Environmental distribution and concentration of tetrabromobisphenol A bis- (2-hydroxyethyl) ether (TBBPA-DHEE) and tetrabromobisphenol A mono- (hydroxyethyl) ether (TBBPA-MHEE), are obscure due to the lack of available analytical methods. Here two novel immunoassays were established to systematically investigate their distributions in Taizhou, Eastern China. Five monoclonal antibodies against pollutants were generated with two designed haptens through animal immunization. After matched with different coating antigens/antibodies, ELISAs were established (LOD for TBBPA-DHEE, 0.12 ng/mL, based on OVA-M3/mAb-D4G6; LOD for TBBPA-MHEE, 0.79 ng/mL, based on OVA-M3/mAb-D2G6) and applied for investigation of their occurrences at a typical e-waste recycling area after 2-year samples collection, where the total 33 water, 32 soil and 16 biological samples were collected with the highest concentrations of 3.46 ng/mL, 2.76 ng/g (dry weight, dw) and 5.01 ng/g (dw), respectively. Meanwhile, our study also indicated that at the centralizing e-waste recycling sites the serious pollution for both chemicals still existed despite of various efforts. Besides, obvious improvements were observed at an abandoned e-waste recycling region treated and remedied for many years by the local Chinese government. These findings highlight the importance of policy decisions in treatment of pollutants to reduce organic pollutant-related health risks.

One-step immunochromatographic assay (ICA) has been developed using colloidal gold-labeled monoclonal antibody probe for the rapid detection of lead ions in water samples. The ICA was based on the theory of competitive reactivity, and the results can be easily judged based on the presence or absence of a red colored test line with visual detection. Under optimal conditions, this method shows high detecting sensitivity with a LOD (limit of detection) of 50 ng/ml. Stability test indicates that the immunochromatographic strips are stable for 8 weeks at room temperature. During practical application, nanometer TiO2 is used to enrich the lead ions in water samples. The ICA is successfully applied in the measurement of lead ion concentrations in local water samples, and the results are highly consistent with that of ICP-MS. Detecting lead ions with ICA can be done within 4 min and is very useful for the rapid onsite testing.

The invasive mussel Xenostrobus securis was recorded for the first time in the Galician Rias Baixas (NW Spain) in 2007, within an area characterized by intense commercial culture of Mytilus galloprovincialis. The main aims of this study were to evaluate whether an immunological assay can be used to detect larvae of this species in field samples of plankton and to determine whether the distribution of larvae matched that of adults. The ability of two monoclonal antibodies to recognize the bivalve was tested by immunofluorescence. Only the M22.8 antibody recognized X. securis larvae. The staining pattern distinguished X. securis from M. galloprovincialis larvae in both laboratory cultures and field samples of plankton. The distribution of larvae did not match that of adults. This tool may prove very useful for monitoring the presence of this invasive species in the plankton, allowing rapid and specific recognition.

Neonicotinoid insecticides are widely used in agriculture for pest control, but the pesticide residues in environmental and agricultural products were a big threat to the health of non-target organisms. In this study, a new immunochromatographic strip test was established for the rapid detection of imidacloprid residue, a neonicotinoid insecticide, based on up-conversion nanoparticles (UCNPs) coupled with the monoclonal antibody against imidacloprid. Under optimal conditions, the half inhibitory concentration (IC₅₀), detection limit, and the linear range of this strip were 8.37 ng/mL, 0.45 ng/mL, and 0.97–250 ng/mL. The strip test could be completed in 30 min. The average recoveries of imidacloprid spiked in water, Chinese cabbages, cucumber, honey, and tea samples were 70.1~101.8%, with coefficient of variations less than 18.9%. The strip was used to test real samples and verified by UPLC-MS/MS method with the good agreement (R² was 0.9825), indicating this novel strip immunoassay is accurate and reliable.

