Effects of temperature, salinity and irradiance on the growth of the red tide dinoflagellate Gyrodinium instriatum Freudenthal et Lee

The effects of temperature, salinity and irradiance on the growth of the red tide dinoflagellate Gyrodinium instriatum Freudenthal et Lee were examined in the laboratory. Exposed to 45 different combinations of temperature (10–30 °C) and salinity (0–40) under saturating irradiance, G. instriatum exhibited its maximum growth rate of 0.7 divisions/day at a combination of 25 °C and a salinity of 30. Optimum growth rates (>0.5 divisions/day) were observed at temperatures ranging from 20 to 30 °C and at salinities from 10 to 35. The organism could not grow at ≤10 °C. In addition, G. instriatum burst at a salinity of 0 at all temperatures, but grew at a salinity of 5 at temperatures between 20 and 25 °C. It is noteworthy that G. instriatum is a euryhaline organism that can live under extremely low salinity. Factorial analysis revealed that the contributions of temperature and salinity to its growth of the organism were almost equal. The irradiance at the light compensation point (I₀) was 10.6 μmol/(m² s) and the saturated irradiance for growth (I_s) was 70 μmol/(m² s), which was lower than I_s for several other harmful dinoflagellates (90–110 μmol/(m² s)).     

The effect of environmental factors on the growth rate of Karenia brevis (Davis) G. Hansen and Moestrup

The red tide dinoflagellate Karenia brevis (Davis) G. Hansen and Moestrup is noted for causing mass mortalities of marine organisms in the Gulf of Mexico. Most research has focused on culture isolates from the eastern Gulf of Mexico. In this investigation, we examine the effects of light, temperature and salinity on the growth rate of K. brevis from the western Gulf of Mexico. Growth rates of K. brevis were determined under various combinations of irradiance (19, 31, 52, 67, and 123 μmol m⁻² s⁻¹), salinity (25, 30, 35, 40 and 45), and temperature (15, 20, 25, and 30 °C). Maximum growth rates varied from 0.17 to 0.36 div day⁻¹ with exponential growth rates increasing with increasing irradiance. Little or no growth was supported at 19 μmol photons m⁻² s⁻¹ for any experiment. Maximum growth rates at 15 °C were much lower than at other temperatures. Maximum growth rates of the Texas clone (SP3) fell within the range of Florida clones reported in the literature (0.17–0.36 div day⁻¹ versus 0.2–1.0 div day⁻¹). The Texas clone SP3 had a very similar light saturation point compared to that of a Florida isolate (Wilson's clone) (67 μmol m⁻² s⁻¹ versus 65 μmol m⁻² s⁻¹), and light compensation (20–30 μmol m⁻² s⁻¹1). The upper and lower salinity tolerance of the Texas clone was similar than that of some Florida clones (45 versus 46 and 25 versus 22.5, respectively). In our study, the Texas clone had the same temperature tolerance reported for Florida clones (15–30 °C). While individual clones can vary considerably in maximum growth rates, our results indicate only minor differences exist between the Texas and Florida strains of K. brevis in their temperature and salinity tolerance for growth. While the literature notes lower salinity occurrences of K. brevis in nearby Louisiana, our isolate from the southern Texas coast has the higher salinity requirements typical of K. brevis in the eastern Gulf of Mexico.     

Characterization of the isolated calcium-binding vesicles from the green alga Mougeotia scalaris,and their relevance to chloroplast movement

The calcium-binding vesicles from the green alga Mougeotia scalaris were isolated and characterized after staining in vivo by neutral red or rhodamine B. They were found to possess, a protonated group with a pK_a-9.9, typifying phenolic hydroxyl groups; upon titration, both, phenolic compound(s) and vital dye were concomitantly released from the vesicular matrix. A shift in peak absorbance from 450 nm to 540 nm of the vitally stained vesicles indicated that the neutral form of neutral red was bound to the vesicular, matrix as an intermediate form, stabilized via intermolecular hydrogen bonds to the phenolic compound(s). Up to 8.5.10⁹ dye molecules were calculated to be adsorbed to a mean-size vesicle. Analysis of Langmuir adsorption isotherms, indicated that there were two binding sites each for both neutral red and rhodamine B. The isolated vesicles were devoid of calcium, probably because vesicular calcium, bound to the vesicle matrix, was displaced upon dye binding. Dye adsorption to the vesicles in vivo results in substantial inhibition of the reorientational movement of the Mougeotia chloroplast and is explained by dye-mediated disorder of the cellular calcium homoeostasis.Abbreviations NR neutral red - RB rhodamine B - SDS sodium dodecyl sulfate This paper is part of the Ph.D. thesis of F. Grolig at Justus-Liebig-Universität Giessen, FRG     

