Ulva lactuca as bioindicator of metal contamination in intertidal waters in Hong Kong

The use of Ulva lactuca L. as an indicator of metal contamination was assessed by analysing the levels of Mn, Fe, Ni, Cu, Zn, Cd and Pb in the alga collected from 24 intertidal sites around the Island of Hong Kong. Twelve of the sites are in the rural southern parts of the Island where the coastal waters are relatively clean. The remaining 12 sites are located in the north and within Victoria Harbour which receives, apart from industrial effluents, untreated domestic sewage from a population of some 3.5 million. The mean levels of Mn, Fe, Ni, Cu, Zn and Pb in Ulva from the urban sites were respectively 4.0, 4.6, 1.8, 2.3, 2.4 and 4.6 folds those from the rural sites. However, similar levels of Cd were found in the alga amongst all the sites. Locations of high levels of metal contamination, particularly to the eastern end of the Harbour, have been identified. Preliminary results indicate that Ulva is a good indicator of Mn, Fe, Cu, Zn and Pb contaminations.     

Ecological studies on Ulva lactuca L. from Veraval,India

Ulva lactuca L., at Veraval (20° 54N and 70° 22E) on the western coast of India, grows in the intertidal belt from June to late February. During the summer months of March, April and May, the species dries up, leaving rhizoidal fragments for perennation. High values of density, phytomass, frequency, weight, and volume have been recorded during September and January. Accumulation of phytomass appears to be chiefly density dependent. High dissolved oxygen content and low temperatures of the surface sea water favoured enhanced growth and, consequently, more phytomass.     

Chemical and physical-chemical characteristics of dietary fibres from Ulva lactuca (L.) Thuret and Enteromorpha compressa (L.) Grev.

Dietary fibres from Ulva lactuca (L.) Thuret (sea lettuce) and Enteromorpha compressa (L.) Grev. (A.O. nori) were measured according to a ‘standard’ method and a ‘physiological’ protocol simulating the gastric and intestinal environments. U. lactuca contained 15.8–8.0% soluble and 24.2–32.6% insoluble fibres according to the ‘standard’ and ‘physiological’ methods, respectively. For E. compressa, these values were 14.9–15.9 and 21.6–28.7%, respectively. For both algae, the composition suggests that the soluble fibres were xylorhamnoglycuronans sulphates and insoluble fibres were essentially composed of glucans. No marked chemical compositional variation was observed between soluble fractions extracted under the simulated gastric and intestinal conditions. Fibres in both algae are hydrophilic but the water holding capacities were higher after extraction of soluble fibres (5.5–9.5 g g⁻¹ for the dry algae; 14.0–16.0 g g⁻¹ for the standard insoluble fibres). Water soluble fibres demonstrated low intrinsic viscosities at 37 °C in buffers, particularly those from E. compressa (36.0–36.5 ml g⁻¹), and was affected by pH for those of U. lactuca (147.5 ml g⁻¹ at pH 3.0 and 175.0 ml g⁻¹ at pH 7.3).     

Perturbation of monovalent cation composition in Ulva lactuca by cadmium, copper and zinc

Discs of thallus cut from the macroalga Ulva lactuca were incubated in filtered seawater containing cadmium, zinc, copper or cobalt (30 m). The metal uptake rates differed for each metal in the order Cu > Zn > Cd > Co. Exposure of the macroalga to metals resulted in a disruption of intracellular monovalent cation composition. Intracellular potassium was irreversibly lost and sodium was accumulated by cadmium- or copper-treated U. lactuca, which was assumed to indicate irreversible disruption of the plasmalemma. Exposure to zinc caused an increase in sodium concentrations, whereas potassium concentrations were not significantly different from the controls, suggesting that the integrity of the plasmalemma had been maintained at the zinc concentration used. Intracellular magnesium was also lost from copper-treated algae, which again indicated a loss of integrity of the cell membrane.     

