Do hummingbirds have a sweet-tooth? Gustatory sugar thresholds and sugar selection in the broad-billed hummingbird Cynanthus latirostris

Nectar is a solution of mainly three sugars: sucrose, glucose and fructose. Studies have demonstrated that pollinators have preferences according to the sugar composition presented in their diet, and these preferences may be caused by sugar assimilation capacities. However, sugar flavor could also play an important role for sugar preferences of nectar-feeding animals. We evaluated the sugar gustatory thresholds of the broad-billed hummingbird Cynanthus latirostris for sucrose, glucose, fructose and a 1:1 mixture of glucose-fructose. We presented eight C. latirostris to paired feeders containing either a sugar solution or pure water. Additionally, we conducted sugar preference tests at three different concentrations (146, 730 and 1022 mmol L^− 1), to relate sugar preferences with sugar gustatory thresholds. C. latirostris had different gustatory thresholds for the three different sugars tested. At low sugar concentrations (146 mmol L^− 1), sugar selection followed the gustatory thresholds. Hummingbird sugar preference patterns can be affected by different mechanisms, both pre- and post-ingestive. At low concentrations gustatory thresholds may play an important role to determine sugar selection. However, at intermediate and high concentrations, sugar assimilation rates, and velocity of food processing generated by osmotic constraints, can be the mechanisms that explain the sugar selection of these animals.     

Variation in fruit sugar composition of Lycium barbarum L. and Lycium chinense Mill. of different regions and varieties

In this study, the contents of fructose, glucose, sucrose, Lycium barbarum polysaccharide (LBP) and total sugar were determined in the fruits of different cultivated varieties of L. barbarum and of the congeneric species Lycium chinense from different regions. The results showed that the sugar accumulated mainly in the form of hexoses (glucose & fructose). Significant correlations among metabolism-related sugars, LBP and total sugar in the fruit of Ningqi No.1 implied the important role of sugar metabolism in the syntheses of LBP and total sugar. The LBP content of Ningqi No.1 in Ningxia region was the highest among all the cultivated varieties and species. The sugar accumulation was significantly affected by ecological factors such as HCO₃⁻, Na⁺, Ca²⁺, Mg²⁺ and Cl⁻ in soil total salt, pH, soil organic matter and available nitrogen. These findings have profound implications for proper fertilization management and any attempt to grow L. barbarum in new sites.     

Swimming in the sea hare Aplysia brasiliana: Cost of transport, parapodial morphometry, and swimming behavior

The energetics and behavior of the parapodial-swimming Aplysia brasiliana were investigated in order to compare net cost of transport (COT_net) between swimming and crawling, and to compare transport costs with other swimmers. Oxygen consumption (V_O2) increased with increasing animal mass for resting, crawling, and swimming animals. Slopes of the regressions of log V_O2 on log mass were 0.90, 0.91, and 0.89 for resting, crawling, and swimming, respectively. The regression for resting V_O2 on mass was significantly lower than regressions of crawling and swimming on mass, which fell into a statistically homogenous subgroup. During 4-h swimming bouts, parapodial beat frequency dropped by less than 10% of starting values after 2 h and then stabilized for the remainder of the trial, whereas velocity steadily decreased to about 70% of starting values over the 4-h period. Initial beat frequency (at the start of a swimming bout) was negatively related to body mass, varying from 1.1 beat s^− 1 for a 34 g individual to 0.7 beats s^− 1 for a 500 g individual. Final beat frequency (at the end of a swimming bout) was also negatively related to body mass, but had a significantly lower intercept than initial beat frequency. Neither initial swimming velocity nor final swimming velocity was related to mass, but final velocity was significantly lower than initial velocity. A 250 g A. brasiliana swam at 345 m h^− 1 and crawled at 7 m h^− 1. Swimming COT_net (0.1 ml O₂ kg^− 1 m^− 1) for a 250 g A. brasiliana was 50 times less than crawling COT_net (5.3 ml O₂ kg^− 1 m^− 1). While the crawling COT_net for A. brasiliana fell within the range of other marine gastropods, swimming COT_net was less than that of swimming crustaceans, and much less than another gastropod, Melibe leonina, that uses lateral bending to swim.     

Spermatozoa concentration influences cryopreservation success in sea trout (Salmo trutta m. trutta L.)

