Assessment of soil-quality index based on microarthropods in corn cultivation in Northern Italy

The aim of this paper is to assess of the stability and repeatability of the biological soil-quality (BSQ) index over time and at short/long distances in agricultural soils of the Po Valley, Northern Italy, to support its correct use in soil-quality monitoring and is based on soil microarthropods representative of the structure, function and composition of ecological systems, but avoiding classification at the species level. Microarthropods are separated from the soil and weighted using the biological form approach to evaluate their level of adaptation to the soil environment. The field test was organized to assess time and scale effects in a landscape within the coordinates 45°00′ to 45°16′N and 9°50′ to 10°15′E for an estimated area of 200 km². The BSQ had, in different soils cropped with corn and fertilized with sewage sludge, an average value of 76.4 ± 23.5; it appears to be reproducible both over short distances as well as over short periods of time, but no significant differences (p > 0.05) between years nor between farms were observed. In the landscape conditions tested, it appears reproducible and stable at a field and basin scale, though very sensitive to seasonal climatic variations. From a practical point of view, the information reported in this paper contributes to the standardization of the application of BSQ and provides reference values for this index in soils of the Northern Italy cropped with corn.     

Formation of di-d-fructofuranose-1,2′:2,1′-dianhydride by three novel inulin fructotransferases from the Nocardiaceae family

In this work, three novel genes encoding di-d-fructofuranose-1,2′:2,1′-dianhydride (DFA I)-forming inulin fructotransferases (IFTases) from Nocardiaceae family, including Nocardioides luteus, Nocardioides sp. JS614, and Nocardioidaceae bacterium Broad-1, were cloned and expressed in Escherichia coli. The recombinant IFTases from N. luteus (NoluIFTase), Nocardioides sp. JS614 (NospIFTase), and N. bacterium Broad-1 (NobaIFTase) were purified, identified, and characterized. SDS-PAGE analysis showed that they had molecular weights of approximately 41–42 kDa, while gel filtration analysis indicated that their native molecular weights ranged from 50 to 62 kDa, suggesting that the three enzymes may be monomers. Their optimum pH values ranged from 5.5 to 6.0, similar to other DFA I-forming IFTases or di-d-fructofuranose-1,2′:2,3′-dianhydride (DFA III)-forming IFTases. NoluIFTase, NospIFTase, and NobaIFTase exhibited maximal activities at 55 °C, 50 °C, and 45 °C and were stable at 70 °C (for 15 min), 70 °C (187 min), and 55 °C (239 min), respectively. Furthermore, by comparing with our previously reported DFA I-forming IFTase, namely CcIFTase, a probable mechanism for the formation of DFA I by the three new enzymes was speculated, and CcIFTase will be selected for future structural resolution to illustrate the catalytic mechanism of DFA I-forming IFTases toward inulin.     

Variability of sea-surface temperature and sea-ice cover in the Fram Strait over the last two millennia

A sediment core located on the West Spitzbergen margin in the Fram Strait (78°54.931′N, 6°46.005′E, water depth: 1497 m) was analyzed for its dinocyst content in order to reconstruct hydroclimatic variations of the last 2500 years. The relative abundance of dinocyst taxa and principal component analysis show a major transition at about 300 cal. years BP. It is characterized by the disappearance of thermophilic taxa Spiniferites mirabilis-hyperacanthus and Impagidinium sphaericum and the increase of polar–subpolar taxa Impagidinium pallidum and Pentapharsodinium dalei. Sea-surface temperature (SST) estimates suggest warmer conditions than present (anomaly～+2 °C) averaging at 7 °C in summer until 300 cal. years BP, although cooling pulses are recorded around 1700, 1500, 1200 and 800 cal. years BP. The last 300 years were marked by a cooling from 7.6 to 3.5 °C and sea-ice cover increasing up to 7 months/yr. The results demonstrate that the Fram Strait area is sensitive to hydroclimatic variations, notably with respect to sea-ice and SSTs, which are linked to the relative strength of northward flow of North Atlantic waters to the East and southward outflow of cold and fresh waters from the Arctic Ocean. Based on our data, the warmest part of our record around 1320 cal. years BP is the only interval of the last 2500 years that provides a possible analogue for the modern post-AD 2000 interval, which is characterized by sea-ice free conditions.     