A rapid and sensitive immunoassay for the simultaneous detection of imidacloprid and thiacloprid was developed by using magnetic nanoparticles (MNPs) and upconversion nanoparticles (UCNPs). The UCNPs of NaYF₄:Yb, Er and NaYF₄:Yb, Tm were synthesized and conjugated with anti-imidacloprid monoclonal antibody (mAb) and anti-thiacloprid mAb as signal labels, while the MNPs were conjugated with antigens of thiacloprid and imidacloprid as separation elements. The fluorescence intensities of Yb/Er- and Yb/Tm-doped UCNPs were detected simultaneously in 544 nm and 477 nm under the excitation of NIR light (980 nm). The amounts of mAb-conjugated UCNPs that were separated by antigen-conjugated MNPs were determined based on competitive immunoassays. Under the optimal conditions, the 50% inhibiting concentration (IC₅₀) and limit of detection (LOD, IC₁₀) were 5.80 and 0.32 ng/mL for imidacloprid and 6.45 and 0.61 ng/mL for thiacloprid, respectively. The immunoassay exhibited negligible cross-reactivity with analogs of imidacloprid and thiacloprid except imidaclothiz (86.2%). The average recoveries of imidacloprid and thiacloprid in environmental and agricultural samples, including paddy water, soil, pears, oranges, cucumbers, and wheat, ranged from 78.4 to 105.9% with relative standard deviations (RSDs) of 2.1–11.9% for imidacloprid and ranged from 82.5 to 102.3% with RSDs of 1.0–16.5% for thiacloprid. In addition, the results of the immunoassay correlated well with high-performance liquid chromatography for the detection of the authentic samples.

We demonstrated the use of a new organometallic nanoprobe for competitive surface-enhanced Raman scattering (SERS) immunoassay devoted to the detection of polycyclic aromatic hydrocarbons (PAH) such as benzo[a]pyrene (BaP) in seawater. The nanoprobes are gold nanoparticles (GNPs) labeled by a Raman reporter, the 5,5′-dithiobis(succinimidyl-2-nitrobenzoate) (DSNB) and functionalized with monoclonal antibodies anti-BaP. The antibodies are bound with a high specificity to the analyte while the GNPs enhanced the Raman scattering of the DSNB. This type of immunoassay involved the grafting of BaP onto a sensing surface. Thus, NH₂-terminated self-assembled monolayer is formed on the surface of gold substrate using cysteamine. Amines finally reacted with 6-formylbenzo[a]pyrene. So, this SERS detection involves four steps: (i) the nanoprobes are incubated with the sample; (ii) a drop of the mixture is then put onto the substrate; (iii) the surface is rinsed; and (iv) the surface is analyzed by Raman spectroscopy. To synthesize the nanoprobes, firstly, we prepared GNPs according to Frens’ method. Then, GNPs were spontaneously labeled by the DSNB Raman reporter, thanks to a strong gold-sulfur interaction. Thereafter, BaP antibodies were cross-linked to the DSNB labeled GNPs by reaction of proteins primary amino groups with N-hydroxyl succinimide (NHS). Before use in SERS detection, their activity was controlled by surface plasmon resonance technique. The present method allows us to detect BaP at trace concentration (2 nmol/L). The results demonstrate that the proposed method has a great potential for application in the monitoring of seawater.

A total of 54 raw wastewater samples collected from three urban treatment plants and two slaughterhouses in Tehran, Iran, were assessed for the presence of the Giardia cysts using immunofluorescence with monoclonal antibodies. To characterize the cysts at the molecular level, the three genetic loci were amplified and sequenced. The assemblages A (37.5 %) and E (58.3 %) were detected in livestock wastewater samples. Assemblage A, which is composed of only G. duodenalis genotype, was detected in 100 % of urban wastewater samples. The subassemblages A2, A3, A-I, A-II, and E3 were identified with β-giardin, triose phosphate isomerase, and glutamate dehydrogenase genes. This study is the first to report on G. duodenalis genotypes in aquatic environmental samples in Iran.

Diethylstilbestrol (DES) is used as a kind of animal feed additive and affects people’s health through the food chain. The purpose of this study is to detect the residue level of DES in 576 human urine samples directly. DES-BSA was used to immunize Balb/c mice. The monoclonal antibody was produced by hybridoma that was screened through cell fusion techniques. Finally, we developed the indirect competitive ELISA method to analyze 576 human urine samples from Zhejiang Province, China. The IC₅₀ of this method was 3.33 ng/mL. The LOD and LOQ were 0.16 and 0.54 ng/mL. Linear range of the standard curve was from LOD to 12.50 ng/mL. There was no cross-reactivity with two kinds of estrogens and two structural analogs with DES. Five hundred seventy-six urine samples were analyzed by the indirect competitive ELISA method, and the detection rate was 98.78%. The mean concentration and geometric mean were 4.70 and 3.50 ng/mL. The indirect competitive ELISA method based on monoclonal antibody was sensitive and reliable for the detection of DES in human urine samples. The results warned us to pay more attention to human health and food safety.