Variation among individuals is substantial for spermatozoa concentration in fresh milt in sea trout (Salmo trutta m. trutta L.). The objective of the present study was to examine effects of spermatozoa concentration in this species on subsequent cryopreservation success. Milt with high spermatozoa concentration was diluted with seminal plasma to obtain concentrations ranging between 6 and 24 × 10⁹ mL⁻¹ with steps of 2 × 10⁹ mL⁻¹. Diluted milts were cryopreserved in 0.25-mL straws with extender (0.3 M glucose) containing 10% methanol and 10 % (vol/vol) supplement of hen egg yolk. The dilution ratio was 1:3 (milt:cryomedium). Cryopreservation efficacies were assessed according to evaluation of motility of frozen/thawed spermatozoa and quantification of fertilizing ability. Percentage of motility of frozen/thawed spermatozoa was influenced by spermatozoa concentration in the cryomedium (P < 0.05). The highest motility was observed in samples with 3.0 to 4.0 × 10⁹ spermatozoa per mL of cryomedium, which corresponds to 12 to 16 × 10⁹ spermatozoa per mL in fresh milt. Higher sperm concentrations and lower sperm concentrations in cryomedium reduced the effectiveness of cryopreservation when compared with the optimum. Cryopreservation success measured according to fertilization rate was in agreement with results for motility of frozen/thawed spermatozoa, but the optimum could not be determined with statistical precision because of differences in fertilization rate among individual donor males. However, a significant positive correlation was found between postthaw motility and fertilization rate and between cryopreserved spermatozoa velocity and fertilization rate (P < 0.05). In sea trout, cryopreservation efficiency is influenced by spermatozoa concentration in cryomedium. Individual adjustment of the dilution ratio, based on initial spermatozoa density, is recommended in the freezing protocol. Maximum cryoresistance of the cell was obtained when spermatozoa concentration in cryomedium ranged from 3.0 to 4.0 × 10⁹ mL⁻¹.     

Patch dynamics of the Mediterranean seagrass Posidonia oceanica: Implications for recolonisation process

Patch dynamics of the Mediterranean slow-growing seagrass Posidonia oceanica was studied in two shallow sites (3–10 m) of the Balearic Archipelago (Spain) through repeated censuses (1–2 year⁻¹). In the sheltered site of Es Port Bay (Cabrera Island), initial patch density (October 2001) was low: 0.05 patches m⁻², and the patch size (number of shoots) distribution was bimodal: most of the patches had less than 6 shoots or between 20 and 50 shoots. Mean patch recruitment in Es Port Bay (0.006 ± 0.002 patches m⁻² year⁻¹) exceeded mean patch loss (0.001 ± 0.001 patches m⁻² year⁻¹), yielding positive net patch recruitment (0.004 ± 0.003 patches m⁻² year⁻¹) and a slightly increased patch density 3 years later (July 2004, 0.06 patches m⁻²). In the exposed site of S’Estanyol, the initial patch density was higher (1.38 patches m⁻², August 2003), and patch size frequency decreased exponentially with size. Patch recruitment (0.26 patches m⁻² year⁻¹) and loss (0.24 patches m⁻² year⁻¹) were high, yielding a slightly increased patch density in the area 1 year later (October 2004, 1.40 patches m⁻²). Most recruited patches consisted of rooting vegetative fragments of 1–2 shoots. Seedling recruitment was observed in Summer 2004 at both sites. Episodic, seedling recruitment comprised 30% and 25% of total patch recruitment in Es Port Bay and S’Estanyol, respectively. Patch survival increased with patch size and no direct removal was observed among patches of 5 shoots or more. Most patches grew along the study, shifting patch distribution towards larger sizes. Within the size range studied (1–150 shoots), absolute shoot recruitment (shoots year⁻¹) increased linearly with patch size (R² = 0.64, p < 4 × 10⁻⁵, N = 125), while specific shoot recruitment was constant (about 0.25 ± 0.05 year⁻¹), although its variance was large for small patches. Given the slow growth rate and the high survival of patches with 5 or more shoots, even the low patch recruitment rates reported here could play a significant role in the colonisation process of P. oceanica.     

