Kappaphycus alvarezii (Rhodophyta, Areschougiaceae) cultivated in subtropical waters in Southern Brazil

Four strains of Kappaphycus alvarezii were cultivated in the subtropical waters of Florianopolis, Santa Catarina State, Brazil (27 degrees 29`19 `` S/48 degrees 32` 28 `` W), from February 2009 to February 2010. Seaweeds were cultivated on floating raft near of mussel farms. Salinity ranged from 29 to 36 psu and temperature from 17.1 to 28.5 degrees C. Higher growth rates (5.12-4.29% day(-1)) were measured in summer and autumn, showing a positive correlation between growth rate and water temperature. Lower growth rates (0.54-0.32% day(-1)) occurred in winter, resulted mainly by biomass loss. Significant differences were observed among the strains in spring and the brown tetrasporophytic strain was the only one which failed to recover, being excluded of the experiments. The effect of cultivation periods (36, 42, and 97 days) on carrageenan yield, gel strength, and viscosity were analyzed. Carrageenan yields were higher for plants kept 42 days in the sea (28%), against 25% for 36 and 97 days. There were no significant differences in carrageenan yield among the strains analyzed. Viscosity increased with the increase of cultivation period, while gel strength seemed to vary at random. Tetrasporangia and cystocarps were not observed, and lost fragments did not attach outside the raft. In general, dissolved inorganic nitrogen concentration decreased around the cultivation area as compared to the mussel farm. Results show that cultivation of K. alvarezii is technically feasible in subtropical waters and can be associated with local mussel farms, mitigating the eutrophication and, eventually, increasing the economic return of the farmers.

Callus induction and micropropagation improved by colchicine and phytoregulators in Kappaphycus alvarezii (Rhodophyta, Solieriaceae)

Tissue culture techniques were applied for micropropagation of the red alga Kappaphycus alvarezii in order to select the best strain and experimental system for in vitro culture. Five strains were tested: brown (BR), green (GR) and red (RD) tetrasporophytes, brown female gametophyte (BFG), and a strain originating from tetraspore germination (""Edison de Paula"", EP). The effects of three culture media were tested on callus formation, regeneration from explants and from callus in the three tetrasporophytic and EP strains: seawater enriched with half-strength of von Stosch`s (VS 50) and Guillard & Ryther`s (F/2 50) solutions, plus synthetic ASP 12-NTA medium, with or without gelling agent. Explants of the EP strain were treated with glycerol and the phytoregulators indole-3-acetic acid (IAA); 2,4-diclorophenoxyacetic acid (2,4-D); and benzylaminopurine (BA), alone or in combination. The effects of colchicine (0.01%) during 24, 48, 72 hours and 14 days were analyzed in the BFG and EP strains. The EP strain showed the highest percentage of explants forming callus and regeneration from explants in VS 50, indicating its high potential for micropropagation in comparison to the other strains. Regeneration from callus was very rare. Treatments with glycerol and IAA:BA (5:1 mg L(-1)) stimulated the regeneration from explants. Significant differences were observed in the percentages of regeneration of EP strain explants treated with colchicine for 14 days. Our results indicate that IAA and BA stimulated the regeneration process, and that colchicine produced explants with high potential for regeneration, being useful for improving the micropropagation of K. alvarezii.

Nutrients removed by Kappaphycus alvarezii (Rhodophyta, Solieriaceae) in integrated cultivation with fishes in re-circulating water

The potential of the red alga Kappaphycus alvarezii to remove nutrients was tested to treat effluents of Trachinotus carolinus fish cultivation, and the production of carrageenan in this condition was analyzed. Experiments were conducted in four tanks of 8000 L with approximately 1200 fishes of 30 g each integrated with three tanks of 100 L with 700 g of K. alvarezii, as initial biomass per tank. Seawater was re-circulated between tanks with seaweed and with fish. As a control, three tanks with seawater circulating in an open system were utilized. Seawater samples were collected daily for 10 days and concentrations of nitrate, nitrite, ammonium and phosphate were determined in the inflow and outflow water of the tanks. Significant differences between both collecting points were considered as nutrient removal by the seaweed. Growth rates and carrageenan yields were also analyzed in seaweed cultivated in seawater and in effluents. Growth rates of seaweed cultivated in tanks were lower than those obtained in open sea and in laboratory cultivation. Effluents had concentrations of nitrate and nitrite ca. 100 times higher than in the control. Maximum values of nutrient removal on effluents were: nitrate= 18.2%; nitrite =50.8%; ammonium =70.5% and phosphate =26.8%. All plants survived throughout the experimental period, but some developed ""ice-ice"", a disease associated with physiological stress. After the experimental period, some plants selected and cultivated in open sea presented higher growth rates in 40 days, indicating nutrient storage. No significant differences between carrageenan yields of K alvarezii cultivated in seawater and in the effluents were observed. Our results show that K. alvarezii can be utilized as a biofilter for fish cultivation effluents, reducing the eutrophication process and can also be processed for carrageenan production, which provides an additional benefit to the fisheries. (C) 2008 Elsevier B.V. All rights reserved.

