Changes in lipid class content and composition of Isochrysis sp. (T-Iso) grown in batch culture

© 2015, Springer International Publishing Switzerland. For decades, the microalgae Isochrysis spp. have been widely utilised as a live feed in aquaculture practices. This species possesses a number of favourable characteristics, notably its long-chain omega-3 polyunsaturated fatty acid (LC n-3 PUFA) content; primarily docosahexaenoic acid (DHA, 22:6n-3). This article describes the lipid class content and composition of this microalga grown in batch culture, covering the entirety of lag, log and stationary growth phases. The total lipid was highest in the lag phase (27 pg/cell). Total lipid significantly decreased in the exponential growth (7 pg/cell), then steadily increasing for the remainder of growth. The increase in total lipid was due to the accumulation of neutral lipid in the form of triacylglycerides. The DHA content (pg/cell) of the neutral lipid remained relatively unchanged for the duration of growth, with the influx of fatty acids being primarily myristic and palmitic acids. DHA (pg/cell) was found at relatively uniform amounts across all lipid classes. However, the DHA content as a percentage differed greatly between classes. The polar lipid class had a significantly higher DHA content, which peaked at 38 % of all polar lipid in log growth. The primary PUFA species present in the glycolipid class was stearidonic acid (18:4n-3). This work gives an overview of the lipid content and composition of Isochrysis sp. (T-Iso) over the entirety of its growth under batch culture. The lipid profile for this species at different stages of culture provides a basal data set that is useful for comparative studies using this organism.

Preserved algae as food for Penaeus monodon larvae

Uni-algal cultures of C. calcitrans, S. costatum, T. chui and Isochrysis sp. obtained from the laboratory were harvested. The harvest, preserved by either freezing or sun-drying, was fed to the larvae of P. monodon . Among the test algal species, Chaetoceros and Tetraselmis were used in larval feeding trials with frozen food while Chaetoceros, Tetraselmis and Isochrysis species were utilized in feeding experiments with sun-dried algae. Their relative effects on larval survival and development were assessed. Results showed that, except the alum-flocculated cells, both frozen Chaetoceros and Tetraselmis can support survival at the zoea stage. Best survival of 68% was atained with dried Chaetoceros followed by Tetraselmis at 44%. Dried Isochrysis did not perform as well, a significantly low survival of only 25% was obtained.

Enrichment of bioactive compounds in microalgae for aquaculture

Microalgae are promising microorganisms for the production of food and fine chemicals. Several species of microalgae are used in aquaculture with the purpose of transfer bioactive compounds up to the aquatic food chain. The main objective of this project was to develop a stress–inducement strategy in order to enhance the biochemical productivity of Nannochloropsis gaditana, Rhodomonas marina and Isochrysis sp. for aquaculture purposes having in account their growth and organizational differences. In this regard, two experiments were design: the first one consisted on the alteration of overall nutrient availabilities in growth medium; and the second one comprised changes in nitrogen and sulfur concentrations maintaining the concentrations of the other nutrients present in a commercial growth medium (Nutribloom plus), which is frequently used in aquaculture. Microalgae dried biomass was characterized biochemically and elemental analysis was also performed for all samples. In first experimental design: linear trends between nutrient availability in growth media and microalgae protein content were obtained; optimum productivities of eicosapentaenoic (EPA) and docosahexaenoic acids (DHA) were attained for both R. marina and N. gaditana in growth media enriched with 1000 L L-1 of nutrient solution whereas for Isochrysis sp. the double of Nutribloom plus was needed; the decrease of glucans and total monosaccharides with nutrient availability for R. marina and Isochrysis sp. showed the occurrence of a possible depletion of carbohydrates towards lipids and proteins biosynthesis. Second experimental desing: N. gaditana exhibited the highest variation in their biochemical composition against the applied perturbation; variations observed for microalgae in their biochemical composition were reflected in their elemental stoichiometry; in N. gaditana the highest nitrogen concentrations lead to overall maximum productivities of the biochemical parameters. The results of the present work show two stress-inducement strategies for microalgae that may constitute a base for further investigations on their biochemical enhancement.