Transmembrane cholesterol migration in planar lipid membranes measured with Vibrio cholerae cytolysin as molecular tool

The rate of transbilayer movement (flip-flop) of cholesterol was estimated using planar bilayers with defined initial asymmetry, formed by the opposing monolayers technique. Vibrio cholerae cytolysin (VCC) was utilized as a molecular tool for measuring the cholesterol concentration in the cis leaflet of asymmetric bilayers. To quantify cholesterol flip-flop in planar lipid bilayers, a mathematical model was developed. It considers both the lateral diffusion rate of cholesterol within each monolayer and the flip-flop rate. The difference in initial and steady-state cholesterol contents in bilayer leaflets was used as a start point. Assuming the lateral diffusion coefficient to be of 1 × 10⁻⁸ cm² s⁻¹, the characteristic time of cholesterol flip-flop at 25 ± 2 °C was estimated as <10 s.     

Kinetic and thermodynamic properties of the folding and assembly of formate dehydrogenase

The folding mechanism and stability of dimeric formate dehydrogenase from Candida methylica was analysed by exposure to denaturing agents and to heat. Equilibrium denaturation data yielded a dissociation constant of about 10⁻¹³ M for assembly of the protein from unfolded chains and the kinetics of refolding and unfolding revealed that the overall process comprises two steps. In the first step a marginally stable folded monomeric state is formed at a rate (k₁) of about 2 × 10⁻³ s⁻¹ (by deduction k₋₁ is about10⁻⁴ s⁻¹) and assembles into the active dimeric state with a bimolecular rate constant (k₂) of about 2 × 10⁴ M⁻¹ s⁻¹. The rate of dissociation of the dimeric state in physiological conditions is extremely slow (k₋₂ ∼ 3 × 10⁻⁷ s⁻¹).     

Vertical tubular photobioreactor for semicontinuous culture of Cyanobium sp

We evaluated the kinetic culture characteristics of the microalgae Cyanobium sp. grown in vertical tubular photobioreactor in semicontinuous mode. Cultivation was carried out in vertical tubular photobioreactor for 2 L, in 57 d, at 30 °C, 3200 Lux, and 12 h light/dark photoperiod. The maximum specific growth rate was found as 0.127 d⁻¹, when the culture had blend concentration of 1.0 g L⁻¹, renewal rate of 50%, and sodium bicarbonate concentration of 1.0 g L⁻¹. The maximum values of productivity (0.071 g L⁻¹ d⁻¹) and number of cycles (10) were observed in blend concentration of 1.0 g L⁻¹, renewal rate of 30%, and bicarbonate concentration of 1.0 g L⁻¹. The results showed the potential of semicontinuous cultivation of Cyanobium sp. in closed tubular bioreactor, combining factors such as blend concentration, renewal rate, and sodium bicarbonate concentration.     

Ingestion rate of the deposit-feeder Hydrobia ulvae (Gastropoda) on epipelic diatoms: effect of cell size and algal biomass

The effects of cell size of epipelic diatoms and sediment Chl a content (as an index of algal biomass) on the ingestion rate of Hydrobia ulvae adults and juveniles were investigated in experimental microcosms. Results showed that both adults and juveniles ingested small and large diatoms without exhibiting cell size selection behaviour. The functional response of H. ulvae, juveniles (<4 mm) and adults (>4 mm), over a wide range of sediment Chl a content, was characterized by an increase of the ingestion rate according to a power law. Ingestion rate varied from 0.75 to 10 ng Chl a ind⁻¹ h⁻¹ for juveniles and from 1 to 52 ng Chl a ind⁻¹ h⁻¹ for adults, in the range ca. 10-100 μg Chl a (g dry weight sediment)⁻¹. The ingestion rate was about three times higher for adults than for juveniles. Based on these experimental results, we further proposed a mechanistic approach, using an individual based-model, to identify simple feeding mechanisms that might be involved in H. ulvae functional response.     

