Declining body size has been suggested to be a universal response of organisms to rising temperatures, manifesting at all levels of organization and in a broad range of taxa. However, no study to date evaluated whether deforestation-driven warming could trigger a similar response. We studied changes in fish body size, from individuals to assemblages, in streams in Southeastern Amazonia. We first conducted sampling surveys to validate the assumption that deforestation promoted stream warming, and to test the hypothesis that warmer deforested streams had reduced fish body sizes relative to cooler forest streams. As predicted, deforested streams were up to 6 °C warmer and had fish 36% smaller than forest streams on average. This body size reduction could be largely explained by the responses of the four most common species, which were 43–55% smaller in deforested streams. We then conducted a laboratory experiment to test the hypothesis that stream warming as measured in the field was sufficient to cause a growth reduction in the dominant fish species in the region. Fish reared at forest stream temperatures gained mass, whereas those reared at deforested stream temperatures lost mass. Our results suggest that deforestation-driven stream warming is likely to be a relevant factor promoting observed body size reductions, although other changes in stream conditions, like reductions in organic matter inputs, can also be important. A broad scale reduction in fish body size due to warming may be occurring in streams throughout the Amazonian Arc of Deforestation, with potential implications for the conservation of Amazonian fish biodiversity and food supply for people around the Basin. The impacts of deforestation on tropical stream fishes have been relatively well-documented, especially those mediated by siltation, and changes in physical habitat and food resources ; 6; 7; 8Temperature is one of the most important environmental factors affecting the performance and distribution of organisms, their interactions, and the functioning of ecological communities ; 15. ForA general, negative effect of increased temperatures on body sizes could manifest across levels of biological organization from communities to individuals . That isAmong natural biological systems, aquatic ecosystems in tropical regions possess a set of characteristics that increase their potential to be impacted by warming-induced reductions in body size. One possibility requiring further validation is that temperature-size responses are greater in aquatic than terrestrial species due to the lower oxygen availability and greater effort required to increase oxygen uptake in water . The hypConsidering that 7.8 million hectares of native habitats are deforested every year (average for 1990–2015), most of which is in mega-diverse agricultural frontiers around the wet tropics , warmingWe conducted this study in the region of the headwaters of the Xingu River in southeastern Amazonia, one of the largest southern tributaries of the Amazon River. Regional climate is “Aw” according to Köppen´s classification, i.e., savanna tropical climate with distinct dry (May-September) and wet (October-April) seasons. Average annual precipitation is 1900 mm, two-thirds of which is concentrated between December and March . Our surveys were conducted at Tanguro Ranch , in the municipality of Querência, in the eastern part of the state of Mato Grosso, Brazil. At the time of sampling (2013), 60% percent of the 83,000 ha of Tanguro Ranch were covered with evergreen seasonal transitional forest between the ombrophilous rainforests in the north and the woody cerrados in the south of the Xingu Basin . The remWe sampled three first-order streams draining forested watersheds  and three streams draining deforested watersheds . None of the first-order streams we sampled were tributaries of the same second-order stream , we measured water temperatures at hourly intervals (from 9 to 19 hours) in 10 sites regularly distributed within the first 2 km of each stream . The order of stream and site sampling was varied haphazardly in each sampling date so as not to bias the measurements. All measurements were taken with a hand-held temperature electrode (ECTestr—Eutech Instruments) midway through the water column in the middle of the channel, except in impounded reaches , where temperature was measured one meter from the margin.In addition, we characterized stream habitat of the 50m-long reaches where fish sampling was performed in October and November 2013 (early rainy season). Replicated measurements of 10 environmental variables were taken at 6 perpendicular transects crossing stream reaches equidistantly (each 10 m). Measured variables were: channel width and depth, proportional substratum cover , canopy cover, and water conductivity and pH. Channel width was measured as the transversal distance between stream margins, and channel depth was measured at mid-channel, and between mid-channel and each margin. Proportional substratum cover was measured at mid-channel, at each margin, and between mid-channel and each margin. Canopy cover was estimated by analysis of photos taken close to water level in mid-channel towards canopy . Conductivity and pH were measured with electrodes (Eutech Instruments) midway through the water column in the middle of the channel.To test the community body size shift hypothesis and the population body size shift hypothesis, we sampled fish assemblages in three 50m-long stream sections regularly spaced over the first two kilometers of each stream . After extensive field trials in the study area, dipnetting was found to be the only sampling methodology appropriate for fish sampling in the narrow, shallow streams typically rich in coarse woody debris, where the low water conductivity (~10 uS/cm) prevented effective electrofishing, and gill and seine netting were impossible. Fish sampling was done by two people, who thoroughly sampled each stream reach with dipnets (1mm mesh size) for a period of 50 minutes in October and November 2013. All fish caught were euthanized with benzocaine just after capture, fixed in 10% formalin solution for 72 hours and then preserved in 70% ethanol. In the lab, specimens were identified to species level, individually measured, and weighed on a semi-analytical balance . All necessary permits were obtained for the described study, which complied with all relevant regulations .Melanorivulus zygonectes  was selected as our model species for being small, experimentally tractable, and the dominant fish species in headwater streams in the region  [M. zygonectes in deforested streams.To isolate the effects of temperature on fish body size, we conducted a laboratory experiment exposing a native species to temperatures corresponding the mean minus one standard deviation of the measures taken in forest streams at the coolest time of day (24 °C), and the mean plus one standard deviation of the measures taken in deforested streams at the warmest time of day (32 °C). llected) . This spllected) ; 35, in ad libitum with ground TetraMin Fish Flakes  until the beginning of the experiment in 18-February-2014 (~90 days). We conducted a fully factorial experiment crossing population of origin (n = 6) and temperature , with ten replicates per treatment combination  aquariums filled with filtered water to a depth of 20 cm  and fed The experimental setup consisted of two water baths 170 X 60 X 10 cm placed in a climatic chamber set at 17 °C, and containing a thermostat to raise the water temperature and a HOBO data logger recording actual temperatures every 30 minutes. Actual temperatures during the experiment were 24.1 ± 0.9 and 32.2 ± 2.3 °C (mean ± standard deviation). Each water bath housed three 50 X 40 X 10 cm aquariums divided in twenty 10 X 10 X 10 cm individual cells . Water in the water bath continuously circulated around and underneath all three aquaria by means of a submersible pump .We selected 20 individuals from each of the six sampled populations aiming to reduce as much as possible interpopulation variation in mean body mass. Mean ± 1 standard deviation (range) mass of selected individuals from each population were APPM: 168.7 ± 53.28 mg (80–273mg); APP2: 141 mg ± 35.72 (94–201 mg); APP2A: 143.95 ± 32.47 mg (80–191 mg); TAN1: 139.85 ± 43.71 mg (94–248 mg); TAN2: 141.22 ± 39.83 mg (82–217 mg); TAN3: 118.25 ± 31.75 mg (88–217mg). Selected fish were randomly placed in ten individual cells at each of the two manipulated temperatures . Three times a week, the fish were fed ground TetraMin Fish Flakes on a 15 mg per capita daily ration. This feeding regime always exceeded the daily consumption rates for all experimental groups, and at the same time minimized remnants of food which could impair water quality. In these occasions, mortality was registered if it occurred and dead fish were removed. Water in the aquariums was changed weekly. The experiment lasted 60 days, and by the end all survived fishes were weighed individually. We used individual growth  as main response variable, and for quality control purposes, potential reductions in survival over time were also assessed.Water temperature measurements were grouped by land use type regardless of stream, sampling date or time, and difference in mean values were tested with a non-parametric Mann-Whitney test using SigmaPlot 11.0 software . Differences among forested and deforested streams in each of the ten environmental characteristics we measured were tested by permutational multivariate analyses of variance (PerMANOVA) with a Monte Carlo permutation procedure  done using the PRIMER-E software . These ac showed that the third option best captured the random variation in the data  a random intercept on stream, 2) random intercept on stream and species, 3) random intercept on stream, random intercept on species, and random slope on species. Model selection among these models using AICthe data . This reer model .Melanorivulus zygonectes, Hyphessobrycon mutabilis, Pyrrhulina australis, Aequidens michaeli, Moenkhausia phaenota, and Hyphessobrycon loweae). These species composed more than 90% of the total fish abundance of this study, so incorporated a vast portion of the fish communities. Here we again used mixed effects models, although independently for each species. These models included a random effect on the intercept to account for multiple observations of each species within a given stream. To evaluate the significance of land use on fish body size for each species, we again compared models with a fixed effect of land use to those with only an intercept using AICc and likelihood ratio tests.The above models included all species (n = 29) collected across all streams and land use types. The random effects structure suggested that the effect of land use on body size varied across species, however. Therefore, we performed an additional analysis to evaluate variation at the population level. We asked first, if individual species varied in body size between land uses, and, second, how variable these responses were. To do so, we restricted our comparisons to six species for which we collected at least ten individuals in each land use type  and/or environmental changes associated with land use  explained observed shifts in fish body size. We used two approaches to get at this question with the field study data. First, we used an Analysis of Similarity (ANOSIM) with the Bray-Curtis similarity index and a permutation procedure  to test whether the relative abundance of species differed significantly between land uses. We paired this examination with a Similarity Percentage Analysis (SIMPER) to determine which taxa were primarily responsible for observed differences between groups . These tc). Initially, four models with the same fixed effect structure, but different random structures were formulated to determine the most appropriate way to include “stream of origin” as a random variable in the final models. We tested the following random effects structures: an effect of origin on the intercept, an effect of origin influencing