Heat coma temperature,relative contents of saturated/unsaturated fatty acids and reproductive maturation in the oceanic sea skaters Halobates micans

This study was performed to clarify how the relative volume of saturated/unsaturated lipid and reproductive maturation relate to resistance to high temperature in the oceanic sea skaters, Halobates micans. Heat coma temperature (HCT) was measured in H. micans adults collected from a fixed sampling location (12°00′N, 135°00′E) in the western tropical Pacific Ocean. After measuring HCT, the specimen were dissected to measure the testes size and to determine the presence and number of oocytes in females. Bodies of the specimen were assessed by lipid analysis to evaluate saturated and unsaturated lipid content. A negative trend was seen between heat coma temperature and percentage of a saturated fatty acid, myristic acid (ratio of carbon number to number of double bonds = 14:0) (Pearson's correlation test: r = − 0.520, p = 0.101). In contrast, a positive trend was detected between heat coma temperature and percentage of an unsaturated fatty acid, palmitoleic acid (16:1) (r = 478, p = 0.137). Young males with small testes showed lower heat coma temperatures, whereas females that showed relatively high heat coma temperatures of 36–40 °C tended to have fewer mature oocytes in their ovaries than those that showed low heat coma temperatures of 30–34 °C. As Halobates appears to exhibit embryonic diapause rather than adult diapause, males of H. micans may develop both testes and resistance to high temperature in the parallel as they grow. In females, a trade-off may occur between heat tolerance function and oogenesis in the oceanic sea skaters.     

An example of the consequences of human activities on the evolution of subalpine landscapes

A pollen study at Survilly (2235m asl, 06° 49′ 12″ E, 45° 59′ 24″ N), a small peatbog located on the Anterne mountain (Upper-Arve Valley, French north-western Alps) highlights the local role of human activities in Holocene vegetation dynamics of the currently treeless subalpine belt and the consecutive resumption of erosion. As early as 8890 cal. years BP (± 122), Pinus cembra grew close to the site. Grasslands without shrubs were established at around 4624 ± 86 cal. years BP. Due to human activities, spruces extended little after 3600 cal. BP. The intense grazing that resulted in the current alpine meadows goes back to 1436 cal. years BP (± 81). After 4624 cal. BP three clay layers show that from this period, the erosion became as active as during the first steps of the colonization of the vegetation prior to 10,050 cal. BP. During peat growth only a millimetre of clay at the end of the 9400–9050 cal. BP climatic event was recorded.     

Characterization of DNA substrate specificities of apurinic/apyrimidinic endonucleases from Mycobacterium tuberculosis

Apurinic/apyrimidinic (AP) endonucleases are key enzymes involved in the repair of abasic sites and DNA strand breaks. Pathogenic bacteria Mycobacterium tuberculosis contains two AP endonucleases: MtbXthA and MtbNfo members of the exonuclease III and endonuclease IV families, which are exemplified by Escherichia coli Xth and Nfo, respectively. It has been shown that both MtbXthA and MtbNfo contain AP endonuclease and 3′ → 5′ exonuclease activities. However, it remains unclear whether these enzymes hold 3′-repair phosphodiesterase and nucleotide incision repair (NIR) activities. Here, we report that both mycobacterial enzymes have 3′-repair phosphodiesterase and 3′-phosphatase, and MtbNfo contains in addition a very weak NIR activity. Interestingly, depending on pH, both enzymes require different concentrations of divalent cations: 0.5 mM MnCl₂ at pH 7.6 and 10 mM at pH 6.5. MtbXthA requires a low ionic strength and 37°C, while MtbNfo requires high ionic strength (200 mM KCl) and has a temperature optimum at 60 °C. Point mutation analysis showed that D180 and N182 in MtbXthA and H206 and E129 in MtbNfo are critical for enzymes activities. The steady-state kinetic parameters indicate that MtbXthA removes 3′-blocking sugar-phosphate and 3′-phosphate moieties at DNA strand breaks with an extremely high efficiency (k_cat/K_M = 440 and 1280  μM^-1∙min⁻¹, respectively), while MtbNfo exhibits much lower 3′-repair activities (k_cat/K_M = 0.26 and 0.65 μM^-1∙min⁻¹, respectively). Surprisingly, both MtbXthA and MtbNfo exhibited very weak AP site cleavage activities, with kinetic parameters 100- and 300-fold lower, respectively, as compared with the results reported previously. Expression of MtbXthA and MtbNfo reduced the sensitivity of AP endonuclease-deficient E. coli xth nfo strain to methylmethanesulfonate and H₂O₂ to various degrees. Taken together, these data establish the DNA substrate specificity of M. tuberculosis AP endonucleases and suggest their possible role in the repair of oxidative DNA damage generated by endogenous and host- imposed factors.     