Germination and survival of tetraspores of Kappaphycus alvarezii var. alvarezii (Solieriaceae, Rhodophyta) introduced in subtropical waters of Brazil

The carrageenophyte Kappaphycus alvarezii was introduced in 1995 and vegetatively propagated in Ubatuba, Sao Paulo State, Brazil, for the purpose of commercial cultivation. This species produces tetraspores mainly in the austral summer and fall. Tetraspore germination and survival were studied under different conditions of temperature, photon flux density, and photoperiod in the laboratory. Field experiments were also carried out. Although tetraspores of K. alvarezii germinated, they had low survival rates, most dying after 20 days. Recruitment of K. alvarezii tetraspores did not occur in experiments conducted in the field. The results indicated that the establishment of K. alvarezii via spore production in the natural environment of the south-east coast of Brazil is rather remote.

Kinetic study of the plastoquinone pool availability correlated with H2O2 release in seawater and antioxidant responses in the red alga Kappaphycus alvarezii exposed to single or combined high light, chilling and chemical stresses

Under biotic/abiotic stresses, the red alga Kappaphycus alvarezii reportedly releases massive amounts of H2O2 into the surrounding seawater. As an essential redox signal, the role of chloroplast-originated H2O2 in the orchestration of overall antioxidant responses in algal species has thus been questioned. This work purported to study the kinetic decay profiles of the redox-sensitive plastoquinone pool correlated to H2O2 release in seawater, parameters of oxidative lesions and antioxidant enzyme activities in the red alga Kappaphycus alvarezii under the single or combined effects of high light, low temperature, and sub-lethal doses of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) and 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB), which are inhibitors of the thylakoid electron transport system. Within 24 h, high light and chilling stresses distinctly affected the availability of the PQ pool for photosynthesis, following Gaussian and exponential kinetic profiles, respectively, whereas combined stimuli were mostly reflected in exponential decays. No significant correlation was found in a comparison of the PQ pool levels after 24 h with either catalase (CAT) or ascorbate peroxidase (APX) activities, although the H2O2 concentration in seawater (R = 0.673), total superoxide dismutase activity (R = 0.689), and particularly indexes of protein (R = 0.869) and lipid oxidation (R = 0.864), were moderately correlated. These data suggest that the release of H2O2 from plastids into seawater possibly impaired efficient and immediate responses of pivotal H2O2-scavenging activities of CAT and APX in the red alga K. alvarezii, culminating in short-term exacerbated levels of protein and lipid oxidation. These facts provided a molecular basis for the recognized limited resistance of the red alga K. alvarezii under unfavorable conditions, especially under chilling stress. © 2006 Elsevier B.V. All rights reserved.

Optimization of the extraction of carrageenan from Kappaphycus alvarezii using response surface methodology

This study aims to optimize an alternative method of extraction of carrageenan without previous alkaline treatment and ethanol precipitation using Response Surface Methodology (RSM). In order to introduce an innovation in the isolation step, atomization drying was used reducing the time for obtaining dry carrageenan powder. The effects of extraction time and temperature on yield, gel strength, and viscosity were evaluated. Furthermore, the extracted material was submitted to structural analysis, by infrared spectroscopy and nuclear magnetic resonance spectroscopy (¹H-NMR), and chemical composition analysis. Results showed that the generated regression models adequately explained the data variation. Carrageenan yield and gel viscosity were influenced only by the extraction temperature. However, gel strength was influenced by both, extraction time and extraction temperature. Optimal extraction conditions were 74 ºC and 4 hours. In these conditions, the carrageenan extract properties determined by the polynomial model were 31.17%, 158.27 g.cm-2, and 29.5 cP for yield, gel strength, and viscosity, respectively, while under the experimental conditions they were 35.8 ± 4.68%, 112.50 ± 4.96 g.cm-2, and 16.01 ± 1.03 cP, respectively. The chemical composition, nuclear magnetic resonance spectroscopy, and infrared spectroscopy analyses showed that the crude carrageenan extracted is composed mainly of κ-carrageenan.

