks that integrated variation in head shape as well as gillraker length and number. Trophic position (Tpos) and niche use (α) were estimated from stable isotopes (δ(13)C, δ(15)N). A comparison of head shape was also made with two North American benthic-limnetic species pairs. RESULTS: We found that head shape differed between marine and freshwater sticklebacks, with marine sticklebacks having more upturned mouths, smaller eyes, larger opercula and deeper heads. Size-adjusted gillraker lengths were larger in marine than in freshwater stickleback. Norwegian sticklebacks were compared on the same head shape axis as the one differentiating the benthic-limnetic North American threespine stickleback species pairs. Here, Norwegian freshwater sticklebacks with a more "limnetic head shape" had more and longer gillrakers than sticklebacks with "benthic head shape". The "limnetic morph" was positively associated with deeper lakes. Populations differed in α (mean ± sd: 0.76 ± 0.29) and Tpos (3.47 ± 0.27), where α increased with gillraker length. Larger fish had a higher Tpos than smaller fish. Compared to the ecologically divergent stickleback species pairs and solitary lake populations in North America, Norwegian freshwater sticklebacks had similar range in Tpos and α values, but much less trait divergences. CONCLUSIONS: Our results showed trait divergences between threespine stickleback in marine and freshwater environments. Freshwater populations diverged in trophic ecology and trophic traits, but trophic ecology was not related to the elapsed time in freshwater. Norwegian sticklebacks used the same niches as the ecologically divergent North American stickleback species pairs. However, as trophic trait divergences were smaller, and not strongly associated with the ecological niche, ecological adaptations along the benthic-limnetic axis were less developed in Norwegian sticklebacks.

Crater lake cichlids individually specialize along the benthic-limnetic axis.

A common pattern of adaptive diversification in freshwater fishes is the repeated evolution of elongated open water (limnetic) species and high-bodied shore (benthic) species from generalist ancestors. Studies on phenotype-diet correlations have suggested that population-wide individual specialization occurs at an early evolutionary and ecological stage of divergence and niche partitioning. This variable restricted niche use across individuals can provide the raw material for earliest stages of sympatric divergence. We investigated variation in morphology and diet as well as their correlations along the benthic-limnetic axis in an extremely young Midas cichlid species, Amphilophus tolteca, endemic to the Nicaraguan crater lake Asososca Managua. We found that A. tolteca varied continuously in ecologically relevant traits such as body shape and lower pharyngeal jaw morphology. The correlation of these phenotypes with niche suggested that individuals are specialized along the benthic-limnetic axis. No genetic differentiation within the crater lake was detected based on genotypes from 13 microsatellite loci. Overall, we found that individual specialization in this young crater lake species encompasses the limnetic-as well as the benthic macro-habitat. Yet there is no evidence for any diversification within the species, making this a candidate system for studying what might be the early stages preceding sympatric divergence. A common pattern of adaptive diversification in freshwater fishes is the repeated evolution of open water (limnetic) species and of shore (benthic) species. Individual specialization can reflect earliest stages of evolutionary and ecological divergence. We here demonstrate individual specialization along the benthic-limnetic axis in a young adaptive radiation of crater lake cichlid fishes.

Tooth microwear formation rate in Gasterosteus aculeatus.

Tooth microwear feature densities were significantly increased in a population of laboratory-reared three-spined stickleback Gasterosteus aculeatus in four days, after they were transferred from a limnetic feeding regime to a benthic feeding regime. These results show that even in aquatic vertebrates with non-occluding teeth, changes in feeding can cause changes in tooth microwear in just a few days, as in mammals.

Crater lake habitat predicts morphological diversity in adaptive radiations of cichlid fishes.

Adaptive radiations provide an excellent opportunity for studying the correlates and causes for the origin of biodiversity. In these radiations, species diversity may be influenced by either the ecological and physical environment, intrinsic lineage effects, or both. Disentangling the relative contributions of these factors in generating biodiversity remains a major challenge in understanding why a lineage does or does not radiate. Here, we examined morphological variation in body shape for replicate flocks of Nicaraguan Midas cichlid fishes and tested its association with biological and physical characteristics of their crater lakes. We found that variability of body elongation, an adaptive trait in freshwater fishes, is mainly predicted by average lake depth (N = 6, P < 0.001, R(2) = 0.96). Other factors considered, including lake age, surface area, littoral zone area, number of co-occurring fish species, and genetic diversity of the Midas flock, did not significantly predict morphological variability. We also showed that lakes with a larger littoral zone have on average higher bodied Midas cichlids, indicating that Midas cichlid flocks are locally adapted to their crater lake habitats. In conclusion, we found that a lake's habitat predicts the magnitude and the diversity of body elongation in repeated cichlid adaptive radiations.

Genomic architecture of ecologically divergent body shape in a pair of sympatric crater lake cichlid fishes.