A chemically modified lipase preparation for catalyzing the transesterification reaction in even highly polar organic solvents

Acylation of Pseudomonas cepacia lipase with Pyromellitic dianhydride to modify 72% of total amino groups was carried out. Different organic solvents were screened for precipitation of modified lipase. It was found that 1,2-dimethoxyethane was the best precipitant which precipitated 97% protein and complete activity. PCMC (protein coated microcrystals), CLPCMC (crosslinked protein coated microcrystals), EPROS (enzyme precipitated and rinsed with organic solvents) and pH tuned preparations of modified and unmodified lipase were prepared and used for carrying out transesterification reaction with n-octane and dimethyl formamide (DMF) as reaction medium. In n-octane, among all the preparations, CLPCMC of modified lipase gave highest rate (1970 nmol min⁻¹ mg⁻¹) as compared to unmodified pH tuned lipase (128 nmol min⁻¹ mg⁻¹). In DMF, with both 1% (v/v) and 5% (v/v) water content, CLPCMC showed highest initial rate of 0.72 and 7.2 nmol min⁻¹ mg⁻¹, respectively. Unmodified pH tuned lipase showed no activity at all in DMF with both 1% and 5% (v/v) water content.     

Effects of current speed, shell length and type of sediment on the erosion and transport of juvenile softshell clams (Mya arenaria)

The erosion and transport of juvenile softshell clams (Mya arenaria) was studied in a laboratory flume in relation to free-stream velocity (0, 7, 16, 29 and 35 cm s^− 1), shell length (0-5, 5-10, 10-15, 15-20 mm) and type of sediment (mud, sandy-mud, sand and gravel). Our results showed that these factors interact together on the erosion of clams from the sediment. Juveniles were eroded in great numbers in sand while mud retained them more easily. Bedload transport was initiated at speeds of 16 cm s^− 1. Most of the clams were eroded in sandy sediments at speeds of 29 and 35 cm s^− 1. The smallest individuals were highly vulnerable to erosion compared to the other size classes studied. A results-based model using the logistic regression statistics was proposed. This allowed the estimation of erosion probabilities for a given hydrosedimentary environment. A field validation of the model was then carried out. Field results confirmed the importance of free-stream velocity, shell length and type of sediment on the erosion rate of clams. The differences observed between predicted and field results suggest that the model underestimated the erosion rate in the field. Results are discussed in the context of hydrosedimentary environments found off the eastern coast of Canada.     

The use of Fourier transform infrared spectroscopy to assay for urease from Pseudomonas aeruginosa and Canavalia ensiformis

A novel assay method was investigated for urease (EC 3.5.1.5) from Pseudomonas aeruginosa and Canavalia ensiformis by Fourier transform infrared spectroscopy. This enzyme catalyzed the hydrolysis of urea in phosphate buffer in deuterium oxide (²H₂O). The intensities of the bicarbonate bands maxima at 1625 and 1365 cm⁻¹ and of the amide I band at 1605 cm⁻¹ were measured as a function of time to study the kinetics of urea hydrolysis. The extinction coefficients ε of urea and bicarbonate were determined to be 0.72, 0.48, and 0.56 mM⁻¹ cm⁻¹ at 1625, 1605, and 1365 cm⁻¹, respectively. The initial velocity is proportional to the enzyme concentration by using the ureases from both C.ensiformis and P. aeruginosa. The kinetic constants (V_max, K_m, and K_cat) determined by Lineweaver-Burk plot were 532.2  U mg⁻¹ protein, 6.4 mM, and 806.36 s⁻¹, respectively. These data are in agreement with the results obtained by a spectrophotometric method using a linked assay based on glutamate dehydrogenase in aqueous media. Therefore, this spectroscopic method is highly suited to assay for urease activity and its kinetic parameters by using either cell-free extracts or purified enzyme preparations with an additional advantage of performing a real-time measurement of urease activity.     

An Hg-sensitive channel mediates the diffusional component of glucose transport in olive cells

