A heated biuret-Folin protein assay which gives equal absorbance with different proteins

This protein assay requires heating the sample with biuret reagent for 100 min at 100°C before addition of Folin. Different proteins give almost identical optical densities on a nitrogen basis with this procedure. The absorbance per microgram-atom of protein nitrogin is linear at or below 0.400. A convenient final concentration of protein in this assay is about 0.2 μg-atom of protein nitrogen per ml. Most of the absorbance is due to amino acid combinations other than tyrosine and trytophan in the protein. The specificity appears similar to that of the bluret where few compounds of biological importance interfere significantly in the assay at normal cellular concentrations.     

Nutritional value of detritus and algae in blenny territories on the Great Barrier Reef

The salariin blenny, Salarias patzneri, primarily ingests detrital aggregates and inorganic sediment, with small quantities of filamentous algae. Samples of particulate matter <125 μm and filamentous algae were collected from the territories of S. patzneri and the nutritional value of these resources assessed using organic content and protein/energy ratios calculated from protein, carbohydrate and lipid content. Samples were collected throughout the day, during both summer and winter, to enable temporal comparisons of nutritional value. The particulates <125 μm, which were predominantly amorphic detritus, accounted for 41±2% and 44±3% of the organic matter in the epilithic algal matrix (EAM) during the summer and winter, respectively, whilst filamentous algae accounted for 37±3% and 41±3% of the organic matter in the summer and winter, respectively. Carbohydrate and lipid concentration in the algal samples was significantly greater than in particulate matter <125 μm, although no significant difference was detected in protein levels of algal samples and particulates <125 μm. Consequently, the protein/energy ratio of particulates <125 μm in the summer (11±1 mg kJ⁻¹) and winter (10±1 mg kJ⁻¹) was similar to that of filamentous algae in the summer (9±1 mg kJ⁻¹) and winter (8±1 mg kJ⁻¹). These results suggest detrital aggregates are at least comparable to filamentous algae as a nutritional resource. The large contribution of high quality detritus to organic matter in the epilithic algal matrix collected from S. patzneri territories throughout the day and between seasons provides strong evidence that detritus is a valuable component of small territorial fish diets and is an integral part of coral reef food webs.     

Copepod feeding behavior and the measurement of grazing rates in vivo and in vitro

In this paper some aspects of the use of the gut fluorescence method for estimating ingestion rates have been, examined. One assumption is that gut turnover time in feeding copepods is equal to the gut clearance time in filtered seawater. When arctic Pseudocalanus were pre-fed on Thalassiosira weisflogii, and then given a trace addition of the same C¹⁴-labelled culture, or were transferred to filtered seawater, results suggested that this assumption was probably justified. In another experiment in which Pseudocalanus were fed at the same concentration of either melted ice algae, or pelagic under ice algae, there were significant differences in both gut clearance times and gut pigment levels in the two cases.Pigment: biogenic silica ratios in epontic algae were higher than those in faecal pellets produced by Pseudocalanus feeding on the algae, suggesting that pigment destruction was occurring during grazing. In a 28 hr time course experiment ingestion rates determined by rate of disappearance of particulate chlorophyll were higher than those simultaneously determined by the gut fluorescence method, which also supports the idea of pigment destruction in copepods guts.     

A novel method for detection of viable zoospores of Pythium in irrigation water

A membrane filtration test has been developed for the detection of viable zoospores of Pythium species. Zoospore suspensions were filtered through 5 (m nitrocellulose membranes and the membranes incubated overnight in 0.07 m glucose, rifamycin (30 mg litre^-1) and pimaricin (100 mg litre^-1). Zoospore germlings were detected using a polyclonal antiserum, raised to mycelial surface washings of five Pythium spp., and visualised with Sigma fast red. The assay gave positive results for all Pythium spp. tested and also to zoospores of Phytophthora cryptogea. Of 10 fungal species isolated from commercial irrigation water, two were detected by the polyclonal antiserum in ELISA tests but only one produced detectable zoospore germlings. The latter isolate was later identified as a Pythium sp. Irrigation water samples collected from commercial UK nurseries yielded zoospores of both Pythium and Phytophthora spp. which, using the assay, were positively identified. Results indicated greater sensitivity than was seen with conventional plating methods. This is a test which could be adapted for on-site use in commercial nurseries.     

