Seasonal and spatial distribution of ciliates in the sandy hyporheic zone of a lowland stream

The seasonal and spatial distribution of abundance and biomass as well as the taxonomic composition of ciliates inhabiting the sandy hyporheic zone of a lowland stream were studied. The mean abundances varied between 0 and 895 cells ml⁻¹ sediment, and the mean ciliate biomass ranged between 0 and 5.3 μg C ml⁻¹ sediment. Ciliate numbers and biomasses were greatest at the sediment surface and declined significantly with increasing sample depth. Abundance and biomass varied seasonally, with maximum values in late autumn and early winter and minimum values in early summer. The community was dominated by small representatives of the Hymenostomatia and Peritrichia. Ciliate community composition changed with depth from a very diverse community at the sediment surface to a less diverse one at greater sediment depths. Ciliate abundance and biomass were two orders of magnitude lower in the channel water than in the hyporheic zone. Although representatives of all sediment taxa could also be found in the channel water, the greatest concentrations of Peritrichia and Suctoria were in the hyporheic zone. The species of the sandy Ladberger Mühlenbach sediment were ubiquitous; there was no single ciliate fauna that proved to be typical for this kind of freshwater biotope.     

Microbial communities involved in biogas production from wheat straw as the sole substrate within a two-phase solid-state anaerobic digestion

Microbial communities involved in biogas production from wheat straw as the sole substrate were investigated. Anaerobic digestion was carried out within an up-flow anaerobic solid-state (UASS) reactor connected to an anaerobic filter (AF) by liquor recirculation. Two lab-scale reactor systems were operated simultaneously at 37 °C and 55 °C. The UASS reactors were fed at a fixed organic loading rate of 2.5 g L⁻¹ d⁻¹, based on volatile solids. Molecular genetic analyses of the bacterial and archaeal communities within the UASS reactors (digestate and effluent liquor) and the AFs (biofilm carrier and effluent liquor) were conducted under steady-state conditions. The thermophilic UASS reactor had a considerably higher biogas and methane yield in comparison to the mesophilic UASS, while the mesophilic AF was slightly more productive than the thermophilic AF. When the thermophilic and mesophilic community structures were compared, the thermophilic system was characterized by a higher Firmicutes to Bacteroidetes ratio, as revealed by 16S rRNA gene (rrs) sequence analysis. The composition of the archaeal communities was phase-separated under thermophilic conditions, but rather stage-specific under mesophilic conditions. Family- and order-specific real-time PCR of methanogenic Archaea supported the taxonomic distribution obtained by rrs sequence analysis. The higher anaerobic digestion efficiency of the thermophilic compared to the mesophilic UASS reactor was accompanied by a high abundance of Firmicutes and Methanosarcina sp. in the thermophilic UASS biofilm.     

Bacteriocidal effects of brevetoxin on natural microbial communities

The sensitivity of bacteria to the marine neurotoxins, brevetoxins, produced by the dinoflagellate Karenia brevis and raphidophytes Chattonella spp. remains an open question. We investigated the bacteriocidal effects of brevetoxin (PbTx-2) on the abundance and community composition of natural microbial communities by adding it to microbes from three coastal marine locations that have varying degrees of historical brevetoxin exposure: (1) Great Bay, New Jersey, (2) Rehoboth Bay, Delaware and (3) Sarasota Bay, Florida. The populations with limited or no documented exposure were more susceptible to the effects of PbTx-2 than the Gulf of Mexico populations which are frequently exposed to brevetoxins. The community with no prior documented exposure to brevetoxins showed significant (p = 0.03) changes in bacterial abundance occurring with additions greater than 2.5 μg PbTx-2 L⁻¹. Brevetoxin concentrations during K. brevis blooms range from ∼2.5 to nearly 100 μg L⁻¹ with typical concentrations of ∼10–30 μg L⁻¹. In contrast to the unexposed populations, there was no significant decrease in bacterial cell number for the microbial community that was frequently exposed to brevetoxins, which implies variable sensitivity in natural communities. The diversity in the bacterial communities that were sensitive to PbTx-2 declined upon exposure. This suggests that the PbTx-2 was selecting for or against specific species. Mortality was much higher in the 200 μg PbTx-2 L⁻¹ treatment after 48 h and >37% of the species disappeared in the bacterial communities with no documented exposure. These results suggest that toxic red tides may play a role in structuring bacterial communities.     

