Proteomic fingerprinting enables quantitative biodiversity assessments of species and ontogenetic stages in Calanus congeners (Copepoda, Crustacea) from the Arctic Ocean.

Species identification is pivotal in biodiversity assessments and proteomic fingerprinting by MALDI-TOF mass spectrometry has already been shown to reliably identify calanoid copepods to species level. However, MALDI-TOF data may contain more information beyond mere species identification. In this study, we investigated different ontogenetic stages (copepodids C1-C6 females) of three co-occurring Calanus species from the Arctic Fram Strait, which cannot be identified to species level based on morphological characters alone. Differentiation of the three species based on mass spectrometry data was without any error. In addition, a clear stage-specific signal was detected in all species, supported by clustering approaches as well as machine learning using Random Forest. More complex mass spectra in later ontogenetic stages as well as relative intensities of certain mass peaks were found as the main drivers of stage distinction in these species. Through a dilution series, we were able to show that this did not result from the higher amount of biomass that was used in tissue processing of the larger stages. Finally, the data were tested in a simulation for application in a real biodiversity assessment by using Random Forest for stage classification of specimens absent from the training data. This resulted in a successful stage-identification rate of almost 90%, making proteomic fingerprinting a promising tool to investigate polewards shifts of Atlantic Calanus species and, in general, to assess stage compositions in biodiversity assessments of Calanoida, which can be notoriously difficult using conventional identification methods.

The role of oceanographic conditions and colony size in shaping the spatial structure of Pyrosoma atlanticum in the NW Mediterranean Sea.

This study investigates the role of winter oceanographic conditions on the horizontal and vertical spatial structure of Pyrosoma atlanticum at different ontogenetic stages. Data were obtained on two oceanographic cruises (February 2017 and 2018) in the NW Mediterranean. Small colonies were exceptionally abundant in 2017, linked to an earlier development of spring conditions and the subsequent seasonal phytoplankton bloom. The mesoscale distribution of P. atlanticum differed depending on the colony size. Large colonies (≥7 mm) were found on the slope all along the density front, whereas small (<4 mm) and medium colonies (4-6.9 mm) extended their distribution over the shelf because of instabilities of the front, and were mostly absent in the cold, low-salinity coastal waters. The analysis of their vertical distribution showed that at night colonies of all sizes remained close to the surface, where chlorophyll-a levels were high, whereas during the day they migrated to deeper layers, reaching greater depths as the colony size increased. The migratory behaviour started when colonies were 4-6.9 mm long. The relative importance of the species in the downward carbon transport is discussed. Our results highlight the need to further study the ecology of this efficient filter feeder in the Mediterranean.

Studies on chill coma recovery in the ladybird, Harmonia axyridis: Ontogenetic profile, effect of repeated cold exposures, and capacity to predict winter survival.

The harlequin ladybird, Harmonia axyridis, is one of the most successful invasive insect species worldwide. We investigated whether (i) chill coma recovery time (CCRt) changes during the ontogenetic development of this species, (ii) CCRt varies in response to repeated cold shocks, and (iii) CCRt could be a good predictor of winter survival ability in adults. CCRt decreased during larval development, the lowest CCRt values were observed in teneral adults (one and four days old), and significantly higher values were observed for older adults (16 and 32 days old). Repeated cold shocks (two hours at -3 °C), interrupted by short (30 min) warm periods (22 °C) resulted in decreased CCRt after the second cold shock, probably depicting an acclimation response, but then CCRt increased with additional cold shocks, likely revealing the accumulation of chill injuries. The CCRt of pre-overwintering individuals was not correlated with their winter survival. This indicates that CCRt is not a reliable measure of cold tolerance in H. axyridis. However, this result could be partially affected by the experimental setup - the use of laboratory-reared individuals who experienced standardized conditions and thus the variability in CCRt of tested animals could be much lower than the variability present in nature. The substantial variation observed over the ontogenetic development of H. axyridis poses important methodological implications for future studies, as animals of the same stage/age should be compared with each other. The observed U-shaped response to repeated cold shocks indicates that the expectation of linearity between cold exposure and insect response is an oversimplification of real situations.