Holocene environmental changes as recorded by mineral magnetism of sediments from Anguli-nuur Lake,southeastern Inner Mongolia Plateau,China

Two cores, one 1141-cm long (An-S) and the other 885-cm long (An-A), were retrieved from Anguli-nuur Lake (41°18′–24′N, 114°20′–27′E, ～ 1315 masl), one of the largest lakes in the transition zone between a semi-humid and semi-arid climate parallel to the present limit of the southeast monsoon along the southeastern Inner Mongolia Plateau in north China. Mineral-magnetic parameters (χ_lf, ARM, IRM_300mT, SIRM and IRM_? 300mT) were measured on An-S and two additional parameters (χ_ARM and HIRM) and four inter-parametric ratios (χ_ARM/SIRM, IRM_300mT/SIRM, IRM_? 300mT/SIRM and SIRM/χ_lf) were calculated. Potential sources of these lake sediments (catchment soils and dune materials close to the lake and in a distant sand plain) were sampled, and the magnetic properties of the surface-material specimens were measured. A chronological model was developed for An-S by comparing and combining AMS¹⁴C dates of An-S with ¹³⁷Cs, ²¹⁰Pb and AMS¹⁴C dates of An-A. With the help of surface-material magnetism, the magnetic data of An-S in combination with particle size, TOC and C/N and pollen analyses indicate the environmental changes during the last ～ 10,000 years around this lake. Conditions began to ameliorate at 10,900 cal. yr BP (9600 ¹⁴C yr BP) and thus relatively wet and warm environments prevailed during 10,900–8900 cal. yr BP (9600–8000 ¹⁴C yr BP). The Holocene optimum or the wettest and warmest conditions, was during 8900–7400 cal. yr BP (8000–6500 ¹⁴C yr BP). The environment began to deteriorate from 7400 cal. yr BP (6500 ¹⁴C yr BP) and the driest and coolest conditions occurred during 2200–480 cal. yr BP. There may have been a minor amelioration after 480 cal. yr BP. The inferred changes in palaeoenvironmental conditions around Anguli-nuur Lake are broadly in agreement with those around most other sites on the Inner Mongolia Plateau.     

Seasonal shifts in seagrass bed primary producers in a cold-temperate estuary: Dynamics of eelgrass Zostera marina and associated epiphytic algae

The seasonal dynamics of seagrass and epiphytic algal primary production were measured in an eelgrass (Zostera marina) bed in the Akkeshi-ko estuary, Hokkaido, Japan (43°02′N, 144°52′E). During spring and early summer, eelgrass biomass increased, with a high production (maximum: 2.89 g C m⁻² day⁻¹), but the production and biomass of epiphytic algae remained low. In contrast, epiphytic algae bloomed in August, with a high production (5.21 g C m⁻² day⁻¹), but eelgrass production ceased and its biomass subsequently decreased. Therefore, the major primary producers in this eelgrass bed switched seasonally from eelgrass in spring and early summer to epiphytic algae in late summer and autumn. Epiphytic algae maintained similar productivity because of the change of photosynthetic kinetics and the dominant epiphytic diatom changed from highly adhesive species to less adhesive or filamentous small species during the bloom. This suggests that the change of epiphyte density and biomass was due to change of its loss rate, possibly due to herbivorous grazing rate. Moreover, competition between epiphytic algae and eelgrass for nutrients and light may also affect the dramatic seasonal changes in the major primary producers.     

Effects of different temperatures on the life history of Evania appendigaster L. (Hymenoptera: Evaniidae), a solitary oothecal parasitoid of Periplaneta americana L. (Dictyoptera: Blattidae)

The influence of temperatures on the life parameters of the solitary oothecal parasitoid Evania appendigaster, was investigated in the laboratory. Parasitized oothecae of Periplaneta americana were left to develop under seven constant temperatures: 15, 17, 20, 25, 30, 35, and 40 °C. At the end, we found that: (i) E. appendigaster was able to complete development within the temperature range of 17–34 °C; (ii) mean adult longevity decreased as temperature increased, with the temperature of 40 °C being fatal in a matter of hours; (iii) males lived longer than females between 15 and 30 °C; (iv) adult emergence rate was the highest at 25 °C, and (v) no wasps emerged at 15 or 40 °C. Non-emerged oothecae contained either unhatched eggs or dead larvae. We determined the theoretical lower developmental threshold and thermal constant for the complete development as 12.9 °C and 584.8 day-degrees for males, and 13.1 °C and 588.2 day-degrees for females, respectively. A good balance between faster development, maximum adult longevity and good egg viability was obtained between 25–30 °C, and that would be the best temperature range for rearing E. appendigaster.     