Enrichment of bioactive compounds in microalgae for aquaculture

Microalgae are promising microorganisms for the production of food and fine chemicals. Several species of microalgae are used in aquaculture with the purpose of transfer bioactive compounds up to the aquatic food chain. The main objective of this project was to develop a stress–inducement strategy in order to enhance the biochemical productivity of Nannochloropsis gaditana, Rhodomonas marina and Isochrysis sp. for aquaculture purposes having in account their growth and organizational differences. In this regard, two experiments were design: the first one consisted on the alteration of overall nutrient availabilities in growth medium; and the second one comprised changes in nitrogen and sulfur concentrations maintaining the concentrations of the other nutrients present in a commercial growth medium (Nutribloom plus), which is frequently used in aquaculture. Microalgae dried biomass was characterized biochemically and elemental analysis was also performed for all samples. In first experimental design: linear trends between nutrient availability in growth media and microalgae protein content were obtained; optimum productivities of eicosapentaenoic (EPA) and docosahexaenoic acids (DHA) were attained for both R. marina and N. gaditana in growth media enriched with 1000 L L-1 of nutrient solution whereas for Isochrysis sp. the double of Nutribloom plus was needed; the decrease of glucans and total monosaccharides with nutrient availability for R. marina and Isochrysis sp. showed the occurrence of a possible depletion of carbohydrates towards lipids and proteins biosynthesis. Second experimental desing: N. gaditana exhibited the highest variation in their biochemical composition against the applied perturbation; variations observed for microalgae in their biochemical composition were reflected in their elemental stoichiometry; in N. gaditana the highest nitrogen concentrations lead to overall maximum productivities of the biochemical parameters. The results of the present work show two stress-inducement strategies for microalgae that may constitute a base for further investigations on their biochemical enhancement.

Riesgo ecológico del sulfato de bario

Se desconoce el efecto del sulfato de bario en los ecosistemas acuáticos donde se realizan actividades hidrocarburíferas y que vienen incrementándose a nivel nacional. Por tal motivo, se evaluó el riesgo ecológico del sulfato de bario empleando la respuesta ecotoxicológica de doce organismos no destinatarios a fin de conocer los posibles efectos que este compuesto pudiera estar ocasionando a los organismos relacionados a los ecosistemas marinos y epicontinentales donde se desarrollan actividades hidrocarburíferas. Las pruebas ecotoxicológicas incluyeron a las microalgas Isochrysis sp., Chlorella sp., las plantas terrestres Medicago sativa y Zea mays, los crustáceos Daphnia sp., Emerita analoga y Apohyale sp., al equinodermo Tetrapygus niger, al insecto acuático Chironomus calligraphus, y a los peces Odontesthes regia regia, Poecilia reticulata y Paracheirodon innesi. Las mediciones de los parámetros y protocolos para las pruebas como la determinación del riesgo ecológico siguieron las pautas y recomendaciones de la USEPA y otros autores. De los principales resultados ecotoxicológicos con sulfato de bario y sus formas solubles, se obtuvo un efecto negativo del sulfato de bario sobre el crecimiento celular de la microalga epicontinental Chlorella sp. (96 h), que registró una concentración de inhibición media (CI50) de 0,1 g/L y una concentración efectiva no observable (NOEC) de 0,02 g/L. Así mismo, se obtuvo un efecto negativo del bario sobre el crecimiento foliar de la planta terrestre monocotiledónea Z. mays (10 d) que registró una concentración efectiva media (CE50) de 0,0011 g/L y una NOEC de 0,0002 g/L. Finalmente, se concluye que existe alto riesgo ecológico (RQ) del sulfato de bario (RQ = 1,224) y sus formas solubles (RQ = 37 500) empleando la respuesta ecotoxicológica de doce organismos no destinatarios.