Use of manure concentrations of ash or specific minerals and nitrogen to estimate loss of volatile nitrogen from manure incubated under laboratory conditions

Changes in the ratio of volatile N to non-volatile specific minerals or total ash in manure can potentially be used to estimate losses of N from manure. The aim of this study was to evaluate the accuracy of using specific minerals (P, K, Ca, Mg, and Na) or total ash as markers to estimate volatile N losses from incubated manure slurries. Holstein cows were fed diets with the forage as predominantly corn silage (CS) or alfalfa silage (AS) or a diet identical to the CS diet except 5 g/kg urea was added (CSU). Total output of urine and feces were measured and used to prepare two types of slurry mixtures (all slurry mixtures contained 1200 g of manure per tray): (1) as excreted (AsEx) and (2) AsEx + sand (SAND). The SAND slurry was designed to mimic manure from barns that used sand as a bedding material. Feces and urine were mixed in the same proportion as excreted by the cows for both slurry types and 240 g of sand were added to the SAND slurries. Initial and final (3 d incubation period) slurry weights and concentrations of N, ash, and specific minerals were measured to calculate loss of volatile N. Losses of volatile N from the slurries were estimated as: (N intake ? N milk) ? [N/ash × (ash intake ? ash milk)], where N and ash intake = g/d consumed by the cow; N and ash milk = g/d secreted in milk; and N/ash = ratio of N and ash concentrations in slurry samples at 3 d. To evaluate specific minerals as markers, the same equation was used except that Ca, K, Mg, Na or P replaced the ash term. Measured N losses from the AsEx slurries were 0.44, 0.62, and 1.03 g/3 d for the AS, CS, and CSU treatments and estimated N losses (using N/ash) were 0.29, 0.49, and 1.14 g/3 d. Estimated losses were less than measured losses for cows with negative N balance and greater than measured losses for cows in positive N balance. For AsEx slurries, the use of N to specific mineral ratios was generally less accurate than using the N/ash ratio. Estimated N losses from SAND slurries were extremely inaccurate for all markers. The use of the N/ash ratio to estimate N volatilization from manure shows promise but markers that are in appreciable concentrations in bedding material will not be accurate.     

Effect of by-product gypsum application rate to grass pasture,grass hay,and corn silage on fermentation by rumen microorganisms in continuous culture

A dual-flow continuous culture fermenter system was used to investigate ruminal fermentation in response to increased by-product gypsum application rate of three forages. The treatments included 0, 22, 45, and 90 tonnes/ha by-product gypsum applied to grass plots and 0, 22, and 45 tonnes/ha by-product gypsum applied to corn plots. Forage was harvested to represent grass pasture (GP), grass hay (GH), and corn silage (CS), dried, ground, and fed to fermenters at a rate of 60 g dry matter (DM)/day. Organic matter (OM) and neutral detergent fiber (aNDF) digestibilities, rumen pH, total volatile fatty acid (VFA) production, and N metabolism were not affected by gypsum application rate for all forage types. The GH had greater sulfur content than recommended as the maximum tolerable level by the National Research Council (NRC). The results of this study indicate that ruminal fermentation was not compromised when by-product gypsum was applied to GP, GH, or CS at rates up to 90 tonnes/ha. By-product gypsum application to pastures and crops shows promise as an economical soil amendment to reduce dissolved phosphorus loss in runoff, although potential animal health issues should be further evaluated.     

