Comparison of three gracilarioids: growth rate, agar content and quality

Three gracilarioid species, Gracilariopsis bailiniae and Gracilaria tenuistipitata from Vietnam and Gracilaria gracilis from Russia, were studied in order to determine whether Gracilaria gracilis might be a superior species for cultivation in brackish-water ponds for agar production compared with the Vietnamese species. The effects of different salinity levels on the growth rate and agar production as well as agar properties of three gracilarioid species were compared in controlled laboratory experiments. Gracilaria tenuistipitata and G. gracilis were tolerant to low salinity (∼10‰), whereas Gp. bailiniae died under these conditions. G. tenuistipitata showed superior growth among the three species examined. Gracilaria gracilis had the highest agar content [36.8–46.6% dry weight (dw)]. Agar yield from Vietnamese gracilarioids did not exceed 30% dw. Gel strength of native agar from Gracilaria gracilis was two-fold higher that from Vietnamese species (278 g cm⁻² vs 130 g cm⁻²). Alkali pretreatment increased gel strength significantly for Gracilaria gracilis (1.4-fold), and G. tenuistipitata and Gp. bailiniae (2.3-fold) compared with native agar. The results suggest that Gracilaria gracilis may be a suitable species for production of reasonably good quality agar.     

Phycological research in the development of the Chinese seaweed industry

The term seaweed industry is employed in a broad sense and includes production both of commercial seaweeds such as Laminaria and Porphyra by phycoculture and of processed seaweed products, such as algin, agar and carrageenan.Before the founding of the People's Republic, China had a very insignificant seaweed industry, producing small quantities of the purple laver Porphyra and the glueweed Gloiopeltis by the primitive rock-cleaning method and the kelps Laminaria and Undaria by the primitive stone-throwing method, both aiming at enhancing the growth of the wild seaweeds. Also, a small quantity of agar was manufactured by the traditional Japanese method of gelling, freezing, thawing and drying the product. The small production was not sufficient to meet the demand of the Chinese people who for ages have appreciated seaweeds and their products for food. Therefore, large quantities of seaweeds and seaweed products had to be imported from various countries, for instance, Eucheuma and Gracilaria from Indonesia and other southeastern Asian countries, Laminaria and agar from Japan, even Porphyra from the USA. Annual Laminaria import from Japan generally amounted to over 10 000 tons and in some years approached 20 000–30 000 tons. Some quantities of the glueweed Gloiopeltis and the vermifuge weed Digenea simplex were exported, mainly to Japan.Since the founding of the People's Republic of China in October, 1949, China has exerted efforts to build up a self-supporting seaweed industry. Now after a lapse of 30-some years, a sizable seaweed industry has been developed. China is now able to produce by phycoculture more than one million tons of fresh seaweeds, including Laminaria, Undaria, Porphyra, Eucheuma, Gracilaria etc. and several thousand tons of seaweed extracts, including algin, agar, carrageenan, mannitol and iodine. At present, China still imports some quantities of seaweeds and seaweed products from various countries but is able to produce sufficient quantities to meet the people's need and even to export some quantities of the seaweeds Laminaria, Undaria and Porphyra and the seaweed products algin and mannitol.At the Tenth International Seaweed Symposium, I presented a paper on the Marine Phycoculture of China, in which I emphasized on the methods of cultivation (Tseng 1981b). Therefore I would like to take this opportunity to supplement the last lecture by presenting a paper on the role of phycological research in the development of China's seaweed industry.     

Agar properties of two species of Gracilariaceae from the Gulf of California,Mexico