the effect of temperature on growth, an effect of origin influencing the effect of land use form on growth, and an effect of origin influencing the effect of land use on growth plus an effect of origin influencing the effect of temperature on growth. This analysis indicated that stream of origin should be included in the model as a random factor influencing only the intercept  and complete model (R2c) were reported according to Nakagawa and Schielzeth [To test the effects of temperature on fish growth in the laboratory experiment we also used linear mixed effects models and the model selection approach through the Akaike selection criterion  disregarding the stream-specific populations. This analysis was done using SigmaPlot 11.0 software .Finally, to analyze fish survival during the lab experiment, we performed a Kaplan-Meier survival analysis with a Log-rank general significance test, and a Holm-Sidak Mean water temperature in deforested streams (28.5 °C) was 3.5 °C higher and markedly more variable  than in forest streams  . During the warmest hours of the day , mean water temperature in deforested streams was up to 6 °C higher than in forest streams , and theIn addition to higher water temperatures, deforested streams also tended to be deeper, have higher water conductivity, and very low canopy cover , which directly affected stream habitat characteristics (as measured as the distribution of stream substrate types). The mean percent cover of coarse benthic organic matter (CBOM) declined from 51% in forest streams to 11% in deforested streams, while the mean percent cover of grasses and macrophytes increased from zero in forest streams to 49% and 14%, respectively, in deforested streams .We collected a total of 2504 specimens of 29 species (representing 17 families and 6 orders), of which 801 specimens of 20 species (representing 12 families and 6 orders) were collected in forest streams, and 1703 specimens of 25 species (representing 15 families and 5 orders) were collected in deforested streams. Mean fish body mass in assemblages from deforested streams was 36% smaller than in assemblages from forest streams ; Table 2Melanorivulus zygonectes, Hyphessobrycon mutabilis, Moenkhausia phaeonota, and Hyphessobrycon loweae, had mean individual mass 46%, 43%, 49% and 55% lower in populations from deforested streams relative to populations from forest streams  in deforested streams, but did not differ significantly by land use , and that the taxa primarily responsible for the observed difference was M. zygonectes . This species is one of the smallest in our dataset, and its increased mean relative abundance in deforested streams (from 46% in forest streams to 54% in deforested streams) accounted for 27% of the total dissimilarity between groups. The six most abundant species together accounted for nearly 82% of the total dissimilarity among land uses, while all 22 remaining species accounted for the remaining 18%.Another factor that could influence average body mass is a turnover in community composition toward more small-bodied species. We found that the relative abundance of fish species differed significantly between land uses (ANOSIM 2m) and that had the lowest AICc included temperature, canopy cover, coarse benthic organic matter (CBOM), and conductivity as main effects .Growth of t 9.5 mg . The modt 9.5 mg  had an iOur results support the hypothesis that deforestation and the resulting increase in stream temperatures in a rapidly developing region of southern Amazonia are important factors influencing fish body size at the assemblage, population, and individual levels. While our field results show that other factors associated with deforestation in addition to temperature, such as canopy openness and changes in organic matter inputs also influence fish body size, our experimental results demonstrate that temperature alone has a substantial effect on the growth and size of fish from this region. The findings of our study have important implications for conservation of fish diversity in response to changing climate and land use. Body size influences virtually every aspect of organism’s biology ; 21, andSalvelinus alpines) [Several observational and experimental studies have shown that increases in temperature can reduce body size of fish and other ectothermic animals e.g., ; 47; 48); 4847; 4alpines) ; 54, butalpines) .M. zygonectes). This increase in abundance was related to a higher availability of shallow areas, the preferred habitat of M. zygonectes, in riparian floodplains of deforested streams . However, in addition to making up a larger proportion of the assemblage, individuals of this species were also significantly smaller in deforested streams. Actually, among the six most abundant species in our study area, four (including M. zygonectes) had mean body sizes that were smaller (43–55%) in populations from deforested streams. Thus, body size reductions at population level in these streams were the most likely driving factor underlying the body size reductions observed at assemblage level. Our model results support this assessment: we found that a model with a random effect of “species” on the slope described the patterns in fish body size better than one not accounting for differential species effects  and could be a source of large-sized individuals to lotic stream reaches [The only species whose populations from deforested streams had mean body size larger than forest populations was ribution ; 57 or iribution ; 59; 60. reaches .A reduced mean body size at the population level might be a result of an increase in the proportion of young individuals and/or a decrease in individuals’ body size relative to age . HoweverIn our study, the inclusion of both canopy cover and coarse benthic organic matter (CBOM) in the final models suggests that, in addition to temperature, alterations in resource availability could also be important for explaining the changes in fish body size observed in the field. Average canopy cover and CBOM declined from 86% and 51% in forest streams to 10% and 11% in deforested streams, respectively. The inputs of organic matter derived from adjacent forests are one of the main sources of carbon to the food web in forested headwater streams, and consequently, deforestation and lower organic matter inputs can reduce fish body size . It is pAustrolebias wolterstorffi [Austrolebias nigrofasciatus [M. zygonectes at 32 °C shows that temperatures similar to those occurring in the warmer hours of the day in deforested stream channels are sufficient to cause growth inhibition and reduce body size in fishes, even in species that seem to be relatively tolerant to warming, such as M. zygonectes. A recent review [M. zygonectes was reduced by just 2.6% for each one degree increase. The physiological mechanisms underlying such body size reductions were not assessed in our study, but several processes could have been involved, including increased energy expenditure in body maintenance at high temperature, reduced food intake rates and/or reduction of food assimilation efficiency [Other studies have suggested that even water temperatures below what we observed in the field might be detrimental to other Brazilian fish. For example, 28 °C was found to impair the growth of rstorffi  and 25 °asciatus . The mast review  investigficiency ; 73; 74.We also found compelling evidence for differences in the ability of populations reared under different environmental conditions/land uses to cope with warming in our lab experiment. Fishes originating from forest populations suffered a more pronounced mass loss and survival reduction at 32°C than fishes originating from deforested streams populations. Such different responses between populations which were experiencing similar temperatures in the experiment suggest that warming promote genetic adaptations and/or non-genetic effects  in deforested stream populations towards a lower sensitivity to warming. Which of these mechanisms underlie the results of our study is unknown, and remains to be investigated in future studies. Understanding the capacity and the means by which organisms may cope with warming is critical to predicting their future persistence under environmental warming driven by climate changes ; 76, andIn conclusion, our study suggests that deforestation-driven stream warming is likely to negatively affect fish body size in tropical stream fish assemblages and support the hypothesis that declining body size is a general biological response to warming. In addition, our study also suggests that other changes in stream conditions driven by deforestation, like canopy openness and changes in organic matter inputs, can affect fish body size. Declining fish size may, in the long term, negatively affect global biodiversity and ecosystem services, including food supplies for nearly a billion people who depend on fish as their primary protein source . In the S1 Tablec score. All models had n = 2426. K, number of parameters in the model; AICc, corrected Akaike Information Criteria; ΔAICc, difference in AICc between current and better model. Model Random Structures: RISt, Random Intercept on Stream; RISp, Random Intercept on Species; RSSt, Random Slope on Stream; RSSp, Random Slope on Species; RIStSp, Random Intercept on Stream and Species; RISSp, Random Intercept and Slope on Species.Results of models evaluating the random effects structure for assemblage model. The best models were considered those with lowest AIC(DOCX)Click here for additional data file.S2 Tablec score. All models had n = 101. Origin, stream of origin; Mass0, initial mass of experimental individuals; temp, temperature; K, number of parameters in the model; AICc, corrected Akaike Information Criteria; ΔAICc, difference in AICc between current and better model. Model Random Structures: RIO, Random Intercept on Origin; RSO, Random Intercept and Slope on Origin; RST, Random Slope on Temperature.Results of models evaluating the most appropriate way to include “stream of origin” as a random variable in the final models of fish growth in the lab experiment. The best models were considered those with lowest AIC(DOCX)Click here for additional data file.S3 TableEstimates and standard errors of the model coefficients for best environmental model. CBOM, coarse benthic organic matter.(DOCX)Click here for additional data file.S4 Tablepost-hoc tests for comparisons between groups for the survival of Melanorivulus zygonectes in the laboratory experiment. *represents significant difference .Statistical results of Holm-Sidak (DOCX)Click here for additional data file.S1 FigThe inset shows the place of Tanguro Ranch in the southeastern region of the Amazonian Arc of Deforestation, with green representing closed canopy forests, yellow representing deforested areas and uncolored areas representing native savannas. From South to North, streams sampled are APPM, APP2A, APP2, TAN1, TAN2, and TAN3.(TIF)Click here for additional data file.S2 FigTwo water baths 170 x 60 x 10 cm (width x length x depth) were placed in a climatic chamber set at 17°C, and containing a thermostat to raise the water temperature to 24 and 32 °C. Each water bath housed three 50 x 40 x 10 cm aquariums divided in twenty 10 x 10 x 10 cm individual cells . Water in the water bath continuously circulated around and underneath all three aquaria by means of a submersible pump. Ten individuals from six streams/populations were randomly placed at each of the two manipulated temperatures .(TIF)Click here for additional data file.S3 FigMelanorivulus zygonectes from forest (circles and solid lines) and deforested (triangles and dashed lines) streams populations reared at 24 °C (filled symbols) and 32 °C (blank symbols) during 60 days of experiment. Symbols represent mean values  and error bars represent standard errors.Survivorship of (TIF)Click here for additional data file.S1 Data(XLSX)Click here for additional data file.S2 Data(XLSX)Click here for additional data file.S3 Data(XLSX)Click here for additional data file.S4 Data(XLSX)Click here for additional data file.