Immune responses,ROS generation and the haemocyte damage of scallop Chlamys farreri exposed to Aroclor 1254

The toxic effects of Aroclor 1254 (0.05, 0.5, 5 and 50 μg l^?1) on scallop (Chlamys farreri) immune system in vivo were studied. The results showed that Aroclor 1254 had significant toxic effect on the parameters tested in this paper (P < 0.05). The total number of haemocytes, the proportion of granulocytes, phagocytosis in all groups as well as the lysosomal membrane stability (LMS) in 5, 50 μg l^?1 and bacteriolytic activity 0.5, 5, 50 μg l^?1 treatments decreased significantly, while the proportion of hyalinocytes and the production of O₂^- in all treatments remarkably increased during the sampling time and tended to be stable gradually after 6–15 d. The bacteriolytic activity in 0.05 μg l^?1 treatments, LMS in 0.05, 0.5 μg l^?1 groups and the DNA damage (comet ratios and arbitrary values) in all treatments increased at the beginning of exposure and reached their peaks on day 1, day 1, day 6 and day 3, following that they all decreased gradually and became stable after 9–15 d. When the indices reached stability, except for DNA damage was higher than controls, the others were all significantly lower than those of controls (P < 0.05). Thus, Aroclor 1254 has evident toxic effects on scallop immune system, which supports the view that a relationship exists between pollution and immunomodulation in aquatic organisms. Also it supports the speculation that the PCBs pollution is one of the important reasons of the mass mortality of the C. farreri.     

Simultaneous decrease in ammonia and hydrogen sulfide inhibition during the thermophilic anaerobic digestion of protein-rich stillage by biogas recirculation and air supply at 60 °C

We had proposed a novel method to reduce ammonia inhibition during thermophilic anaerobic digestion via recirculation of water-washed biogas into the headspace (R1 system) or liquid phase (R2 system) of reactors. The feasibility of reducing the ratio of recirculated biogas to biogas produced (called the biogas recirculation ratio) was investigated in the present study. Thermophilic anaerobic digestion at 53 °C and 60 °C with a biogas recirculation ratio of 150 facilitated stable digestion performance and biogas production at a higher organic loading rate of 7 g/L/d in the R1 system, while the ammonia removal efficiency increased 1.23-fold when the temperature increased from 53 °C to 60 °C. At 60 °C, the biogas recirculation ratios in the R1 and R2 systems decreased to 50 and 10, and the ammonia absorption rates were 6.1 and 8.3 mmol/L/d, respectively, without decreasing the anaerobic digestion performance. The ammonia absorption rate of 8.3 mmol/L/d in the R2 system was higher than the rate of 7.8 mmol/L/d at the biogas recirculation ratio of 150 in the R1 system. The hydrogen sulfide content in the biogas was reduced to less than 50 ppm by supplying air at 3% of the amount of biogas produced into the reactor.     

Effects of hybrid,kernel maturity,and storage period on the bacterial community in high-moisture and rehydrated corn grain silages

This study evaluated changes in the bacterial community in high-moisture and rehydrated corn grain silage, and their correlation with fermentation quality attributes in distinct corn hybrids, the storage period, and kernel maturity at plant harvest. Most silages achieved good fermentation (pH < 4.2). Rehydrated corn had a higher pH across all storage periods evaluated and increased dry matter losses. Leuconostoc and Lactococcus were the dominant genera in fresh material, while Lactobacillus and Acetobacter were prevalent in silages. Clostridium and Enterococcus prevailed in rehydrated corn after 120 days storage, and Clostridium was highly and positively correlated with acetone, butyric acid, and 2,3-butanediol contents. The storage period and kernel maturity were the most important factors responsible for changes in the bacterial community of silages. Results confirmed the existence of a specific bacterial microbiome that was unique for each maturity and storage time. Variations in these factors also affected the fermentation quality through influencing the bacterial community.     