Effects of the multiple stressors high temperature and reduced salinity on the photosynthesis of the hermatypic coral Galaxea fascicularis

This study investigates the physiological responses in the hermatypic coral Galaxea fascicularis exposed to salinity stress (from 37 ppt to 15 ppt) for 12 h, combined effects of reduced salinity (from 37 ppt to 20 ppt) and two temperatures (26 °C and 32 °C) for 12 h and combined effects of reduced salinity (from 37 ppt to 25 ppt) and two temperatures (26 °C and 29.5 °C) for 10 d. The results demonstrate that the coral is tolerant to 12 h exposure to extremely low salinity (15 ppt). The study also shows that combined effects of temperature and low salinity aggravate the damage on the photosynthesis of the symbiotic dinoflagellates in 12 h exposure to 20 ppt sea water. This study suggests that high temperature (29.5 °C) aggravates the damage of trivially low salinity (30 ppt) on the holobiont (the coral and its symbiotic dinoflagellates) in 10 d exposure. However, high temperature (29.5 °C) may have an antagonistic effect between temperature and low salinity (25 ppt) on metabolism of the holobiont. Based on the above results, we suggest that (1) the true mechanism of corals exposed to combined effects of low salinity and high temperature is complicated. This calls for more studies on different corals. Future studies should aim at investigating long-term low-level stress in order to simulate in situ conditions more accurately; (2) when corals exposed to extremely severe combined stressors for short-term or trivially severe stressors for relative long-term, the combined effects of two stressors (such as low salinity and high temperature) may be negative, otherwise, the effects may be additive.     

Haemocyte response associated with induction of shell regeneration in the deep-sea vent mussel Bathymodiolus azoricus (Bivalvia: Mytilidae)

This study reports on the haemocyte responses after induction of shell regeneration in the hydrothermal mussel Bathymodiolus azoricus. Haemolymph was drawn from live mussels collected at Menez Gwen hydrothermal vent site (850 m depth) at the Mid Atlantic Ridge (MAR) and was compared with those collected following laboratory acclimatisation (1 atm and Ca-rich algal diet) and also with induced specimen for up to 30 days. Simultaneously, histological changes in mantle micro-morphology with the histochemical detection of Ca mobilisation in tissues were conducted.On the basis of light- and transmission electron microscopy, it is concluded that the physiological equipment involved in shell regeneration in the deep sea bivalve closely resembles that in littoral mytilids, a group that B. azoricus is closely related. This in spite of previously alleged molecular and cellular adaptations to extreme conditions typical at deep sea hydrothermal vents. Three types of blood cells were identified sharing various morphological similarities with those in many non-vent bivalves. Significant increase in the number of circulating haemocytes was detected from day 5 after induction shell regeneration. It is suggested that the increase may be a result of migration of haemocytes from the connective tissue, probably to the shell growth frontline. It is alleged that a first peak in haemocyte number is a non-specific immune response related wound healing, which renders changes in the pallial fluid that are favourable for CaCO₃ deposition. The conspicuous presence of an unidentified, acid soluble, highly refractive structure in the haemolymph of induced mussels was detected, which may play a role in Ca nucleation.This study has set the stage for investigations underway on the influence of hydrostatic pressure on shell biomineralisation in B. azoricus subjected to post-capture hyperbaric simulations.     