Agar properties of two potentially commercial important seaweeds from the Gulf of California were studied. Maximum yield in Gracilaria vermiculophylla (45.7%) occurred during the summer months, coinciding with high water temperatures (31°C) whereas minimum yields (11.6%) were obtained during the coldest months of the year when populations of this species diminish in the bay. Gracilariopsis longissima showed two yield peaks, one in spring and another in fall, before the maximum and minimum seawater temperatures. Gel strength in native agar from the two species was low (<22.5 g cm⁻²) for most of the year. G. vermiculophylla native agar showed a slight increase in gel strength from June to August, which were the hottest months. Maximum value was 85 g cm⁻¹ in August. Maximum gel strength in G. longissima was observed in October (91 g cm⁻¹), and an unusual native agar with no detectable gel strength was observed in March and April samples. Gelling temperatures range from 27.7 to 36.5°C in G. vermiculophyla and from 26.6 to 34.9°C in G. longissima, meanwhile melting points were 73.9 – 53.5°C and 75.5 – 56.6°C, respectively. Sulfate content was high, 6.3–13.9% in G. vermiculophylla and 1.9–11.9% in G. longissima, and on the other hand 3,6 anhydrogalactose content was low 12.1–26.7% and 9.1–23%, respectively compared to other species. Results obtained showed that mean native agar yields of Gracilaria vermiculophylla and Gracilariopsis longissima from the Gulf of California are comparable to other tropical Gracilaria. However, the low gel strength, high sulfate content and low 3,6 anhydrogalactose content observed in the native agar extracted from these species make this an agaroid, thus alternative methods of extraction should be used to evaluate the possibility of commercial utilization of both species.     

Effects of temperature and salinity on the growth of Gracilaria verrucosa and G. chorda, with the Potential for mariculture in Korea

Effects of temperature and salinity on the growth of the two agarophytes, Gracilaria verrucosa (Hudson) Papenfuss and Gracilaria chorda Holmes were examined in Korea. Both species grew over a wide range of temperatures (10–30 ^∘C) and salinities (5–35‰), and grew well at 17–30 ^∘C and a salinity of 15–30‰. In culture, G. verrucosa grew faster than G. chorda and their maximum growth rates were 4.95% day⁻¹ (30 ^∘C, 25‰) and 4.47% day⁻¹ (at 25 ^∘C, 25‰), respectively. In the field population the maximum growth and fertility of G. chorda were observed in summer. The growth rate of G. verrucosa was slightly higher than that of G. chorda for 2 weeks on the cultivation rope and in culture but it was much lower after being contaminated with epiphytes. The biomass of the epiphytes was 0.82 g dry wt. per host plant in G. verrucosa and 0.001 g in G. chorda. G. chorda exhibited resistance to epiphytism and grew 7 times in length and the dry weight increased 15 times after 55 days. In conclusion, G. chorda appears to be a good agarophyte with a fast growth rate and resistance to epiphytesm, and compared with G. verrucosa, has good potential for commercial cultivation.     

Influence of external salinity on the osmolality of xylem sap,leaf tissue and leaf gland secretion of the mangrove<Emphasis Type="Italic"> Laguncularia racemosa</Emphasis> (L.) Gaertn

This study assessed if mature leaves of Laguncularia racemosa were able to demonstrate salt secretion, and if the magnitude of secretion was a function of soil salinity. Thus, salinity influence on the osmolality of leaf tissue, xylem sap and leaf secretion was assessed in field and glasshouse experiments. As salinity increased, solutes were accumulated in sufficient quantity to decrease osmotic potential over the whole range of water potential. In the field, xylem osmolality (mol m^–3) increased with salinity from 32.4±2.9 at 17 to 38.2±0.6 at 28. Similarly, in the glasshouse, xylem sap osmolality (mol m^–3) increased from 33.4±1.8 (15) to 40.6±1.5 (30). Changes in Na⁺ concentration explained about 51–58% of increase in xylem osmolality. Rates of secretion (mmol m^–2 day^–1) in the field increased from 0.80±0.12 (17) to 1.16±0.14 (28), and in the glasshouse the secretion increased from 0.73±0.07 (15) to 1.25±0.07 (30). The Na⁺ accounted for 40–53% of total secretion. This study presented evidence of the capability of mature leaves of L. racemosa to secrete salt for the first time, and that the rates of secretion were enhanced as soil salinity increased.     