Anaerobic digestion of harvested aquatic weeds: water hyacinth (Eichhornia crassipes), cabomba (Cabomba Caroliniana) and salvinia (Salvinia molesta)

The aim of this study was to explore the potential of three aquatic weeds, water hyacinth, cabomba, and salvinia, as substrates for anaerobic digestion. A set of four pilot-scale, batch digestions were undertaken to assess the yield and quality (% methane) of biogas from each plant species, and the rate of degradation. A set of 56 small-scale (100 mL) biological methane potential (BMP) tests were designed to test the repeatability of the digestions, and the impact of drying and nutrient addition.The results of the pilot-scale digestions show that both water hyacinth and cabomba are readily degradable, yielding 267 L biogas kg^?1 VS and 221 L biogas kg^?1 VS, respectively, with methane content of approximately 50%. There is evidence that the cabomba fed reactor leaked midway through the digestion therefore the biogas yield is potentially higher than measured in this case. Salvinia proved to be less readily degradable with a yield of 155 L biogas kg^?1 VS at a quality of 50% methane.The BMPs showed that the variability was low for water hyacinth and cabomba but high for salvinia. They also showed that the addition of nutrient solution and manure did not significantly increase the biogas yields and that drying was detrimental to the anaerobic degradability of all three substrates.Based on these results treatment of both water hyacinth and cabomba by anaerobic digestion can be recommended. Anaerobic digestion of Salvinia cannot be recommended due to the low biogas yields and high variability for this substrate.     

Zinc protects Ceratophyllum demersum L. (free-floating hydrophyte) against reactive oxygen species induced by cadmium

Evidence for Zn protection against Cd-induced reactive oxygen species in the free-floating hydrophyte Ceratophyllum demersum L. is presented in this paper. Metal treatments of 10 μmol/L Cd, 10 Cd μmol/L supplemented with Zn (10, 50, 100 and 200 μmol/L) and Zn-alone treatments of the same concentrations were used. Using 5,5 dimethyl pyrroline-N-oxide as the spin-probe, electron spin resonance spectra indicated a drastic increase in hydroxyl radicals (OH) in Cd-10 μmol/L treatments, which was closely correlating with the enhanced formation of hydrogen peroxide (H₂O₂) and generation of superoxide radical (O₂^?) triggered by the oxidation of NADPH. The supplementation of adding Zn (10–200 μmol/L) to the Cd-10 μmol/L treatments significantly decreased the production of free radicals especially by eliminating the precursors of OH through inhibition of NADPH oxidation. Cd-enhanced ROS production which substantially increased the oxidative products of proteins measured as carbonyls was effectively inhibited by Zn supplementation.     

Evaluation of techniques to detach particle-associated microorganisms from rumen contents

Effects of the most commonly used treatments to detach particle-associated microorganisms from rumen contents were investigated using rumen particles of different sizes. Particles were obtained before feeding from ruminally cannulated sheep. The extent of microorganism dissociation was determined using ¹⁵N as an external marker. The first experiment studied the effect of anaerobiosis on efficiency of 1 g/l methylcellulose, pH 8, and chilling (4 °C). Due to poor detachment, the anaerobic procedure was discarded. The following factors, separately or in combination, were then examined in aerobic conditions on two classes of particles obtained from whole ruminal contents (large: >400 μm; small: 100–400 μm) being: (a) stomacher pummelling (5 min); (b) Waring blender homogenisation (3×1 min); (c) chilling (4 °C); (d) pH 8; (e) 1 g/l methylcellulose; (f) 10 ml/l methanol and 10 ml/l tertiary butanol. Samples were incubated for 5 h, except for treatments (a) and (b), and washed after treatment for 2 min under running tap water in a 25 μm gauze. Blending proved to be the most effective treatment (from 50 to 57% removal). Combinations of treatments did not improve detachment. N losses from treated samples were linearly related to ¹⁵N removal. The percentage of particles removed was calculated using incubated and/or washed particles as the control, according to treatments. Results suggest that caution is needed when evaluating the effectiveness of treatments, because results are dependent on the type of particles chosen as the control.     

