Rapid screening of monoclonal antibodies against porcine circovirus type 2 using colloidal gold-based paper test.

A proof of concept for using paper test as a suitable method in the production of monoclonal antibodies (MAbs) is reported. The paper test which detects antibodies against porcine circovirus type 2 (PCV2) using colloidal gold-labelled capsid protein as the antigen probe was applied exclusively in the screening of anti-PCV2 MAbs. It allowed the detection of 118 single cell clones within 30 min using naked eyes. MAbs with specific binding to authentic epitopes on the virus were selected using a blocking strategy in which the antibody was pre-incubated with PCV2 viral sample before applying to the test paper. Five hybridomas secreting MAbs against the capsid protein were obtained, with only three of them capable of binding to PCV2. The results were validated and confirmed using enzyme-linked immunosorbent assay and immunofluorescence assay. The paper test is simple, rapid, and independent on professional technicians and proves to be an excellent approach for the screening of MAbs against specific targets.

Development of an immunochromatographic lateral flow test strip for detection of beta-adrenergic agonist Clenbuterol residues.

A rapid immunochromatographic lateral flow test strip was developed in a competitive format with the gold-conjugated monoclonal antibody to specifically determine the residues of Clenbuterol (CL), a beta-adrenergic agonist. The test strip is made up of a sample pad, a conjugate reagent pad, a test membrane containing a control line and a test line, and an absorbent pad. CL standard samples of 0, 0.1, 0.3, 0.9, 2.7, 8.1 ng/ml in swine urine were determined by the test strip. It was shown that detection limit of the test strip was as low as 0.1 ng/ml of CL and that the half of maximal inhibition concentration (IC50) in relative optical density was calculated to be 1.78+/-0.17 ng/ml under an optical density scanner. The sensitivity by eye measurement was 1.0 ng/ml. It takes 10 min to accomplish a test. Parallel analysis of urine samples from pigs fed with CL showed comparable results obtained from the test strip and GC-MS. Therefore, the test strip is very useful as a screening method for quantitative, semi-quantitative or qualitative detection of CL residues in swine urine.

Nitrate distribution and denitrification in the saturated zone of paddy field under rice/wheat rotation.

Nitrate concentration in well water collected from the wells near farm houses was investigated in the Taihu Lake basin (TBL) of China. Nitrate-N content of the well water ranged from 0.1 to 23 mgNl(-1), and 41% exceeded the criteria (10 mg Nl(-1)). It was found that the difference in well conditions, especially the depth of the well, was the main cause of the difference in the nitrate concentration of well water, i.e. it was higher in shallow well and lower in deeper well. A recommendation was made for local farmers to drill wells deeper than 10 m in order to reduce the risk of high ingestion of nitrate-N in their drinking water. Nitrate distribution and denitrification in the saturated zone of a paddy field under rice/wheat rotation in the TBL were studied. Porous pipes were installed in triplicate at depths of 1.5, 2.0, 2.5, 3.5 and 5 m respectively to collect the soil solution samples. Results showed that nitrate was the predominant N form in soil solution of saturated zone, and it increased from 1.5 to 2.5 m depth, and decreased from 2.5 to 5 m depth. N2O captured in the soil solution was very high comparing with N2O content in air. N2O content was positively correlated with nitrate concentrations in the soil profile. These results indicate that nitrate leached into saturated zone was mainly transformed via denitrification processes. Comparing the sum of inorganic nitrogen with the total nitrogen in soil solution samples collected from those wells at the field, some soluble organic nitrogen was found about 1-2 mg N l(-1) in average.

The environmental consequences of altered nitrogen cycling resulting from industrial activity, agricultural production, and population growth in China.

Human activities exerted very little effect on nitrogen (N) cycling in China before 1949. Between 1949 and 1999, however, rapid economic development and population growth led to dramatic changes in anthropogenic reactive N, inputted recycling N, N flux on land, N2O emission, and NH3 volatilization. Consequently, these changes have had a tremendous impact on the environment in China. In the current study, we estimated the amount of atmospheric wet N deposition and N transportation into water bodies from the watersheds and major valleys in China. Additionally, we addressed issues on leaching and accumulation of NO3- in the farmland under different climate zones, land use, and cropping systems as well as the potential influence of NO3- on underground water in China.

[Free radical scavenging and antifatigue activities of Saussurea involucrata polysaccharides].

The polysaccharides of Saussurea involucrata Kar et Kir was first isolated and identified by us. The polysaccharides scavenged superoxide anions by nitroblue tetraazolium colorimetric method with a median scavenging concentration of 22 micrograms.ml-1 and 95% confidence limit was 19.9-24.1 micrograms.ml-1. The polysaccharides inhibited the formation of thiobarbituric acid reaction substance in mouse liver homogenate and its IC50 was 2.3 mg/g fresh liver and 95% confidence limit was 2.05-2.55 mg/g fresh liver. By ip 25 mg.kg-1.d-1 x d in mice, the polysaccharides decreased oxygen consumption by 34.4% and ip same dosage x 6 d, the polysaccharide prolonged swimming time by 1.69-fold.