Responses of neotropical mangrove seedlings grown in monoculture and mixed culture under treatments of hydroperiod and salinity

We investigated the combined effects of salinity and hydroperiod on seedlings of Rhizophora mangle and Laguncularia racemosa grown under experimental conditions of monoculture and mixed culture by using a simulated tidal system. The objective was to test hypotheses relative to species interactions to either tidal or permanent flooding at salinities of 10 or 40 g/l. Four-month-old seedlings were experimentally manipulated under these environmental conditions in two types of species interactions: (1) seedlings of the same species were grown separately in containers from September 2000 to August 2001 to evaluate intraspecific response and (2) seedlings of each species were mixed in containers to evaluate interspecific, competitive responses from August 2002 to April 2003. Overall, L. racemosa was strongly sensitive to treatment combinations while R. mangle showed little effect. Most plant responses of L. racemosa were affected by both salinity and hydroperiod, with hydroperiod inducing more effects than salinity. Compared to R. mangle, L. racemosa in all treatment combinations had higher relative growth rate, leaf area ratio, specific leaf area, stem elongation, total length of branches, net primary production, and stem height. Rhizophora mangle had higher biomass allocation to roots. Species growth differentiation was more pronounced at low salinity, with few species differences at high salinity under permanent flooding. These results suggest that under low to mild stress by hydroperiod and salinity, L. racemosa exhibits responses that favor its competitive dominance over R. mangle. This advantage, however, is strongly reduced as stress from salinity and hydroperiod increase.     

Long-term changes in the population of Eurytemora affinis (Copepoda,Calanoida) in the Gironde estuary (1978–1992)

Long-term changes in the population of Eurytemora affinis were investigated during 15 years (1978–1992) at a fixed station in the Gironde estuary (South West France). Total numbers, sex-ratio, % ovigerous females, proportion of copepodites and clutch-size were taken into account and their long-term distribution was related to temperature, river flow, salinity, suspended matter and chlorophyll a concentrations after the seasonal effect was removed for all variables.There was a great decrease of the river flow during the period of investigation, due to a general deficit in pluviosity. On the contrary, the general trend for salinity was an increase from 1978 to 1992. The same pattern was observed for chlorophyll, indicating an intrusion of marine phytoplankton. Water temperature increased significantly during the study period, following the general increase in air temperature observed in the area. For the suspended particulate matter (SPM), an increase was observed between 1978 and 1981–1982, then a very sharp decrease occurred from 1984 onwards.Numbers of E. affinis were inversely correlated with temperature and salinity, and positively correlated with the river flow. E. affinis was negatively correlated with chlorophyll concentration because of the covariation with salinity. No clear long-term trend was observed for the sex-ratio. A significant correlation was found between females carrying egg-sacs and SPM concentration, probably due to a decrease of the predation pressure in very turbid waters. The percentages of copepodites tended to decrease with time and were inversely correlated with temperature. Clutch-size significantly decreased during the 15-year period. This trend was mainly explained by temperature and salinity.It is concluded that, in the absence of strong human alteration in the Gironde estuary the long-term distribution of zooplankton can be explained by the natural environmental variability. The dominant factors are the river flow which governs the movement of the populations along the estuary and the temperature which influences the reproductive processes. Any change in climate will have consequences on the pelagic community.     

Aspects of the Cryobiology of the Intertidal Harpacticoid CopepodTigriopus brevicornis(O. F. Müller)

Cold-resistance studies of marine invertebrates have concentrated on intertidal sedentary organisms, which are often subjected to subzero air temperatures in winter. Mobile rock pool inhabitants have been rarely studied because such habitats are thought to buffer environmental variation. However, it is not uncommon for small upper-shore rock pools (2 by 1 cm) to become completely frozen. Such supralittoral habitats are subject to extreme physicochemical fluctuations especially in salinity (0 to 300‰) and temperature (−1 to +32°C) due to evaporation and dilution. The dominant invertebrate in such habitats is the harpacticoid copepodTigriopus brevicornis.Aspects of the cryobiology ofT. brevicorniswere investigated using differential scanning calorimetry (DSC). Thermograms obtained from DSC allowed determinations of freeze-onset (supercooling point, SCP), melt-onset, and melt-peak (melting point, MP) temperatures, together with estimation of the proportion of water freezing in the samples. The effects of acclimation salinity, temperature, starvation, and reproductive state on these cryobiological parameters were investigated. Acclimation to increasing salinity depressed the SCP, with the highest salinity (70‰) producing the lowest SCP, melt-onset, and MP temperatures at −27.5, −15.2, and −9.5°C respectively. The highest acclimation temperature (20°C) produced the lowest SCP (−23.4°C). Starvation significantly increased the SCP, melt-onset, and MP temperatures in comparison to fed individuals acclimated to the same salinity. The presence of eggs or ovaries in individual copepods elevated the SCP compared to nongravid females and males. LT₅₀studies showed that acclimation to high salinity improved the ability ofT. brevicornisto survive in frozen seawater. Seventy parts per thousand acclimated individuals had an LT₅₀of 64.9 h compared with just 1.4 h for 5‰ acclimated individuals in frozen seawater at −5°C. The study shows that the cold-resistance capabilities ofT. brevicorniscan be affected by several different factors, and the link between the osmoconforming nature of this species and its cold resistance is discussed.     