The population dynamics of some freshwater carideans (Crustacea: Decapoda) in Hong Kong,with special reference to Neocaridina serrata (Atyidae)

During a 15-month investigation of the population dynamics of three caridean shrimp species in the Lam Tsuen River, New Territories, Hong Kong, the numbers of the commonest species, Neocaridina serrata (Stimpson)(Atyidae), were reduced as water temperatures fell. By contrast, Caridina lanceifrons Yu became more numerous during the winter. The relatively scarce Macrobrachium hainanense (Parisi)(Palaemonidae) tended to be more abundant during the summer, and the abundance of this species was directly correlated with that of N. serrata. The population size of these two species was positively correlated with water temperatures prevailing one and two months prior to the collection of samples, but there was no significant relationship between C. lanceifrons abundance and water temperature.All species exhibited similar growth patterns with an inflection at the attainment of sexual maturity, occurring after the (presumed) 14th moult for N. serrata and in the (presumed) 13th and 18th stadium for C. lanceifrons and M. hainanensis respectively. Ovigerous shrimps of all species were only recorded when water temperatures exceeded 20 °C and the % incidence of ovigerous N. serrata was positively correlated with prevailing temperatures. It is suggested that restriction of freshwater caridean breeding periods by low temperatures may be common in the subtropics while perennial breeding is more likely to be typical of tropical regions.The mean carapace length of N. serrata populations was negatively correlated with prevailing temperatures, but more strongly correlated with temperatures recorded one and two months prior to sampling when gametogenesis may have taken place. Smallest mean sizes were recorded in late summer upon the cessation of juvenile recruitment. N. serrata has a maximum longevity of approximately 12 months and reproduction occurs in the 7th month of life or later. The hatchlings are well developed and clutch size is not related to the size of the brooding female. An annual life cycle, as seen in N. serrata, may be typical of many small freshwater carideans.Department of Zoology, The University of Hong Kong     

Mycorrhizal associations in Hong Kong Fagaceae

Polyphenols histochemically detected in fresh uninfected roots of Quercus, Castanopsis and Lithocarpus growing in Hong Kong and shown to be condensed tannins were found mainly as intracellular material in the cells of the root cap, the epidermal layer and the endodermis. The cell walls of the outer cortex and the endodermis also contained suberin. Following invasion by compatible ectomycorrhizal symbionts, condensed tannins disappeared from cells of the root cap and the epidermal layer but hyphae were prevented from colonizing the cortex presumably due to suberin barriers. In vitro experiments indicated that a number of broad-host ectomycorrhizal fungi could utilise various polyphenolic compounds, including tannins found in the root exudates of the host trees, with different degrees of efficiency.     

Eutrophication and macroalgal blooms in temperate and tropical coastal waters: nutrient enrichment experiments with Ulva spp.

Receiving coastal waters and estuaries are among the most nutrient‐enriched environments on earth, and one of the symptoms of the resulting eutrophication is the proliferation of opportunistic, fast‐growing marine seaweeds. Here, we used a widespread macroalga often involved in blooms, Ulva spp., to investigate how supply of nitrogen (N) and phosphorus (P), the two main potential growth‐limiting nutrients, influence macroalgal growth in temperate and tropical coastal waters ranging from low‐ to high‐nutrient supplies. We carried out N and P enrichment field experiments on Ulva spp. in seven coastal systems, with one of these systems represented by three different subestuaries, for a total of nine sites. We showed that rate of growth of Ulva spp. was directly correlated to annual dissolved inorganic nitrogen (DIN) concentrations, where growth increased with increasing DIN concentration. Internal N pools of macroalgal fronds were also linked to increased DIN supply, and algal growth rates were tightly coupled to these internal N pools. The increases in DIN appeared to be related to greater inputs of wastewater to these coastal waters as indicated by high δ¹⁵N signatures of the algae as DIN increased. N and P enrichment experiments showed that rate of macroalgal growth was controlled by supply of DIN where ambient DIN concentrations were low, and by P where DIN concentrations were higher, regardless of latitude or geographic setting. These results suggest that understanding the basis for macroalgal blooms, and management of these harmful phenomena, will require information as to nutrient sources, and actions to reduce supply of N and P in coastal waters concerned.     