Exposure to toxic Alexandrium minutum activates the detoxifying and antioxidant systems in gills of the oyster Crassostrea gigas

Harmful algal blooms of Alexandrium spp. dinoflagellates regularly occur in French coastal waters contaminating shellfish. Studies have demonstrated that toxic Alexandrium spp. disrupt behavioural and physiological processes in marine filter-feeders, but molecular modifications triggered by phycotoxins are less well understood. This study analyzed the mRNA levels of 7 genes encoding antioxidant/detoxifying enzymes in gills of Pacific oysters (Crassostrea gigas) exposed to a cultured, toxic strain of A. minutum, a producer of paralytic shellfish toxins (PST) or fed Tisochrysis lutea (T. lutea, formerly Isochrysis sp., clone Tahitian (T. iso)), a non-toxic control diet, in four repeated experiments. Transcript levels of sigma-class glutathione S-transferase (GST), glutathione reductase (GR) and ferritin (Fer) were significantly higher in oysters exposed to A. minutum compared to oysters fed T. lutea. The detoxification pathway based upon glutathione (GSH)-conjugation of toxic compounds (phase II) is likely activated, and catalyzed by GST. This system appeared to be activated in gills probably for the detoxification of PST and/or extra-cellular compounds, produced by A. minutum. GST, GR and Fer can also contribute to antioxidant functions to prevent cellular damage from increased reactive oxygen species (ROS) originating either from A. minutum cells directly, from oyster hemocytes during immune response, or from other gill cells as by-products of detoxification.

Caracterizaçäo bioquímica de microalgas produzidas industrialmente em aquacultura

Dissertação de mest. em Aquacultura, Unidade de Ciências e Tecnologias dos Recursos Aquáticos, Univ. do Algarve, 1997

Use of micro-algae concentrates for hatchery culture of sandfish, Holothuria scabra

Three Instant Algae® (Reed Mariculture Inc., Campbell, CA, USA, 95008) products: (1) mono-cultured Isochrysis sp. (Haptophyceae) (Isochrysis 1800®); (2) mono-cultured Pavlova sp. (Haptophyceae) (Pavlova 1800®); and (3) mono-cultured Thalassiosira weissflogii (Bacillariophyceae) (TW 1200®) were used to feed sandfish (Holothuria scabra) larvae both singly and in ternary combination to assess their nutritional efficacy. Two-day auriculariae were held at a starting density of 0.3 mL-1 and were initially fed a daily ration equivalent to the dry weight of 10,000 cells mL-1 of Isochrysis 1800®. This ration was increased by the dry weight equivalent of 1000 cells mL-1 of Isochrysis 1800® per day as larval development proceeded. Post-settled larvae fed TW 1200® were significantly larger than those fed the ternary diet, Isochrysis 1800® or Pavlova 1800®. There were significant differences in the mean (±SE) survival of auriculariae and post-settled larvae between treatments and survival to settlement was significantly higher (P < 0.05) for larvae fed TW 1200® (13.7 ± 0.7%) alone. Laval development, competency and survival were significantly correlated with dietary levels of total protein, lipid and nitrogen-free extract (NFE), and with total polyunsaturated fatty acid (PUFA) content of the diets, and the levels of some specific fatty acids (FA). The proportion of late auriculariae with hyaline spheres (day 13), numbers of competent doliolariae (day 15) and the total length of post-settled larvae (day 21) were all positively correlated with dietary NFE and palmitic acid (16:0) contents, as well as dietary EPA: DHA ratio. This study is the first comprehensive assessment of the nutritional value of micro-algae concentrates for sandfish larvae based on their nutrient compositions. Our study confirms the feasibility of using commercially available microalgae concentrates as a sole food source for hatchery culture of sandfish, and is the first to report successful hatchery culture of H. scabra without using live micro-algae. All micro-algae concentrates used in this study proved nutritious for H. scabra larvae and supported normal growth and development and relatively high survival, through settlement. Use of commercially available micro-algae concentrates as a replacement for live micro-algae in sandfish hatcheries supports development of cheaper, simpler larval rearing protocols for this species. Statement of relevance: Hatchery culture of sandfish using micro-algae concentrates, without the use of live micro-algae, is a significant finding that supports development of cheaper, simpler larval rearing protocols for this species. Our study provides detailed nutritional compositional data for the products used and on this basis we were able to deduce new information relating to key nutrients for sea cucumber larvae.

Development of hyaline spheres in late auriculariae of sandfish, Holothuria scabra : is it a reliable indicator of subsequent performance?