Determining the genetic bases of adaptations and their roles in speciation is a prominent issue in evolutionary biology. Cichlid fish species flocks are a prime example of recent rapid radiations, often associated with adaptive phenotypic divergence from a common ancestor within a short period of time. In several radiations of freshwater fishes, divergence in ecomorphological traits - including body shape, colour, lips and jaws - is thought to underlie their ecological differentiation, specialization and, ultimately, speciation. The Midas cichlid species complex (Amphilophus spp.) of Nicaragua provides one of the few known examples of sympatric speciation where species have rapidly evolved different but parallel morphologies in young crater lakes. This study identified significant QTL for body shape using SNPs generated via ddRAD sequencing and geometric morphometric analyses of a cross between two ecologically and morphologically divergent, sympatric cichlid species endemic to crater Lake Apoyo: an elongated limnetic species (Amphilophus zaliosus) and a high-bodied benthic species (Amphilophus astorquii). A total of 453 genome-wide informative SNPs were identified in 240 F2 hybrids. These markers were used to construct a genetic map in which 25 linkage groups were resolved. Seventy-two segregating SNPs were linked to 11 QTL. By annotating the two most highly supported QTL-linked genomic regions, genes that might contribute to divergence in body shape along the benthic-limnetic axis in Midas cichlid sympatric adaptive radiations were identified. These results suggest that few genomic regions of large effect contribute to early stage divergence in Midas cichlids.

Morphogenesis and molecular phylogeny of a new freshwater ciliate, Notohymena apoaustralis n. sp. (Ciliophora, Oxytrichidae).

The live morphology, infraciliature, and morphogenesis of a new oxytrichid ciliate, Notohymena apoaustralis n. sp. collected from a freshwater pond in Qingdao (Tsingtao), China, were studied in vivo and after protargol impregnation. Notohymena apoaustralis n. sp. is characterized as follows: undulating membranes in Notohymena-pattern; cortical granules yellow-green, grouped around the marginal cirri and dorsal bristles, and in short irregular rows elsewhere in the cell; single contractile vacuole positioned at anterior 1/3 of the body length; two macronuclear nodules and one micronucleus; about 39 adoral membranelles; 18 frontoventral transverse cirri in typical Oxytricha-pattern; one right and one left marginal row, almost confluent posteriorly; dorsal ciliature in typical Oxytricha-pattern; 8-10 caudal cirri arranged in three rows, one each at the posterior end of dorsal kineties 1, 2, and 4, indistinguishable from marginal cirri in life. The morphogenetic process in N. apoaustralis n. sp. is consistent with that of the type species, Notohymena rubescens Blatterer and Foissner, 1988. Phylogenetic analyses based on small subunit rDNA sequence data suggest a sister relationship between N. apoaustralis n. sp. and Paraurostyla weissei, which cluster in a clade with Rubrioxytricha ferruginea.

Experimental evidence for trait utility of gill raker number in adaptive radiation of a north temperate fish.

North temperate fish in post-glacial lakes are textbook examples for rapid parallel adaptive radiation into multiple trophic specialists within individual lakes. Speciation repeatedly proceeded along the benthic-limnetic habitat axis, and benthic-limnetic sister species diverge in the number of gill rakers. Yet, the utility of different numbers of gill rakers for consuming benthic vs. limnetic food has only very rarely been experimentally demonstrated. We bred and raised families of a benthic-limnetic species pair of whitefish under common garden conditions to test whether these species (i) show heritable differentiation in feeding efficiency on zooplankton, and (ii) whether variation in feeding efficiency is predicted by variation in gill raker numbers. We used zooplankton of three different size classes to investigate prey size dependency of divergence in feeding efficiency and to investigate the effect strength of variation in the number of gill rakers. Our results show strong interspecific differences in feeding efficiency. These differences are largest when fish were tested with the smallest zooplankton. Importantly, feeding efficiency is significantly positively correlated with the number of gill rakers when using small zooplankton, also when species identity is statistically controlled for. Our results support the hypothesis that a larger number of gill rakers are of adaptive significance for feeding on zooplankton and provide one of the first experimental demonstrations of trait utility of gill raker number when fish feed on zooplankton. These results are consistent with the suggested importance of divergent selection driven feeding adaptation during adaptive radiation of fish in post-glacial lakes.

