Production of microalgal concentrates by flocculation and their assessment as aquaculture feeds

A novel technique was developed for the flocculation of marine microalgae commonly used in aquaculture. The process entailed an adjustment of pH of culture to between 10 and 10.6 using NaOH, followed by addition of a non-ionic polymer Magnafloc LT-25 to a final concentration of 0.5 mg L-1. The ensuing flocculate was harvested, and neutralised giving a final concentration factor of between 200- and 800-fold. This process was successfully applied to harvest cells of Chaetoceros calcitrans, C. muelleri, Thalassiosira pseudonana, Attheya septentrionalis, Nitzschia closterium, Skeletonema sp., Tetraselmis suecica and Rhodomonas salina, with efficiencies >=80%. The process was rapid, simple and inexpensive, and relatively cost neutral with increasing volume (cf. concentration by centrifugation). Harvested material was readily disaggregated to single cell suspensions by dilution in seawater and mild agitation. Microscopic examination of the cells showed them to be indistinguishable from corresponding non-flocculated cells. Chlorophyll analysis of concentrates prepared from cultures of Concentrates of T. pseudonana prepared using pH-induced flocculation gave better growth of juvenile Pacific oysters (Crassostrea gigas) than concentrates prepared by ferric flocculation, or centrifuged concentrates using a cream separator or laboratory centrifuge. In follow up experiments, concentrates prepared from 1000 L Chaetoceros muelleri cultures were effective as supplementary diets to improve the growth of juvenile C. gigas and the scallop Pecten fumatus reared under commercial conditions, though not as effective as the corresponding live algae. The experiments demonstrated a proof-of-concept for a commercial application of concentrates prepared by flocculation, especially for use at a remote nursery without on-site mass-algal culture facilities.

Profiling of carotenoids and antioxidant capacity of microalgae from subtropical coastal and brackish waters

Carotenoids are associated with various health benefits, such as prevention of age-related macular degeneration, cataract, certain cancers, rheumatoid arthritis, muscular dystrophy and cardiovascular problems. As microalgae contain considerable amounts of carotenoids, there is a need to find species with high carotenoid content. Out of hundreds of Australian isolates, twelve microalgal species were screened for carotenoid profiles, carotenoid productivity, and in vitro antioxidant capacity (total phenolic content (TPC) and ORAC). The top four carotenoid producers at 4.68-6.88 mg/g dry weight (DW) were Dunaliella salina, Tetraselmis suecica, Isochrysis galbana, and Pavlova salina. TPC was low, with D. salina possessing the highest TPC (1.54 mg Gallic Acid Equivalents/g DW) and ORAC (577 μmol Trolox Equivalents/g DW). Results indicate that T. suecica, D. salina, P. salina and I. galbana could be further developed for commercial carotenoid production.

Diferenças intra e interespecíficas na atividade da fosfatase alcalina em microalgas marinhas costeiras selecionadas