Adaptive Responses of the Larvae of Cirripede Barnacle Peltogasterella gracilis to Changes in Seawater Temperature and Salinity

The responses of the larvae of the cirripede barnacle Peltogasterella gracilis (Crustacea: Cirripedia: Rhizocephala) that parasitizes the hermit crab Pagurus pectinatus to different combinations of seawater temperature (25, 22, 20, 16, and 12°C) and salinity (from 34 to 8) were studied in a laboratory. The nauplii of P. gracilis completed the entire cycle of development at 22 to 12°C in a narrow range of salinity (from 34 to 28), which agrees well with the environmental conditions of the crab hosts' habitat. At favorable temperatures (22–20°C) and salinity (34–28), the nauplii reached the cypris stage in 88 ± 2 h, while at 12°C and 34–30, the naupliar development took 156 ± 5 h. The cypris larvae appeared more resistant compared with the nauplii, in terms of changes in both the temperature and salinity of seawater. They actively swam at all experimental temperatures and in the salinity range of 34–18. At temperatures (22–16°C) and salinities (34–24) favorable for the cyprids, their longevity in plankton equaled 6–10 days. Thus, the nauplii of P. gracilis is the more vulnerable stage of development in the life cycle of this parasitic barnacle. The tolerance against changes in environmental factors is due to the adaptive capabilities of parasitic larvae and the environmental conditions in the habitats of its host, a typical marine crustacean. The insignificant parasitization rate of the hermit crab by its rhizocephalan parasite may be explained by the death of the nauplii of P. gracilis, which occurs when they enter to the surface water layer.     

Infection with the protozoan parasite Bonamia ostreae modifies in vitro haemocyte activities of flat oyster Ostrea edulis

Bonamia ostreae is an intracellular protozoan parasite, infecting haemocytes of the European flat oyster Ostrea edulis. Oyster defence mechanisms mainly rely on haemocytes. In the present study in vitro interactions between parasites and flat oyster haemocytes were investigated using flow cytometry and light microscopy.Haemocyte parameters including: non specific esterase activity, reactive oxygen species (ROS) production and phagocytosis were monitored using flow cytometry after 2 h cell incubation with live and dead B. ostreae. Two ratios of parasites per haemocyte were tested (5:1 and 10:1), haemocytes alone were used as controls and the experiment was carried out three times. Flow cytometry revealed a decrease of non specific esterase activities and ROS production by haemocytes after incubation with live parasites, while there was little difference in phagocytosis activity when compared with controls. Similarly, dead parasites induced a decrease in haemocyte activities but to a lesser extent compared to live parasites. These results suggest that B. ostreae actively contributes to the modification of haemocyte activities in order to ensure its own intracellular survival.     

Photosynthetic and respiratory responses of Gracilaria vermiculophylla (Ohmi) Papenfuss collected from Kumamoto, Shizuoka and Iwate, Japan

The photosynthetic and respiratory responses to irradiance, salinity and temperature of the red alga, Gracilaria vermiculophylla, collected from Kumamoto, Shizuoka and Iwate in Japan were studied using an electronic Dissolved Oxygen sensor. The parameters derived from the photosynthesis versus irradiance relationship indicated the potential to acclimate to broad irradiance variations in all of the populations of G. vermiculophylla collected from these three sites. In addition, the light-saturated photosynthesis rate (P _max) and the dark respiration rate of all populations increased with increasing temperature up to 20–30°C, while the P _max decreased at 35°C. All populations also showed a broad variation of photosynthetic responses to salinity changes in the range from 10 to 30 psu. On the other hand, the population from Iwate showed high photosynthetic efficiency, especially in the temperature range of 5–10°C, and showed low values of saturation irradiance compared to the populations from Shizuoka and Kumamoto. These results suggest that there is greater potential to acclimate to low irradiance and low temperature in the population from Iwate compared to those from the Shizuoka and Kumamoto populations. However, the P _max of the populations from Iwate and Shizuoka was reached at 20°C and 25°C, respectively, while the Kumamoto population reached P _max at 30°C. This implies that the latter population has greater potential to tolerate higher temperatures than the former. Such characteristics in photosynthesis and respiration of G. vermiculophylla collected from the three locations probably indicate an acclimation to prevailing environmental conditions in their respective habitats.     