Cell wall structure of the agarophytes Gracilaria tikvahiae and G. cornea (Rhodophyta) and penetration by the epiphyte Ulva lactuca (Chlorophyta)

Use of light, transmission, and scanning electronmicroscopes revealed that the epidermal cell wall ofthe red algal agarophytes Gracilaria tikvahiaeMcLachlan and G. cornea J. Agardh consists of adecklamelle and outer and inner wall layers. The twospecies differed, with G. cornea having asignificantly thicker outer wall and a more diffusedecklamelle. After induction, the zooids of Ulvalactuca would attach to glass slides and the twospecies of Gracilaria via an adhesion pad. Within a few days, 3–5 celled germlings penetrated thedecklamelle and outer wall layer of both basiphytes. By the time the epiphyte germlings reached the 15celled stage, they had penetrated the inner walllayer. The differences in epidermal cell wallconstruction between the two basiphytes may play arole in the ability of zooids of U. lactuca toattach in nature where epiphytization of G.cornea is infrequent.     

Growth of the seaweed Ulva rigida C. Agardh in relation to biomass densities, internal nutrient pools and external nutrient supply in the Sacca di Goro lagoon (Northern Italy)

Growth of the seaweed Ulva rigida C. Agardh was investigated in relation to biomass densities, internal nutrient pools and external nutrient supply. Research was carried out from 23 March to 5 July 1994 in the Sacca di Goro (Po Delta, Northern Italy), whose south-eastern part was covered by extensive mats of Ulva rigida. Two types of field experiments were conducted by incubating Ulva thalli inside large cages. In the first experiment, beginning on 23 March, 100 g of wet thalli were placed into the cages, allowed to grow for two weeks, then collected and replaced. This procedure was repeated 8 times over the study period. In the second experiment, Ulva thalli were left inside the cages and collected at selected time intervals (14, 27, 41, 64 and 76 days) in order to simulate the effects of increased density on growth and nutrient storage.We recorded specific growth rates (NGR) ranging from 0.025 to 0.081 d^–1 for a period up to two months in the repeated short-term experiments performed at relatively low initial algal densities (300–500 g AFDW m^–3). These NGR resulted significantly related to dissolved inorganic nitrogen (DIN) in the water column. Tissue concentrations of total Kjeldahl nitrogen (TN) were almost constant, while extractable nitrate decreased in a similar manner to DIN in the water column. Total phosphorus showed considerable variation, probably linked to pulsed freshwater inflow.In the long-term incubation experiment, NGR of Ulva was inversely related to density. Internal concentrations of both total P and TN reached maximum values after one month; thereafter P concentration remained almost constant, while TN decreased below 2% w/w (by dry weight). The TN decrease was also accompanied by an abrupt decrease in nitrate tissue concentration. The biomass incubated over the two month period suffered a progressive N limitation as shown by a decreasing NY ratio (49.4 to 14.6). The reciprocal control of Ulva against biogeochemical environment and viceversa is a key factor in explaining both resource competition and successional stages in primary producer communities dominated by Ulva. However, when the biomass exceeds a critical threshold level, approximately 1 kg AFDW m^–3, the macroalgal community switches from active production to rapid decomposition, probably as a result of selfshading, biomass density and development of anaerobic conditions within the macroalgal beds.     

Mineral composition of Phragmites australis in Scottish lochs as related to eutrophication. I. Seasonal changes in organs

Monthly levels of nitrogen, phosphorus, potassium, sodium, calcium, magnesium and iron in the organs (root, rhizome, stem, leaf and panicle) of Phragmites australis were determined from March to November, 1975 in Forfar Loch (polytrophic), Balgavies Loch (eutrophic) and Loch of the Lowes (mesotrophic), Scotland. Generally no consistent seasonal changes in the mineral levels were detected for the root, rhizome and panicle. For the stem and leaf, the levels of nitrogen, phosphorus, potassium, calcium and magnesium rose to a peak before declining for the rest of the season. Consistently higher levels of tissue nitrogen, phosphorus, potassium and calcium were found in the Forfar plants over those of Balgavies and Lowes, reflecting the very eutrophic condition in Forfar Loch. The mineral compositions in the reeds in the present study were compared with similar studies in other areas.     