A incorporação de fósforo (P) foi avaliada através da técnica de marcador enzimático fluorescente a fim de determinar a atividade da fosfatase alcalina (PA) em dois clones de Phaeodactylum tricornutum (Bohlin), Ub3 e Ub7, isolados de Ubatuba (SP), em Tetraselmis aff. chui (Butcher) e Prorocentrum minimum (Pavillard) J. Schiller, isoladas da Baía de Guanabara (RJ) e na Comunidade Natural da Baía de Guanabara (RJ). A fosfatase alcalina (PA) é uma enzima extracelular associada à membrana que catalisa a hidrólise de compostos orgânicos de fósforo em resposta à limitação de fosfato. Sua análise, a partir do marcador ELF-97, proporciona uma avaliação individual e, portanto, determina as condições nutricionais de fósforo inorgânico em células fitoplanctônicas. Os clones de P. tricornutum apresentaram diferenças no desenvolvimento quando incubados no tratamento P-repleto. O clone Ub7 de P. tricornutum apresentou a maior atividade enzimática quando comparado às demais espécies testadas, em condições P-repletas. Enquanto P. minimum apresentou a maior atividade da fosfatase alcalina em condições P-limitadas. Entre as espécies T. aff. chui e P. minimum, a maior atividade enzimática ocorreu durante a fase estacionária de desenvolvimento, entretanto diferenças foram observadas somente nas menores concentrações de fosfato. P. tricornutum, T. aff. chui e P. minimum, ao longo dos experimentos, utilizaram duas estratégias para incorporação de fosfato, aumentando a atividade da fosfatase alcalina, assim como alterando o biovolume ou a máxima dimensão linear para manter a relação S/V estável. Em P. tricornutum os sítios da atividade enzimática ocorreram na membrana celular, em T. aff. chui encontrados intracelularmente, enquanto em P. minimum observados tanto nas membranas, quanto no interior das células. No experimento realizado com a comunidade natural, houve predomínio das diatomáceas entre todos os grupos e tratamentos; as espécies foram agrupadas nas estratégias adaptativas C e R e classificadas principalmente como R-estrategistas. Os dinoflagelados da Ordem Prorocentrales utilizaram a incorporação do fósforo orgânico como estratégia para obtenção de fósforo em condições limitantes. Entretanto, as diatomáceas apresentaram tal estratégia de forma mais variável. Quanto às prasinofíceas, embora Tetraselmis sp. tenha apresentado baixa atividade enzimática nos experimentos unialgais, as concentrações de fosfato ao longo do experimento não resultaram na utilização de P orgânico para o grupo. Os resultados destacaram as diferenças intra e interespecíficas na atividade da fosfatase alcalina, e, consequentemente, na incorporação de fósforo orgânico, uma vez que as espécies testadas regularam a atividade enzimática de acordo com as diferentes concentrações externas de fosfato.

O Fitoplâncton e a dialética da balneabilidade de praias: um estudo de caso na Praia de Itacoatiara, Niterói, RJ.

A praia de Itacoatiara, área de proteção ambiental, localizada no município no Niterói- RJ, tem importante contribuição para o turismo ecológico da região. Busca a certificação ambiental do programa Bandeira Azul e este trabalho buscou cumprir determinação de monitoramento ambiental como um dos critérios do programa. O objetivo foi avaliar a proliferação de microalgas marinhas como sensor da qualidade da balneabilidade diagnosticada através de um bioindicador, o fitoplâncton. As coletas foram realizadas em dois pontos nos meses de novembro de 2008 e de janeiro a março de 2009. Foram encontradas 52 unidades taxonômicas, sendo que 33 pertencem às diatomáceas (Bacillariophyceae), 15 dinoflagelados (Dinophyceae), 1 às cianobactéria (Cyanophyceae), 1 euglenofícea (Euglenophyceae), 1 crisofícea (Chrysophyceae) e 1 prasinofícea (Prasinophyceae) e fitoflagelados. A metodologia seguida foi de sedimentação em cubetas e analisadas em microscópio invertido. A praia de Itacoatiara está caracterizada por sofrer influência de ambientes adjacentes com massas dágua comprometidas promovendo proliferações algais monoespecíficas com densidades consideradas de Bloom. As espécies dominantes foram Tetraselmis sp (Prasinophyceae) considerada potencialmente nociva por apresentar contaminação visual e as Cianobactérias que podem produzir toxinas. Os parâmetros vento, ondulação e maré foram apontadas como os principais carreadores das massas dágua comprometidas até a região estudada. Por cumprir o objetivo geral, conclui-se que o fitoplâncton marinho pode ser utilizado como parâmetro para a caracterização e qualidade da balneabilidade de praias.

