Ammonia oxidizers and denitrifiers in response to reciprocal elevation translocation in an alpine meadow on the Tibetan Plateau

Purpose

Global climate change, in particular temperature variation, is likely to alter soil microbial abundance and composition, with consequent impacts on soil biogeochemical cycling and ecosystem functioning. However, responses of belowground nitrogen transformation microorganisms to temperature changes in high-elevation terrestrial ecosystems are not well understood.

Materials and methods

Here, the effects of simulated cooling and warming on the abundance and community composition of ammonia-oxidizing archaea (AOA) and bacteria (AOB), as well as the abundance of denitrifiers, were investigated using quantitative polymerase chain reaction and clone library approaches, on the basis of a 2-year reciprocal elevation translocation experiment along an elevation gradient from 3,200 to 3,800 m above sea level on the Tibetan Plateau.

Results and discussion

We found that, compared with the temperature variations caused by elevation translocation, the soil origin exerted a much stronger influence on AOA abundance. There were significant effects of both soil origin and elevation translocation on AOB abundance, which was particularly decreased by elevation-enhanced (simulated cooling) and increased by elevation-decreased (simulated warming) treatments. Altered temperature affected the abundance of nirK rather than nirS and nosZ genes, and the latter two seemed to be associated tightly with the soil origin. Furthermore, the results showed that temperature changes had obvious influences on the community structure and diversity of AOB, but not AOA. More apparent response of AOB to warming than in other studies on grassland and forest ecosystems may be attributed to higher elevation and lower mean annual temperature in this study.

Conclusions

Our findings thus suggest that, in comparison with AOA and denitrifying populations, AOB may respond more sensitively to natural temperature variation caused by elevation translocation in this alpine grassland ecosystem on the Tibetan Plateau.     

Behavioral responses of bottlenose dolphins, Tursiops truncatus, to gillnets and acoustic alarms

Along the east coast of the United States, by-catches of bottlenose dolphins, Tursiops truncatus, in gillnet fisheries exceed removal levels set under the US Marine Mammal Protection Act. One measure proposed to reduce this mortality is the use of acoustic alarms, or pingers, which have proven effective in reducing by-catches of other small cetaceans, but have not been tested with bottlenose dolphins. We examined the responses of bottlenose dolphins to a commercial gillnet equipped with functional (active) and non-functional (control) Dukane NetMark® 1000 alarms near Fort Macon, NC. Between 5 April and 10 May 2001 we used a theodolite to track 59 groups of dolphins around the net. Choice of treatment was random each day and the two shore-based observers were unaware of whether alarms were active (13 days) or controls (9 days). There were no significant differences in the number of groups observed (P=0.315; 1−β=0.835) or in the closest observed approach to the net (P=0.307; 1−β=0.828) between treatments. However, dolphins entered a circular buffer approximately 100 m around the net more frequently with control than active alarms (P=0.015). We conclude that some dolphins responded to the alarms by avoiding the net, but caution that the potential efficacy of alarms is confounded by dolphin behavior. Most dolphins were aware of the net, regardless of the status of alarms, and some dolphins fed on fish in the net or discarded by the fishing vessel. We believe that it would be unwise to use pingers in these fisheries because of the limited behavioral responses we observed in our experiment. Furthermore, the responses we observed are likely to diminish or change over time as dolphins habituate or sensitize to these alarms. Further research is required to understand the behavior responsible for entanglement.     

Odor-active headspace components in fermented red rice in the presence of a monascus species

Headspace components from rice and agar with (experimental) and without (control) inoculation with a Monascus spp. were investigated. Kinetics studies were carried out. Using rice as a substrate, 10 and 19 compounds were found for the control and the experimental groups, respectively, at day 14. Experimental group compounds were composed mainly of alcohols, ketones, and esters, whereas control group compounds were composed of aldehydes and ketones. With agar as a substrate, only five and three components were found in the control and experimental groups, respectively. Five alcohols, four esters, two ketones, and one furan with odor activity values (OAV) >1 dominated the overall flavor of the product. With liquid inoculation, the first six components with high OAVs were in the following order: 3-methyl-1-butanol (17) > ethanol (14) > ethyl acetate (10) > 2-methyl-1-propanol (15) > ethyl butanoate (11) > 3-methylbutyl acetate (13). Kinetic studies showed that most compounds reached their maximum concentrations at 10-12 days. Many compounds identified in the model red rice were reported in commercial red sufus, and several appeared to contribute solely by red rice.     

Low compliance with recommendations on folic acid use in relation to pregnancy: is there a need for fortification?

OBJECTIVE: As a means to prevent neural tube defects (NTDs), women planning pregnancy in Denmark are recommended to take a dietary supplement of 400 microg folic acid daily during the periconceptional period. We examined compliance with this recommendation in a national survey. DESIGN: Cohort study on pregnant women in Denmark. SETTING: The Danish National Birth Cohort (DNBC). SUBJECTS: From November 2000 to February 2002, 22,000 pregnant women were recruited for DNBC. Use of dietary supplements was recorded at enrollment. Compliance with the recommendation was related to an information campaign that took place during the second half of 2001, and to lifestyle factors provided in a telephone interview by the end of the first trimester of pregnancy. RESULTS: An increase was seen in the proportion of women complying with the recommendation in the study period and this coincided with the information campaign events. However, even at the end of the period, only 22.3% of the women who had planned their pregnancy fully complied with the recommendation. No increase at all was seen in periconceptional folic acid use among women with unplanned pregnancies. Young age, low education and smoking were identified as factors that determined non-compliance. CONCLUSIONS: Alternative and more effective strategies are needed if the Danish population is to benefit fully from the knowledge that folic acid prevents NTDs. Future strategies should not only target vulnerable groups, such as the less educated and the young, but also women who get pregnant without planning this. The only possible way to reach the last group may be through fortification of foods with folic acid.     