In several organisms solute transport is mediated by the simultaneous operation of saturable and non-saturable (diffusion-like) uptake, but often the nature of the diffusive component remains elusive. The present work investigates the nature of the diffusive glucose transport in Olea europaea cell cultures. In this system, glucose uptake is mediated by a glucose-repressible, H⁺-dependent active saturable transport system that is superimposed on a diffusional component. The latter represents the major mode of uptake when high external glucose concentrations are provided. In glucose-sufficient cells, initial velocities of d- and l-[U-¹⁴C]glucose uptake were equal and obeyed linear concentration dependence up to 100 mM sugar. In sugar starved cells, where glucose transport is mediated by the saturable system, countertransport of the sugar pairs 3-O-methyl-d-glucose/d-[U-¹⁴C]glucose and 3-O-methyl-d-glucose/3-O-methyl-d-[U-¹⁴C]glucose was demonstrated. This countertransport was completely absent in glucose-sufficient cells, indicating that linear glucose uptake is not mediated by a typical sugar permease. The endocytic inhibitors wortmannin-A and NH₄Cl inhibited neither the linear component of d- and l-glucose uptake nor the absorption of the nonmetabolizable glucose analog 3-O-methyl-d-[U-¹⁴C]glucose, thus excluding the involvement of endocytic mediated glucose uptake. Furthermore, the formation of endocytic vesicles assessed with the marker FM1-43 proceeded at a very slow rate. Activation energies for glucose transport in glucose sufficient cells and plasma membrane vesicles were 7 and 4 kcal mol^− 1, respectively, lower than the value estimated for diffusion of glucose through the lipid bilayer of phosphatidylethanolamine liposomes (12 kcal mol^− 1). Mercury chloride inhibited both the linear component of sugar uptake in sugar sufficient cells and plasma membrane vesicles, and the incorporation of the fluorescent glucose analog 2-NBDG, suggesting protein-mediated transport. Diffusive uptake of glucose was inhibited by a drop in cytosolic pH and stimulated by the protein kinase inhibitor staurosporine. The data demonstrate that the low-affinity, high-capacity, diffusional component of glucose uptake occurs through a channel-like structure whose transport capacity may be regulated by intracellular protonation and phosphorylation/dephosphorylation.     

Investigation of the interaction of gold(III)-alkyldiamine complexes with l-histidine and imidazole ligands by H and C NMR, and UV spectrophotometry

Reactions of Au(III)-alkyldiamine complex with l-histidine and imidazole were carried out and monitored time-dependant by ¹H and ¹³C NMR. Kinetics for the [Au(en)Cl₂]⁺ reaction with l-histidine was determined by initial rate method at constant pH and 25 °C using UV-Vis absorption technique, and found to be first order with respect to each component, with a pseudo second order rate constant of 39 ± 3 M⁻¹ s⁻¹. Reaction rates of l-histidine and imidazole reactions with the [Au(en)Cl₂]⁺ complex was found to be strongly dependant on pD. The pD also has profound effect on the stability of the complex. It was observed that concurrent redox reactions also take place in solution in which Au(III) is reduced to metallic Au(0), while l-histidine and imidazole are oxidized to oxy and hydroxyl products. The optimization of the structure of [(His)Au(en)]³⁺ complex was carried out by gaussian03 at the RB3LYP level that showed a distorted square pyramid with the histidine carboxyl group at the pyramid top.     

Structural characterization and protective effect against murine sepsis of fucogalactans from Agaricus bisporus and Lactarius rufus

Fucogalactans from edible Agaricus bisporus (RFP-Ab) and wild Lactarius rufus (RFP-Lr) mushrooms were obtained on aqueous extraction followed by purification. RFP-Ab had M_w 43.8 × 10⁴ g mol⁻¹ and RFP-Lr M_w 1.4 × 10⁴ g mol⁻¹. RFP-Lr had a (1 → 6)-linked α-d-Galp main-chain partially substituted at O-2 by nonreducing end-units of α-l-Fucp (29%). While RFP-Ab had a similar main chain, it was partially substituted at O-2 by nonreducing end-units of α-l-Fucp (2.8%) and β-d-Galp (14.5%), and partially methylated at HO-3. Both RFP-Lr and RFP-Ab were tested in mice against polymicrobial sepsis. Lethality rate, myeloperoxidase (MPO) activity and cytokine levels were determined. It was observed a reduction in late mortality rate by 62.5% and 50%, respectively, prevention of neutrophil accumulation in ileum and decreasing in TNF-α and IL-1β serum levels.     