Growth performances of three microalgal species in filtered brackishwater with different inorganic media

The growth of three microalgae species, viz., Nannochloropsis oculata, Tetraselmis chui and Chaetoceros muelleri which are commonly used in aquaculture, was investigated using three different inorganic nutrient media: (i) Modified Guillard's f/2 medium (ii) Rix Mix medium and (iii) BFRI medium. Each microalgae species was cultured for 24 days in small- scale with initial inoculation density of 17xl04 cell /ml in the three media with triplicates. N. oculata cultured in modified Guillard's f/2 medium showed superior growth with a mean peak density of 221 ±4.24 x 104 cell/ ml, to Rix Mix medium (141 ± 10.54xl04 cell/ml) and BFRI medium (47±4.94 x 104 cell/ml) on the 16th day of culture at stationary phase. Considering the increase in cell density for 20 days of culture in Rix Mix medium, C. muelleriwas significantly (P<0.05) highest than in other two media. N. oculata cultured in BFRI medium resulted in the poorest growth with a mean peak increase in density of 84±9.19 x 104 cell/ml in 12 days of culture. However, with an increase in cell density, growth of T. chui (182 ± 6.26 x 104 cell/ml) was significantly (P<0.05) higher in BFRI medium than in modified Guillard's f/2 medium. The results of the present study suggest that N. oculata and C. muelleri can be grown very well in both the modified Guillard's f/2 medium and Rix Mix medium. Better growth of T. chui can be obtained while culturing either in BFRI and Rix Mix medium. These three nutrient media used in the present study may be useful for microalgae species culture for establishing green-water culture for suitable target zooplankton, and fish and crustacean larvae in marine and brackishwater hatcheries.

Growth and reproductive performance of locally isolated brackish water rotifer (Brachionus plicatilis) feeding different micro algae

Population growth and reproductive capacity of brackishwater rotifer, Brachionus plicatilis, were evaluated, for a period of 8 days in a temperature controlled ( =25°C) microalgallaborarory, under three different algal feeding regimens. The algal species that were tested are: (i) Chlorella sp. (T1), Tetraselmis chui (T2), Nannochloropsis oculata (T 3). The feeding density of each algal species was maintained similar as of 4.5xW6 ceHs mi. The rotifer fed on T. chui showed the highest (p<0.05) population growth (131.5 ind./ml), compared to that fed on Chlorella sp (45.67 ind./ml) and N oculata (43.44 ind./ml). The abundance of egg bearing rotifers was also higher (35.77%) with T. chuithan with Chlorella sp (27.76%) and N oculara (24.60%). The results of the present study indicate that T. chui could be the most suitable algal food for the stock culture of locally isolated rotifer B. plicatilis.

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.

Survey of food supplementary (Pseudomonas fluorescence) and live food (Algae and brine shrimp) on Penaeus indicus larval stages

In this study ,the effects of Pseudomonas fluorescence obtained from generator pond water of Kolahi as supplementary and four algae consisting of : Chaetoceros sp, Chlorella and Skeletonema sp and Tetraselmis sp, three types of artemia as live food larval states from zoa to postlarvae (PL4 ) Penaeus indicus were investigated. The results indicate that Pseudomonas fluorescence has positive effect on Penaeus indicits larvae growth and their living food. Effective ranges at minimum and maximum were estimated. In most cases optimum dosage was approximately determined. Optimum dosage is between 50 -150 milligrams per liter for living food and Penaeus larval More than 200 milligram per liter resulted in a negative effect on the growth and survival. Also the results indicate Uromiana artemia. Requires a higher concentration of the bacteria the imported artemia. As a conclusion it is recommended to introduce Pseudonionas fluorescence as a new medium for the growth of some mentioned algae .

Allelopathic interactions between the macroalga Ulva pertusa and eight microalgal species

HETEROSIGMA-AKASHIWO RAPHIDOPHYCEAE; CENTRAL VENICE LAGOON; ALEXANDRIUM-TAMARENSE; RED-TIDE; COASTAL LAGOONS; PHYTOPLANKTON; GROWTH; BAY; DINOFLAGELLATE; COMPETITION

Microalgae taxonomy and breeding

Microalgae are a diverse group of organisms that form the basic component of many eco­- systems. The systemic classification of algae is primarily based on their pigment composition and is divided into nine classes. The largest groups are Chlorophyceae (green algae), Phaeophyceae (brown algae), Pyrrophyceae (dinoflagellates), Chrysophyceae (golden brown algae), Bacillariophyceae (diatoms) and Rhodo­ phyceae (red algae). It has been estimated that between 22,000 and 26,000 species exist (Norton et al., 1996), of which only a few species have been identified to be useful for commercial application, such as Spirulina, Chlorella, Haematococcus, Dunaliella, Botryococcus, Phaeodactylum and Porphyridium. Several other species that are also cultivated commercially for the hatcheries in the aquaculture field include Chaetoceros, Crypthecodinium, Isochrysis, Nannochloris, Nitzschia, Schizo­chytrium, Tetraselmis and Skeletonema (Raja et al., 2008).