Polyamines regulate phosphorylation-dephosphorylation kinetics in a crustacean gill (Na+, K+)-ATPase.

Aiming to clarify the mechanism of inhibition of (Na+, K+)-ATPase activity by polyamines, we examined the effects of exogenous putrescine, spermidine, and spermine on the kinetic behavior of phosphoenzyme-linked partial reactions using a microsomal gill (Na+, K+)-ATPase from juvenile and adult M. amazonicum, a freshwater palaemonid shrimp. The time course of phosphointermediate formation is greater (0.089 ± 0.006 s-1) in adults than in juveniles (0.053 ± 0.003 s-1) for spermidine, but similar to juveniles (0.059 ± 0.004 s-1) for putrescine. Maximum phosphointermediate formation for the (Na+, K+)-ATPase from juveniles decreased by 46% and 32% with spermidine and putrescine, respectively. In adults, maximum phosphointermediate levels decreased by 50% and 8%, respectively. For both spermidine and putrescine, dephosphorylation rates were higher for adults than for juveniles, and were higher than in controls without polyamines. Spermine had a negligible effect (<10%) on phosphorylation/dephosphorylation rates of both juvenile and adult enzymes. This is the first report on the effects of polyamines on phosphoenzyme-linked partial reactions in juvenile and adult M. amazonicum gill (Na+, K+)-ATPases. Our findings suggest that the phosphorylation/dephosphorylation steps of this gill enzyme may be regulated by polyamines during ontogenetic development.

Regression models for estimating leaf area of seedlings and adult individuals of Neotropical rainforest tree species / Modelos de regressão para estimativa da área foliar de indivíduos jovens e adultos de espécies arbóreas de floresta neotropical

Abstract Individual leaf area (LA) is a key variable in studies of tree ecophysiology because it directly influences light interception, photosynthesis and evapotranspiration of adult trees and seedlings. We analyzed the leaf dimensions (length – L and width – W) of seedlings and adults of seven Neotropical rainforest tree species (Brosimum rubescens, Manilkara maxima, Pouteria caimito, Pouteria torta, Psidium cattleyanum, Symphonia globulifera and Tabebuia stenocalyx) with the objective to test the feasibility of single regression models to estimate LA of both adults and seedlings. In southern Bahia, Brazil, a first set of data was collected between March and October 2012. From the seven species analyzed, only two (P. cattleyanum and T. stenocalyx) had very similar relationships between LW and LA in both ontogenetic stages. For these two species, a second set of data was collected in August 2014, in order to validate the single models encompassing adult and seedlings. Our results show the possibility of development of models for predicting individual leaf area encompassing different ontogenetic stages for tropical tree species. The development of these models was more dependent on the species than the differences in leaf size between seedlings and adults.

Resumo Área foliar individual (AF) é uma variável chave em estudos sobre a ecofisiologia de arbóreas, porque influencia diretamente a interceptação de luz, a fotossíntese e a evapotranspiração das árvores adultas e das mudas. Foram analisadas as dimensões foliares (comprimento - C e largura - L) de indivíduos adultos e de mudas de sete espécies arbóreas de florestas neotropicais (Brosimum rubescens, Manilkara maxima, Pouteria caimito, Pouteria torta, Psidium cattleyanum, Symphonia globulifera e Tabebuia stenocalyx), com o objetivo de testar a viabilidade de modelos de regressão linear para estimar a AF de indivíduos adultos e mudas. No sul da Bahia, Brasil, um primeiro conjunto de dados foi coletado entre março e outubro de 2012. A partir das sete espécies analisadas, apenas duas (P. cattleyanum e T. stenocalyx) apresentaram relações muito semelhantes entre e AF e CL, em ambos os estádios ontogenéticos. Para estas duas espécies, um segundo conjunto de dados foi coletado em agosto de 2014, a fim de validar os modelos únicos que englobam folhas de indivíduos adultos e mudas. Nossos resultados mostram a possibilidade de desenvolvimento de modelos para a predição da área foliar, abrangendo diferentes estádios ontogenéticos para espécies arbóreas tropicais. O desenvolvimento destes modelos foi mais dependente das espécies do que das diferenças entre o tamanho das folhas de mudas e de indivíduos adultos.