Composição taxonômica da assembleia de larvas de peixes no reservatório de Rosana, Rio Paranapanema, Brasil / Taxonomic composition of the assemblage of fish larvae in the Rosana reservoir, Paranapanema River, Brazil

Em reservatórios, são escassos os estudos que consideram as fases iniciais do ciclo de vida de peixes, sendo que informações sobre a ecologia de larvas de peixes são úteis para o entendimento da biologia das espécies, visto que os estágios iniciais representam um período crítico no ciclo de vida dos peixes. Neste contexto, o conhecimento a respeito das assembleias de larvas pode ser valioso para a predição e o entendimento do recrutamento, auxiliando desta maneira, no manejo do ambiente. Desta forma, o presente trabalho pretende verificar a composição da assembleia de larvas de peixes em diferentes regiões do reservatório. As coletas foram realizadas trimestralmente (março, junho, setembro e dezembro) no ano de 2002, nas regiões limnética e litorânea das zonas fluvial, transição e lacustre do reservatório de Rosana. No laboratório, o material foi triado e as larvas foram separadas e identificadas ao menor nível taxonômico possível. Os táxons que tiveram maior contribuição na região limnética foram Plagioscion squamosissimus (Perciformes), Hypophthalmus edentatus, Pimelodus maculatus (Siluriformes), Astyanax spp. e Roeboides descalvadensis (Characiformes), enquanto na região litorânea Bryconamericus stramineus e Apareiodon affinis (Characiformes). As regiões limnética e litorânea do reservatório, assim como suas respectivas zonas apresentaram diferenças na composição de espécies de larvas e a segregação das assembleias pode estar sendo influenciada pela preferência por habitats ou por questões evolutivas que envolvem as estratégias reprodutivas das espécies.

There are few studies in reservoirs that consider the Early Life Stages (ELS) of fish. Information on the ecology of fish larvae are useful for understanding the biology of the species, since ELS represent a critical period in fish life cycle. In this context, knowledge about the assemblages of larvae can be valuable for the prediction and understanding of recruitment, thus assisting in the management of the environment. Therefore, this study intends to investigate the composition of the assemblage of fish larvae in different regions of Rosana reservoir. Samples were collected quarterly (March, June, September and December) in 2002, in the limnetic and littoral regions of the zones fluvial, transition and lacustrine of reservoir. In the laboratory, the material was sorted and larvae were separated and identified to lowest possible taxonomic level. The taxa that were the major contributors in the limnetic region were Plagioscion squamosissimus (Perciformes), Hypophthalmus edentatus, Pimelodus maculatus (Siluriformes), Astyanax spp. and Roeboides descalvadensis (Characiformes), whereas in the coastal region Bryconamericus stramineus and Apareiodon affinis (Characiformes) prevailed. Limnetic and littoral regions of the reservoir, as well as their respective zones showed differences in larvae species composition and segregation of assemblages can be influenced by a preference for habitats or evolutionary questions involving reproductive strategies of species.

Fitoplancton del río Tehuantepec, Oaxaca, México y algunas relaciones biogeográficas / Phytoplankton from Tehuantepec River, Oaxaca, Mexico and some biogeographical relationships

Phytoplankton species collected from the limnetic to euryhaline sections of Tehuantepec River were identified, classified and compared with regional information from Mexico and South America. We collected 15 samples every three months from July 1997 through August 1998 with a 20 microm net and a Van Dorn bottle. Indicator values and a code checklist are included. A total of 58 families, 121 genera, 273 species, one subspecies, 75 varieties, 13 forms and one morphotype were identified in the taxa Bacillariophyta (42.0 %), Chlorophyta (29.0 %), Cyanoprocaryota (18.0 %), Euglenophyta (5.0 %), Dinophyta (3.0 %), Cryptophyta (2.0 %) and Chrysophyta (1.0 %). The predominant families were Scenedesmaceae (24 species), Oocystaceae (22), Bacillariophyceae (21), Chaetocerotaceae (15) and Euglenaceae (14). Five families, eight genera, 72 species, 45 varieties and eight forms are first records for Mexico. The species Chroococcus turgidus, Microcystis flosaquae and Pseudanabaena limnetica (which produce massive blooms or red tides) are important in this river of moderate water quality.

El fitoplancton recolectado en el río Tehuantepec con características limnéticas a eurihalinas se determinó, clasificó y comparó con información regional de México y Suramérica. Se incluyen valores indicadores y un código del listado florístico para facilitar el manejo de estas algas. Se determinó un total de 58 familias, 121 géneros, 273 especies, una subespecie, 75 variedades, 13 formas y un morfotipo, pertenecientes a las divisiones Bacillariophyta (42.0 %), Chlorophyta (29.0 %), Cyanoprocaryota (18.0 %), Euglenophyta (5.0 %), Dinophyta (3.0 %), Cryptophyta (2.0 %) y Chrysophyta (1.0 %). Las familias mejor representadas fueron Scenedesmaceae (24 especies), Oocystaceae (22), Bacillariophyceae (21), Chaetocerotaceae (15) y Euglenaceae (14). Se establecen nuevos registros para México: cinco familias, ocho géneros, 72 especies, 45 variedades y ocho formas. En este río, que presenta una moderada calidad del agua, sobresalieron las especies Chroococcus turgidus, Microcystis flosaquae y Pseudanabaena limnetica (como productoras de mareas rojas).