Large subunit rDNA sequences from Alexandrium catenella strains isolated during harmful algal blooms in Hong Kong

The partial sequences of the large subunit (24S) ribosomal RNA gene(LsurRNA) of three Alexandrium catenella strains isolatedduring the 1998 and 1989 red tide incidents in Hong Kong were compared,togetherwith the sequences of other Asian A. catenella speciescomplex. The sequences of two A. catenella strainsisolatedin 1998 were identical, despite being isolated at different time (March andApril) and different locations (eastern and southern Hong Kong). In cladisticanalysis, the two A. catenella HK98A & B strains, theA. catenella HK1989 strain, the Chinese strain clade andtheKorean/Japanese A. catenella form a clade distinct fromother A. catenella strains. Interestingly, the twoA. catenella HK98A & B strains only form a clade withtheA. catenella HK1989 strain/ChineseA. catenella (CCMP1493) with low bootstrap values (49%).     

Burrow Architecture of the Ghost Crab Ocypode ceratophthalma on a Sandy Shore in Hong Kong

The ghost crab Ocypode ceratophthalma (Pallas) creates burrows of variety shapes at different ages. Juveniles (mean carapace length 11 mm) produced shallow J-shaped burrows, which incline vertically into the substratum (mean depth 160 mm). Larger crabs (17–25 mm carapace length) have Y-shaped and spiral burrows (mean depth 361 mm). These Y-shaped burrows have a primary arm, which extends to the surface forming the opening, and a secondary arm which terminates in a blind spherical ending. The two arms join in a single shaft and end with a chamber at the base. The secondary arms and chambers are believed to be used for mating or as a refuge from predation. The spiral burrows have spiral single channel ending in a chamber. Older crabs (mean carapace length 32.6 mm) had simple, straight single tube burrows, which inclined into the substratum at mean of 73° and had a mean depth of 320 mm. During summer daytime periods, the burrows shelter the crabs from heat and desiccation stress. The sand surface temperature at the burrow opening was ~48 °C but temperatures inside the burrows can drop to 32 °C at a depth of 250 mm. Variation in the burrow architecture with crab age appears to be related to the crab’s behaviour. Juvenile crabs have smaller gill areas and move out of the burrows regularly to renew their respiratory water and, as a result, they do not need a deep burrow. Larger crabs, in contrast, can tolerate prolonged periods without renewing their respiratory water and therefore create deeper and more complex burrows for mating and refuges.     

Phenology of Sargassum spp. in Tung Ping Chau Marine Park, Hong Kong SAR, China

Eight species of the brown alga Sargassum have been recorded from Tung Ping Chau Marine Park in Hong Kong and the phenology of four of these was monitored from 1996 to 2000. All four species followed a typical growth cycle of Sargassum species reported elsewhere but with some annual variations. For S. hemiphyllum, the maximum mean (±SD) plant length, ranging from 38.5 ± 10.5 to 61.9 ± 19.9 cm, was recorded in January to March. The peak reproductive season was also mainly in February to March with up to 89% of the plants being reproductive. Some plants, however, remained reproductive until May. For. S. henslowianum, the maximum mean plant length, ranging from 45.5 ± 25.5 to 77 ± 24.8 cm, was recorded mainly in November to January. The peak reproductive season was in November to February with up to 100% of the plants being reproductive. For S. siliquastrum, the maximum mean plant length, ranging from 48.2 ± 29.9 to 63.4 ± 22.1 cm, was also recorded mainly in January. The reproductive plants were found mainly between late December and mid February with up to 98% of the plants being reproductive. For. S. patens, the maximum mean plant length ranging from 87.6 ± 62.4 to 118.7 ± 41.3 cm was recorded in January to March. Reproduction of this species was not monitored. Changes in water temperature over seasons were likely to be critical in affecting the phenological patterns of these species.     

Gracilaria and its epiphytes: 4. The response of two Gracilaria species to Ulva lactuca in a bacteria-limited environment

The responses of Gracilaria lemaneiformis, an easily epiphytized host,and the relatively resistant G. cornea mutant, to the green alga Ulva lactuca were studied using biculture experiments with and withoutantibiotics. Both Gracilaria species grown with and without U.lactuca showed different levels of growth rate, release of hydrogenperoxide and of halogenated hydrocarbons. These quantitative differencesled to a successful response against Ulva lactuca in the case of G.cornea mutant and to a failure in response in the case of G.lemaneiformis. The response of each Gracilaria species to U.lactuca was qualitatively similar to its response to bacteria. This suggeststhe involvement of oligosaccharide elicitors produced in the presence ofepiphytes and bacteria. A clear Gracilaria inhibition was demonstratedwith extracts of the culture medium. It appears that hydrogen peroxide,halogenated hydrocarbons and oligosaccharides may be components of theinhibitory activity of the extracts. The responses of Gracilaria speciesto the presence of U. lactuca suggest the characterization of adefence response.     

