Validation of the in Vivo Iodo-nitro-tetrazolium (INT) Salt reduction method as a proxy for plankton respiration

Knowledge of the magnitude and variability of plankton respiration is a crucial gap in our understanding of marine carbon cycling. In order to validate the INT reduction method as a proxy for plankton respiration, we have compiled and analysed a dataset (n = 376) of concurrent measurements of dissolved oxygen consumption (CRO2) and in vivo reduction of 2-para (iodophenyl)-3(nitrophenyl)-5(phenyl) tetrazolium chloride tetrazolium salt (INT) spanning a wide range of oceanic regions and physicochemical conditions. Data were randomly divided into two independent subgroups: two thirds of the data were used to derive a regression conversion between dissolved oxygen consumption and INT reduction ("training" dataset) and one third of the data was used to validate the regression ("test" dataset). There was a significant relationship between the log-transformed dissolved oxygen consumption rates and the log-transformed INT reduction rates (INTT) with the "training" dataset (logCRO2 = 0.72logINTT + 0.44, R2 = 0.69, n = 249, p 8 °C and chlorophyll-a concentrations >0.2 μg L-1 and reduced at lower temperatures and chlorophyll-a concentrations. The results of this study endorse the adequate performance and reliability of the INT method for natural plankton communities

INT reduction is a valid proxy for eukaryotic plankton respiration despite the inherent toxicity of INT and differences in cell wall structure

The reduction of 2-para (iodophenyl)-3(nitrophenyl)-5(phenyl) tetrazolium chloride (INT) is increasingly being used as an indirect method to measure plankton respiration. Its greater sensitivity and shorter incubation time compared to the standard method of measuring the decrease in dissolved oxygen concentration, allows the determination of total and size-fractionated plankton respiration with higher precision and temporal resolution. However, there are still concerns as to the method’s applicability due to the toxicity of INT and the potential differential effect of plankton cell wall composition on the diffusion of INT into the cell, and therefore on the rate of INT reduction. Working with cultures of 5 marine plankton (Thalassiosira pseudonana CCMP1080/5, Emiliania huxleyi RCC1217, Pleurochrysis carterae PLY-406, Scrippsiella sp. RCC1720 and Oxyrrhis marina CCMP1133/5) which have different cell wall compositions (silica frustule, presence/absence of calcite and cellulose plates), we demonstrate that INT does not have a toxic effect on oxygen consumption at short incubation times. There was no difference in the oxygen consumption of a culture to which INT had been added and that of a replicate culture without INT, for periods of time ranging from 1 to 7 hours. For four of the cultures (T. pseudonana CCMP1080/5, P. carterae PLY-406, E. huxleyi RCC1217, and O. marina CCMP1133/5) the log of the rates of dissolved oxygen consumption were linearly related to the log of the rates of INT reduction, and there was no significant difference between the regression lines for each culture (ANCOVA test, F = 1.696, df = 3, p = 0.18). Thus, INT reduction is not affected by the structure of the plankton cell wall and a single INT reduction to oxygen consumption conversion equation is appropriate for this range of eukaryotic plankton. These results further support the use of the INT technique as a valid proxy for marine plankton respiration

Biogeochemical cycling of dissolved zinc along the GEOTRACES South Atlantic transect GA10 at 40°S

The biogeochemical cycle of zinc (Zn) in the South Atlantic, at 40°S, was investigated as part of the UK GEOTRACES program. To date there is little understanding of the supply of Zn, an essential requirement for phytoplankton growth, to this highly productive region. Vertical Zn profiles displayed nutrient-like distributions with distinct gradients associated with the watermasses present. Surface Zn concentrations are among the lowest reported for theworld’s oceans (<50 pM). A strong Zn-Si linear relationshipwas observed (Zn (nM)= 0.065 Si (μM), r2=0.97, n = 460). Our results suggest that the use of a global Zn-Si relationship would lead to an underestimation of dissolved Zn in deeper waters of the South Atlantic. By utilizing Si* and a new tracer Zn* our data indicate that the preferential removal of Zn in the Southern Ocean prevented a direct return path for dissolved Zn to the surface waters of the South Atlantic at 40°S and potentially the thermocline waters of the South Atlantic subtropical gyre. The importance of Zn for phytoplankton growth was evaluated using the Zn-soluble reactive phosphorus (SRP) relationship. We hypothesize that the low Zn concentrations in the South Atlantic may select for phytoplankton cells with a lower Zn requirement. In addition, a much deeper kink at ~ 500m in the Zn:SRP ratio was observed compared to other oceanic regions

Spatial extent and historical context of North Sea oxygen depletion in August 2010

Prompted by recent observations of seasonal low dissolved oxygen from two moorings in the North Sea, a hydrographic survey in August 2010 mapped the spatial extent of summer oxygen depletion. Typical near-bed dissolved oxygen saturations in the stratified regions of the North Sea were 75–80 % while the well-mixed regions of the southern North Sea reached 90 %. Two regions of strong thermal stratification, the area between the Dooley and Central North Sea Currents and the area known as the Oyster Grounds, had oxygen saturations as low as 65 and 70 % (200 and 180 µmol dm-3) respectively. Low dissolved oxygen was apparent in regions characterised by low advection, high stratification, elevated organic matter production from the spring bloom and a deep chlorophyll maximum. Historical data over the last century from the International Council for the Exploration of the Sea oceanographic database highlight an increase in seasonal oxygen depletion and a warming over the past 20 years. The 2010 survey is consistent with, and reinforces, the signal of recent depleted oxygen at key locations seen in the (albeit sparse) historical data

