Enhanced degradation of emerging contaminants by Far-UVC photolysis of peracetic acid: Synergistic effect and mechanisms.

Krypton chloride (KrCl*) excimer lamps (222 nm) are used as a promising irradiation source to drive ultraviolet-based advanced oxidation processes (UV-AOPs) in water treatment. In this study, the UV222/peracetic acid (PAA) process is implemented as a novel UV-AOPs for the degradation of emerging contaminants (ECs) in water. The results demonstrate that UV222/PAA process exhibits excellent degradation performance for carbamazepine (CBZ), with a removal rate of 90.8 % within 45 min. Notably, the degradation of CBZ in the UV222/PAA process (90.8 %) was significantly higher than that in the UV254/PAA process (15.1 %) at the same UV dose. The UV222/PAA process exhibits superior electrical energy per order (EE/O) performance while reducing resource consumption associated with the high-energy UV254/PAA process. Quenching experiments and electron paramagnetic resonance (EPR) detection confirm that HO• play a dominant role in the reaction. The contributions of direct photolysis, HO•, and other active species (RO• and 1O2) are estimated to be 5 %, 88 %, and 7 %, respectively. In addition, the effects of Cl-, HCO3-, and humic acid (HA) on the degradation of CBZ are evaluated. The presence of relatively low concentrations of Cl-, HCO3-, and HA can inhibit CBZ degradation. The UV222/PAA oxidation process could also effectively degrade several other ECs (i.e., iohexol, sulfamethoxazole, acetochlor, ibuprofen), indicating the potential application of this process in pollutant removal. These findings will propel the development of the UV222/PAA process and provide valuable insights for its application in water treatment.

Sequential combination of pre-chlorination and powdered activated carbon adsorption on iodine removal and I-THMs control in drinking water.

Combining pre-oxidation with activated carbon adsorption was explored as an ideal approach for removing iodine from water source to eliminate the formation of Iodinated trihalomethanes (I-THMs). Compared with permanganate and monochloramine, chlorine is more suitable as pre-oxidant to obtain higher active iodine species (HOI/I2). Active iodine species adsorption using both powdered activated carbon (PAC) and granular activated carbon (GAC) can be well fitted the pseudo-second-order kinetic model indicating that chemical adsorption was the dominant mechanism for HOI/I2 adsorption. The average pore size of activated carbons was the most strongly correlated with the adsorption capacity (R2 > 0.98), followed by methylene blue (R2 > 0.76), pore volume (R2 > 0.70) and iodine number (R2 > 0.67). Moreover, three models, including intraparticle diffusion, Byod kinetic, and diffusion-chemisorption were used to illustrate the mechanisms of HOI/I2 adsorption. Chemical adsorption was the dominant mechanism for HOI/I2 adsorption. In summary, at the molar ratio of [NaClO] and [I-] as 1.2, pre-chloriantion time of 5 min, subsequently dosage of 15 mg/L of PAC E with 20 min adsorption can remove 79.8% iodine. In addition, the combined process can eliminate 61%-87.2% of I-THMs in the subsequent chlor(am)ination. The results indicate that pre-chlorination combined with PAC can effectively removed HOI/I2 and attenuate I-THMs formation in the subsequent disinfection process.

Enhanced coagulation and oxidation by the Mn(VII)-Fe(III)/peroxymonosulfate process: Performance and mechanisms.

To improve the performance of the conventional coagulation process, a permanganate (Mn(VII)) pre-oxidation combined with Fe(III)/peroxymonosulfate (PMS) coagulation process (Mn(VII)-Fe(III)/PMS) that can significantly improve the removal of dissolved organic carbon (DOC), turbidity, and micropollutants is proposed in this study. Compared with conventional Fe(III) coagulation, the Mn(VII)-Fe(III)/PMS process can also significantly enhance the removal of iohexol and sulfamethoxazole in raw water. During this process, the primary reduction product, Mn(IV), after Mn(VII) pre-oxidation was adsorbed on the floc surfaces and involved in the Fe(III)/PMS process. The natural organic matter (NOM) in raw water mediated the redox cycle of iron. The synergistic effect of NOM, Fe, and Mn facilitated the redox cycle of Mn(III)/Mn(IV) and Fe(III)/Fe(II) to promote the activation of PMS. The sulfate radical (SO4•-) played an important role in the degradation of micropollutants. The formation potential of the detected volatile disinfection by-product (DBP) during the subsequent chlorination was reduced by 21.9% after the Mn(VII)-Fe(III)/PMS process. This study demonstrated the promising application of the Mn(VII)-Fe(III)/PMS process for coagulation and micropollutant control and illustrated the reaction mechanism. This study provides guidance for improving conventional drinking water treatment processes.

Micropollutant removal and disinfection byproduct control by sequential peroxymonosulfate-UV treatment in water: A case study with sulfamethoxazole.