Critical problems with extraction of ATP for bioluminescence assay of plankton biomass

Recovery of ATP by boiling tris extraction was 90–95 percent greater in 1 liter grab samples than in concentrated net samples. ATP losses were attributed to insulating effects promoted by accumulation of detritus on filters. A series of extractions over a concentration range of whole or size-segregated plankters and cultured algae was made to determine volume of water to be filtered for optimum extraction efficiency. Accuracy of ATP assays was optimized by: (i) using large diameter (i.e. 47 mm) acetate filters; (2) limiting sample volume filtered to 50 ml when particulate organic carbon (POC) exceeded 0.4 mg l^–1; and (3) performing extractions in boiling tris maintained initially on a laboratory hot plate at 400°C as opposed to hot water bath at 100°C.Additional problems were encountered in using published cellular carbon: ATP ratios for conversion of ATP data to biomass as carbon. Ratios of POC: ATP in cultures of sheathed blue-green algae reached 550 : i, while non-sheathed forms yielded ratios near values previously reported for plankton communities. Difficulties in applying a uniform conversion factor may be expected in plankton communities containing significant volumes of sheathed blue-greens.     

THE EFFECT OF SEAWEED DIETS ON GROWTH OF GREEN ABALONE,HALIOTIS FULGENS,FROM BAJA CALIFORNIA,MEXICO

Two abalone species: green Haliotis fulgens and yellow Halioti corrugata represent nearly 97% of the total production in the Mexican abalone fishery. It has been assumed that abalone feed on the kelp algae Macrocystis pyrifera. Regional hatcheries use this species as a main source of natural food. M. pyrifera does not occur at the southern limit of the distribution of abalone species along the Baja California Peninsula. In this study, growth rates of juveniles H. fulgens, 17.3 ± 2.2 mm shell length and 0.4 ± 0.2 g body weight, were evaluated. Juveniles were fed with common species in the benthic environments inhabited by abalone along the western coast of Baja California during 191 days. Three diets were based on algae: palm kelp, Eisenia arborea, giant kelp, M. pyrifera and Gelidium robustum, and one on seagrass, Phyllospadix torreyi. Shell length and body growth rates varied between 21.5 μm day^?1 and 2.2 mg day^?1 for E. arborea and between 45.9 μm day^?1 and 6.7 mg day^?1 for M. pyrifera. Higher specific growth rates (SGR) in length and weight were determined for M. pyrifera: 0.2% and 0.7% day^?1. Significant differences between values of juveniles fed M. pyrifera with the rest of the diets were found. The highest mortality (21%) was in juveniles fed the red algae G. robustum.     

Photosynthesis of Marine Macroalgae from Antarctica: Light and Temperature Requirements

The photosynthetic performance of macroalgae isolated in Antarctica was studied in the laboratory. Species investigated were the brown algae Himantothallus grandifolius, Desmarestia anceps, Ascoseira mirabilis, the red algae Palmaria decipiens, Iridaea cordata, Gigartina skottsbergii, and the green algae Enteromorpha bulbosa, Acrosiphonia arcta, Ulothrix subflaccida and U. implexa. Unialgal cultures of the brown and red algae were maintained at 0°C, the green algae were cultivated at 10°C. I_K values were between 18 and 53 μmol m^?2 s^?1 characteristic or low light adapted algae. Only the two Ulothrix species showed higher I_K values between 70 and 74 μmol m^?2 s^?1. Photosynthesis compensated dark respiration at very low photon fluence rates between 1.6 and 10.6 μmol m^?2 s^?1. Values of α were high: between 0.4 and 1.1 μmol O₂ g^?1 FW h^?1 (μmol m^?2 s^?1)^?1 in the brown and red algae and between 2.1 and 4.9 μmol O₂ g^?1 FW h^?1 (μmol m^?2 s^?1)^?1 in the green algal species. At 0°C P_max values of the brown and red algae ranged from 6.8 to 19.1 μmol O₂ g^?1 FW h^?1 and were similarly high or higher than those of comparable Arctic-cold temperate species. Optimum temperatures for photosynthesis were 5 to 10°C in A. mirabilis, 10°C in H. grandifolius, 15°C in G. skottsbergii and 20°C or higher in D. anceps and I. cordata. P: R ratios strongly decreased in most brown and red algae with increasing temperatures due to different Q₁₀ values for photosynthesis (1.4 to 2.5) and dark respiration (2.5 to 4.1). These features indicate considerable physiological adaptation to the prevailing low light conditions and temperatures of Antarctic waters. In this respect the lower depth distribution limits and the northern distribution boundaries of these species partly depend on the physiological properties described here.     