Distribution Patterns of Isomorphic Cold-Water Dinoflagellates (Scrippsiella/Woloszynskia Complex) Causing ‘red tides’ in the Baltic Sea

During the latest years medium-sized (15–30 μm), single-celled dinoflagellates have been reported to form blooms in the northern Baltic Proper and the Gulf of Finland in winter and spring. Recent studies (Kremp et al., 2003. Proceedings of the 7th International conference of Modern and Fossil Dinoflagellates, September 21–25, Nagasaki, Japan, 66 pp.) indicate that those blooms are caused by two isomorphic species – Scrippsiella hangoei (Schiller) Larsen, and a new species, tentatively belonging to the genus Woloszynskia. Until now there has been no report on how widely distributed these phytoplankton species are in the Baltic Sea. In this study, the occurrence of Scrippsiella/Woloszynskia complex in the entire Baltic Sea was investigated, by using monitoring data from 1997 to 2003. The species occurred in a salinity range from 2 to 8 PSU. Highest concentrations were observed at salinity 4.5–6.5 PSU. Maximum cell densities of Scrippsiella/Woloszynskia complex in the water column were mainly obtained in April or in the beginning of May by the water temperature <3 °C prior to stratification was formed. In the central Gulf of Finland, the second maximum was found in 1999 and 2002 by the temperature >6 °C. Bloom formations in the Baltic Proper and in the Gulf of Finland may not only be explained by optimum temperature and salinity, but also with other factors e.g. high nutrient concentrations and good seeding conditions from the sediments.     

Effects of environmental parameters on net photosynthesis of a free-living brown seaweed,Cystoseira barbata formarepens: determination of optimal photosynthetic culture conditions

The net photosynthesis of the Mediterranean brown seaweedCystoseira barbata f.repens is measured according to irradiance, temperature and salinity. There is not only, a good utilization of low light intensities (light-shade adaptation), but also a specific ability to use a broad range of irradiance, which corresponds in the photosynthesis-irradiance curves to a high initial slope and an extended light saturation level from 300 to 1500 mol photon m^–2 s^–1; only very high irradiances induce photoinhibition. Maximum net photosynthesis occurred at temperatures ranging from 20 °C to 30 °C. The alga tolerates not only a low level of salinity, but also a slight increase in salinity; however, at more than 47.5 g 1^–1 NaCl, oxygen exchange is significantly reduced.Light, temperature and salinity requirements are discussed, taking into account ecological considerations. Yields and quality of alginic acid are presented according to the irradiance and yearly evolutionin situ in order to aid future cultivation of this species.     

Distribution Patterns of Strongylocentrotus Sea Urchins along the Coast of Eastern Kamchatka

The distribution patterns of three sea urchin species of the genus Strongylocentrotus in relation to depth, type of substrate, surf action, and to some hydrological parameters were studied along the coast of eastern Kamchatka and in adjacent waters in 1984–1996. The sea urchin Strongylocentrotus polyacanthus dominated the open coast on rocky subintertidal sites with increased surf action and standard oceanic salinity. S. droebachiensis prevailed in shoals with lower salinity, higher water temperature and lower turbulence (enclosed and semienclosed bays). S. pallidus dominated at depths corresponding to the position of the cold intermediate layer deeper than 30–50 m with normal oceanic salinity and lower temperature.     