Molecular analysis of the bacterial community in digestive tract of rabbit

This work aimed to study the stability over time of the bacterial community in cæcum and fæces of the rabbit (diversity index and structure) without experimental disturbance and to evaluate its relationships with environmental parameters. Soft and hard fæces of 14 rabbits were sampled for 5 weeks while cæcal content was sampled on the 3rd week (by surgery) and the 5th week (at slaughter). Bacterial communities were assessed by studying CE-SSCP profiles of 16S rRNA genes fragments. Redox potential, pH, NH3-N concentration and volatile fatty acid concentrations were measured in the cæcum. Data showed that bacterial communities of soft and hard fæces barely differed from that of the cæcum (ANOSIM-R < 0.25; p < 0.05). Without disturbance, the bacterial communities of fæces were stable over time (ANOSIM-R < 0.25; p < 0.001). However, the bacterial communities of cæcum and fæces were affected by the surgery (ANOSIM-R = 0.22–0.33; p < 0.001). The cæcal content was an acidic (pH = 6.03 ± 0.33) and an anaerobic environment (redox potential = ?160 ± 43 mV). Only the redox potential was correlated with the diversity index of the bacterial community of the cæcum (R² = 0.35; p < 0.05) and no environmental parameters were correlated to its structure.     

The effect of iron and zinc supplementation and discontinuation of this practice on iron and zinc level in tissues in rats fed deficient diets

The effect of iron and iron/zinc supplementation on their levels in tissues of rats fed initially one of the three following regimen: C – control AIN-93 diet, D – iron deficient diet and R – diet with 50% reduction of all vitamins and minerals was investigated. The study was conducted on 6-week male Wistar rats, in 3 stages: (1) 4-week adaptation to the diets (C, D or R); (2) 4-week supplementation with the same regimen enriched with 10-times more iron (CSFe, DSFe, RSFe) or iron/zinc (CSFeZn, DSFeZn, RSFeZn); (3) 2-week post-supplementation period (the same diets as the stage I). Iron and zinc content in serum, the initial segment of intestine, liver and kidney were measured using FAAS method. After supplementation period (stage II) the content of iron in the intestine, liver and kidney in groups of rats fed DSFe and DSFeZn-diet were significantly higher (all p-values ≤ 0.05) than in rats fed D-diet (intestine: DSFe = 50.1 ± 9.0 μg/g wet weight, DSFeZn = 43.0 ± 9.9 μg/g vs. D = 16.5 ± 2.1 μg/g; liver: DSFe = 149 ± 30 μg/g, DSFeZn = 152 ± 25 μg/g vs. D = 56 ± 13 μg/g; kidney: DSFe = 74.0 ± 8.1 μg/g, DSFeZn = 72.7 ± 6.6 μg/g vs. D = 59.3 ± 9.5 μg/g). The same significant associations (all p-values ≤ 0.05) were observed in R rats in the intestine and liver (intestine: RSFe = 60.8 ± 6.6 μg/g, RSFeZn = 54.8 ± 6.6 μg/g vs. R = 31.5 ± 8.2 μg/g; liver: RSFe = 161 ± 10 μg/g, RSFeZn = 166 ± 21 μg/g vs. R = 136 ± 24 μg/g). After post-supplementation period the statistically significant differences between supplemented and non-supplemented rats fed D- and R-diets were still observed. There was not found the effect of applied treatments on zinc status. In conclusion, iron or iron/zinc supplementation increased similarly iron level in tissues of rats fed D-diet or R-diet with prolonged effect after supplementation discontinuation.     

High levels of plasma selenium are associated with metabolic syndrome and elevated fasting plasma glucose in a Chinese population: A case-control study