DIURNAL FLUCTUATION OF NITRATE REDUCTASE ACTIVITY IN THE MARINE RED ALGA GRACILARIA TENUISTIPITATA (RHODOPHYTA)1

The biochemical characteristics and diurnal changes in activity of the enzyme nitrate reductase (NR; EC 1.6.6.1) from the marine red alga Gracilaria tenuistipitata var. liui Zhang et Xia are described. Different assay conditions were tested to determine the stability of NR. The crude extract of G. tenuistipitata has a NR specific activity of 10.2 U.mg⁻¹, which is higher than the NR activities found for other algae, plants, and fungi. This NR is highly active at a slightly alkaline pH and is stable over a wide range of temperature, with an optimal activity at 20° C. The apical portions of the thallus contain 64.9 ± 6.6% of the total NR specific activity. The apparent Michaelis-Menten (K_m) constant found for KNO₃ was 197 μM, and it was 95 μM for NADH. The NR from G. tenuistipitata can be included in the NADH-specific group, because no activity was found when NADPH was used as an electron donor. In extracts of algae grown under either continuously dim light or a light-dark cycle, the activity of NR exhibits a daily rhythm, peaking at the middle of the light phase, when activity is 30-fold higher than during the night phase.     

Cell Lysis of Cyanobacteria and Its Implications for Nutrient Dynamics

The dynamics of nutrients, such as phosphorus, nitrogen, and carbohydrates, during cyanobacteria cell lysis was investigated under darkness incubation in the laboratory. The cell lysis rate of cyanobacteria sampled from Lake Taihu was measured using an esterase assay. Based on particulate esterase activity, the calculated cyanobacteria lysis rate was 0.094 d^–1. During 30 days of darkness incubation, Chlorophyll a concentration decreased from 56 μg L^–1 to 2.0 μg L^–1. Parallel to this, total particulate carbohydrate concentration decreased rapidly. The fluctuation of dissolved organic carbon concentration was a function of the production of non‐carbohydrate by cyanobacteria and the decomposition of carbohydrate by bacteria. Total dissolved carbohydrates and dissolved polysaccharides concentrations showed a similar pattern, declining at the beginning of the experiment and keeping relatively stable, thereafter. In contrast, the concentration of dissolved monosaccharides remained constant during the entire process. The concentrations of NH₄⁺ and PO₄^3– increased at the early stage, and then decreased afterwards. A gradual decrease in NO₃^– concentration after day 8 indicated that anaerobic conditions might be produced during the cell lysis process. The present results demonstrated cyanobacteria cell lysis has a big influence on the nutrient status of the surrounding water. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

High tyrosine-specific protein kinase activity in normal human peripheral blood lymphocytes

Tyrosine-specific protein kinase (ATP: protein phosphotransferase, EC 2.7.1.37) activity was measured in normal human nonadherent peripheral blood lymphocytes using synthetic peptide substrates having sequence homologies with either pp60^src or c-myc. A high level of tyrosine-specific protein kinase activity was found associated with the cell particulate fraction (100 000 × g pellet). High-pressure liquid chromatography and phosphoamino acid analysis of the synthetic peptide substrates substantiated the phosphorylation of tyrosine residues by the particulate fraction enzyme. The human enzyme was also capable of phosphorylating a synthetic random polymer of 80% glutamic acid and 20% tyrosine. Enzyme activity was half-maximal with 22 μM Mg·ATP and had apparent K_m values for the synthetic peptides from 1.9 to 7.1 mM. The enzyme preferred Mg²⁺ to Mn²⁺ for optimal activity and was stimulated 2–5-fold by low levels (0.05%) of some ionic as well as non-ionic detergents including deoxycholate, Nonidet P-40 and Triton X-100. The enzyme activity was not stimulated by N⁶;O^2′-dibutyryl cyclic AMP (100 μM), N⁶;O^2′-dibutyryl cyclic GMP (100 μM), Ca²⁺ (200 μM), insulin (1 μg/ml) or homogeneous human T-cell growth factor (3 μg/ml) under the conditions used. Alkaline-resistant phosphorylation of particulate proteins in vitro revealed protein bands with M_r 59 000 and 54 000 suggesting that there are endogenous substrates for the human lymphocyte tyrosine protein kinase.     