Influence of Gamma Radiation on the Transport of Indolyl-3-Acetic Acid in Bark Tissue from Apple Trees

The influence of gamma radiation on the polarity of IAA translocation was investigated. Pieces of apple tree shoots, taken from 3-year-old trees, were irradiated in a dark irradiation chamber BK-⁶⁰Co source, 10 kCi. Doses from 20 to 150 krad were given. Strips of bark were peeled off both irradiated and control shoots. The middle parts of the bark strips were placed on agar blocks loaded with IAA-1-¹⁴C. The radioactivity of basal and acropetal segments was determined after about 20 h and results expressed as polarity quotient. It has been shown that irradiation with 20 krad (200 Gy) decreases the polarity quotient remarkably (polarity quotient of control equals 7; for 20 krad—2.9; 100 krad—1.6). The decrease in polarity results from inhibition of the basipetal translocation of IAA. Irradiated tissue shows a decrease in IAA uptake. Possible mechanisms of gamma irradiation effects on polar translocation of IAA are discussed.     

Commercial potential of seaweeds from St Lawrence Island,Alaska

This study assessed the possibility of using drift and subtidal seaweeds from St Lawrence Island, Alaska (lat. 63°N) for sale by the native population after simple processing. Over 125 km of coastline were surveyed for distribution of both drift and subtidal seaweeds. Drift seaweed wet weight densities ranged from 0.2 to over 9 kg m⁻², with an average of over 4 t km⁻¹ in the areas sampled. Attached, benthic seaweed densities ranged from 0.15 to 0.32 kg m⁻². Thirty and 35% of the biomass was composed of Agarum cribrosum and species of Laminaria, respectively, both as drift and as benthic seaweed. Data from tagged Laminaria indicated that growth rates were relatively slow for most of the year. The drift seaweed resource on the coasts south and west of the city of Gambell appeared to have good potential for a small-scale commercial harvest. (*author for correspondence)     

The Influence of Salinity on the Rate of Dark Respiration and Structure of the Cells of Brown Algae Thalli from the Barents Sea Littoral

We studied the changes in the rate of dark respiration (DR) and structure of the cells in Ascophyllum nodosum and Fucus vesiculosus thalli during the incubation at 40, 34, 20, 10, and 2 salinity for 14 days. The changes in salinity affected the rate of DR and the structure of the thallus apical cells: the organelles swollen and were destroyed later. The effect of hyposalinity on the algae was more pronounced as compared to hypersalinity. The stress intensity directly increased with the degree of desalination. Further adaptation of the algae to low salinity enhanced DR and, hence, was an energy-dependent process. Despite higher DR rates (during the stress and adaptation) in F. vesiculosus as compared to A. nodosum, the seaweeds had similar pattern of adaptation to the variation of salinity. Different primary response of the seaweeds to 20 salinity was an important exception; apparently, the salinity around 20 is the limit of these species distribution in desalination zones.     

Effect of salinity and pH on growth and agar yield of Gracilaria tenuistipitata var. liui in laboratory and outdoor cultivation

Acclimation responses of the red alga Gracilaria tenuistipitata var. liui collected on the northwest coast of Philippines were determined in laboratory setups and outdoor cultivation tanks in Haifa, Israel. Growth under laboratory conditions was influenced by all three variables studied, namely, temperature (20 or 30 ^°C), salinity (20, 30 or39‰) and seawater pH (6.5, 7.0, 8.0 or ≥ 9.0). In 250 mL flasks lacking pH control growth was influenced by temperature only at 20 ‰, whereas at 39 ‰, growth rates were similar at 20 or 30 ^°C. In 500 mL cylinders in which pH was controlled, growth rates were significantly different at a pH of 6.5 and 7.0 for all salinities, with maximal rates occurring in 39 ‰. At pH 8.0, and above, growth rates between salinities were similar and reduced to approximately 50% at a pH of 9.0 compared to rates at a pH of 6.5. Photosynthesis responses generally resembled growth responses both, in 250 mL and 500 mL cultures. In 40-L outdoor tanks, weekly growth and agar yields were apparently enhanced by increasing light intensities (up to full sunlight) and nutrient concentrations (up to 0.2 mM PO₃ ^2- and 2.0 mM NH₄ ⁺), and rates averaged four times higher than rates determined in the smaller flask cultures. This study shows broad salinity tolerance of G. tenuistipitata var. liui and its ability to sustain growth rates that are among the highest measured for Gracilaria spp. in outdoor cultures. This revised version was published online in August 2006 with corrections to the Cover Date.     