Co-stressors chilling and high light increase photooxidative stress in diuron-treated red alga Kappaphycus alvarezii but with lower involvement of H(2)O(2)

Diuron is one of the most commonly found N-phenylurea herbicides in marine/estuarine waters that promotes toxic effects by inhibiting photosynthesis and affecting the production of reactive oxygen species (ROS) in autotrophs. Since photo- and thermoacclimation are also ROS-mediated processes, this work evaluates a hypothetical additive effect of high light (HL) and chilling (12 degrees C) on 50 nM diuron toxicity to the highly-photosynthetically active apices of the red alga Kappaphycus alvarezii. Additive inhibition of photosynthesis was mainly evidenced by significant decreases of quantum yield of photosystem II and electron transfer rates upon co-stressors exposure to diuron-treated algae. Under extreme 12 degrees C/HL/diuron conditions, unexpected lower correlations between H(2)O(2) concentrations in seawater and radical-sensitive protein thiols were concomitantly measured with the highest indexes of photoinhibition (parameter beta). Altogether, these data support the hypothesis that co-stressors chilling/HL additively inhibit photosynthesis in diuron-exposed K. alvarezii but with less involvement of H(2)O(2) in injury effects than with only chilling or HL. (C) 2010 Elsevier Inc. All rights reserved.

Co-stressors chilling and high light increase photooxidative stress in diuron-treated red alga Kappaphycus alvarezii but with lower involvement of H2O2

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

Effects of Natural Radiation, Photosynthetically Active Radiation and Artificial Ultraviolet Radiation-B on the Chloroplast Organization and Metabolism of Porphyra acanthophora var. brasiliensis (Rhodophyta, Bangiales)

We undertook a study of Porphyra acanthophora var. brasiliensis to determine its responses under ambient conditions, photosynthetically active radiation (PAR), and PAR+UVBR (ultraviolet radiation-B) treatment, focusing on changes in ultrastructure, and cytochemistry. Accordingly, control ambient samples were collected in the field, and two different treatments were performed in the laboratory. Plants were exposed to PAR at 60 mu mol photons m(-2) s(-1) and PAR+UVBR at 0.35 W m(-2) for 3 h per day during 21 days of in vitro cultivation. Confocal laser scanning microscopy analysis of the vegetative cells showed single stellate chloroplast in ambient and PAR samples, but in PAR+UVBR-exposed plants, the chloroplast showed alterations in the number and form of arms. Under PAR+UVBR treatment, the thylakoids of the chloroplasts were disrupted, and an increase in the number of plastoglobuli was observed, in addition to mitochondria, which appeared with irregular, disrupted morphology compared to ambient and PAR samples. After UVBR exposure, the formation of carpospores was also observed. Plants under ambient conditions, as well as those treated with PAR and PAR+UVBR, all showed different concentrations of enzymatic response, including glutathione peroxidase and reductase activity. In summary, the present study demonstrates that P. acanthophora var. brasiliensis shows the activation of distinct mechanisms against natural radiation, PAR and PAR+UVBR.

Cultivo ecosostenible de Kappaphycus alvarezii (Doty) Doty ex P. C. Silva en Panama

[ES] Kappaphycus alvarezii (Doty) Doty ex Silva ha sido cultivada en Cativá, localizado en la zona costera del caribe panameño, al noreste del Canal de Panamá, desde el 2004 con intereses comerciales. El cultivo fue desarrollado en dos partes: el cultivo in situ (en el mar y en tanques) y el cultivo in vitro (en el laboratorio y en aclimatación en tanques y acuarios). Ambas cultivos tienen como objetivo obtener un desarrollo sostenible en la zona costera y un producto atractivo para la industria que dé oportunidades económicas a los cultivadores. Los resultados muestran que la tasa de crecimiento observada (4.0 a 5.0 %) permanece constante durante todo el año y es independiente de los sitios seleccionados, de las temporadas: lluviosa y seca, y de los parámetros ambientales. Ya que menos de una hectárea ha sido cultivada y existen 40 disponibles, esto multiplica grandemente la posibilidad de explotación. En cuanto a los sistemas de plantación, las estructuras fijas causan menos impacto, son menos costosas y son beneficiosas para el crecimiento. Además, Kappaphycus alvarezii puede ser mantenida en tanques con tasas de crecimiento similares a las naturales, por lo tanto puede ser usada como biomasa para semillas. El secador usado, dio un cantidad estimada de 36 sacos de 11 Kg. de biomasa seca cada uno, materia prima por polígono (300 m2) año-1. La producción de carragena mantiene un patrón constante (ca. 35%) en relación a la producción y al peso molecular aceptado por la industria (1.6 105 Da). La producción de diferentes fracciones, iota y kappa, varía con el tiempo de cultivo, siendo mayor en talos jóvenes para la primera y menor para en los más viejos para la segunda. Las técnicas de los cultivos in vitro permiten el mantenimiento y propagación de stocks de fragmentos de explantos en el laboratorio y aclimatados en tanques y acuarios. La aclimatación ex vitro ha demostrado ser factible para la recuperación y futura propagación de los explantos mantenidos en el laboratorio a sitios en el mar.