BackgroundSelenium is important for human health and involved in various metabolic processes. Deficiency of selenium associates with increased risk for cancer and cardiovascular diseases. There has been an increase use of selenium supplements for the treatment of autoimmune thyroid conditions. However, the potential biological effects of selenium overload arouse the public concern. The aim of this study was to investigate the associations of plasma selenium concentrations of adults with metabolic syndrome (MS) in Chinese population.MethodsA matched case-control study including 204 metabolic syndrome patients and 204 healthy controls was conducted in 2012. The MS cases were defined according to the criteria of Chinese Diabetes Society (CDS). Healthy controls without abnormality of metabolic components were matched with cases in age, gender and region. Plasma concentrations of selenium were determined by graphite furnace atomic absorption spectrometry (GFAAS). Fasting plasma glucose (FPG), total cholesterol (TC), triglycerides (TG), high density lipoprotein cholesterol (HDL), and low density lipoprotein cholesterol (LDL) were detected by automatic biochemical analyzer.ResultsThe median levels of plasma selenium in MS group were 146.3 (107.3–199.4) μg/L, which were significantly higher than that in the control group (127.4: 95.7–176.0) μg/L; Plasma levels of selenium were related to the risk of MS in dose-response manner. Risk of MS was significantly higher in subjects with plasma selenium in the highest tertile (T3: ≥176.0 μg/L) compared to those in the lowest tertile (T1: <95.7 μg/L) [odds ratio (OR) = 2.416 (95% CI: 1.289–4.526)]. The plasma levels of selenium were positively correlated with fasting plasma glucose (FPG) (r_s = 0.268, P < 0.001). Plasma selenium at the median (T2: 95.7–176.0 μg/L) or upper tertile (T3: ≥176.0 μg/L) was associated with increased risk of elevated FPG (defined by FPG ≥ 6.1 mmol/L) as compared with the lowest tertile (T1: ≤95.7 μg/L) [T2 vs. T1, OR = 3.487 (1.738–6.996); T3 vs. T1, OR = 6.245 (3.005–12.981)].ConclusionsHigher levels of plasma selenium might increase the risk of metabolic syndrome and elevated fasting plasma glucose. Selenium supplements should be used with prudence for CVD and cancer prevention.     

Celery oil modulates DEHP-induced reproductive toxicity in male rats

The objective of the study was to investigate the protective effect of Apium graveolens (AP) against di-(2-ethylhexyl) phthalate (DEHP)-induced testes injury in rats. Adult rats were divided into nine groups: (1) control group (no treatment); (2) corn oil (60 μg/kg body weight – bwt); (3) AP (50 μg/kg bwt); (4) 300 mg DEHP/kg bwt; (5) 500 mg DEHP/kg bwt; (6) 1000 mg DEHP/kg bwt; (7) 300 mg DEHP/kg bwt + AP; (8) 500 mg DEHP/kg bwt + AP; and (9) 1000 mg DEHP/kg bwt + AP. Oral administration of treatments was performed daily for 6 weeks. DEHP decreased (p < 0.01) body weight, testis weight and serum concentrations of testosterone, cholesterol and total proteins. Moreover, DEHP increased (p < 0.001) total antioxidant capacity in the testis and plasma DEHP level. In addition, DEHP decreased mRNA expression of two testicular steroidogenic enzymes: 3β-hydroxysteroid dehydrogenase and 17β-hydroxysteroid dehydrogenase. DEHP also caused atrophy, vacuolar degeneration and aspermia of the seminiferous tubules. AP administered concurrently with DEHP effectively alleviated most of the DEHP-induced effects. In conclusion, in male rats, DEHP had adverse effects on the testis including inhibition of androgen production. A concurrent administration of A. graveolens (celery oil) protected the testis against DEHP-induced toxicity.     

Determination of p-phenylenediamine and its metabolites MAPPD and DAPPD in biological samples using HPLC-DAD and amperometric detection

A sensitive and selective HPLC method using a diode array detector (DAD) and an electrochemical detector (ECD) in series has been developed and validated for the quantitative measurement of p-phenylenediamine and its acetylated metabolites N-acetyl-p-phenylenediamine (MAPPD) and N,N′-diacetyl-p-phenylenediamine (DAPPD) in biological samples. The separation was carried out on a hydrophilic modified AQUA C₁₈ column and the mobile phase was composed of acetonitrile: ammonium acetate solution (5:95, 25 mM, v/v). Spectrophotometric detection was performed at 240 or 255 nm and amperometric detection was carried out using a positive oxidation potential of 400 mV. The quantification of the three analytes was validated in the range of 0.05–50 μM and the established limits of determination were 0.5 μM for PPD and MAPPD and 1 μM for DAPPD. The standard deviations (N = 9) were lower than 7.5% at a concentration of 1 μM. The samples were stabilised with ascorbic acid to prevent PPD from oxidizing. Pretreatment of samples or analyte enrichment before sample injection is not required. The method proved to be accurate, sensitive and sufficiently specific. It was applied to the ecotoxicological study of the kinetics of the PPD N-acetylation in cell lysates in two different media.     