First report of paralytic shellfish poisoning (PSP) caused by Alexandrium tamiyavanichii in Kuantan Port,Pahang, East Coast of Malaysia

Harmful algal bloom (HAB) is a proliferation of algae, which naturally produce biotoxins and cause harmful effects to humans, the environment and organisms associated with it. Paralytic shellfish poisoning (PSP) was reported for the first time in Kuantan Port, Pahang, Malaysia, in November 2013, followed by a second episode in August 2014. The toxicity level reported during the second event was as high as 3500 μg of STX equiv./100 g shellfish. Ten people were hospitalized with PSP symptoms after consuming contaminated shellfish. This study was conducted at Kuantan Port to identify the organisms responsible for these events. Water samples were collected monthly for a period of 12 months beginning in September 2014. HAB species were identified based on their morphology using light and fluorescence microscopes, and their classification was supported by molecular evidence based on internal transcribed spacer (ITS) sequences. Monthly cell abundance of Alexandrium tamiyavanichii was measured at four sampling stations. Toxin production by three strains isolated from the area was determined using HPLC. Our results revealed the presence of several HAB species, including the PSP‐producing species A . tamiyavanichii . The highest cell density of A . tamiyavanichii was 840 cells L^?1. The presence of GTX components was detected in these strains. However, other toxin components could not be determined. This study reported, for the first time, the presence of PSP‐producing A . tamiyavanichii on the Pahang coast of east Peninsular Malaysia and confirmed that the PSP events in Kuantan Port were attributable to this species. The presence of this species further indicates that several safety measures need to be considered to safeguard public health, particularly in Pahang coastal waters.     

LIGHT AND ELECTRON MICROSCOPY OF GRAZING BY POTERIOOCHROMONAS MALHAMENSIS (CHRYSOPHYCEAE) ON A RANGE OF PHYTOPLANKTON TAXA1

Grazing of fluorescent latex beads, bacteria, and various species of phytoplankton by Poterioochromonas malhamensis (Pringsheim) Peterfi (about 8.0 μm in diameter) was surveyed. The alga ingested fluorescent beads and various live or killed and nomnotile or motile organisms including bacteria, blue-green algae, green algae, diatoms, and chrysomonads. The size range of grazed prey was from 0.1 to 6.0 μm for latex beads and from 1.0 μm (bacteria) to about 21 μm (Carteria inverse) for organisms. As many as 17 latex beads (2.0 μm) or more than 10 Microcystis cells (5–6 μm) were ingested by a single P. malhamensis cell. Following such grazing, the cell increased in volume by up to about 30-fold. The range of cell volume of ingested prey was from 0.52 μm³ (bacteria) to about 3178 μm³(Carteria inversa). This study demonstrates for the first time that P. malhamensis is capable of grazing algae 2–3 times larger in diameter than its own cell and of grazing intact motile algae. Poterioochromonas malhamensis is an omnivorous grazer. Food vacuole formation and digestion processes were examined. The membrane that was derived from the plasma membrane and surrounded the prey disappeared sometime after ingestion. The food vacuole was then formed by successive fusion of numerous homogeneous vesicles accumulated around the prey. The prey was enclosed in a single membrane-bound food vacuole and then digested.     