Treatment meals included dinner (day 1), breakfast, lunch, and snack (day 2). Continuous glucose monitoring provided incremental area under the curve (iAUC) and mean amplitude of glycaemic excursion (MAGE) and 10 h indirect calorimetry  (day 2) provided energy expenditure and substrate oxidation. Our results demonstrated that the low GI diet resulted in lower 24 h glucose iAUC  and lower 24 h glycaemic variability . Simultaneously, 10 h respiratory quotient increased more during high GI (p = 0.014) and fat oxidation was higher after low GI breakfast (p = 0.026), lunch (p < 0.001) and snack (p = 0.013). This indicates that lower GI mixed meals incorporating isomaltulose are able to acutely reduce the glycaemic response and variability and promote fat oxidation.Low glycaemic index (GI) foods minimize large blood glucose fluctuations and have been advocated to enhance fat oxidation and may contribute to weight management. We determined whether the inclusion of isomaltulose compared to sucrose in a low/high GI meal sequence can modulate the glycaemic response and substrate oxidation in an Asian population. Twenty Chinese men (body mass index (BMI): 17–28 kg/m Asia has the unenviable reputation as being the epicenter for type 2 diabetes as the Asian phenotype has been shown to be more susceptible to diabetes than Caucasians ,2. More There is increasing interest in the use of low GI foods in the management of type 2 diabetes and obesity . OverweiNumerous ingredients have been reported to reduce glycaemic response of foods; these include beta-glucan, polyols, fiber, polyphenols, and isomaltulose ,21,22,23In this study, we simultaneously measured the dynamic response in both glycaemic response and substrate oxidation up to 42 h during an acute randomized trial to investigate the metabolic effects of the inclusion of isomaltulose or sucrose in a daily meal sequence in a free living Asian population. The study population was homogeneous with regards to ethnicity (Chinese) and gender , though representing a wide range of BMI. In the present study, the glycaemic load of the entire meal was modulated by adding sucrose or isomaltulose to create a high or low GI diet. We hypothesize that the inclusion of isomaltulose to develop low GI meals will lower the glycaemic response and variability and will result in higher fat oxidation.2, normal blood pressure (<140/80 mmHg), not intentionally restricting food intake, and fasting blood glucose lower than 6 mmol/L. All subjects followed both the treatment diets and none dropped out or were excluded from the analysis.Twenty healthy Chinese male subjects were recruited by a variety of methods, which included flyers, online advertisement, and personal communication around the University campus. Subjects underwent an initial screening and measurements, included anthropometry , fat percentage via air displacement plethysmography , blood pressure, resting heart rate, fasting blood glucose, and %HbA1C. Additionally, questionnaires on general health, physical activity  and eatiThe study was conducted at the Clinical Nutrition Research Centre (CNRC), Singapore. Written informed consent was obtained from all eligible participants before commencement and the research procedures and trial protocols were followed in accordance to the good clinical practice guidelines and with the ethical standards in concordance to the Declaration of Helsinki, 1983. Ethical approval of all procedures involving human subjects was obtained from the National Healthcare Group Domain Specific Review Board (NHG DSRB). This trial was registered as NCT 03031886 .The study consisted of two dietary treatments in a randomized double-blinded, controlled cross-over design: a low glycaemic index (LGI) and high glycaemic index (HGI) diet. Subjects attended both sessions where they followed a 24 h LGI or HGI diet depending on the randomization. Manipulated treatment meals included: dinner on day 1 and breakfast, lunch, and an afternoon snack on day 2; dinner on day 2 would be standardized over the two dietary treatments. The two test sessions of three consecutive days each were separated by a wash-out period of at least five days. Each test session would start at 17:00 on day 1 and end on day 3 at 11:00 consisting of a 42 h continuous glucose measurement (CGM) spanning the three days and a 10 h measurement of energy expenditure and substrate oxidation in a whole body calorimetry (WBC) room on day 2. On days 1 and 2, subjects were allowed to leave the research centre at 18:00 and on Day 3 at 11:00. The study consisted of a low GI and high GI treatment diet and treatment meals included: dinner on day 1 and breakfast, lunch, and an afternoon snack on day 2 . The gly®Ultra®2, LifeScan, Inc., Milpitas, CA, USA). A cross-over design with a minimum of eight subjects would be sufficient to detect a 15% change in area under the glucose curve (24 h) with a power of 0.85 at a significance level of 0.05 [Continuous glucose monitoring (CGM)  was used to measure glycaemic response, defined as the primary outcome. The insertion of the sensor was performed on day 1 at 17:00 and the sensor was removed on day 3 at 10:00. Data was collated and processed using online software (Medtronic Diabetes CareLink iPro) . The dat of 0.05 .3, furnished with a single-bed, a foldable chair, a bureau with a built-in sink, deep-freeze toilet , a colour television, an alarm clock, a radio, a telephone, a laptop, WIFI connection, and an automated intercom for communication between the researcher and the participant. It is built to mimic a normal room with two windows for visual contact between the researcher and participant.Energy expenditure, respiratory quotient (RQ) and substrate oxidation were measured using a dual room WBC facility based on the system described by Schoffelen et al. ,35,36. E2) and carbon dioxide (CO2) concentrations. Oxygen concentration was measured using a paramagnetic O2 analyser , while carbon dioxide concentration was measured using an infrared photometer . The air samples are measured in an automated sequence and alternated with a calibration span gas  and zero (100% nitrogen) gases [2 and CO2 measurements in our WBC facility were: O2 = 100.6% ± 0.5% (chamber 1) and 100.9% ± 0.4% (chamber 2), and CO2 = 99.2% ± 0.5% (chamber 1) and 99.7% ± 0.5% (chamber 2), while the coefficient of variation was 3.0% (n = 21) for repeated 30-minute RMR measurements with our WBC facility.During the test session, gaseous exchanges were measured continuously as oxygen consumption and carbon dioxide production through differences between the inlet and outlet oxygen (On) gases . Gaseous2 consumption (VO2) and CO2 production (VCO2) using the Weir equation [®, Simport Scientific, Beloeil, QC, Canada). The total volume of urine over 10 h was measured and a representative urine sample was stored for nitrogen analysis. Nitrogen content (%) was measured using the copper catalyst Kjeldahl method . Protein oxidation (g/min) was calculated by multiplying 10 h urinary nitrogen (g) by 6.25 and converted to per minute values. CHO oxidation and fat oxidation were calculated by using the following equations based on the volumes of O2 consumed and CO2 produced in oxidation of glucose, fat, and protein as published by Frayn [Energy expenditure was calculated based on volume of Oequation . Substraequation . Urine sby Frayn :CHO oxiThe participants and researchers involved in the measurements and analysis and of the data were blinded to the randomized treatment sequence until the end of the study period. All statistical analyses were performed using SPSS version 23 . Data and figures were processed in a Microsoft Excel . Values were presented as means ± standard deviation (SD), unless otherwise stated. Prior to statistical analysis, the normality of the data was assured using the Shapiro-Wilks test.t-test was performed to test the differences in the glycaemic response (one-tailed), MAGE, and substrate oxidation (two-tailed) between LGI and HGI treatment diets, and alpha (α) was set at 0.05 for statistical analyses.The primary outcome of this study was to determine how the inclusion of isomaltulose (LGI) compared to sucrose (HGI) in a diet sequence  impacts 24 h glycaemic response and variability and energy regulation. First, the baseline glucose value for each subject was determined from the two-hour average CGM interstitial glucose readings before basal metabolic rate (BMR) measurement in a fasted state on day 2 and used to calculate the change in glucose level for all 42 h. Glycaemic response was expressed as the incremental area under the curve (iAUC) and calculated using the trapezoidal rule and the change in glucose above baseline ,40. TotaSubject characteristics at baseline are presented in p = 0.002) with a lower postprandial glucose iAUC after breakfast and lunch (p < 0.001). Moreover, the low GI diet treatment resulted in lower 24 h glycaemic variability .Low and high GI treatment resulted in a significantly different response in most glycaemic parameters as presented in p = 0.014), a significantly higher 10 h increase in carbohydrate oxidation (p < 0.001) and a trend for a larger 10 h decrease in fat oxidation compared to low GI (p = 0.052) (p = 0.026) and lunch (p < 0.001) compared to the high GI diet treatment (The high GI diet treatment resulted in a significantly higher 10 h increase in RQ (= 0.052) . Meal-spreatment .The change in carbohydrate and fat oxidation compared to the meal-specific baseline is presented in The current study investigated the effects of a 24 h low vs. high glycaemic index diet on the glycaemic response and substrate metabolism in normal and overweight Chinese subjects. The novelty of this study was the simultaneous and continuous measurement of these two metabolic parameters in healthy Chinese subjects to investigate the acute effect of including isomaltulose to mixed meals compared to sucrose. The addition of isomaltulose at every meal event is a unique way in which the glycaemic response of the foods may be significantly diminished. A low GI diet proved to be able to lower the blood glucose response and glycaemic variability while at the same time favouring higher fat oxidation. The effects were most pronounced after breakfast and lunch. The analogous response reflects a high likelihood that insulin is modulated by a lower GI diet and stimulates higher fat oxidation.Our study showed that the addition of isomaltulose or sucrose to a low or high GI meal was able to acutely lower 24 h glycaemic response and 24 h glycaemic variability. This attenuation of glucose response was also observed during the 10 h period in the whole body calorimeter. These results are in line with shorter acute trials showing an attenuated glucose and insulin response after isomaltulose co-ingestion compared to sucrose in Caucasian subjects ,27. AddiFor the first time in an Asian population, substrate oxidation during a low and high GI diet consumption was measured using and dual room whole body calorimeter unit simultaneous to the continuous glucose monitoring. As expected, there is a meal induced increase in carbohydrate oxidation and decrease in fat oxidation compared to baseline and as hypothesized, the 10 h respiratory quotient increased more during high GI, indicating relatively lower fat oxidation and higher carbohydrate oxidation. During the 10 h whole body calorimeter measurement, a higher increase in carbohydrate oxidation and a trend for a larger decrease in fat oxidation were seen during the sucrose containing high GI diet, which is line with current literature . Meal spStevenson and colleagues have shown that meals composed of low GI carbohydrates were able to reduce postprandial plasma glucose and increase fat oxidation during subsequent exercise ,16. StudSome researchers found that the GI of mixed meals was unable to modify fuel partitioning in sedentary obese women ,57. DiazA minor limitation was that the protein oxidation could not be measured over a continuous 24 h period and did not provide detailed hourly changes. It was, therefore, necessary to estimate the average protein oxidation based on the 10 h cycle in the WBC. The rate of protein oxidation was assumed to be constant during the time in the WBC . AlthougUnique to our study was the simultaneous measurement of blood glucose and substrate oxidation using a whole-body calorimeter when subjects were fed low or high GI meals modulated as such by including isomaltulose or sucrose. This study demonstrates that including isomaltulose to develop low-GI mixed meals is able to acutely moderate glycaemic response while promoting fat oxidation over carbohydrate oxidation during relative inactivity in Chinese subjects when compared to high GI meals. Whilst the link between blood glucose levels and fat oxidation has been demonstrated in this study, further research is necessary to quantitate how insulin and other hormones may influence tissue oxidation in humans. The consistent observation that Asians living on a high glycaemic, high carbohydrate diets are susceptible to weight gain and obesity needs some explanation. It is likely that the nature of obesity and adipose tissue accretion seen in Asians may be triggered by the consumption of high-GI diets and suggests that the drivers of obesity may not be confined to the consumption of high-fat diets alone. Our observations provide substantial public health support for the encouragement of consuming low GI meals in Asians and suggest that replacement of added sucrose in foods with isomaltulose can be a unique way in which the glycaemic response of the foods may be significantly attenuated. Thus far, the role of low GI meals has focused attention on its impact of glycaemia, though now, our study shows that it may also have an important impact on fat oxidation and possibly facilitate weight management in Asians. Our results further suggest that the use and application of isomaltulose as a part of our diet in Asia may be an even more pressing need.