Biochemical characteristics of phytases from fungi and the transformed microorganism

Five sources of phytases were used to study their biochemical characteristics. Phytase E was from an original Escherichia coli (E. coli), phytase PI and PG from the transformed Pichia pastoris (P. pastoris) with phytase gene of E. coli, phytase B and R from Aspergillus niger (A. niger). The results showed that the relative phytase activities had no significant changes when temperature was below 60 °C (P>0.05), and then decreased significantly with temperature increasing (P<0.01). The fungal phytase with the phytase gene from A. niger had the higher thermostability than the bacterial phytase with the phytase gene from E. coli; i.e. at 70 °C, 27–58% of phytase activity (compared with 30 °C) was retained for the bacterial phytase, and 73–96% for the fungal phytase; at 90 °C, 20–47% was retained for the bacterial phytase, and 41–52% for the fungal phytase, especially for the most thermostable phytase R (P<0.01). The optimum pH ranges were 3.0–4.5 for the bacterial phytases and 5.0–5.5 for the fungal phytases (P<0.01). When pH levels were 1, 7 and 8, only 3–7% of phytase activity (compared with the maximum phytase activity at a pH point) was retained for both bacterial and fungal phytases. The amount of inorganic P released from soybean meal was significantly increased when the levels of phytase activity in the soybean meal increased from 0 to 1.0 U/g soybean meal (P<0.01), except for phytase PI. The maximum P released was obtained at 1 U/g soybean meal for all five kinds of phytases (P<0.01). The most economical phytase concentration for P released was 0.25 U/g for phytase PI and B, and 0.50–1.0 U/g for phytase PG, E and R. In addition, the linear and non-linear regression models were established to estimate phytase activity and its characteristics very easily and economically.     

Physiological responses of Ostreopsis ovata to changes in N and P availability and temperature increase

Ostreopsis ovata is a benthic dinoflagellate that produces palytoxin and ovatoxins. Blooms of O. ovata causing human health problems and mortality of benthic fauna have been reported from many tropical and temperate marine waters. In the present study we examined the combined effects of temperature and different nutrient conditions on the biochemical composition, growth, toxicity and carbohydrate production of an O. ovata strain originating from the Tyrrhenian Sea. O. ovata cultures with N:P ratios of 1.6, 16 and 160 (N deficient, NP sufficient and P deficient, respectively) were grown at 20 °C and 30 °C. Biomass accumulation, growth rates, cell volumes, biochemical composition, cell toxicity and carbohydrate production in each treatment were studied. Results indicated that under nutrient sufficiency O. ovata biomass accumulation increased significantly compared to N and P deficiency and also that N limitation severely affected growth. The highest growth rates were recorded at 30 °C. Cellular contents and the atomic ratios of C, N and P were higher in the cells grown at 20 °C than in those grown at 30 °C. O. ovata cell volumes increased at 20 °C. N deficiency significantly increased cell toxicity. Toxicity per cell was higher at 20 °C, but per carbon was highest at 30 °C. The highest carbohydrate production was found in conditions of N deficiency and at the lower temperature.Our study suggests that temperature increases due to global warming and nutrient enrichment of coastal waters stimulate the proliferation of O. ovata, particularly for the strains that have become adapted to warm temperate waters.     

Purification and characterization of an extracellular laccase from a Pseudomonas sp. LBC1 and its application for the removal of bisphenol A

The Pseudomonas sp. LBC1 produced extracellular laccase when grown in the nutrient broth. The enzyme was purified using acetone precipitation and an anion-exchange chromatography. The molecular weight of the purified laccase was estimated as 70 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis. An enzyme showed maximum substrate specificity towards o-tolidine than other substrates of laccase including 2,2′-azinobis, 3-ethylbenzothiazoline-6-sulfonic acid, hydroquinone, N,N′-dimethyl phenylene diamine, syringic acid and veratryl alcohol. The optimum pH and temperature for the laccase activity were 4.0 and 40 °C, respectively. Cyclic voltammogram revealed the redox potential of purified enzyme as 0.30 V. The laccase was stable up to 40 °C and within pH range 6.0–8.0. Sodium azide and EDTA strongly inhibited laccase activity. The purified laccase completely degraded the higher concentration of bisphenol A within 5 h. Biodegradation metabolites of bisphenol A were characterized by using FTIR, HPLC and GC–MS.     