MICROALGAL LIGHT-HARVESTING IN EXTREME LOW-LIGHT ENVIRONMENTS IN MCMURDO SOUND, ANTARCTICA1

Microalgal pigment composition, photosynthetic characteristics, single-cell absorption efficiency (Qa_(λ)) spectra, and fluorescence-excitation (FE) spectra were determined for platelet ice and benthic communities underlying fast ice in Mc Murdo Sound, Antarctica, during austral spring 1988. Measurements of spectral irradiance (E_(λ)) and photosynthetically active radiation (PAR) as well as samples for particulate absorption measurements were taken directly under the congelation ice, within the platelet layer, as profiles vertically through the water column, and at the benihic surface. Light attenuation by.sea ice, algal pigments, and particulates reduced PAR reaching the platelet ice layer to 3%(9–33 fimol photons m-²-^?s-¹) of surface values and narrowed its spectral distribution to a band between 400 and 580 nm. Attenuation by the water column further reduced PAR reaching the sea floor (28–m depth) to 0.05% of surface levels (< 1 μmol photons m-² s-¹), with a spectral distribution dominated by 470–580–nm wavelengths. The photoadaptive index (I) for platelet ice algae (5.9–12.6 μmol photons m-^2.s-¹) was similar to ambient PAR, indicating that algae had acclimated to their light environment (i.e. the algae were light-replete). Maximum Qa_(λ) at the blue absorption peak (440 nm) was 0.63, and enhanced absorption was observed from 460–500 nm and was consistent with observed high cellular chlorophyll (chi) c:chl a and fucoxanthin: chl a molar ratios (0.4 and 1.2, respectively). Benthic algae were light-limited despite the maintenance of very low I_k values (4–11 μmol photons.m-^2.s-¹). Extremely high fucoxanthin: chi a ratios (1.6) in benthic algae produced enhanced green light absorption, resulting in a high degree of complementation between algal absorption and ambient spectral irradiance. Qa_(λ) values for benthic algae were maximal (0.9) between 400 and 510 nm but remained >0.35 even at absorption minima. Strong spectral flattening, a characteristic of intense pigment packaging, was also apparent in the Qa_(λ) spectra for benthic algae. FE and Qa_(λ) spectra were similar in shape for platelet ice algae, indicating that the efficiency at which absorbed energy was transferred to photosystem II (PSII) was independent of wavelength. Fluorescence emission by benthic algae was greatest for the 500–560–nm excitation wavelengths, suggesting that most energy absorbed by accessory pigments was transferred to PSII. These results suggest that under ice algae employ complementary pigmentation and maximize absorption efficiency as adaptive strategies to low-light stress. Regulating the distribution of absorbed energy between PSI and PSII may be an adaptive response to the restricted spectral distribution of irradiance.     

Green algae in tundra soils affected by coal mine pollutions

Green algal communities were investigated in clean and pollution-impacted tundra soils around the large coal mine industrial complex of Vorkuta in the E. European Russian tundra. Samples were collected in three zones of open-cast coal mining with different degrees of pollution-impacted soil transformation. A total of 42 species of algae were found in all zones. The species richness decreased from 27 species in undisturbed zones to 19 species in polluted zones. Under open-cast coal mining impacts the community structure simplified, and the dominant algae complexes changed. Algae that are typical for clean soils disappeared from the communities. The total abundance of green algae (counted together with Xanthophyta) ranged between 100–120 × 10³ (cells/g dry soils) in undisturbed zones and 0.5–50 × 10³ in polluted zones. Soil algae appear to be better indicators of coal mine technogenic pollution than flowering plants and mosses. Presented at the International Symposium Biology and Taxonomy of Green Algae V, Smolenice, June 26–29, 2007, Slovakia.     

Daphnia population growth but not moulting is a substantial phosphorus drain for phytoplankton

SUMMARY 1. Negative effects of zooplankton on the availability of phosphorus (P) for phytoplankton as a result of the retention of nutrients in zooplankton biomass and the sedimentation of exoskeletal remains after moulting, have been recently proposed. 2. In a mesocosm study, the relative importance of these mechanisms was tested for the freshwater cladoceran Daphnia hyalina×galeata. A total of 13 mesocosm bags was suspended in a mesotrophic German lake during summer 2000 and fertilised with inorganic P in order to obtain a total nitrogen to total P ratio closer to the Redfield ratio. D. hyalina×galeata was then added at a logarithmically scaled density gradient of up to 40 ind. L^?1. Zooplankton densities, dissolved inorganic, particulate organic (seston <100 μm), as well as total nutrient concentrations were monitored. Additionally, nutrient concentrations of sediment water removed from the bottom of the mesocosm bags via a manual pump were determined. 3. Seston carbon (C), seston P and total P were significantly negatively correlated with Daphnia densities. The amount of particulate P (～5–6 μg P L^?1) sequestered from the seston compartment by Daphnia corresponded roughly to the increase of zooplankton biomass (population growth). Soluble reactive phosphorous (SRP) was at all times high (～25–35 μg P L^?1) and possibly unavailable to phytoplankton as a result of P adsorption to calcite during a calcite precipitation event (whiting). P concentrations determined in sediment water were generally <60 μg P m^?2 and thus never exceeded 1% of the total amount of P bound in particulate matter of the overlying water column. 4. Seston C : P ratios followed a polynomial second‐order function: At Daphnia densities <40 ind. L^?1 a positive linear relationship was evident, which is explained by the stronger reduction of P compared with C in seston, and transfer of seston P to zooplankton. Highest seston C : P ratios of ～300 : 1 were observed at Daphnia densities of ～30–50 ind. L^?1, which is in agreement with proposed threshold values limiting Daphnia reproductive growth. At Daphnia densities >40–50 ind. L^?1 C : P ratios were decreased because of the strong reduction of seston C at close to constantly low seston P‐values of ～3–4 μg P L^?1. 5. At least for Daphnia, it may be concluded that – unlike population growth – the sedimentation of faecal pellets and carapaces after moulting seem negligible processes in pelagic phosphorus dynamics.     