This retrospective study aims to investigate the efficacy and safety of a combined posterior lateral and anteromedial approach in the treatment of terrible triad of the elbow (TTE).TTE patients who received a combination of posterior lateral and anteromedial approach or other conservative treatments were included in the present study. The postoperative functions of the elbow and the severity of traumatic arthritis were assessed using the Mayo Elbow Performance Score (MEPS) and visual analog scale (VAS). Extension–flexion of elbow joint and rotation of forearm were also measured.A combined posterior lateral and anteromedial approach or other conservative treatments showed significant improvements in the activity of the elbow, MEPS, VAS, the excellent rate, and x-ray results. The postoperative healing time and complication rate of patients who received a combined posterior lateral and anteromedial approach significantly decreased compared to those who received other conservative treatments.Patients with TTE who received a combined posterior lateral and anteromedial treatment had an increased fracture healing rate, showed improved recovery of elbow functions and had fewer complications. Generally, TTE patients suffer from complications such as joint stiffness, ulnar nerve complications, or posttrauma arthritis.,3 The main treatments of TTE include conservative or surgical treatments such as the Kocher/lateral or anteromedial approaches. Historically, difficulty in dealing with these injuries has been ascribed to insufficient information regarding elbow stabilization and the lack of appropriate surgical techniques. Therefore, it is important to study the efficacy and safety of the joint KLA and anteromedial in the treatment of TTE.“ Terrible triad of the elbow ” (TTE) consists of elbow dislocation, coronoid process fracture, or radial head fracture. Posterolateral exposure of the elbow is a beneficial technique for addressing complex fractures of the radial head. However, an extensive and deep dissection is generally required in the lateral approach. This may further damage the already traumatized soft tissues in the region and the surgical wound is at a risk of other complications such as deep infection. Additionally, the anteromedial approach facilitates repair of the anterior joint capsular, coronal head, and internal collateral ligamentous injuries.,10 When the lateral approach is used in combination with the anteromedial approach, it completely exposes these injuries and can help to restore elbow stability and reconstruct fractures of radial head and lateral ligament complex. This in turn contributes to the restoration and fixation of the coronoid fracture process. Therefore, in this paper, a combined posterior lateral and anteromedial approach and other conservative treatments were compared in the treatment of TTE.The KLA approach aims to repair radial head fracture and lateral collateral ligaments.22.1The present study was performed in accordance with the guidelines established by the Medicine Ethics Review Committee at the Affiliated YiWu Hospital of Wenzhou Medical University. All patients signed written informed consents.2.2 there were 18 cases of type I fractures, 15 cases of type II fractures, and 11 cases of type III fractures. According to the Mason classification standard on radial head fractures, there were also 12 type I fractures, 21 type II fractures, and 11 type III fractures. The inclusion criteria were as follows: patients conformed to the diagnostic criteria of TTE; patients were over 18 years old and were compliant in terms of treatment and observation; patients could tolerate the surgery ; patients had completed medical records; and patients had normal functioning of the elbow joint and no prior fractures. The exclusion criteria were as follows: patients suffered from previous fractures or pathologic fractures; patients had metabolic, endocrine or autoimmune fractures; patients suffered from mental illnesses or severe primary diseases in the heart, brain, liver, kidney, or hematopoietic system; and patients had incomplete medical records. Thirty-seven patients in the experiment group underwent surgical treatments 1 to 9 days after injury (average of 5.5 days).A total of 44 patients with closed TTE were selected from the Affiliated YiWu Hospital of Wenzhou Medical University from January 2009 to 2014. Thirty-seven cases were randomly chosen for the joint therapy (experiment group) and the remaining 7 cases received conservative treatments consisting of closed reduction and plaster fixation for 8 weeks (control group). The sample contained 25 males and 19 females aged between 33 and 65 years with a mean age of 37.7 ± 4.8 years. There were 21 cases of left-side fractures and 23 cases of right-side fractures. The cause of injury included falls (25 cases) and traffic accidents (19 cases). According to the O’Driscoll classification standard on ulna coracoid process fracture,2.3Before surgery, patients underwent preoperative computed tomography (CT) scans to determine the location of fracture displacement. After admission to the hospital, all patients were treated with manual reduction and the elbow joint was supported by external fixation until the situation became stable and suitable for surgery. If required, patients received brachial plexus anesthesia or general anesthesia. After preoperative preparation, 44 patients with closed fractures were operated on 5.5 days (on average) after injury. Antibiotics were taken 20 to 30 minutes before surgery and an x-ray machine was used to monitor the surgery. Patients with type II or type III radial head fractures were treated using the Kirschner wire or cannulated screw fixation. If necessary, artificial radial head replacement was also used in type III fractures. And the avulsion of the complexities of the lateral ulnar collateral ligament from the lateral epicondyle on a stop point must be operative management with the radial head surgery preserved and the connection between the humeroradial joint maintained. The method of coronoid process fracture fixation: a type I nonabsorbable suture was used to fix fracture mass as a base for the coronoid process. For type II or III fractures, the ulna was fixed using 1 or 2 titanium cannulated lag screws from back to front after reduction of postoperative. The joint capsule, lateral collateral ligament and medial accessory ligament was repaired and stability of the elbow joint was examined. If the valgus was unstable before lateral collateral ligament repair, hinged external fixtures were used to assistant and stabilize the bone joint. In the control group, 7 patients under conservative treatments were fixed with closed reduction and plaster for 8 weeks.2.4o using a plaster cast. During the perioperative period (3 days), routine antibiotics were prescribed for infection prevention (IP). During this period, wound care and acral blood circulation was given extra attention. Patients were told to start appropriate exercises for joint movement. Wounds were kept dry and the appropriate pressure and stitches were removed after 2 weeks. Two and 3 weeks after surgery, patients started to do passive exercise such as elbow joint flexion–extension and rotation of the forearm. The plaster cast was removed after 4 weeks and patients started to do active exercises by themselves such as elbow joint flexion–extension and rotation of forearm while avoiding an elbow flexion of above 150o for the first 6 weeks.After surgery, elbow position of patients was fixed with a bend at 902.5 The maximum score is 100 points. A final score of over 90 points was regarded as excellent, 75 to 89 as good, 60 to 74 as acceptable, and a score below 60 as poor. Elbow joint flexion–extension and rotation of the forearm was measured. The fracture site (FS) of patients was observed using an x-ray. Heterotopic ossification (HO) reduction and the complication rate were also recorded.The Mayo Elbow Performance Score (MEPS) was used to evaluate elbow joint stability, activity, and pain of patients.2.6All the patients received a follow-up check once a month for the first 3 months after surgery. Afterward 3 months, all patients received follow-ups once every 3 months. All 44 patients received follow-up checks for 6 to 15 months after surgery (10 months was the median). The follow-ups included: a lateral x-ray of the elbow joint; elbow joint flexion–extension and rotation of the forearm measurements; and (3) a MEPS.2.7t tests. Measurement data are expressed as mean ± standard deviation (SD) and the t test was used for verification. Categorical data were verified using the χ2 test. P <.05 indicates that a difference is statistically significant.All data were analyzed using SPSS 16.0 software. The median was used as the dividing line for the survival analysis and the average values were compared using 33.1P >.05, Table The study subjects included 25 males and 19 females aged between 33 and 65 years old, mean age of 37.7 ± 4.8 years. There were 21 cases of left-side fractures and 23 cases of right-side fractures. The causes of fractures included falls (25 cases) and traffic accidents (19 cases). The age, mean age, smoking status, alcohol consumption, cause of fracture, FS, body mass index (BMI), mean duration of injury, and length of stay (LOS) were not statistically different between the experiment and control groups . Complications in this experiment included superficial or deep infection, skin necrosis, and elbow dislocation. Thirty-seven cases had no complications, however, 3 TTE patients who received conservative treatments suffered from elbow pain, discomfort, and limited range of motion.The healing time and complication rate in the experiment and control groups Table  were sta3.3In the experiment group (A), all FSs healed. There was proper fixation, no flexibility in the FS x-ray, and there was no elbow joint dislocation or HO. The CT scan also revealed significant fracture improvements. However, in the control group (B) FSs did not heal as well , 11 cases considered good, 7 cases considered average, and 2 cases considered poor. In the control group, 4 cases were considered poor and 3 cases were considered average. Of the 7 patients in the control group who received conservative treatments , the rate of cases that were considered excellent was statistically different to the experiment group . In the experiment group before treatment, the average elbow joint flexion was between 85° and 125°, and after treatment it was 116.5°. Before treatment, the average rotation of the forearm was between 95° and 140°, and after treatment it was 125.5°. In the control group before treatment, the elbow joint flexion was 70° to 95°, and after treatment it was 79.5°. Before treatment, the average rotation of the forearm was between 80o and 