Seasonal changes in plankton respiration and bacterial metabolism in a temperate shelf sea

The seasonal variability of plankton metabolism indicates how much carbon is cycling within a system, as well as its capacity to store carbon or export organic matter and CO2 to the deep ocean. Seasonal variability between November 2014, April 2015 and July 2015 in plankton respiration and bacterial (Bacteria+Archaea) metabolism is reported for the upper and bottom mixing layers at two stations in the Celtic Sea, UK. Upper mixing layer (UML, >75 m in November, 41 - 70 m in April and ~50 m in July) depth-integrated plankton metabolism showed strong seasonal changes with a maximum in April for plankton respiration (1.2- to 2-fold greater compared to November and July, respectively) and in July for bacterial production (2-fold greater compared to November and April). However UML depth-integrated bacterial respiration was similar in November and April and 2-fold lower in July. The greater variability in bacterial production compared to bacterial respiration drove seasonal changes in bacterial growth efficiencies, which had maximum values of 89 % in July and minimum values of 5 % in November. Rates of respiration and gross primary production (14C-PP) also showed different seasonal patterns, resulting in seasonal changes in 14C-PP:CRO2 ratios. In April, the system was net autotrophic (14C-PP:CRO2 > 1), with a surplus of organic matter available for higher trophic levels and export, while in July balanced metabolism occurred (14C-PP:CRO2 = 1) due to an increase in plankton respiration and a decrease in gross primary production. Comparison of the UML and bottom mixing layer indicated that plankton respiration and bacterial production were higher (between 4 and 8-fold and 4 and 7-fold, respectively) in the UML than below. However, the rates of bacterial respiration were not statistically different (p > 0.05) between the two mixing layers in any of the three sampled seasons. These results highlight that, contrary to previous data from shelf seas, the production of CO2 by the plankton community in the UML, which is then available to degas to the atmosphere, is greater than the respiratory production of dissolved inorganic carbon in deeper waters, which may contribute to offshore export

Dissolved Organic Nitrogen Inputs from Wastewater Treatment Plant Effluents Increase Responses of Planktonic Metabolic Rates to Warming

Increased anthropogenic pressures on coastal marine ecosystems in the last century are threatening their biodiversity and functioning. Global warming and increases in nutrient loadings are two major stressors affecting these systems. Global warming is expected to increase both atmospheric and water temperatures and increase precipitation and terrestrial runoff, further increasing organic matter and nutrient inputs to coastal areas. Dissolved organic nitrogen (DON) concentrations frequently exceed those of dissolved inorganic nitrogen in aquatic systems. Many components of the DON pool have been shown to supply nitrogen nutrition to phytoplankton and bacteria. Predictions of how global warming and eutrophication will affect metabolic rates and dissolved oxygen dynamics in the future are needed to elucidate their impacts on biodiversity and ecosystem functioning. Here, we experimentally determine the effects of simultaneous DON additions and warming on planktonic community metabolism in the Baltic Sea, the largest coastal area suffering from eutrophication-driven hypoxia. Both bacterioplankton community composition and metabolic rates changed in relation to temperature. DON additions from wastewater treatment plant effluents significantly increased the activation energies for community respiration and gross primary production. Activation energies for community respiration were higher than those for gross primary production. Results support the prediction that warming of the Baltic Sea will enhance planktonic respiration rates faster than it will for planktonic primary production. Higher increases in respiration rates than in production may lead to the depletion of the oxygen pool, further aggravating hypoxia in the Baltic Sea

UBE1L is a retinoid target that triggers PML/RARα degradation and apoptosis in acute promyelocytic leukemia

All-trans-retinoic acid (RA) treatment induces remissions in acute promyelocytic leukemia (APL) cases expressing the t(15;17) product, promyelocytic leukemia (PML)/RA receptor α (RARα). Microarray analyses previously revealed induction of UBE1L (ubiquitin-activating enzyme E1-like) after RA treatment of NB4 APL cells. We report here that this occurs within 3 h in RA-sensitive but not RA-resistant APL cells, implicating UBE1L as a direct retinoid target. A 1.3-kb fragment of the UBE1L promoter was capable of mediating transcriptional response to RA in a retinoid receptor-selective manner. PML/RARα, a repressor of RA target genes, abolished this UBE1L promoter activity. A hallmark of retinoid response in APL is the proteasome-dependent PML/RARα degradation. UBE1L transfection triggered PML/RARα degradation, but transfection of a truncated UBE1L or E1 did not cause this degradation. A tight link was shown between UBE1L induction and PML/RARα degradation. Notably, retroviral expression of UBE1L rapidly induced apoptosis in NB4 APL cells, but not in cells lacking PML/RARα expression. UBE1L has been implicated directly in retinoid effects in APL and may be targeted for repression by PML/RARα. UBE1L is proposed as a direct pharmacological target that overcomes oncogenic effects of PML/RARα by triggering its degradation and signaling apoptosis in APL cells