This study assessed the influence of diet composition on hyaline sphere (HS) development in auricularia larvae of the sandfish, Holothuria scabra, and the subsequent relationships between the presence and size of hyaline spheres and competency through settlement and early juvenile performance. Two-day old larvae were fed one of three commercially available micro-algae diets that varied in their nutrient compositions: (1) Isochrysis sp. (Haptophyceae); (2) Pavlova sp. (Haptophyceae); and (3) Thalassiosira weissflogii (Bacillariophyceae) or a ternary combination of the three. There were positive significant correlations between HS development in late auriculariae on days 10, 11, 12 and 13 post-fertilisation, and the proportion of competent doliolariae on day 15, post-settlement size (day 21) and post-settlement survival (day 25). The dietary components that most strongly influenced these relationships were carbohydrates and the n-3 polyunsaturated fatty acids arachidonic acid (ARA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Our result confirm the strong relationship between HS formation in late auriculariae of sandfish and subsequent larval competency through settlement. As such, the presence and size of HS is a reliable indicator of subsequent performance for sandfish. Given that HS development was influenced by the nutrients available to sandfish auriculariae, there is clear opportunity for development of more appropriate diets for hatchery culture of this species that will improve HS formation and larval performance supporting improved hatchery production. Statement of relevance Our results have confirmed that for sandfish larvae, the presence and size of HS at the end of the auricularia stage, is indeed a reliable indicator of subsequent performance. Our results have also confirmed that HS formation and size is significantly influenced by the nutrient composition of the auricularia diet, and that there are significant positive correlations between HS formation and dietary levels of carbohydrate, EPA:DHA and EPA:ARA ratios. This information provides a strong basis for development of more appropriate diets for sandfish larvae that will improve HS formation and larval performance during hatchery culture and in turn improve hatchery production of this species.

Electronic states of trans-polyacetylene, poly(p-phenylene vinylene) and sp-hybridised carbon species in amorphous hydrogenated carbon probed by resonant Raman scattering

Inclusions of sp-hybridised, trans-polyacetylene [trans-(CH)x] and poly(p-phenylene vinylene) (PPV) chains are revealed using resonant Raman scattering (RRS) investigation of amorphous hydrogenated carbon (a-C:H) films in the near IR – UV range. The RRS spectra of trans-(CH)x core Ag modes and the PPV CC-H phenylene mode are found to transform and disperse as the laser excitation energy ћωL is increased from near IR through visible to UV, whereas sp-bonded inclusions only become evident in UV. This is attributed to ћωL probing of trans-(CH)x chain inhomogeneity and the distribution of chains with varying conjugation length; for PPV to the resonant probing of phelynene ring disorder; and for sp segments, to ћωL probing of a local band gap of end-terminated polyynes. The IR spectra analysis confirmed the presence of sp, trans-(CH)x and PPV inclusions. The obtained RRS results for a-C:H denote differentiation between the core Ag trans-(CH)x modes and the PPV phenylene mode. Furthermore, it was found that at various laser excitation energies the changes in Raman spectra features for trans-(CH)x segments included in an amorphous carbon matrix are the same as in bulk trans-polyacetylene. The latter finding can be used to facilitate identification of trans-(CH)x in the spectra of complex carbonaceous materials.

Direct and indirect effects of egg parasitism by Neopolyscystus Girault sp. (Hymenoptera: Pteromalidae) on Paropsis atomaria Olivier (Coleoptera: Chrysomelidae)

Abstract Neopolycystus sp. is the only primary egg parasitoid associated with the pest beetle Paropsis atomaria in subtropical eucalypt plantations, but its impact on its host populations is unknown. The simplified ecosystem represented by the plantation habitat, lack of interspecific competition for host and parasitoid, and the multivoltinism of the host population makes this an ideal system for quantifying the direct and indirect effects of egg parasitism, and hence, effects on host population dynamics. Within-, between- and overall-egg-batch parasitism rates were determined at three field sites over two field seasons, and up to seven host generations. The effect of exposure time (egg batch age), host density proximity to native forest and water sources on egg parasitism rates was also tested. Neopolycystus sp. exerts a significant influence on P. atomaria populations in Eucalyptus cloeziana. plantations in south-eastern Queensland, causing the direct (13%) and indirect (15%) mortality of almost one-third of all eggs in the field. Across seasons and generations, 45% of egg batches were parasitised, with a within-batch parasitism rate of around 30%. Between-batch parasitism increased up to 5–6 days after oviposition in the field, although within-batch parasitism rates generally did not. However, there were few apparent patterns to egg parasitism, with rates often varying significantly between sites and seasons.