110°, and after treatment it was 105.5°. There is a statistically significant difference between the before and after surgery results of the two groups (both P <.05, Table After a combined posterior lateral and anteromedial approach in the treatment of TTE, elbow joint flexion–extension and forearm rotation was significantly improved in the experiment and control groups (both 4 Although conservative treatments can deal with simple elbow dislocations, a more complex elbow fracture or dislocation requires a more intensive treatment. Therefore, it is imperative to find a reasonable and effective surgical treatment for TTE.TTE can be caused by high-energy trauma during a fall, which may also lead to serious elbow instability. Early surgical intervention is paramount in avoiding complications. A combined posterior lateral and anteromedial approach provides sufficient operation space and full exposure of the wound to facilitate the effective repair of soft tissue injury and bone structure fixation damages. In this study, all the elbow FSs achieved solid coalescence after the operation without delay, absent coalescence or complications. Many patients regained elbow stability even after suffering serious TTE injuries. The efficacy of conservative treatments however is quite poor, as demonstrated by the prolonged healing time and increased rate of complications.In this study, a combined posterior lateral and anteromedial approach had significant benefits in regards to healing time and complication rate over traditional conservative methods. Using a combined posterior lateral and anteromedial approach, patients showed no elbow dislocation or other complications. Furthermore, the CT results further confirmed the later rehabilitation improvements. Additionally, patients who were treated with the conservative methods showed significantly limited elbow activity and discomfort. During the surgical treatment period, elbow stability should be restored through brachioradialis connection, lateral collateral ligament repair, or internal fixation of the coronal position. demonstrated that a prerequisite for good elbow function is to replicate exercises, which helps to stabilize the elbow joint and reconstruct the coronoid process. Apart from a smaller incision in the lateral approach, it provides better exposure for radial head fracture fixation and the repair of the lateral collateral ligament than the posterior approach. The anteromedial approach also has a clear exposure of ulnar coronoid process and radial head fractures. This can anatomically restore serious types II and III fractures, achieve good elbow stability, and improve functioning. Moreover, this study shows that the Mayo score in joint operation is significantly improved compared to the conservative approach. de Aquino Santos et al also demonstrated that when functional activity of the elbow is taken into consideration, the results of the surgery were usually satisfactory. This is consistent with our results.It was also found in this study that elbow functioning scores were higher in patients who received a combined posterior lateral and anteromedial approach than in those who received conservative treatments. Interestingly, Kälicke et alThis study aimed to investigate the clinical efficacy and safety of a joint posterior lateral and anteromedial approach in TTE treatment. It was found that the joint approach yielded higher healing rates, fewer complication rates, and is worthy of being promoted for clinical applications. However, because of the complex mechanisms involved in the pathogenesis of TTE, follow-up studies are required.


Concentrations of Ca, K, Mg and Na were enhanced by baseflow contributions and followed closely the electrical conductivity. Al and Fe minerals occurred in insoluble forms, and their pollutographs were similar to those of TMs, whose concentrations mimicked, to some extent, the flux of TSS. The TMs with the highest and lowest particulate fractions were Cr&Pb and Cu&Zn, respectively. The concentrations of total TMs in snowmelt were two to four times higher than those in stormwater, and both sources likely exceeded some of the stormwater effluent limits  proposed in Sweden. Where such concentrations depended on water hardness, the risk of toxicity might be reduced by elevated hardness of the monitored snowmelt and stormwater. Recognizing the good ecological status of the study area receiving water, Lake Storsjön, some protection against polluted runoff and snowmelt may be needed and could be achieved by implementing stormwater management measures controlling TSS and TMs.Inorganic chemicals in urban stormwater and snowmelt runoff originate from catchment geology and anthropogenic activities. The occurrence, partitioning and mobility of six minerals and six trace metal (TM) indicators of anthropogenic activities were studied in stormwater, snowmelt and baseflow in four urban catchments, and the sampling of inorganics was supplemented by measurements of electrical conductivity (EC), pH and total suspended solids (TSSs). Minerals occurred at concentrations several orders of magnitude higher (1–10 Chemical characterization of urban stormwater has been reported extensively during the past 50 years, particularly with respect to stormwater impacts on receiving water quality US EPA, . The mosFrom a general perspective, stormwater runoff can be described as a process occurring across the urban landscape and hence reflecting both the local geology and anthropogenic activities  in stormwater and snowmelt originate from traffic  from soils and alteration of their structure. Similar concerns apply to stormwater discharged into (i) substrates of permeable pavements, from which salt-laden inflows may release particulates with adsorbed chemicals and temporarily reduce the facility environmental performance  serving for drinking water supply, recreation and fishing. Lake Storsjön is classified as a water resource with a good ecological status, and as such, the lake water quality has to comply with the EU WFD regulations concerning raw drinking water and bathing water quality , were collected during selected time periods in four urban catchments located in the City of Östersund, Sweden. The city has a population of about 45,000 inh. (2010 data), is located in the middle part of Sweden  and is drained by the traditional curb and gutter system discharging into a separate storm sewer system with multiple outlets. The built-up area of the city can be divided into a number of sewersheds with singular outlets discharging without treatment into Lake Storsjön, which is one of the largest lakes in Sweden  simultaneously to obtain samples from the same events. A total of six rain-driven events and six snowmelt events were sampled between September 2012 and June 2013. The rain-driven runoff events occurred and were sampled during the warmer part of the year and were associated with rainfalls ranging from 2.8 to 11.2 mm, preceded by dry periods varying from 3 to 14 days, allowing accumulation of pollutants. The snowmelt events occurred in late winter/early spring and were characterized by dry weather. Baseflows in sewersheds B and D were sampled on nine occasions in each sewershed during dry weather, between rain and snowmelt periods. Altogether, about 270 samples of stormwater, snowmelt and baseflow were collected during the study period and submitted for analysis of selected inorganics.3 of water passed through the site, depending on the runoff intensity. After withdrawal, the samples were placed in coolers and submitted for analysis.During individual sampling events of various durations, 4 to 15 grab samples were collected by dipping pre-rinsed 2-L polypropylene sample bottles into the sewer flow at the mid-section of the sewer. Discrete grab samples were collected proportionally to a predefined flow volume, using flow rates measured by an area-velocity flow meter (type ISCO 2150) installed in the storm sewer a short distance upstream of the sampling point. The frequency of sample withdrawal varied from catchment to catchment, with consecutive samples withdrawn after 1 to 40 mtot = 0.45 μg/L, Crtot = 15 μg/L, Cutot = 30 μg/L, Nitot = 20 μg/L, Pbtot = 10 μg/L and Zntot = 90 μg/L. Such guidelines were proposed for controlling environmental risks of TM release in stormwater discharges into Swedish receiving waters.The selection of water quality constituents to be studied was based on the study objectives aiming to elucidate differences between the two types of inorganics in urban drainage effluents: (i) inorganics of mostly mineral origin, affected by local soils and geology , pH and TSSs to assist in data assessment and interpretations.All chemical analyses employed in this study were done by a SWEDAC-accredited commercial laboratory (ALS Scandinavia), following the prescribed analytical and QA/QC procedures. For the analysis of inorganics, a subsample of 100 mL was withdrawn in the field from the total sample 2 L) and kept at cool temperatures prior to analysis. Before submission to the laboratory, a 50-mL aliquot was pre-filtered (filter pore size 0.45 μm) and both non-filtered and pre-filtered subsamples were submitted for total and dissolved inorganic content analysis, respectively. The inorganics were determined by inductively coupled plasma (ICP) sector field mass spectrometry  and ICP optical emission spectrometry  according to slightly modified US EPA methods 200.8 and 200.7 US EPA, , respect L and keThe rest of the whole-water sample was analysed for physico-chemical properties in the laboratory of the Östersund municipality. The content of suspended solids was measured by the TSS standard method by extracting and filtering sample aliquots through pre-weighed glass fibre filters (Whatman GF/A filter)  asV) during the sampled portion of the runoff event and subscripts i and n denote the ith sample and the total number of samples, respectively.where the numerator is equal to the total flux of the sampled constituent, the denominator is equal to the total volume (Mean EC and baseflow mean concentrations (measured prior to each event in a single sample) were calculated as arithmetic means. TM concentrations below detection limits were considered in EMC calculations as one half of the reporting limit values.The cluster analysis (CA), employing the distance measures, was used to assess the relationships among uncorrelated groups of parameters, using concentrations in individual samples. Such data were skewed and, therefore, were log-transformed before analysis. The cluster correlation analysis was used for addressing similar temporal behaviour among parameter groups, assuming that similar variation patterns indicate similar sources. Finally, the Pearson correlation analysis was applied to the log-transformed TMs and TSS data to assess