FRACTIONATION OF NUCLEAR,CHLOROPLAST, AND MITOCHONDRIAL DNA FROM POLYSIPHONIA BOLDII (RHODOPHYTA) USING A RAPID AND SIMPLE METHOD FOR THE SIMULTANEOUS ISOLATION OF RNA AND DNA1

Extraction of nucleic acids from red algae is complicated by the presence of phycocolloids. For this reason, methods used for nucleic acid isolation from other organisms are not always amenable to use with red algal preparations; modifications in some cases lead to protocols that are time consuming and complicated, often requiring large amounts of algal tissue for starting material. Here we describe the isolation of both RNA and DNA followed by fractionation and identification of nuclear, chloroplast, and mitochondrial DNAs from a single preparation of Polysiphonia boldii Wynne and Edwards using a simple method that yielded approximately 100 μg of total RNA and 20 μg of total DNA from 1 g of frozen powdered algae. The potent protein denaturant guanidinium thiocyanate and the detergent sarkosyl were used to gently lyse the cells and organelles and immediately inhibit nuclease activity in the extract. The nucleic acids were isolated by ultracentrifugation into a dense solution of CsCl; the RNA was recovered as a pellet and the DNA as a band within the CsCl solution. Agarose gel electrophoresis of the total RNA showed discrete ribosomal RNA bands, indicating little nonspecific degradation. The nuclear, chloroplast, and mitochondrial DNAs were fractionated by density gradient ultracentrifugation in the presence of the DNA binding dye, bisbenzimide H (Hoechst 33258), which binds preferentially to DNA with a high A + T:G + C ratio, thus altering its density to a greater degree than it does that of DNA with a lower nucleotide ratio. The three fractions were identified by Southern blot analysis using heterologous gene probes specific for the different genomes. The protocol should be applicable to different types of algae. The simple nucleic acid isolation step can be performed on multiple samples simultaneously without subsequent fractionation of DNA, allowing comparison of DNA from different individuals, populations, or species.     

Effects of indole‐3‐butyric acid on growth,pigments and UV‐screening compounds in Nostoc linckia

In the present research, the effect of indole‐3‐butyric acid (IBA) on the growth, and the production of some primary and secondary metabolites was studied in Nostoc linckia. In this respect, algae cultures were supplied with 0, 0.01, 0.1, 1, 10, and 100 μM IBA for 14 days. IBA at concentrations of 10 and 100 μM induced algal growth expressed as fresh weight in N. linckia. Treatment with IBA at all concentrations stimulated heterocyst formation. In addition, low concentrations of IBA (0.01, 0.1, and 1 μM) had a stimulatory effect on chlorophyll a and carotenoids accumulation. In contrast, higher concentrations of IBA induced the accumulation of phycocyanin, allophycocyanin, and phycoerythrin in the treated algae. In this case, IBA at the concentration of 10 μM was more effective. A significant decrease in protein content was observed in the algae treated by 0.01 μM IBA. All concentrations of IBA caused a decrease in sugar content, but lower concentrations were more effective. IBA application in all of the concentrations except 100 μM increased oligosaccharide‐linked mycosporine‐like amino acids (OS‐MAAs) content. Lower concentrations had a more significant effect on increasing OS‐MAAs content. However the concentrations of 10 and 100 μM IBA decreased scytonemin content. These results indicated the stimulatory impact of IBA on weight, heterocyst formation, and photosynthetic pigments in N. linckia.     