Nitrate and phosphate uptake kinetics of the harmful diatom Eucampia zodiacus Ehrenberg,a causative organism in the bleaching of aquacultured Porphyra thalli

The diatom Eucampia zodiacus Ehrenberg is a harmful diatom which indirectly causes bleaching of aquacultured Nori (Porphyra thalli) through competitive utilization of nutrients during bloom events. In the present study, we experimentally investigated the nitrate (N) and phosphate (P) uptake kinetics of E. zodiacus, Harima-Nada strain. Maximum uptake rates (ρ_max), which were obtained by short-term experiments, were 0.777 and 0.916 pmol cell^?1 h^?1 for nitrate and 0.244 and 0.550 pmol cell^?1 h^?1 for phosphate at 9 and 20 °C, respectively. The half-saturation constants for uptake (K_s) were 2.59 and 2.92 μM N and 1.83 and 4.85 μM P at 9 and 20 °C, respectively. Although the maximum specific uptake rate (V_max; V_max = ρ_max/Q₀, Q₀; minimum cell quota) and V_max/K_s for nitrate at 9 °C are about 1/2 of those obtained at the optimum temperature (20 °C), they are still higher than those obtained for many other phytoplankton at their optimum temperature conditions for uptake. These results suggest that E. zodiacus utilizes nitrogen efficiently at low water temperature, and it is one of the important factors causing the serious damage to Porphyra thalli by bleaching due of this species. For phosphate, the K_s values of E. zodiacus were higher than those reported for other species; the V_max and V_max/K_s values were much lower than those of other diatoms such as Skeletonema costatum (Greville) Cleve. These results suggest that E. zodiacus is disadvantaged compared to other diatom species during competitive utilization of phosphate.     

Microbial surface displaying formate dehydrogenase and its application in optical detection of formate

In the present work, NAD⁺-dependent formate dehydrogenase (FDH), encoded by fdh gene from Candida boidinii was successfully displayed on Escherichia coli cell surface using ice nucleation protein (INP) from Pseudomonas borealis DL7 as an anchoring protein. Localization of matlose binding protein (MBP)-INP-FDH fusion protein on the E. coli cell surface was characterized by SDS-PAGE and enzymatic activity assay. FDH activity was monitored through the oxidation of formate catalyzed by cell-surface-displayed FDH with its cofactor NAD⁺, and the production of NADH can be detected spectrometrically at 340 nm. After induction for 24 h in Luria-Bertani medium containing isopropyl-β-d-thiogalactopyranoside, over 80% of MBP-INP-FDH fusion protein present on the surface of E. coli cells. The cell-surface-displayed FDH showed optimal temperature of 50 °C and optimal pH of 9.0. Additionally, the cell-surface-displayed FDH retained its original enzymatic activity after incubation at 4 °C for one month with the half-life of 17 days at 40 °C and 38 h at 50 °C. The FDH activity could be inhibited to different extents by some transition metal ions and anions. Moreover, the E. coli cells expressing FDH showed different tolerance to solvents. The recombinant whole cell exhibited high formate specificity. Finally, the E. coli cell expressing FDH was used to assay formate with a wide linear range of 5–700 μM and a low limit of detection of 2 μM. It is anticipated that the genetically engineered cells may have a broad application in biosensors, biofuels and cofactor regeneration system.     