the common pathways of these water quality constituents in stormwater and snowmelt runoff.The study results are presented under five subheadings addressing stormwater and snowmelt export patterns of minerals and TMs, correlation patterns among the constituents studied, pollutographs of minerals and TMs during discrete events, partitioning of TMs and correlations with TSS and an environmental assessment of the mineral inorganics and TMs studied.The EMCs of minerals , TMs  and TSS and the EC readings are presented in Table Minerals and TM EMCs were significantly higher in snowmelt runoff, with ECs and TSS concentrations up to two and five times higher, respectively, compared to rain runoff. Higher mineral and TSS concentrations in snowmelt were attributed to the grit materials applied as traction agents in winter road maintenance. pH of stormwater runoff from rain and snowmelt varied between 6.9–8.4 and 6.2–8.6, respectively. Such values are within the limits recommended by the European Commission (EC) for fresh waters supporting fish life EC, .When comparing concentrations of minerals and TMs in rain runoff, snowmelt and baseflow, the highest concentrations of dissolved minerals were observed in baseflow, followed by snowmelt and rain runoff, and this ranking was identical to that of respective EC measurements. Such a similarity in ranking is expected, because EC measures total dissolved solids (TDSs) in water. Two minerals occurring predominantly in insoluble forms, Al and Fe, exhibited transport patterns similar to those of trace metals.Trace metals were transported mostly as a particulate load with TSS, and hence, their concentrations followed those of TSS. TSS in stormwater reflected the strength of sources (snowmelt > rain runoff >> baseflow), with transport capacities of these three flows being comparable. The measured major ion concentration ranges  during rainfall runoff Table  were comThe higher ion concentrations in snowmelt were explained by smaller volumes of snowmelt, compared to rainfall runoff events, and high supplies of grit applied in the studied catchments. Furthermore, in catchments B and D, baseflow originating from the ingress of local groundwater . These events were selected so that they produced single-peak hydrographs during rainfall  and similar runoff volumes during snowmelt . Furthermore, Ca and Fe exhibited the highest concentrations among the minerals studied, in the dissolved and particulate fraction, respectively. Thus, the featured pollutographs demonstrate various flux patterns for typical examples of minerals and TMs studied. The dissolved constituent (Ca) behaved differently from minerals in the particulate fraction. Ca started in both catchments at relatively elevated concentrations (22–41 mg/L), which were quickly diluted as runoff flow increased. In catchment A, Ca flux followed a first flush pattern, and towards the end of the event, concentrations stabilized at low values. In the residential catchment B with baseflow, the concentrations followed a U shape; the initially high Ca concentrations (from baseflow) became diluted by stormwater but, towards the end of runoff, rose again as baseflow took over the control of Ca concentrations. Hence, in this case, the baseflow affected concentration variations during the event.Pollutographs (i.e. graphs of concentration vs. time) of two minerals, Ca occurring prevalently in the dissolved fraction and Fe occurring mostly in the particulate fraction, and two TMs, Cr and Pb, occurring preferentially in the particulate fraction, are displayed in Figs. The remaining three elements, Fe, Cr and Pb, exhibited similar mobility patterns driven by transport of solids in sewers. At the start of hydrographs, during low flows, concentrations were rather low, increased with increasing flow (transporting more solids) and then declined with diminishing flow and supply of solids on the catchment surface. These patterns were similar in both catchments, in spite of differences in runoff flow discharges (the peak flow in the more impervious catchment B was about three times greater).Qmax = 2 L/s) in both catchments, and Ca followed the same pattern as for rainfall runoff, with initial and end values influenced by baseflow, and the middle section reflecting dilution by snowmelt. Baseflow enrichment with Ca reflects supplies from sulphide-rich glacial soils  produced much smaller runoff flow rates  were almost identical, because of a negligible presence of Pbdiss (2–12%); (ii) both Pbtot and Pbpart rain pollutographs displayed a weak first flush with concentration variations reflecting TSS transport, and exhaustion of TSS sources towards the end of the event; and (iii) in snowmelt, Pb concentrations were relatively small, because of low TSS transport capacity of the generated runoff flow (Q < 1 L/s).The pollutographs showed the following tendencies: (i) the graphs of total and particulate Pb , among the six TMs studied, Cu and Zn exhibited the highest dissolved fractions, with the upper values exceeding 50% in at least three of the four studied catchments, and the relative magnitudes of dissolved fractions could be arranged, in a descending order, as follows:Such relatively high dissolved Cu and Zn concentrations agree with the previous findings of Gasperi et al.  and HubeIn snowmelt runoff, the dissolved TM fractions were about one half of those in stormwater and reflected the abundance of TSS in snowmelt, compared to rain runoff. The relative magnitudes of dissolved TM fractions in snowmelt could be arranged in the descending order as follows:p < 0.05) shown in Table Total concentrations of TMs were highly correlated to TSS during both rain and snowmelt events, with statistically significant correlation coefficients  equal to 0.65–1.3 and SAR equal to 10–18  contributed to higher hardness of stormwater and snowmelt and thereby would provide some buffering of toxic effects of such waters. The measured mineral inorganics could be safely discharged into green infrastructure facilities, because of their low salinity (EC < 0.58 dS/m) and SAR (<0.6).The observed trace metal total concentrations in stormwater runoff were comparable to the literature values, except for higher Zn values indicating additional catchment-specific sources. Total TM concentrations in snowmelt were higher than those in rain runoff for most of the TMs studied, possibly because of long periods of accumulation of TMs in snowpacks and their storage in high TSS burdens resulting from grit applications in winter road maintenance.Particulate TM concentrations correlated well with TSS concentrations . Thus, measurements of TSS could be used as surrogates for TMs and Al and Fe.The assessment of the observed TM concentrations against the proposed Swedish stormwater effluent limits  indicated the risk of exceedance in all the catchments studied for Zn and for Cd in central catchment C, and potential risks of exceedances for Cu in catchments C and D.good ecological status of the study area receiving water, Lake Storsjön, some protection against polluted runoff and snowmelt from the study area, and other contributing sewersheds, may be needed and could be planned, after increasing the robustness of the dataset on stormwater and snowmelt characterization. Reduction of pollution discharges into Lake Storsjön would be achieved by implementing stormwater management measures suitable for controlling TSS and TMs.Recognizing the The occurrence, mobility and partitioning of six abundant inorganics of primarily mineral origin  and six TM indicators of anthropogenic activities  were studied in stormwater and snowmelt runoff from four urban catchments in cold climate. In the context of study limitations, with respect to the number of events sampled and the number of samples, the following conclusions were drawn:


The results of the ordination analysis indicated that 81.2 and 84.3% of the cumulative variance for the species composition of AOA and AOB communities could be explained by the climate, land use and local factors. The climate and local environments played important roles in shaping AOA communities, whereas catchment agriculture and water chlorophyll-a concentration were key influencing factors of AOB communities. Our findings suggest that the composition and structure of sediment ammonia-oxidizing communities in Yangtze lakes are strongly influenced by different spatial scale factors.Ammonia-oxidizing archaea (AOA) and bacteria (AOB) play important roles in regulating the nitrification process in lake ecosystems. However, the relative effects of climate, catchment land use and local conditions on the sediment ammonia-oxidizing communities in lakes remain unclear. In this study, the diversity and abundance of AOA and AOB communities were investigated in ten Yangtze lakes by polymerase chain reaction (PCR), clone library and quantitative PCR techniques. The results showed that the abundances of both AOA and AOB in bare sediments were considerably but not significantly higher than those in vegetated sediments. Interestingly, AOB communities were more sensitive to changes in local environmental factors and vegetation characteristics than were AOA communities. Amongst climate and land use variables, mean annual precipitation, percentage of agriculture and percentage of vegetation were the key determinants of AOB abundance and diversity. Additionally, total organic carbon and chlorophyll-The online version of this article (doi:10.1186/s13568-017-0479-x) contains supplementary material, which is available to authorized users. Nitrification plays an important role as a link between nitrogen (N) inputs from anthropogenic sources and N losses by denitrification and anaerobic ammonium oxidation , clone library and quantitative PCR techniques. The climate, catchment land uses and local factors  were also measured. The aims of the present study were (1) to explore which factors are significantly correlated with the diversity and abundance of AOA and AOB in lake sediments and (2) to identify the total and relative contributions of each factor to the species composition of sediment AOA and AOB communities.2 surface area) in the Yangtze River basin, China  for water quality analyses were collected at approximately 0.5 m above the lake bottom before sampling sediments.At each sampling site, triplicate sediment samples were collected randomly within an area of approximately 20 m3‒), and ammonium (NH4+) were measured in situ using a YSI 6920 multi-parameter water quality probe . The total organic carbon (TOC) concentration of the filtered water samples was analysed by an elemental analyser . Water total phosphorus (TP) was determined by an acid digest followed by the colorimetric assay with the molybdenum blue method. The chlorophyll-a (Chl-a) concentration was measured by filtering 