Cultivation and carrageenan yield and quality ofKappaphycus alvarezii in the waters of Vietnam

The carrageenan-producing red algaKappaphycus alvarezii (Doty) Doty was brought to Vietnam from Japan in 1993. Branch fragments of this species were cultivated in a pond, lagoon, inlet and offshore in Vietnam for the first time. The best daily growth rate (DGR) of plants grown in the lagoon area attained 9–11 % day^–1 in May to June (cold season). The water temperature and salinity in this area ranged from 27.2–32.4 °C and 31.4–33.7 °C, respectively. DGR of plants grown in the inlet ranged from 7 to 9% day^–1 in June. Grazing by fish has been observed to occur in this area. The DGR of plants grown in the pond ranged from 5–6% in January–July, but decreased to less than 4% day^–1 in August (hot season). K. alvarezii in Vietnam showed a carrageenan yield of 18.8–24.6% and gel strength of 1566–1712 g cm^–2. These values are similar ones obtained fromK. alvarezii cultivated in the Philippines and Indonesia.     

Seasonal Occurrences of Epiphytic Algae on the Commercially Cultivated Red Alga Kappaphycus Alvarezii (Solieriaceae, Gigartinales, Rhodophyta)

Common problems faced in farming of the red algal genus Kappaphycus/Eucheuma are “ice-ice disease” and the occurrence of epiphytes. Considerable work has been documented on “ice-ice disease” and it's mode of infection but limited information is available on the emergence of epiphytes. The present study addresses the phenomenon of epiphyte infection, its prevalence in commercially cultivated red alga, Kappaphycus alvarezii, and their variability associated with seasonality. Cultured seaweed became susceptible to epiphytes in the dry seasons (1) between March – June and (2) September – November. Findings revealed Neosiphonia savatieri (Hariot) M. S. Kim et I. K. Lee, as the dominant infecting epiphyte, representing up to 80–85% of the epiphyte present during peak seasons. Besides N. savatieri, Neosiphonia apiculata, Ceramium sp., Acanthophora sp. and Centroceras sp. were observed in smaller quantities. SEM (Scanning Electron Microscope) micrographs revealed the epiphyte's attachment to the host. Further histological study showed the extent of penetration of epiphytes into the host's cortex tissues and condition of its surrounding tissues. The outbreak of epiphytic filamentous red algae also correlated with drastic changes in seawater temperature and salinity during March– June and September – November.     

Amphora margalefii Tomàs var. lacustris P. Sánchez var. nova,a new brackish water diatom

The rotifer Brachionus plicatilis can grow in a wide range of salinities and temperatures, but rapid shifts in both salinity and temperature may result in immobilized, non-swimming rotifers. The goal of this study was to examine the effect of perturbations in temperature and salinity on the swimming pattern of the rotifer.Only slight changes in mobility were observed when rotifers were exposed to changes in temperature (from 20 °C to 8–30 °C) and to an increase in salinity (from 20% to 30%). When the salinity was reduced to 15% and 5%, the proportion of mobile rotifers was reduced to 50% and 5%, respectively. The rotifers were throughout more resistant to perturbations in temperature than to those of salinity.Combined temperature and salinity perturbations compared to perturbations in each factor separately suggested a synergetic effect of temperature and salinity on the rotifers locomotion. Transfer from cultivation conditions to low salinity (5%) and high temperature (28 °C) resulted in very low percent of mobile rotifers (0–10%). However, if the temperature was reduced to 8 °C concomitant with the changes in salinity, the percent of mobile rotifers was 85%.Rotifers use a high share of their metabolic energy for locomotion, and it is therefore not surprising that perturbations in salinity and temperature may result in partial or complete immobilization.     

Susceptibility of Larvae of Galleria mellonella to Infection by Aspergillus fumigatus is Dependent upon Stage of Conidial Germination