Mathematical modeling of Microcystis aeruginosa growth and [D-Leu1] microcystin-LR production in culture media at different temperatures

The effect of temperature (26 °C, 28 °C, 30 °C and 35 °C) on the growth of native CAAT-3-2005 Microcystis aeruginosa and the production of Chlorophyll-a (Chl-a) and Microcystin-LR (MC-LR) were examined through laboratory studies. Kinetic parameters such as specific growth rate (μ), lag phase duration (LPD) and maximum population density (MPD) were determined by fitting the modified Gompertz equation to the M. aeruginosa strain cell count (cells mL⁻¹). A 4.8-fold increase in μ values and a 10.8-fold decrease in the LPD values were found for M. aeruginosa growth when the temperature changed from 15 °C to 35 °C. The activation energy of the specific growth rate (Eμ) and of the adaptation rate (E₁/LPD) were significantly correlated (R² = 0.86). The cardinal temperatures estimated by the modified Ratkowsky model were minimum temperature = 8.58 ± 2.34 °C, maximum temperature = 45.04 ± 1.35 °C and optimum temperature = 33.39 ± 0.55 °C.Maximum MC-LR production decreased 9.5-fold when the temperature was increased from 26 °C to 35 °C. The maximum production values were obtained at 26° C and the maximum depletion rate of intracellular MC-LR was observed at 30–35 °C. The MC-LR cell quota was higher at 26 and 28 °C (83 and 80 fg cell⁻¹, respectively) and the MC-LR Chl-a quota was similar at all the different temperatures (0.5–1.5 fg ng⁻¹).The Gompertz equation and dynamic model were found to be the most appropriate approaches to calculate M. aeruginosa growth and production of MC-LR, respectively. Given that toxin production decreased with increasing temperatures but growth increased, this study demonstrates that growth and toxin production processes are uncoupled in M. aeruginosa. These data and models may be useful to predict M. aeruginosa bloom formation in the environment.     

Novel l-adenosine analogs as cardioprotective agents

Two l-nucleosides, l-3′-amino-3′-deoxy-N⁶-dimethyladenosine (l-3′-ADMdA) 1, previously synthesized in our laboratory, and the novel l-3′-amino-3′-deoxy-N⁶-methyladenosine-5′-N-methyluronamide (l-3′-AM-MECA) 2 were evaluated in an ischemia/reperfusion model on Langendorff perfused mouse heart. l-3′-ADMdA 1 was found to enhance functional recovery from ischemia (32.2 ± 3.7 cm H₂O/s % rate pressure product, compared to 21.3 ± 1.4 for the control and 30.7 ± 3.4 for adenosine) and increase the time to onset of ischemic contracture (14.5 ± 0.9 min, compared to 10.5 ± 1.0 min for the control and 13.6 ± 0.6 min for adenosine) comparable to adenosine. Consistent with the functional recovery data, decreased infarction area was seen in the case of 1 (19.1 ± 8.4, compared to 40.5 ± 7.2% for the control and 11.5 ± 2.1% for adenosine). In contrast, l-3′-AM-MECA 2 did not show significant functional recovery, increased onset of contracture, nor decreased infarction area compared to control. Unlike adenosine, neither 1 nor 2 induced cardiac standstill in mouse heart.     

Cloning,expression, and characterization of a thermostable and pH-stable laccase from Klebsiella pneumoniae and its application to dye decolorization

A thermostable and pH-stable laccase from Klebsiella pneumoniae was cloned and expressed in Escherichia coli. The recombinant laccase (rLac) achieved a specific activity of 7.12 U/mg after purification by Ni-affinity chromatography. Optimal enzyme activity was observed at pH 4.0 and 35 °C for 2,2′-azino-bis (3-ethylbenzthiazoline sulfonic acid) (ABTS) oxidization and pH 8.0 and 70 °C for 2,6-dimethoxyphenol (2,6-DMP) oxidization. Thermostability and pH stability studies showed that the rLac was stable over the range of 30–70 °C and pH 5.0–9.0 using 2,6-DMP as substrate. Circular dichroism analysis suggested that the secondary structure of the rLac mainly consisted of α-helix that played a vital role in maintaining laccase activity and revealed the potential mechanisms for the changes in laccase activity under varying pHs (3.0–11.0) and temperatures (20–90 °C). Finally, the rLac could decolorize the tested dyes with high decolorization efficiency.     