0.1‒0.2 L of water through 0.45-μm glass microfiber filters, followed by extraction with 90% acetone and measurement with a spectrophotometer .Water column pH, dissolved oxygen (DO), nitrate (NO3) of wet sediments. The contents of sediment total carbon (STC) and total nitrogen (STN) were analysed by an elemental analyzer  using air-dried and sieved (100-mesh) sediment samples.Sediment moisture was measured after drying the wet sediments at 80 °C for 2 days, while sediment density was determined by weighing a known volume  in the same areas of sediment sampling. The species richness and fresh biomass of submerged vegetation in each sampling site were determined according to Liu et al. . SubmergThe percentage of agriculture, built-up land, vegetation and water bodies in each lake catchment were calculated in software ArcGIS 10.0 based on 2010 Landsat TM and HJ-1-CCD images Table , 2016. TGenomic deoxyribonucleic acid (DNA) was isolated from approximately 0.2 g of replicate sediment subsamples using the PowerSoil DNA Isolation Kit . The quality of the extracted DNA was checked in 1% TAE-agarose gel stained with Goldview and imaged under UV light. The DNA yield was quantified using NanoDrop 2000 fluorospectrometer .amoA (ammonia monooxygenase subunit A) genes, respectively. The sequences of primers and thermal cycling procedures are shown in Additional file The primers Arch-amoAF/Arch-amoAR  by Sangon Biotech Co., Ltd. After removing sequences of poor-quality or insufficient length by Geneious Pro 8.0.2 software , the remaining sequences were aligned using the MAFFT software , 1 μL of primers (10 μM) and 2 μL of DNA template. The primers and qPCR thermal profiles are listed in Additional file amoA gene fragments were isolated using the SK8191 SanPrep Kit . Standard curves were constructed with serial plasmid dilutions of a known amount of plasmid DNA involving the target genes.The abundances  of the Prior to statistical analysis, the distribution of the data was tested for normality using the Shapiro–Wilk test. Where necessary, data were natural log or square root transformed to achieve normal distributions. The relationships between ammonia-oxidizing communities and environmental factors were examined using Pearson’s correlation analysis with the PASW 19.0 software . Additionally, a T test was used to examine differences in the diversity and abundance of nitrifiers between vegetated and bare sites. Canonical correspondence analysis (CCA) and redundancy analysis (RDA) were applied to quantify the influences of environmental factors on the species composition of AOA and AOB, respectively. A detrended correspondence analysis (DCA) was used to select the appropriate ordination analysis method (CCA or RDA). The statistical significance of the environmental factors was tested using Monte Carlo permutation tests in the software CANOCO 4.5 .The MAP ranged from 1175 mm in Lake Taihu to 1724 mm in Lake Junshanhu, with an average of 1394 mm . Therefore, AOA might play an important role in regulating the ammonia oxidation processes of the Yangtze lakes. The ratio of AOA to AOB in sediments was influenced by several factors, such as pH, organic matter, and NH4+ . In general, group I.1a existed mainly in oceans, except for Nitrosopumilus sp., which was abundant in agricultural soils (Jung et al. Nitrosopumilus group was present in sediments of freshwater lakes in the Yangtze River basin (Fig. Nitrosospira cluster (Fig. Nitrosospira and Nitrosomonas were ubiquitously present in freshwater sediments (Liu et al. Nitrosospira was found to be more advantageous in various habitats such as agricultural soils (Wessen et al. The phylogenetic tree indicated that the sediment AOA community was dominated by the group I.1b cluster Fig. , a findisin Fig. . The phyter Fig. . Althoug4+. The C/N ratio is a direct measure of resource quality and can reflect the N availability in sediments. When the sediments have a high C/N ratio, N demand by ammonia-oxidizing microorganisms is high and AOA and AOB might compete for available NH4+ (Adair and Schwartz 4+ when the NH4+ concentration was relatively low (Yu et al. A number of environmental factors can impact the community composition of AOA and AOB, and their influences are complicated and mutable (Zheng et al. a concentration on the community composition of ammonia-oxidizing microorganisms. Consistent with a previous study conducted in the Derwent Estuary, Australia (Abell et al. a concentration had a significant relationship with the AOB community composition in Yangtze lakes. This result was unexpected, but may be explained by the fact that the overgrowth of algae will change the redox conditions, which may in turn affect the composition of sediment AOB communities.RDA indicated that TP was a key determinant of AOB community composition, which was in agreement with the previous reports (e.g., Zheng et al. In conclusion, by using PCR, clone library and quantitative PCR techniques, we investigated the abundance, diversity and community composition of ammonia-oxidizing microorganisms in 35 sediment samples from ten shallow lakes in the Yangtze River basin. We found that there were no significant differences in the abundance and diversity indices of AOA and AOB between bare and vegetated sediments. The average abundance of AOA was approximately 10 times higher than that of AOB, suggesting that AOA might play an important role in regulating the ammonia oxidation processes of the Yangtze lakes. We also found that the diversity indices of AOB but not AOA were strongly influenced by different spatial scale factors such as MAP and TOC. The ordination analysis indicated that 81.2 and 84.3% of the cumulative variance for the species composition of AOA and AOB communities could be explained by the climate, land use and local factors.


Bioenergy sorghum is targeted for production in water-limited annual cropland therefore traits that improve plant water capture, water use efficiency, and resilience to water deficit are necessary to maximize productivity. A crop modeling framework, APSIM, was adapted to predict the growth and biomass yield of energy sorghum and to identify potentially useful traits for crop improvement. APSIM simulations of energy sorghum development and biomass accumulation replicated results from field experiments across multiple years, patterns of rainfall, and irrigation schemes. Modeling showed that energy sorghum's long duration of vegetative growth increased water capture and biomass yield by ~30% compared to short season crops in a water-limited production region. Additionally, APSIM was extended to enable modeling of VPD-limited transpiration traits that reduce crop water use under high vapor pressure deficits (VPDs). The response of transpiration rate to increasing VPD was modeled as a linear response until a VPD threshold was reached, at which the slope of the response decreases, representing a range of responses to VPD observed in sorghum germplasm. Simulation results indicated that the VPD-limited transpiration trait is most beneficial in hot and dry regions of production where crops are exposed to extended periods without rainfall during the season or to a terminal drought. In these environments, slower but more efficient transpiration increases biomass yield and prevents or delays the exhaustion of soil water and onset of leaf senescence. The VPD-limited transpiration responses observed in sorghum germplasm increased biomass accumulation by 20% in years with lower summer rainfall, and the ability to drastically reduce transpiration under high VPD conditions could increase biomass by 6% on average across all years. This work indicates that the productivity and resilience of bioenergy sorghum grown in water-limited environments could be further enhanced by development of genotypes with optimized VPD-limited transpiration traits and deployment of these crops in water limited growing environments. The energy sorghum model and VPD-limited transpiration trait implementation are made available to simulate performance in other target environments. The tav_amp APSIM function was used to calculate annual average ambient temperature (TAV) and annual amplitude in mean monthly temperature (AMP). The soil depth parameters were adjusted to be reflective of College Station, TX, where energy sorghum root systems rarely extend beyond 1,000 mm below the soil surface. This constraint was implemented by making the 6th soil layer (1200–1500 mm depth) a water table that restricted root growth and water uptake past the 6th soil layer.The daily progression of sorghum biomass accumulation given environmental data and management practices was simulated using Agricultural Production Systems Simulator  per hectare based on production practices described previously  and soil water supply. For a given day, if water supply is not limiting, the amount of radiation intercepted determines biomass accumulation. Daily plant water demand is calculated from radiation energy supply. The required amount of water for transpiration (water demand) needs to be extracted by roots from the soil profile to maximize biomass accumulation. If water available from the soil profile is less than the water required by the plant during canopy gas exchange for maximal biomass accumulation, then water supply reduces daily biomass accumulation. The amount of water utilized by the plant during gas exchange required for biomass accumulation is calculated from plant water demand when soil water supply was sufficient to meet demand, or from available water supply when supply was less than plant water demand.To extend this model, VPD-limited transpiration was introduced into the calculation of potential change in biomass per day by impacting plant water demand as a function of VPD. Daily weather input and biomass accumulation potential were interpolated at hourly timesteps, and for each daytime hour, the daily potential for biomass accumulation restricted by water was calculated as a function of hourly VPD. This effectively introduced the ability for a plant to slow its transpiration rate under conditions of high VPD , in which case the plant would demand less water under conditions of high VPD. Consequently, this slowed the rate of biomass accumulation under conditions of high VPD and reduced the amount of water used in high VPD conditions when transpiration efficiency is low. The calculations are described below, and their implementation in the APSIM sorghum C++ module is made available (see Code Availability).vpdBP, m1, m2} which are typically obtained from experiments quantifying VPD-limited transpiration , the slope of the linear relationship between transpiration rate and VPD for VPDs that are less than the VPD breakpoint (m1), and the slope of the linear relationship between transpiration rate and VPD