The Way Forward for Hong Kong to Combat Vessel Source Emissions in the Pearl River Delta Region

Air pollution has long been of great concern in both Hong Kong and the wider Pearl River Delta (PRD) region, where shipping pollution is a major contributor to poor air quality. This article addresses, first, the current global regulatory framework covering the main sources of vessel source pollution; second, the implementation and enforcement of the rules in Annex VI of the International Convention for the Prevention of Pollution from Ships (MARPOL); and third, the legal and policy measures for combating vessel source emissions in Hong Kong. In conclusion, the article offers suggestions on the way forward for Hong Kong to combat such vessel source emissions in the PRD region.     

Nitrate uptake and storage in the seaweed Ulva rigida C. Agardh in relation to nitrate availability and thallus nitrate content in a eutrophic coastal lagoon (Sacca di Goro, Po River Delta, Italy)

The seasonal cycle of biomass and tissue composition of Ulva rigida C. Agardh, in relation to nitrogen availability in the water column, was studied in 1991-1992 in the Sacca di Goro, a highly eutrophic lagoon in the Po River Delta (Italy). Nitrate uptake rates and storage capacity were also determined in laboratory experiments. The seasonal growth of U. rigida was related to the seasonal trend of nitrogen concentration in the water column. U. rigida biomass increased exponentially during spring and attained peaks of about 300-400 g dry mass (DM) m⁻² in June. As biomass increased, U. rigida depleted nitrate in the water column. Thallus nitrate reserves also declined from 100 μmol N (g DM)⁻¹ to almost undetectable levels, and total thallus nitrogen declined from 4% to 2.5% DM and 1.25% DM in 1991 and 1992, respectively. During summer, U. rigida decomposition increased, and organic nitrogen concentrations in the water column increased. The uptake experiments demonstrated an inverse relationship between thallus nitrate content and nitrate uptake rates. A modified Michaelis-Menten equation that accounts for thallus nitrate fit the uptake data well. U. rigida can accumulate up to about 400-500 μmol nitrate (g DM)⁻¹ in cellular reserves. U. rigida in the Sacca di Goro has higher K_m and lower V_max/K_m ratios for nitrate uptake than other chlorophycean species, indicating a low efficiency of uptake at low nitrate concentrations. This low uptake efficiency, and the ability to exploit N availability by storing cellular nitrate pools in excess of immediate growth needs, may represent a physiological response to an eutrophic environment where nitrate is in large supply for most of the year.     

The reproductive biology of <Emphasis Type="Italic">Nassarius festivus</Emphasis> (Powys, 1835) (Gastropoda: Nassariidae) in relation to seasonal changes in temperature and salinity in subtropical Hong Kong

The reproduction of Nassarius festivus (Gastropoda: Nassariidae) has been studied on three contrasting beaches in Hong Kong. There was no significant difference in shell heights between females and males of N. festivus at three study sites and the sex ratio was always female biased. The highest female sex ratio was recorded from Starfish Bay, followed by Tai Tam Bay and then Tai Mong Tsai. The sexual cycle of N. festivus is completed in 1 year in Hong Kong. It is a winter spawner with a reproductive period ranging from September to May. The gonads progressively mature in autumn with maturity being achieved in winter. Spring and summer are times of gonadal decline and gametogenesis initiation, respectively. The ranges in water temperature and salinity at which mature individuals occur were 13.7–28.0°C and 19.3–29.0‰, respectively. N. festivus produces between 16 and 46 eggs·capsule⁻¹. Over its short life span of 1.21 years, therefore, N. festivus attempts to maximize reproductive output. The eggs hatch as veligers which remain in the plankton for between 33 and 41 days. In terms of its reproductive biology, it is concluded that N. festivus behaves as a typical sub-tropical species with a mix of tropical and temperate characteristics seen in other nassariids from these latitudes.     