The distribution of chlorophyll a in the eastern equatorial Pacific Ocean in boreal autumn

The distribution and size fractions of chlorophyll a (Chl a) concentration in the eastern equatorial Pacific Ocean in boreal autumn were investigated during October and November, 2011. Environmental factors, including hydrology and nutrients, that might affect the distribution and size composition were analyzed. A total of 18 stations including 11 CTD stations and 7 navigation stations were selected which stretch from the northwest coast of South America to the area of the central Pacific Ocean south of the Hawaiian Islands (2.77°S–13.02°N, 84.11–154.02°W). The studied area can be divided into two transects: the 6°N transect (124–148°W) and the154°W transect (10–13°N). Results showed that the surface Chl a concentration was higher in the east near the northwest coast of South America (>0.200 mg/m³) and lower in the west (0.100–0.200 mg/m³), and it presented a highly significant negative correlation with sea surface temperature (p < 0.001). There were some differences between the sectional distribution of Chl a concentration between the 6°N and 154°W transects. The high values of Chl a concentration occurred near the surface along the 6°N transect (0–75 m), while they were relatively deeper along the 154°W transect (50–100 m). Iron might be the factor that limited the growth of phytoplankton in the eastern equatorial Pacific Ocean. Picophytoplankton (Pico) was the dominant taxa in the surveyed area, particularly in the waters along the two transects (>70% of total Chl a). The Pico to total Chl a ratio was higher in the upper layer (>70%) than in the deeper layer.     

Seed storage and germination in Kumara plicatilis,a tree aloe endemic to mountain fynbos in the Boland,south-western Cape,South Africa

Seed storage under appropriate conditions is a relatively inexpensive means of safeguarding plant genetic material for ex situ conservation. Post-storage germination trials are used to determine the viability of stored seeds, and hence the efficacy of the particular storage treatment. Kumara plicatilis (= Aloe plicatilis) is a tree aloe endemic to mountain fynbos in the Boland, south-western Cape. The viability and germination behaviour of K. plicatilis seeds were assessed for seeds stored for four and nine months at − 80 °C, 4 °C, 25 °C and under ambient conditions in a laboratory. Seeds were germinated under controlled conditions and germination rates and percentages determined. Ungerminated seeds were tested for viability using tetrazolium salt. Seed viability was not significantly reduced during storage. Seeds stored at − 80 °C for four and nine months exhibited the fastest germination rate overall (both 5.9 ± 0.3 weeks, mean ± S.E.), and slowest was for seeds stored under ambient conditions for four and nine months (both 7.8 ± 0.4 weeks). All seed lots showed similar percentage germination after four months of storage (78.0–90.4%). The highest percentage germination overall was for seeds stored at − 80 °C for four months (90.4%) and the lowest was for seeds kept at 4 °C and − 80 °C for nine months (39.2 and 39.6%, respectively). Respective percentage viability for ungerminated seeds in these two treatments was 82% and 87%, respectively, indicating the induction of secondary dormancy. Induced dormancy triggered by protracted cold temperatures may be an adaptation that enables seeds to survive prolonged extreme conditions that are unfavourable for germination. Further research on the long-term storage of aloe seeds would be beneficial for developing long-term seed storage and germination testing protocols for ex situ conservation.     

Toxic strains of the Alexandrium ostenfeldii complex in southern South America (Beagle Channel,Argentina)

During phytoplankton monitoring in the Beagle Channel (≈54°52′ S, 67°32′ W) a previously undetected Alexandrium species was observed in coincidence with mouse bioassay toxicity. Detailed thecal plates analysis using epifluorescence and scanning electron microscopy revealed the presence of the Alexandrium ostenfeldii species complex, showing a mixture of the diagnostic features usually used to discriminate between the morphospecies A. ostenfeldii and A. peruvianum. Cells of the A. ostenfeldii complex were commonly observed during spring after the main annual diatom bloom, when temperatures and salinities were respectively around 7.5–10 °C and 30–30.5 psu, and nutrients showed a seasonal decrease. Toxin analysis by liquid chromatography–mass spectrometry revealed the production of 13-desmethyl spirolide C and 20-methyl spirolide G in cell cultures. The cellular contain of spirolides during exponential phase growth was 0.5906 ± 0.0032 and 0.1577 ± 0.0023 pg cell⁻¹ for 13-desMe-C and 20-Me-G, respectively. A third unknown compound, with a structure resembling that of spirolides was also detected in culture. Moreover, an additional compound with a similar m/z (692) than that of 13-desMe-C but presenting a higher retention time (Rt = 40.5 min) was found in high proportions in mussel samples. PSP toxins were present at low concentration in mussels but were not detected in cultures. These results extend the world-wide distribution of toxic strains of the A. ostenfeldii complex to the Beagle Channel (southern South America), where toxic events have been traditionally linked to the presence of Alexandrium catenella. This is the first confirmed occurrence of spirolides in mussels and plankton from Argentina, which highlights the importance of monitoring these toxins and their producing organisms to protect public health and improve the management of shellfish resources.