Physiological responses of Pterocladia and Gelidium (Gelidiales,Rhodophyta) from the Azores,Portugal

Manometric studies were conducted on Pterocladia capillacea, Gelidium latifolium and Gelidium spinulosum from the Azores, Portugal to determine optimal values of temperature, light and salinity for growth. Physiological responses were considered in relation to vertical distribution patterns of these species commonly observed throughout the Azores. Optimal parameters for the growth of Pterocladia capillacea, Gelidium latifolium and G. spinulosum were 17 to 25 °C, a photon flux density between 200 and 300 µmol m^–2 s^–1 and salinities of 25 to 35.     

Laboratory-induced development of the ice-ice disease of the farmed red algae Kappaphycus alvarezii and Eucheuma denticulatum (Solieriaceae,Gigartinales, Rhodophyta)

The most common perception of unfavorable environmental factors causing the ice-ice disease in the farmed seaweeds, Kappaphycus and Eucheuma, was demonstrated in this study for the first time using stressful conditions of abiotic factors in a continuous culture system. Light intensity of less than 50 mol photon m^–2 s^–1 and salinity of 20% or less induced ice-ice whitening characterized by short segments at midbranches which were similar to those observed in the Philippine seaweed farms, while temperatures of up to 33–35 °C resulted in wide-scale whitening leading to complete damage of the branches. These effects were preceded by slow growth rates from an optimum of 3.7% d^–1 to almost –2.0% d^–1. Mechanical stress by wound injury did not result to ice-ice whitening similar to the above. Environmental factors observed to trigger ice-ice in the laboratory, although may not necessarily parallel those in the field, may act synergestically to produce similar effects.     

Comparison of growth rate in culture,dry matter content,agar content and agar quality of two New Zealand red seaweeds,Gracilaria chilensis Bird,McLachlanet Oliveira andGracilaria truncata Kraft

Plant growth rates and agar characteristics were compared for two agarophytes,Gracilaria chilensis (Gracilariales, Rhodophyta) andG. truncata, so that the suitability ofG. truncata for mariculture could be assessed.G. chilensis plants grew steadily in the laboratory culture system at rates of 5–8% day^-1 (mean Relative Growth Rate) throughout the 6 week experimental period, with no decline in health.G. truncata plants grew at rates of 2–4% day^-1 for 5 weeks, but during the sixth week their apical tips became necrotic and growth rates fell to zero. There was no significant difference in the dry matter content (as a percentage of their wet weight) between the two species ofGracilaria, with values falling in the range of 16–19%. Slightly higher agar yields were obtained from alkali-treatedG. chilensis (17–20% dry matter) than from untreatedG. truncata (16–18%) although the agar fromG. truncata formed softer gels from which it was more difficult to recover. The gel strength of untreated agar extracted fromG. chilensis was very low (ca. 100 g cm^-2 for a 1% gel) but when an alkali treatment step was included in the extraction process, it increased to 520 g cm^-2 for a 1% gel. Contrary to an earlier report, untreatedG. truncata agar also had a very low gel strength (ca. 100 g cm^-2 for a 1.5% gel), but it rose to only 167 g cm^-2 after alkaline treatment.     

Seasonal variation of the agar quality and chemical composition of Gracilaria veleroae and Gracilaria vermiculophylla (Rhodophyceae,Gracilariaceae) from Baja California Sur,Mexico

Yield and physico‐chemical properties of agar from Gracilaria veleroae E.Y. Dawson and Gracilaria vermiculophylla (Ohmi) Papenfuss were studied and the chemical composition of the two seaweeds was determined. Samples were collected seasonally from summer 2003 to spring 2005. The agar yield did not vary significantly between seasons for both species. The lowest agar gel strength was obtained from G. veleroae (207.5 g cm^–2) in summer 2003 and the highest from G. vermiculophylla (793.1 g cm^–2) in winter 2004. Melting temperatures and hysteresis were higher in G. vermiculophylla, whereas gelling temperatures and 3,6‐anhydrogalactose content were higher for G. veleroae. Moisture, ash, crude fiber, and ether extract showed no significant seasonal variation for G. veleroae. The chemical composition of G. vermiculophylla showed significant seasonal variation. G. vermiculophylla possesses a better agar quality than G. veleroae and is a species that could be considered as a source of agar for commercial use.     