Use of the green alga Ulva rigida C. Agardh as an indicator species to reassess metal pollution in the Thermaikos Gulf, Greece, after 13 years

The concentrations of metals (Mn, Pb, Fe, Zn, Cu, Cd,Co, Ni, Cr, Na, K, Ca, Mg) were determined in thegreen alga Ulva rigida, in the sediment andseawater of Thermaikos Gulf (Greece) during monthlysamplings in 1994–1995. This Gulf is the recipientof domestic and industrial effluents. Pb, Fe, Cu, Coand Cr concentrations in U. rigida at the studyarea were higher than those 13 years earlier andapparently came from different sources than those forZn, Cd and Ni. The relative abundance of metals inthe alga decreased in the order: Mg > Na > K >Ca > Pb > Fe > Mn > Zn > Cr, Cu > Ni >Co > Cd. Only Cu concentrations in the alga fromKalochori and Cd ones from Viamyl showed significantseasonal changes. Cu and Cd concentrations ingeneral followed the same pattern of variation, withminimum values in winter-spring. This pattern isdiscussed in relation to growth dynamics and tissueage. Only Pb concentrations in the alga showed asignificant positive correlation with concentrationsin the seawater. There were both positive andnegative correlations among some metals in the alga. It is concluded that U. rigida can be used as anindicator species, especially for Pb.     

Recent changes in the contributions of river Rhine and North Sea to the eutrophication of the western Dutch Wadden Sea

From 1955 to the mid 1980s the loads of both nitrogen and phosphorus from the river Rhine to the Dutch coastal area, the Wadden Sea included, increased. Since 1985 the phosphorus loads has decreased significantly, while the nitrogen load remained about the same.Annual primary production in the western Dutch Wadden Sea has increased fromc. 40 g C m^–2 (1950) to 150 (mid 1960s) and over 500 g C m^–2 (1986). The biomass of macrozoobenthos has more than doubled since 1970. Simultaneously, the meat yield of cultured blue mussels (Mytilus edulis), has increased since the 1960s. Previously, it was indicated that the increase in primary production of the phytoplankton over the period 1950 to 1986 was stimulated by the load of dissolved inorganic phosphate from Lake IJssel, a reservoir supplied by Rhine water. Since 1990, however, primary production has been higher than was expected from decreased phosphate loads from Lake IJssel. It is argued that this lack of response may have been caused by increased concentrations of dissolved inorganic phosphate at sea originating from increased inflow from a.o. the Strait of Dover, which compensate for the decrease in phosphate from the rivers, possibly in combination with a significant improvement of the light conditions of the water in the Wadden Sea.     

Growth and nutrient allocation in Phaseolus vulgaris L. colonized with endomycorrhizae or Rhizobium

Two cultivars of Phaseolus vulgaris L., one responsive to colonization with microsymbionts (Mexico 309) and one less-responsive (Rio Tibagi) were grown in Leonard jars containing sand/vermiculite under greenhouse conditions. Bean plants were either left non-inoculated (controls) or were inoculated with the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus etunicatum or a strain of Rhizobium leguminosarum bv. phaseoli (UMR-1899). Plants from the Mexico 309 cultivar maintained a higher growth rate, supported proportionately more nodules and mycorrhizae, and assimilated relatively more N or P when colonized by Rhizobium or Glomus, respectively, than did plants of the Rio Tibagi cultivar. Estimated specific nodule activity for Mexico 309 beans was more than twice that of Rio Tibagi, whereas the specific phosphorus uptake rate (SPUR) was 35% greater in the non-inoculated roots of Rio Tibagi compared to Mexico 309. Colonization by G. etunicatum more than doubled the SPUR for each cultivar compared to control roots. New acid phosphatase isozymes appeared in VAM-colonized roots of both cultivars compared to controls. Acid and alkaline phosphatase activities were significantly higher in G. etunicatum-colonized Mexico 309 roots, but not in Rio Tibagi mycorrhizae, compared to uninfected roots. Polyphosphate hydrolase activity was elevated in mycorrhizae of both cultivars compared to control roots. These results indicate that the dependence of a host on a specific endophyte increases when there are limitations to the supply of a nutrient that the endophyte can provide. The greater the increase in absorption or utilization capacity following colonization by the microsymbiont, the greater the dependence by the host. More importantly, identification of enzymatic activities that influence these plant-microbe associations opens the possibility that the specific genes that code for these enzymes could be targeted for future manipulation.