Evaluation of selenite bioremoval from liquid culture by Enterococcus species

The genus Enterococcus belong to the genera of bacteria that produce lactic acid and can confer health benefits to living organisms. Selenium (Se) is an essential micronutrient for humans and animals. Thirty-six Enterococcus species isolated from dairy products were screened for Se(IV) sorption capacity for use as a probiotics in animal nutrition. Several isolates grew luxuriantly and significantly removed Se(IV) from Se(IV) amended medium. Two isolates, LAB 14 and LAB 18, identified by 16S rRNA gene sequence analysis as Enterococcus faecalis (98% nucleotide sequence similarity) and Enterococcus faecium (97% nucleotide sequence similarity), respectively, were selected for further studies. The two isolates grew optimally and removed selenium at initial pH 7.0. Optimum removal of Se(IV) from the medium was recorded at 25 °C. Time course studies showed that after 8 h of incubation LAB 14 and LAB 18 cultures displayed the highest biomass production and Se(IV) bioremoval and most selenite in culture depleted in 24 h. At initial concentrations of 10 mg L⁻¹ and 60 mg L⁻¹, E. faecium (LAB 18) removed 9.91 mg L⁻¹ and 59.70 mg L⁻¹, respectively after 24 h. Similar Se(IV) bioremoval capacity was recorded with E. faecalis (LAB 14). Substantial amount of Se was detected in biomass of E. faecium (0.4599 mg g⁻¹ of dry weight) and E. faecalis (0.4759 mg g⁻¹ of dry weight). The significant uptake and transformation of Se(IV) by the Enterococcus species observed in this study suggest that they can be used to deliver dietary Se through feed augmentation with Se(IV)-enriched Enterococcus biomass.     

Biodegradation of phenanthrene and pyrene by Ganoderma lucidum

Biodegradation of two polycyclic aromatic hydrocarbons (PAHs), phenanthrene and pyrene, by a white rot fungus, Ganoderma lucidum, in broth cultures was investigated. It was found that the biomass of the organism decreased with the increase of PAH concentration in the cultures. In the cultures with 2 to 50 mg l⁻¹ PAHs, the degradation rate constants (k₁) increased with the PAH concentration, whereas, at the level of 100 mg l⁻¹, the degradation rate constants decreased. In the presence of 20 mg l⁻¹ PAHs, the highest degradation rates of both PAHs occurred in cultures with an initial pH of 4.0 at 30 °C. The addition of CuSO₄, citric acid, gallic acid, tartaric acid, veratryl alcohol, guaiacol, 2,2′-azino-bis-(3- ethylbenzothazoline-6-sulfonate) (ABTS) enhanced the degradation of both PAHs and laccase activities; whereas the supplement of oxalate, di-n-butyl phthalate (DBP), and nonylphenol (NP) decreased the degradation of both PAHs and inhibited laccase production. In conclusion, G. lucidum is a promising white rot fungus to degrade PAHs such as phenanthrene and pyrene in the environment.     

Disruption of Stard10 gene alters the PPARα-mediated bile acid homeostasis

STARD10, a member of the steroidogenic acute regulatory protein (StAR)-related lipid transfer (START) protein family, is highly expressed in the liver and has been shown to transfer phosphatidylcholine. Therefore it has been assumed that STARD10 may function in the secretion of phospholipids into the bile. To help elucidate the physiological role of STARD10, we produced Stard10 knockout mice (Stard10^−/−) and studied their phenotype. Neither liver content nor biliary secretion of phosphatidylcholine was altered in Stard10^−/− mice. Unexpectedly, the biliary secretion of bile acids from the liver and the level of taurine-conjugated bile acids in the bile were significantly higher in Stard10^−/− mice than wild type (WT) mice. In contrast, the levels of the secondary bile acids were lower in the liver of Stard10^−/− mice, suggesting that the enterohepatic cycling is impaired. STARD10 was also expressed in the gallbladder and small intestine where the expression level of apical sodium dependent bile acid transporter (ASBT) turned out to be markedly lower in Stard10^−/− mice than in WT mice when measured under fed condition. Consistent with the above results, the fecal excretion of bile acids was significantly increased in Stard10^−/− mice. Interestingly, PPARα-dependent genes responsible for the regulation of bile acid metabolism were down-regulated in the liver of Stard10^−/− mice. The loss of STARD10 impaired the PPARα activity and the expression of a PPARα-target gene such as Cyp8b1 in mouse hepatoma cells. These results indicate that STARD10 is involved in regulating bile acid metabolism through the modulation of PPARα-mediated mechanism.     