Growth rates and agar properties of three gracilarioids in suspended open-water cultivation in St. Helena Bay, South Africa

Relative growth rates (RGRs), yields and agar characteristics of threegracilarioid isolates (Gracilariopsis sp. from St. Helena Bay, and Gracilaria gracilis isolates from Langebaan Lagoon and Saldanha Bay) weremeasured to assess the suitability of a site in St. Helena Bay for suspendedcultivation. The gracilarioids were grown on polypropylene ropes and`netlon' lines, and the RGRs were 4.0–11.0% d<sup>-1</sup> and 5.0–7.0%d<sup>-1</sup>, respectively. The RGR of the Langebaan isolate of G. gracilis grown on ropes was significantly higher than the RGR of otherisolates. The mean net yield for the Langebaan isolate grown on `netlon'lines was 2.6 ± 0.9 kg wet wt m^-2 30 day^-1. Thecultured gracilarioids were extracted for native and alkali treated agars. Themean native agar yield over the entire period was 39.0% dry wt. Alkalipretreatment reduced the yield by 55%, but significantly increased gelstrength. High gel strengths (>750 g cm^-2) were measured inagars from Gracilariopsis sp. and Saldanha Gracilaria gracilis inmid-summer and winter. The dynamic gelling and melting temperatures ofnative and alkali treated agars varied among the gracilarioids. The meangelling and melting temperatures of agars were about 39.0 ^°C and86.0 ^°C, respectively. The 3,6-AG content ranged from 29% to38% for native agars and 34–45% for alkali treated agars. While theseresults indicate that this site is suitable for gracilarioid cultivation, occasionallow-oxygen events in St. Helena Bay lead to production of hydrogensulphide in the sea water (`black tides'). Such events killed most inshorebiota (including seaweeds) in 1994 and 1998. This frequency (on average1–2 per decade) and duration (maximum 2 weeks) would have to beconsidered in planning commercial seaweed farming in St. Helena Bay.     

Current state of seaweed resources in Spain

Commercial seaweeds in Spain are harvested on the north and northwest coasts. They are mainly agarophytes and carrageenophytes (Gelidium spp. and some Irish moss-like species, respectively), although some Phaeophyceae species (Fucus spp. and Laminaria spp.) are also exploited for alginates. No industrial seaweed cultivation is carried out in Spain at present. Spain's total commercial seaweed harvest can be estimated at 6,528 ± 2,076 t dry wt year^–1. Gelidium spp. are by far the most harvested, attaining 5,135 ± 1,761 t dry wt y^–1. To date, Spain's commercial seaweed harvest is low compared with that of other countries, but data on field standing crops and productivities of commercial taxa suggest that harvesting could be increased greatly.     

Influence of changes in sea water salinity on the growth,photosynthetic pigment content,and cell size of the benthic alga <Emphasis Type="Italic">Attheya ussurensis</Emphasis> Stonik,Orlova et Crawford, 2006 (Bacillariophyta)

The study deals with the effect of changes in salinity from 32 to 4‰ (at an interval of 4‰) on the growth, chlorophyll a and carotenoid contents, and cell size of the benthic alga Attheya ussurensis (Bacillariophyta). A. ussurensis showed high tolerance to reduced salinity and ability to adapt to salinity changes from 16 to 12‰. In this salinity range, the cells restored their shapes, sizes, and physiological functions. The number of cells and photosynthetic pigment content were highest at a salinity reduction to 24‰. At 8‰, algal cells remained alive, but the process of cell division was inhibited; as a result, the number of cells was significantly lower than in the control, the cells did not restore their sizes and shapes and remained deformed until the end of the experiment. A drop in salinity to 4‰ caused a complete loss of cell viability of A. ussurensis within a day of exposure to this factor.     