Development of an ELISA for the detection of the residues of the fungicide iprovalicarb

A competitive indirect enzyme-linked immunosorbent assay was developed for the fungicide iprovalicarb, using a polyclonal antibody produced against a hapten conjugated through the carboxyl group on the benzene ring to keyhole limpet hemocyanin. Under an optimized condition using a heterologous format, an IC(50) of 3.51 ng/mL and the lowest detection limit of 0.065 ng/mL were obtained. When the isopropoxy group was removed from the iprovalicarb structure for the synthesis of a hapten, the resulting hapten was not successful as an immunogen, indicating that the isopropyl moiety was an important epitope, as evidenced by the cross-reactivities of some structurally related compounds. When applied to the real crop and water samples, the recoveries were in the range of 80.52-144.70% (n = 4) and 72.11-100.43% (n = 4), respectively. Accordingly, this ELISA can be used as a useful method for monitoring iprovalicarb residues in crop and water samples.     

Development of an enzyme-linked immunosorbent assay for the detection of the fungicide fenarimol

To develop an enzyme-linked immunosorbent assay for the fungicide fenarimol, two synthesized haptens, haptens-1 and -2, and the purchased 4,4'-DDA were conjugated to carrier proteins (BSA, KLH, and OVA). Polyclonal antibodies raised against hapten-1,2-KLH conjugates in rabbits and the coating antigens of hapten-1,2-BSA conjugates, hapten-2-OVA conjugate, and 4,4'-DDA-BSA conjugate were screened and selected for the homologous and/or heterologous ELISA formats. Two competitive indirect ELISAs were selected: assays I and II. The optimized ciELISAs of assays I and II showed average IC(50) values of fenarimol of 5.4 and 9.4 ng/mL, detection ranges of 1.1-25.9 and 1.1-82.7 ng/mL, and lowest detection limits of 0.3 and 0.3 ng/mL, respectively. The cross-reactivities with several structurally related compounds indicated the importance of the steric fitness in the antigen-antibody interaction. Recoveries of fenarimol from apple and pear samples spiked with the analyte by assay I were in the range of 93-113% by simple extraction, concentration, and dilution. This assay could be a convenient and supplemental analytical tool for monitoring fenarimol residues in environmental and agricultural samples.     

Antimicrobial activity of p-hydroxyphenyl acrylate derivatives

To estimate the antimicrobial effect of p-hydroxyphenyl acrylate (H5) derivatives on the basis of their molecular structure, the hydroxy and acryl groups of p-hydroxyphenyl acrylate were modified. The antimicrobial activity of the resulting compounds was assessed against a Gram-positive bacterium (Staphylococcus aureus), a Gram-negative bacterium (Pseudomonas aeruginosa), and fungi (Aspergillus fumigatus and Penicillium pinphilum) by the halo zone and the shake flask test. The antimicrobial activity of H5 was ascribed mainly to the acryl group. Compounds with acryl or acryloxy groups bound to the phenyl moiety were found to exhibit particularly high antimicrobial activities. The activities of phenyl acrylate and phenyl vinyl ketone were excellent as compared to aliphatic acrylates such as cyclohexyl acrylate and hexyl acrylate, indicating that the stereoelectronic effect of the phenyl group was important to the antimicrobial activity.     

Potent Inhibitory effect of flavonoids in Scutellaria baicalensis on amyloid beta protein-induced neurotoxicity

The free radical scavenging activities of two major flavonoids (baicalein and baicalin) in Scutellaria baicalensis were determined. The antioxidant capacities of baicalein and baicalin were determined by the 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)(*)(-) scavenging assay and showed about 110 and 70% vitamin C equivalent antioxidant capacity, respectively. Because amyloid beta (Abeta) protein is known to increase free radical production and lipid peroxidation in PC12 nerve cells, leading to apoptosis and cell death, treatment with baicalein and baicalin may result in the prevention of cellular damage by the Abeta-induced reactive oxygen species. We found that baicalein and baicalin resulted in a dose-dependent anti-Abeta toxicity by means of three different assays [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction, lactate dehydrogenase release, and trypan blue exclusion assays]. These results suggest that baicalein as well as baicalin can reduce the cytotoxicity of Abeta protein in PC12 cells, possibly by a reduction of oxidative stress, and these flavonoids may be useful in the chemoprevention of Alzheimer's disease.     

Determination of pyrethroid residues in agricultural products by an enzyme-linked immunosorbent assay

To determine cypermethrin and permethrin in agricultural products, a competitive enzyme-linked immunosorbent assay (ELISA) method was employed. The matrix interferences were minimized by direct dilution of the extracts. No further cleanup was needed. A minimum matrix effect with a 1:10 dilution of white wine for cypermethrin and a 1:200 dilution of red and white wines, fruits, and vegetables for permethrin was found when phosphate-buffered saline containing 40% methanol was employed as the diluent. Good recoveries of spiked levels were observed. The mean percentage recoveries of cypermethrin spiked in white wine and permethrin spiked in red and white wines were 99.7, 74, and 78%, respectively. The mean percentage recoveries of permethrin spiked in apple, banana, cucumber, lettuce, onion, and peach were 99.2, 105, 70.2, 97.5, 94.4, and 89.4%, respectively. Validation of the ELISA method with permethrin-spiked lettuce and peach was carried out using gas chromatography with mass spectrometry, resulting in a good recovery and correlation.