Effects of selenium, iron and cobalt addition to growth and yessotoxin production of the toxic marine dinoflagellate Protoceratium reticulatum in culture

The marine dinoflagellate Protoceratium reticulatum has been recently identified as a source for the disulfated polyether toxin, yessotoxin (YTX), and may pose a risk to human health, aquaculture development and coastal environments. The requirements of P. reticulatum for selenium, iron and cobalt were assessed in culture. P. reticulatum was grown in nutrient enriched seawater (1/10 GP medium) without selenium or with 0.003 and 0.0003 μM selenium added; without iron or with 0.076 and 0.0076 μM iron added; and without cobalt or with 0.008 μM cobalt added. Test flasks were monitored for growth rate, cell yield and YTX production. P. reticulatum was found to exhibit a strong requirement for both selenium and iron. Growth rate and cell yield in treatments without added selenium were significantly (P<0.05) reduced to 60.2% (μ=0.15 day⁻¹) and 20.2% (4942 cell ml⁻¹), respectively, of those with selenium added (μ=0.23 day⁻¹ and 24, 387 cell ml⁻¹). YTX production was significantly increased by addition of selenium in two of three transfers tested. Cells of P. reticulatum subjected to medium without selenium added showed morphological changes observable at the light microscope level which included enlarged cell size. The diameter of cells in medium without selenium added were significantly (P<0.05) enlarged to 36.7±0.90 μm compared to cells in the medium with selenium added, 27.5±1.25 μm. Growth rate and cell yield in treatments without added iron were also significantly reduced to 70.1% (μ=0.16 day⁻¹) and 34.2% (8003 cells ml⁻¹), respectively, of those with iron added (μ=0.23 day⁻¹ and 23,416 cells ml⁻¹). No significant effect on YTX production was measured. In contrast to selenium and iron, no limitation of growth or cell yield or differences in YTX production were observed for flasks without cobalt as compared to those with cobalt added. The possibility that harmful algal events of P. reticulatum may be influenced by selenium or iron in neritic waters is discussed.     

The effects of algae species and densities on the population growth of the marine rotifer, Colurella dicentra

Colurella dicentra clones isolated from bay water in the Mississippi Gulf Coast were cultured with artificial seawater. Experiments were conducted to determine the effects of six algae species (Nannochloropsis oculata, Tetraselmis chuii, Chaetoceros gracilis, Rhodomonas salina, Isochrysis galbana, and Prorocentrum micans), six C. gracilis densities, and six N. oculata densities (25,000, 50,000, 100,000, 250,000, 500,000, and 1,000,000 cells ml^− 1) on C. dicentra population growth. Algae type influenced rotifer production (p < 0.0001). C. gracilis treatment (9120 ± 3351SD) produced the highest number of rotifers followed by N. oculata (5760 ±2232SD). P. micans had the lowest number of rotifers, although not significantly different from numbers in T. chuii, R. salina, and I. galbana treatments (p > 0.05).The population growth rate (r) varied with algae species treatment. The highest values were recorded for C. gracilis treatment (0.22 to 0.26 d^− 1), followed by N. oculata (0.21 to 0.24 d^− 1), and the lowest for P. micans (− 0.19 to 0.14 d^− 1). C. gracilis and N. oculata densities had significant effects (p < 0.0001) on C. dicentra population growth. The highest rotifer production was recorded at a C. gracilis density of 100,000 cells ml^− 1, followed by 250,000 cells ml^− 1 and 50,000 cells ml^− 1. Algae densities of 500,000 cells ml^− 1 and above produced the lowest rotifer numbers. Population growth rate (r) varied with C. gracilis densities. The highest values were observed for C. gracilis concentrations of 100,000 cells ml^− 1 (0.17 to 0.19 d^− 1), and the lowest for concentrations of 500,000 cells ml^− 1 and above (− 0.19 to 0.09 d^− 1). The 100,000 cells ml^− 1N. oculata density gave the highest rotifer production followed by 50,000, 250,000, 25,000, and 500,000 cells ml^− 1. Algae densities of 1,000,000 cells ml^− 1 produced the lowest rotifer numbers. Population growth rate (r) varied with N. oculata densities, with the highest values obtained for algae densities of 100,000 cells ml^− 1 (0.35 to 0.40 d^− 1), and the lowest for concentrations of 1,000,000 cells ml^− 1 (0.05 to 0.012 d^− 1). This is the first report of C. dicentra in Mississippi Coastal waters, and perhaps the smallest marine rotifer species (93 by 49 μm) ever cultured successfully.