Regional nutrient thresholds in wadeable streams of New York State protective of aquatic life

Human influence on the landscape has caused nutrients in surface waters to increase to the point where their presence has substantially altered biological communities. Because this is a nationally recognized problem, the United States Environmental Protection Agency (USEPA) tasked each state, tribe, and territory to adopt numeric nutrient criteria. Here we integrate the concept of ecological thresholds with the derivation of effects-based numeric nutrient criteria. Acceptable levels of risk exceeding predefined biocriteria were determined using conditional probability and nonparametric changepoint analysis. We show how certain community metrics exhibit threshold responses to nutrients. Using these thresholds, we suggest nutrient values protective of aquatic life and characterize community composition. Nutrient criteria were suggested for two aggregations of USEPA's nutrient ecoregions in New York State an upland pristine forested region (Ecoregions VIII and XI) and a nutrient-enriched lowland region (Ecoregions VII and XIV). Of 11 biological community metrics evaluated, 5 had a strong response to nutrients (NBI-P, NBI-N, HBI, TRI, and DMA). Maximum probabilities of exceeding the biological impairment thresholds established for these metrics ranged from 81% to 100%. Changepoint analysis conducted on probability outcomes of these metrics resulted in nutrient thresholds at or above USEPA nutrient guidance values, depending on ecoregion and nutrient variable (Ecoregion VIII/XI: 15 μg/L TP, 472 μg/L TN, 150 μg/L NO₃-N, Ecoregion VII/XIV: 17 μg/L TP, 1133 μg/L TN, 356 μg/L NO₃-N). Results of taxonomic similarity percentages (SIMPER) and species contributions indicate that several orders of macroinvertebrates and diatoms exhibit significant shifts in their percent of contributions to sample similarity in response to changes in nutrient concentrations.     

Repeated freezing induces oxidative stress and reduces survival in the freeze-tolerant goldenrod gall fly,Eurosta solidaginis

Freeze tolerant insects must not only survive extracellular ice formation but also the generation of reactive oxygen species (ROS) during oxygen reperfusion upon thawing. Furthermore, diurnal fluctuations in temperature place temperate insects at risk of being exposed to multiple freeze–thaw cycles, yet few studies have examined metrics of survival and oxidative stress in freeze-tolerant insects subjected to successive freezing events. To address this, we assessed survival in larvae of the goldenrod gall fly Eurosta solidaginis, after being subjected to 0, 5, 10, 20, or 30 diurnally repeated cold exposures (RCE) to −18 °C or a single freeze to −18 °C for 20 days. In addition, we measured indicators of oxidative stress, levels of cryoprotectants, and total aqueous antioxidant capacity in animals exposed to the above treatments at 8, 32, or 80 h after their final thaw. Repeated freezing and thawing, rather than time spent frozen, reduced survival as only 30% of larvae subjected to 20 or 30 RCE successfully pupated, compared to those subjected to fewer RCE or a single 20 d freeze, of which 82% pupated. RCE had little effect on the concentration of the cryoprotectant glycerol (4.26 ± 0.66 μg glycerol·ng protein⁻¹ for all treatments and time points) or sorbitol (18.8 ± 2.9 μg sorbitol·mg protein⁻¹ for all treatments and time points); however, sorbitol concentrations were more than twofold higher than controls (16.3 ± 2.2 μg sorbitol·mg protein⁻¹) initially after a thaw in larvae subjected to a single extended freeze, but levels returned to values similar to controls at 80 h after thaw. Thawing likely produced ROS as total aqueous antioxidant capacities peaked at 1.8-fold higher than controls (14.7 ± 1.6 mmol trolox·ng protein⁻¹) in animals exposed to 5, 10, or 20 RCE. By contrast, aqueous antioxidant capacities were similar to controls in larvae subjected to 30 RCE or the single 20 d freeze regardless of time post final thaw, indicating these animals may have had an impaired ability to produce primary antioxidants. Larvae lacking an antioxidant response also had elevated levels of oxidized proteins, nearly twice that of controls (21.8 ± 3.2 mmol chloramine-T·mg protein⁻¹). Repeated freezing also lead to substantial oxidative damage to lipids that was independent of aqueous antioxidant capacity; peroxides were, on average, 5.6-fold higher in larvae subjected to 10, 20 or 30 RCE compared to controls (29.1 ± 7.3 mmol TMOP·μg protein⁻¹). These data suggest that oxidative stress due to repeated freeze–thaw cycles reduces the capacity of E. solidaginis larvae to survive freezing.     