Cultivos ecosostenibles de Kappaphycus alvarezii en las costas del Caribe Panameño y caracterización de la carragena

Valoración de la cantidad y calidad de la carragena de cultivares de Panamá

Caracterización biológica y química de Kappaphycus alvarezii de Panamá

Programa de doctorado Ecología y Gestión de los Recursos Vivos Marinos

Carrageenans from Australian representatives of the family Cystocloniaceae (Gigartinales, Rhodophyta), with description of Calliblepharis celatospora sp. nov., and transfer of Austroclonium to the family Areschougiaceae

Ten Australian representatives from seven of the 10 genera presently constituting the family Cystolcloniaceae have been analyzed for their cell-wall galactans. Included in our survey are the monotypic Australian-endemic genera Austroclonium, Gloiophyllis, Erythronaema, and Stictosporum, one species of Craspedocarpus, three species of Rhodophyllis, and two species of Calliblepharis. As one of the species of the latter genus is endemic to Western Australia and presently undescribed, we illustrate its habit and anatomical features in formally proposing to name it Calliblepharis celatospora Kraft, sp. nov. All the species surveyed essentially produce typical iota (iota)-carrageenans, with the exception of Austroclonium. The sulfated galactans from Austroclonium predominantly contain the repeating units of iota-, alpha (alpha)-, and 6'-O-methylated iota- and alpha-carrageenans; whether these exist as discrete polysaccharides or a complex hybrid structure was not resolved. Thus, Austroclonium carrageenans resemble the polysaccharides from Rhabdonia, Areschougia, and Erythroclonium. Although these latter three genera are currently included in the large gigartinalean family Solieriaceae, all produce significantly different carrageenans from Solieria itself and related genera such as Eucheuma, Kappaphycus, Betaphycus, Sarcodiotheca, Agardhiella, Sarconema, and Callophycus. In consideration of these findings, as well as of significant anatomical similarities, we provisionally recommend reestablishment of the family Rhabdoniaceae Kylin (as the family Areschougiaceae J. Agardh) for Rhabdonia, Areschougia, Erythroclonium, and Austroclonium.

The cell wall galactans from Australian representatives of the genus Meristotheca (solieriaceae, rhodophyta)

The Solieriaceae, has the largest number of genera (16-18) of any family in the carrageenophyte order Gigartinales. One of these genera, Meristotheca, consists of three or four species of foliose, erect to prostrate plants sporadically recorded from the tropics of both hemispheres. The hot-water-soluble polysaccharides from Australian representatives of the type species, M. papulosa, and M. procumbens from Lord Howe Island have been characterized by compositional assays, linkage analysis, and Fourier transform infrared and C-13-nuclear magnetic resonance spectroscopy. The results show that polysaccharides from both species are similar, being predominantly composed of 4-linked 3,6-anhydro-alpha-D-galactopyranose 2-sulphate alternating with 3-linked beta-D-galactopyranose 4-sulphate, as is typical of iota-carrageenan. Small proportions of the 3-linked units occur as the pyruvated residue 4,6-O-(1-carboxyethylidene)-beta-D-galactopyranose, and other minor variations from idealized iota-carrageenan were also detected. The polysaccharides from representatives of Meristotheca are comparable to those of other solieriacean algae analysed to date, but the minor structural variations suggest a closer chemotaxonomic affinity with noneucheumoid genera of the Solieriaceae, such as Sarconema, Solieria, and Tikvahiella, than to the eucheumoid genera Eucheuma, Kappaphycus and Betaphycus (tribe Eucheumatoideae) from which most kappa- and iota-carrageenans are commercially extracted.