STUDIES ON TAXONOMY, DISTRIBUTION, ECOLOGY AND REPRODUCTIVE POTENTIAL OF ROTIFERS FROM SELECTED CENTRES IN COCHIN BACKWATER SYSTEM, KERALA

The studies were conducted in nine stations with varying ecological characteristics along Cochin backwaters and adjoining canals. Many workers opined that the distribution of rotifers is cosmopolitan. The significance of rotifers as first food for early larvae was indicated by Fujita. Aquaculture is a fast growing field in fisheries sector and it is gaining more importance as the fish landings and supply are getting irregular. A consistent supply of fish/shellfish can only be achieved through aquaculture. The success of any culture activity depends on the timely production of seeds of finfishes/shellfishes. The availability of wild seed is seasonal and erratic. So, a dependable source of seed of fishes and shellfishes is possible only through large scale production in hatchery. A successful seed production activity depends on the availability of a variety of suitable live feed organisms in sufficient quantities at the proper time for use in the larval stages. As the live feeds promote high growth rates, easy digestion, assimilation and the quality of not contaminating the culture water when compared to other artificial feeds, make the culture of live feed organisms the principal means of providing food for the larvae of finfishes and shellfishes. Rotifers are considered to be an excellent and indispensable food for larvae of many finfishes and crustaceans. It (1960) was the first to culture Brachionus plicatilis for feeding marine fish larvae, and now it is being extensively used as live feed in hatcheries all over the world. They are a group of microscopic organisms coming under the Phylum Rotifera which comprises of about 2000 species. Their slow swimming habits, ability to tolerate a wide range of salinities, parthenogenetic mode of reproduction and ability to get enriched easily, make rotifers an ideal live feed organism. The major factors such as temperature, salinity and food that influence the reproductive potential and thereby the population size of rotifer, Salinity is one of the most important aspect influencing the reproductive rate of rotifers. The feed type and feed concentration play a vital role in influencing the reproductive rate of rotifers. For culture of rotifers, the commonly used micro algae belong to Chlorella, Nannochloropsis, Isochrysis and Tetraselmis. While some studies have suggested that, algal diet has little effect on reproductive rates in 1979 while using the rotifer, Brachionus plicatilis as feed for the larvae of red sea bream, Pagrus major. It is generally accepted that rotifers play a pivotal role in the successful rearing of marine fish larvae.

Study of photorespiration in marine microalgae through the determination of glycolic acid using hydrophilic interaction liquid chromatography

Determination of organic acids in intracellular extracts and in the cultivation media of marine microalgae aid investigations about metabolic routes related to assimilation of atmospheric carbon by these organisms, which are known by their role in the carbon dioxide sink. The separation of these acids was investigated by hydrophilic interaction liquid chromatography (HILIC) using isocratic elution with a mobile phase composed of 70: 30 v/v acetonitrile/20 mmol/L ammonium acetate buffer (pH 6.8) and detection at 220 nm. HILIC allowed the determinations of glycolic acid, the most important metabolite for the evaluation of the photorespiration process in algae, to be made with better selectivity than that achieved by reversed phase liquid chromatography, but with less detectability. The concentration of glycolic acid was determined in the cultivation media and in intracellular extracts of the algae Tetraselmis gracilis and Phaeodactylum tricornutum submitted to different conditions of aeration: (i) without forced aeration, (ii) aeration with atmospheric air, and (iii) bubbling with N(2). The concentration of glycolic acid had a higher increase as the cultures were aerated with nitrogen, showing higher photorespiratory flux than that occurring in the cultures aerated with atmospheric air.