Determination of erythromycin in urine and plasma using microbore liquid chromatography with tris(2,2′-bipyridyl)ruthenium(II) electrogenerated chemiluminescence detection

Erythromycin is determined in both urine and plasma samples using microbore reversed-phase liquid chromatography with tris(2,2′-bipyridyl)ruthenium(II) [Ru(bpy)₃²⁺] electrogenerated chemiluminescence (ECL) detection. Ru(bpy)₃²⁺ is included in the mobile phase thus eliminating band broadening caused by post-column reagent addition. Extra column band broadening is an important concern in microbore liquid chromatography due to the small peak volumes. Erythromycin was studied in both water and biological samples. The detection limit for erythromycin in standards is 0.01 μM or 50 fmol injected with a S/N of 3 and a linear working range that extends four orders of magnitude. Human urine and blood plasma were also studied. Urine samples were diluted and filtered before injection. Ultrafiltration was used to remove protein from blood plasma samples prior to injection. Erythromycin was selectively detected in the body fluid samples without any further sample preparation. The detection limits obtained for erythromycin in urine and plasma are 0.05 and 0.1 μM, respectively, for 5 μl injected on a 150×1 mm I.D. C₁₈ column.     

RETENTION OF DISSOLVED COMPOUNDS BY MEMBRANE FILTERS AS AN ERROR IN THE 14C METHOD OF PRIMARY PRODUCTION MEASUREMENT1

Membrane filters retain ¹⁴C bicarbonate, ¹⁴C glycollate, and other ¹⁴C labeled substances from filtrates of algal cultures and lake water. By refiltering different volumes of the filtrate from algal cultures and from lake water after incubation with ¹⁴C bicarbonate, it was shown that the labeled material retained was not proportional to volume but showed a saturation effect with increasing volume filtered. When the radioactivity retained by a filter is divided by the volume filtered, decreasing values are obtained with increased volume filtered. This radioactivity may represent a significant addition to the radioactivity in particulate material on the filters, resulting in a similar type of curve when different volumes of lake water or cultures are filtered. Values of radioactivity per milliliter were constant using Chlorella in Chu 10 medium. However, the curve could, be obtained by increasing pH and bicarbonate concentrations in the medium and on resuspending the algae in Lake Ontario (winter) filtrate. The values of cpm/ml retained from filtrates were low in Cryptomonas cultures and the curve was not obtained unless population density was reduced, thus increasing the relative contribution to the radioactivity on the filters. The curve was not always obtained in lake water. It was significant in 10 out of 14 experiments in Lake Ontario and in 2 out of 5 experiments in Grenadier Pond. Changes in lake water rather than in experimental techniques were probably responsible. On 2 occasions when values of cpm/ml were constant in Lake Ontario, addition of sodium bicarbonate without a pH change resulted, in a significant curve. Our experiments do not disprove the possibility of cell damage during Millipore filtration, however, it has been shown that ¹⁴C labeled substances retained from solutions can account for the entire range of decreasing values as a function of volume filtered.     

Use of fluorescently labelled algae to measure the clearance rate of the rotifer Keratella cochlearis

1. Fluorescently labelled algae (FLA) were used to measure clearance rates of the rotifer Keratella cochlearis. The freshwater algae Chlorella vulgaris and Stichococcus bacillaris were labelled with a fluorescent dye, 5-(4,6-dichlorotriazin-2-yl) aminofluorescein (DTAF), following a modified staining procedure. 2. Keratella cochlearis ingested both algal species. Clearance rates on tracer foods varied between 2.4 and 6.9 μl ind^?1 h^?1, which are comparable with those determined using other techniques. 3. The main drawback of the FLA technique was that only a little more than one-third of the total amount of algal cells of both C, vulgaris and S. bacillaris were well stained with the dye (DTAF), despite the use of a higher concentration of dye and a longer staining period than recommended in the literature. 4. The FLA method can be successfully applied in grazing studies involving size selection and competition for food among zooplankton. The method complements existing techniques for measuring the clearance and ingestion rates of filter-feeders.