The ability of conidia of the human pathogenic fungus Aspergillus fumigatus to kill larvae of the insect Galleria mellonella was investigated. Conidia at different stages of the germination process displayed variations in their virulence as measured using the Galleria infection model. Non-germinating (‘resting’) conidia were avirulent except when an inoculation density of 1 × 10⁷ conidia per insect was used. Conidia that had been induced to commence the germination process by pre-culturing in growth medium for 3 h were capable of killing larvae at densities of 1 × 10⁶ and 1 × 10⁷ per insect. An inoculation density of 1 × 10⁵ conidia per insect remained avirulent. Conidia in the outgrowth phase of germination (characterised as the formation of a germ tube) were the most virulent and were capable of killing 100% of larvae after 5 or 24 h when 1 × 10⁷ or 1 × 10⁶ conidia, that had been allowed to germinate for 24 h, were used. Examination of the response of insect haemocytes to conidia at different stages of the germination process established that haemocytes could engulf non-germinating conidia and those in the early stages of the germination process but that conidia, which had reached the outgrowth stages of germination were not phagocytosed. The results presented here indicate that haemocytes of G. mellonella are capable of phagocytosing A. fumigatus conidia less than 3.0 μm in diameter but that conidia greater than this are too large to be engulfed. The virulence of A. fumigatus in G. mellonella larvae can be ascertained within 60–90 h if infection densities of 1 × 10⁶ or 1 × 10⁷ activated conidia (pre-incubated for 2–3 h) per insect are employed.     

Root-zone temperature and salinity: Interacting effects on tillering,growth and element concentration in barley

The effects of root-zone salinity (0, 30, and 60 mmol L^–1 of NaCl) and root-zone temperature (10, 15, 20, and 25°C) and their interactions on the number of tillers, total dry matter production, and the concentration of nutrients in the roots and tops of barley (Hordeum vulgare L.) were studied. Experiments were conducted in growth chambers (day/night photoperiod of 16/8 h and constant air temperature of 20°C) and under water-culture conditions. Salinity and root temperature affected all the parameters tested. Interactions between salinity and temperature were significant (p<0.05) for the number of tillers, growth of tops and roots, and the concentration of Na, K, P in the tops and the concentration of P in the roots. Maximum number of tillers and the highest dry matter were produced when the root temperature was at the intermediate levels of 15 to 20°C. Effect of salinity on most parameters tested strongly depended on the prevailing root temperature. For example, at root temperature of 10°C addition of 30 mmol L^–1 NaCl to the nutrient solution stimulated the growth of barley roots; at root temperature of 25°C, however, the same NaCl concentration inhibited the root growth. At 60 mmol L^–1, root and shoot growth were maximum when root temperature was kept at the intermediate level of 15°C; most inhibition of salinity occurred at both low (10°C) and high (25°C) root temperatures. As the root temperature was raised from 10 to 25°C, the concentration of Na generally decreased in the tops and increased in the roots. At a given Na concentration in the tops or in the roots, respective growth of tops or roots was much less inhibited if the roots were grown at 15–20°C. It is concluded that the tolerance of barley plant to NaCl salinity of the rooting media appears to be altered by the root temperature and is highest if the root temperature is kept at 15 to 20°C.     

Growth and toxin production in batch cultures of a marine dinoflagellate Alexandrium tamarense HK9301 isolated from the South China Sea

Nutritional and environmental conditions were characterized for a batch culture of the marine dinoflagellate Alexandrium tamarense HK9301 isolated from the South China Sea for its growth (cells ml⁻¹), cellular toxin content (Qt in fmol cell⁻¹) and toxin composition (mol%). Under a nutrient replete condition, Qt increased with cell growth and peaked at the late stationary phase. Toxin content increased with the nitrate concentration in the culture while it reached a maximum at 5 μM phosphate. When nitrate was replaced with ammonia, Qt decreased by 4.5-fold. Salinity and light intensity were important factors affecting Qt. The latter increased two-fold over the range of salinity from 15 to 30‰, while decreased 38% as light intensity increased from 80 to 220 μE m⁻² s⁻¹. Toxin composition varied with growth phase and culture conditions. In nutrient replete cultures, toxin composition varied greatly in the early growth phase (first 3 days) and then C1/C2, C3/C4 and GTX1 remained relatively constant while GTX4 increased from 32 to 46% and GTX5 decreased from 28 to 15%. In general, the composition of GTXs was affected in a much greater extent than C toxins by changes in nutrient conditions, salinity and light intensity. This is especially true with GTX4 and GTX5. These data indicate that the cellular toxin content and toxin composition of A. tamarense HK9301 are not constant, but that they vary with growth phase and culture conditions. Use of toxin composition to identify a toxigenic marine dinoflagellate is not always valid. The data also reveal that high salinity and low light intensity, together with high nitrate and low phosphate concentrations, would favor toxin production by this species.