for VPDs that are greater than the VPD breakpoint (m2); these three parameters can be defined in the .xml sorghum file provided by the user. Given these parameters, a daily biomass accumulation t) basis over the course of a day (sunrise to sunset). In order to evaluate VPD-limited transpiration on an hourly basis, VPD and biomass data calculated from radiation energy supply was interpolated hourly with respect to climate data by implementing the piecewise function described by Eccel , based on potential biomass accumulation from radiation energy, Br(t), leaf area index, lai, and an hourly vapor pressure deficit, vpd(t), such that the following can be evaluated (Table VPD-limited transpiration was modeled using the parameters {by Eccel  from sunTv(t) ≥ 0∀t, and T(vpdBP) = m1 × vpdBP − (−(1/4) × m1) so that at VPD less than or equal to (1/4) there is no transpiration  × lai,T(t), leaf area, lai, and transpiration efficiency, Transpiration rate dependent on VPD, radiation energy available, and extractable soil water,www.github.com/MulletLab/sorghum-energy-crop-model/).The files used to simulate energy sorghum crop growth and the code to modify growth based on VPD-limited transpiration can be found on GitHub , leaf number, radiation use efficiency, height, and stem density Table . The sor−2 (~40 Mg hectare−1) of dry shoot biomass and unlimited irrigation plots in 2009 produced ~5 kg m−2 (~50 Mg hectare−1) of dry shoot biomass. Model predictions of biomass accumulation in 2009 with unlimited irrigation tracked field data for most of the season but predicted somewhat lower biomass accumulation late in the growing season, a trend also observed when modeling biomass accumulation in 2008 under limited irrigation. In the targeted management regimes of energy sorghum, under strictly rainfed conditions, the bioenergy sorghum crop model predicted biomass accumulation that was within the range observed in 2008 mechanical harvests of larger plots  mm m−2 of water during a 200 day growing season in College Station cropping environments. In contrast, in the absence of irrigation, energy sorghum was predicted to produce on average 2.7 ± 0.8 (mean ± SD) kg m−2 of biomass and used 485 ± 120 (mean ± SD) mm m−2 of water  decreases with increasing VPD  were simulated using the VPD-limited transpiration modification with no vpdBP (m1 = m2)  accumulated almost twice as much biomass  compared to Tx436 (m1 = 6.62) under these conditions  accumulated less biomass during the first 75 days of development in fully irrigated conditions compared to BTx623 (m1 = 56.3), however biomass accumulation by the two genotypes was similar between 75 and 200 DAS. This occurred because later in the season when the genotypes reached canopy closure and VPD was higher, both genotypes extracted the maximum available soil water each day.Modeling was used initially to investigate genotypes that vary in P Figure . Specifi) Figure . In fulls Figure . A models Figure . Values m1 of 56.3 was predicted to yield approximately 2.5 kg m−2 approximately 50% less biomass than in non-water limiting conditions   accumulated biomass more rapidly in rainfed and fully irrigated environments compared to genotypes with lower m1 values. The rate of LAI increase was similar in all genotypes until ~60 DAS because leaf area development in all genotypes was limited by thermal time in the model. Genotypes with larger m1 values use water more rapidly therefore by 60 DAS the genotype with an m1 = 56.3 (BTx623) had depleted ~50% of available soil water in rainfed environments, more than the other genotypes  accumulated similar amounts of biomass between 150 and 200 DAS on average over the 14 years analyzed as the genotype with a lower m1 In rainfed cropping conditions a genotype with an s Figure . In rain) Figure . While bs Figure . During s Figure . For a gs Figure . Compare Figures . In rain Figures . In rain Figures . LAI decvpdBP where the slope of the transpiration rate response to VPD changes from m1 to m2. BQL41 is characterized by a low vpdBP of 1.17 kPA and the genotype's transpiration response to VPD is postulated to improve performance of grain sorghum in water limited environments  m2 = 7.27, the transpiration rate that was experimentally obtained from the BQL41 genotype, (b) m2 = 0.0, a maximum transpiration rate which reflects how VPD-limited transpiration was modeled previously , (d) m2 = −6.1, and (e) m2 = −12.49 transpiration responses to VPD that were experimentally obtained from SC803 and SC35 genotypes by Gholipoor et al. , 2.31 kg m−2 , 2.29 kg m−2 , 2.28 kg m−2 , and 2.25 kg m−2   marginally outperformed the genotype lacking a vpdBP over a 14 year average by 1.6 %  and accumulate more biomass during the latter part of the season  on average yielded more biomass than a sorghum genotype lacking a vpdBP (m2 = m1 = 20.26) in the water-limited low yield environments . A range of vpdBP values  representing the range bounded by BQL41 and SC35, respectively, and m2 values , the range bounded by SC35 and BTX623, respectively, were explored  accumulate more biomass when soil water resources are not limiting during the early portion of the season, and that the VPD-breakpoint trait is beneficial when water is limiting during the middle of the growing season when VPD is higher. There may exist a theoretical m2 for m1 = 56.3 that would be predicted to increase productivity beyond the 0.2% in the observed parameter space. For an m1 = 56.3, the exploration of a theoretical m2 outside what has been previously observed for VPD-limited-transpiration predicted that an m2 = −290 with a vpdBP = 2.05 would increase biomass by 6.5%, yielding an average of 2.73 kg m−2 in rainfed College Station, TX (2000-2014)  in the VPD-limited transpiration diagram, where as vpdBP increased, the range of m2 parameters that could be combined for beneficial productivity increases. This relationship with vpdBP is likely related to the effect of VPD on transpiration efficiency, TE, which is inversely proportional to VPD through the TEc coefficient: The results of modeling show that genotypes with a high m1 ~5 accumulam1 ~5 accumulaCrop modeling and structure-function trait analyses can be used to help identify constraints that limit plant productivity, assess the benefits of traits, and to increase productivity by optimizing crop deployment across variable environments. Since energy sorghum is a relatively new C4 grass biomass crop with an unusually long vegetative growth phase, crop modeling could help identify traits useful to select in breeding programs. The APSIM bioenergy sorghum model developed in this study was able to replicate the developmental time course of leaf area development, leaf number, plant height and biomass accumulation observed in field trials in College Station, Texas. Field data from other locations/years and from newer energy sorghum hybrids would be useful to further validate and enhance the accuracy of modeling predictions. In addition, collection of detailed information on soil profiles, root system development, architecture, rates of soil water extraction and information on water deficit induced changes in ABA levels would further improve the energy sorghum model and to enable modeling of root traits in conjunction with canopy architecture and VPD-transpiration traits.−1 per year. This model prediction is consistent with the biomass yield of TX08001 (~15–26 Mg hectare−1) grown without irrigation in large plots that were machine harvested in 2008 and 2009  and hot and dry summers where bioenergy sorghum hybrids are currently under selection in a breeding program. In fully irrigated conditions, modeling and field data show that energy sorghum has the genetic potential to accumulate ~50 Mg/hectare per year . However, the results showed that a higher biomass accumulation rate did not influence LAI development, a topic worth further investigation. Second, shifting water use for biomass accumulation from the early portion of the season (0–75 DAS) when average VPD is lower to later in the season (75–150 DAS) when VPD is higher and TE is lower reduces water use efficiency. The negative impact of this shift in the timing of water use can be mitigated in part by VPD-breakpoint limited transpiration traits. However, genotypes with current vpdBP and m2 values still use water at higher VPD in the summer, although at lower rates, compared to water used at lower VPD during the first portion of the planting season. This interpretation is consistent with the predicted benefits of a theoretical m2 that completely shuts off transpiration above the vpdBP. In that hypothetical genotype, transpiration would be restricted to VPD below the breakpoint during the entire growing season mitigating the impact of shifting the seasonal timing of water use. Third, in high rainfall years, VPD-limited transpiration could reduce the use of all available water resources for biomass production, especially when significant rainfall occurs in the last 60 days of the growing season. These trade-offs may help explain why in optimization simulations  were found to accumulate more biomass in the environments analyzed because these parameters allowed maximal biomass accumulation early in the growing season.Prior studies demonstrated that limited transpiration can result in improved grain yield in drought and heat stressed environments by shifting water utilization from the vegetative and booting phases to the anthesis and post anthesis phases where grain yield is more sensitive to water deficit  induction of a VPD-limited transpiration trait (m2 < −200) later in the season when VPD is higher, possibly in response to water deficit. Prior research showed that traits that limit transpiration can be environmentally triggered by changes in VPD  demonstrated that modeling of energy sorghum in the APSIM framework is able to track field growth and biomass accumulation in different environments, (ii) extended the daily APSIM-sorghum model to incorporate hourly VPD-limited transpiration, (iii) found that VPD-limited transpiration can improve crop productivity in water-limiting environments by increasing TE and by maintenance of crop canopies to enable utilization of sporadic rainfall, and (iv) this model is made available for further evaluation of bioenergy sorghum production in other targeted environments and for other traits such as facultative CAM , Office of Science, Biological and Environmental Research (BER), award numbers DE-SC0012629 and DE-FC02-07ER64494 (Great Lakes Bioenergy Research Center), and by the U.S. DOE, Advanced Research Projects Agency-Energy (ARPA-E), award number DE-AR0000596. The views and opinions of authors expressed herein do not necessarily state or reflect those of the United States Government or any agency thereof.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.