The Reaction of the Starfish Asterias amurensisand Patiria pectinifera (Asteroidea) from Vostok Bay (Sea of Japan) to a Salinity Decrease

The reactions of the starfish Asterias amurensis and Patiria pectinifera that live in Vostok Bay at the salinity of 32–33 to a salinity decrease were studied under laboratory conditions. The lower limits of the desalination tolerance range of A. amurensis and P. pectinifera were, respectively, 24 and 20. A. amurensis proved to be less resistant to desalination. Under experimental conditions, all specimens of this species survived the salinity of 22, while those of P. pectinifera tolerated 18. At the same time, A. amurensis responded more actively than P. pectinifera to unfavorable changes in the environment. Turned to their dorsal side and exposed to a salinity of 16 to 32, the former reverted to the normal position within a shorter time than the latter. Being a more euryhaline species, P. pectinifera endured a salinity decrease to 6 or 8 over, respectively, 21 or 28 h. However, only 30–40% of all specimens could recover locomotory activity 12 or 8.5 h after being placed into water of normal salinity.     

Biochemical characteristics of cellulose and a green alga degradation by Gilvimarinus japonicas 12-2T,and its application potential for seaweed saccharification

ABSTRACT

Cellulose is one of the major constituents of seaweeds, but reports of mechanisms in microbial seaweed degradation in marine environment are limited, in contrast to the multitude of reports for lignocellulose degradation in terrestrial environment. We studied the biochemical characteristics for marine cellulolytic bacterium Gilvimarinus japonicas 12-2^T in seaweed degradation. The bacterial strain was found to degrade green and red algae, but not brown algae. It was shown that the bacterial strain employs various polysaccharide hydrolases (endocellulase, agarase, carrageenanase, xylanase, and laminarinase) to degrade seaweed polysaccharides. Electrophoretic analysis and peptide sequencing showed that the major protein bands on the electrophoresis gel were homologous to known glucanases and glycoside hydrolases. A seaweed hydrolysate harvested from the bacterial culture was found useful as a substrate for yeasts to produce ethanol. These findings will provide insights into possible seaweed decomposition mechanisms of Gilvimarinus, and its biotechnological potential for ethanol production from inedible seaweeds.     

Low salinities adversely affect photosynthetic performance of the mangrove,Avicennia marina

Photosynthetic performance of the highly salt tolerant mangrove, Avicennia marina, was compared at two sites differing insubstrate soil salinities. Carbon dioxide exchange and chlorophyll fluorescence weremonitored at a high salinity site in Durban Bay (35) and at a low salinitysite in Beachwood (< 12). Mean CO₂ exchange, conductanceand transpiration were consistently higher at the high salinity site. Carbondioxide response curves indicated that carboxylation efficiency was higherand stomatal limitation lower at the Durban Bay site. PSII quantum yield,electron transport rates (ETR) and intrinsic PSII efficiency(F_v/F_m) were significantly higher at the high salinity site.Quenching analysis indicated a higher degree ofphotoinhibition/photoprotection in leaves at the low salinity site. Predawnand midday leaf water potentials were –1.6 and –3.1 MPa at Beachwood,compared to –2.6 and –3.8 MPa, respectively, at Durban Bay. Leafconcentrations of Na⁺, K⁺, Ca²⁺, Mg²⁺,Cl^- and N were significantly higher at Durban Bay. Photosyntheticperformance is apparently impaired at the low salinity site in Beachwood asa result of K⁺ and N deficiencies in the leaves.     

Abundance of drifting seaweeds in eastern East China Sea

Drifting seaweeds play important ecological roles in offshore waters. Recently, large amounts of drifting seaweed rafts were found in the eastern East China Sea between the continental shelf and the oceanic front of the Kuroshio Current. However, so far there have been no quantitative reports about this particular area. Two research cruises were organized to survey abundance and standing crop of drifting seaweeds in eastern East China Sea in May 2002 and March 2004, using visual census and net sampling of drifting seaweeds. Visual census data were composed of drifting seaweed raft diameter, perpendicular distance from the transect (navigation course of the research vessel) to the raft, and positions. Using these data, we calculated the “effective stripe width” using the DISTANCE software. Drifting seaweed abundance (composed exclusively of Sargassum horneri) in waters located between the continental shelf peripheral area and the Kuroshio oceanic front was estimated to be higher than in any other area within eastern East China Sea in March and May. Abundance means in May 2002 and March 2004 were 6.14 and 29.05 rafts km⁻², respectively, while standing crop reached 126.81 and 20.35 kg km⁻² (wet weight). Mean diameter and drifting seaweed rafts in May 2002 were significantly greater than in March 2004, reflecting seasonal growth of Sargassum horneri.