The main aim of the study was to test for the presence of Legionnaires’ disease-causing microorganisms in air-conditioned buildings in Kuwait using molecular technologies. For this purpose, 547 samples were collected from 38 cooling towers for the analysis of Legionella pneumophila. These samples included those from water (n = 178), air (n = 231), and swabs (n = 138). Out of the 547 samples, 226 (41 %) samples were presumptive positive for L. pneumophila, with L. pneumophila viable counts in the positive water samples ranging from 1 to 88 CFU/ml. Of the Legionella culture-positive samples, 204 isolates were examined by latex agglutination. These isolates were predominately identified as L. pneumophila serogroup (sg) 2–14. Using the Dresden panel of monoclonal antibodies, 74 representatives isolates were further serogrouped. Results showed that 51 % of the isolates belonged to serogroup 7 followed by 1 (18 %) and 3 (18 %). Serogroups 4 (4 %) and 10 (7 %) were isolated at a lower frequency, and two isolates could not be assigned to a serogroup. These results indicate the wide prevalence of L. pneumophila serogroup 7 as the predominant serogroup at the selected sampling sites. Furthermore, the 74 L. pneumophila (sg1 = 13; sg3 = 13; sg4 = 3; sg7 = 38; sg10 = 5; sgX = 2) isolates were genotyped using the seven gene protocol sequence-based typing (SBT) scheme developed by the European Working Group for Legionella Infections (EWGLI). The results show that Legionella isolates were discriminated into nine distinct sequence typing (ST) profiles, five of which were new to the SBT database of EWGLI. Additionally, all of the ST1 serogroup 1 isolates were of the OLDA/Oxford subgroup. These baseline data will form the basis for the development of a Legionella environmental surveillance program and used for future epidemiological investigations.

BACKGROUND, AIM, AND SCOPE: Okadaic acid (OA) and analogues of dinophysistoxin (DTX) are key diarrheic shellfish poisoning (DSP) toxins, which possibly arouse DSP symptoms by consuming the contaminated shellfish. Because of the stable toxicity in high temperature and the long-term carcinogenicity, the outbreaks of DSP related to consumption of bivalve mollusks contaminated by DSP toxins pose a hazard to public health. Therefore, it is worth developing a fast and reliable analytical method for the detection of OA and analogues in shellfish. In this paper, an indirect competitive enzyme-linked immunosorbent assay (ELISA) (icELISA) for detecting OA and DTX-1 in seafood was developed based on monoclonal antibody (McAb). METHODS: The OA was conjugated to human immunoglobulin G (IgG) and bovine serum albumin (BSA) by the active ester method as the immune antigen and the detective antigen. The spleen cells from BALB/c mice immunized with OA-IgG were fused with SP2/0 myeloma cells. A hybridoma cell line, which secreted McAb against OA, was selected by “limiting dilution” cloning. An icELISA was developed based on immobilized conjugate (OA-BSA) competing the McAb with the free OA in seafood sample. RESULTS: A hybridoma cell line, which secreted IgG1 subclass monoclonal antibody (McAb) against OA, was selected. The IC₅₀ of the McAb for OA and dinophytoxin-1 (DTX-1) were 4.40 and 3.89 ng/mL, respectively. Based on the McAb, an indirect competitive ELISA for detection of OA and DTX-1 in seafood was developed. The regression equation was y = 54.713x − 25.879 with a coefficient correlation of R ² = 0.9729. The linear range and the limit of detection were 0.4–12.5 and 0.45 ng/mL, respectively. The average recovery of OA and DTX-1 spiked shellfish was 82.29% with the coefficient of variation of 7.67%. CONCLUSION: The developed icELISA is a fast, sensitive, and convenient assay for detecting of total amount of OA and DTX-1 in seafood.