Advantages of the integrated pig-biogas-vegetable greenhouse system in North China

An integrated pig-biogas-vegetable greenhouse system (PBVGS) was designed and studied in Laiwu, Shandong Province of North China from 2001 to 2002, where 20 groups of PBVGS and their corresponding controls were investigated. The PBVGS involves building a pigsty and a biogas digester in a vegetable greenhouse, putting pig dung into the biogas digester for fermentation, using the biogas for increasing illumination and air temperature in the greenhouse, and using the fermented waste as organic manure. The data indicate that the pig growth, biogas production and vegetable production were effectively improved in PBVGS, and that ecological, economic and social benefits were simultaneously achieved. The average annual net income of a standard PBVGS was 10,900 RMB, with an increase of 58.0% over its traditional non-integrated parts. It could use up 14,000 kg fresh pig dung and produce 10,000 kg organic manure one year for the improvement of soil fertility. The daily net weight increase for a pig in PBVGS averaged 0.82 kg, 227.6% higher than its controls. The average yield per hectare of cucumbers and tomatoes, increased by 18.4 and 17.8% over their controls, respectively. In addition, the biogas produced in the digester increased by 32.4% annually. Based on biogas fermentation, the PBVGS provides a fine ecological cycle from livestock feeding to vegetable production, resulting in a higher conversion efficiency in nutrient cycle and energy flow.     

Body-size spectra of biofilm-dwelling protozoa and their seasonal shift in coastal ecosystems

Community-based assessment of protozoa is usually performed at a taxon-dependent resolution. As an inherent ‘taxon-free’ trait, however, body-size spectrum has proved to be a highly informative indicator to summarize the functional structure of a community in both community research and monitoring programs in aquatic ecosystems. To demonstrate the relationships between the taxon-free resolution of protozoan communities and water conditions, the body-size spectra of biofilm-dwelling protozoa and their seasonal shift and environmental drivers were explored based on an annual dataset collected monthly from coastal waters of the Yellow Sea, northern China. Body sizes were calculated in equivalent spherical diameter (ESD). Among a total of 8 body-size ranks, S2 (19–27 μm), S3 (28–36 μm), S4 (37–50 μm) and S5 (53–71 μm) were the top four levels in frequency of occurrence, while rank S1 (13–17 μm), S2 and S4 were the dominant levels in abundance. These dominants showed a clear seasonal succession: S2/S4 (spring) → S2/S4 (summer) → S4 (autumn) → S2 (winter) in frequency of occurrence; S1 (spring) → S4 (summer) → S2 (autumn) → S1 (winter) in abundance. Bootstrapped average analysis showed a clear seasonal shift in body-size spectra of the protozoa during a 1-year cycle, and the best-matching analysis demonstrated that the temporal variations in frequency of occurrence and abundance were significantly correlated with water temperature, pH, dissolved oxygen (DO), alone or in combination with chemical oxygen demand (COD) and nutrients. Thus, the body-size spectra of biofilm-dwelling protozoa were seasonally shaped and might be used as a time and cost efficient bioindicator of water quality in marine ecosystems.