UV/peroxymonosulfate (UV/PMS) advanced oxidation process has attracted significant attention for removal of micropollutants in water. However, during practical water treatment applications, the PMS treatment must be performed before the UV treatment to achieve full contact. In this study, sulfamethoxazole (SMX) was selected as the target micropollutant. Four different operational approaches, including UV alone, PMS alone, simultaneous UV/PMS and sequential PMS-UV, were compared for their differences in SMX removal and disinfection by-product (DBP) formation potentials during chlorine-driven disinfection. Among the four approaches, UV/PMS and PMS-UV achieved over 90% removal efficiencies for SMX without substantial differences. For raw water, the trichloronitromethane (TCNM) formation potential after treatment with PMS-UV was lower than that after UV/PMS treatment. The time interval over which the PMS-UV process was conducted had little effect on the final removal efficiency for SMX. However, a brief (5 min) pre-PMS treatment significantly reduced the TCNM formation potential and the genotoxicity from DBPs. The formation risk for TCNM during chlorination increased markedly with increasing PMS dosages, and the appropriate dosage under these experimental conditions was suggested to be 0.5-1.0 mmol/L. Under alkaline conditions, PMS-UV treatment can enhance SMX degradation as well as dramatically reduced the formation potentials for haloketones, haloacetonitriles and halonitromethanes. This study suggests that proper optimization of UV/PMS processes can remove SMX and reduce its DBP formation.

Insight into the formation of iodinated trihalomethanes during chlorination, monochloramination, and dichloramination of iodide-containing water.

In this study, the formation of iodinated trihalomethanes (I-THMs) was systematically evaluated and compared for three treatment processes - (i) chlorination, (ii) monochloramine, and (iii) dichloramination - under different pH conditions. The results demonstrated that I-THM formation decreased in the order of monochloramination > dichloramination > chlorination in acidic and neutral pH. However, the generation of I-THMs increased in the dichloramination < chlorination < monochloramination order in alkaline condition. Specifically, the formation of I-THMs increased as pH increased from 5 to 9 during chlorination and monochloramination processes, while the maximum I-THM formation occurred at pH 7 during dichloramination. The discrepancy could be mainly related to the stability of the three chlor (am) ine disinfectants at different pH conditions. Moreover, in order to gain a thorough insight into the mechanisms of I-THM formation during dichloramination, further investigation was conducted on the influencing factors of DOC concentration and Br-/I- molar ratio. I-THM formation exhibited an increasing and then decreasing trend as the concentration of DOC increased from 1 to 7 mg-C/L, while the yield of I-THMs increased with increasing Br-/I- molar ratio from 5:0 to 5:10. During the three processes mentioned above, similar I-THM formation results were also obtained in real water, which indicates that the excessive generation of I-THMs should be paid special attention during the disinfection of iodide-containing water.

Enhanced degradation of emerging contaminants by permanganate/quinone process: Case study with bisphenol A.

Permanganate (Mn(VII)) is widely used as a mild oxidant in water treatment. However, the reaction rates of some emerging contaminants with Mn(VII) are extremely low. In this study, benzoquinone (BQ), a redox mediator with the important component in dissolved organic matter (DOM), enhanced the oxidation of bisphenol A (BPA) by Mn(VII) in a wide pH range of 4.0-10.0. The redox cycle of BQ would produce semiquinone radicals, which could act as ligands to stabilize the formed Mn(III) in the system to promote the oxidation of BPA. Notably, the presence of BQ might promote the formation of MnO2. A novel mechanism was proposed that singlet oxygen (1O2), Mn(III)-ligands (Mn(III)-L) and in-situ formed MnO2 were the main contributors to accelerate BPA degradation in the Mn(VII)/BQ system. Under acidic conditions, the in-situ formed MnO2 involved in the redox reaction and part of the Mn(IV) was reduced to Mn(III), indicating that the electron transfer of BQ promoted the formation of active Mn species and enhanced the Mn(VII) oxidation performance. Semiquinone radicals generated by BQ transformation would couple with the hydrogen substitution products of BPA to inhibit BPA self-coupling and promote the ring-opening reactions of BPA. Mn(VII)/BQ had better effect in raw water than in pure water, indicating that the Mn(VII)/BQ system has high potential for practical application. This study provided insights into the role of DOM in enhancing the Mn(VII) oxidation in water treatment.

The formation, analysis, and control of chlor(am)ination-derived odor problems: A review.

Odors and tastes have become universal problems related to drinking water quality. In addition to the typical odor problems caused by algae or microorganisms, the occurrence of odors derived from drinking water disinfection have attracted attention. The chlor(am)ination-derived odor substances have certain toxicity and odor-causing characteristics, and would enter the tap water through water distribution systems, directly affecting drinking water safety and customer experience. This study provided a comprehensive overview of the occurrence, detection, and control of odor substances derived from drinking water chlor(am)ination disinfection. The occurrence and formation mechanisms of several typical types of disinfection derived odor substances were summarized, including haloanisoles, N-chloroaldimines, iodotrihalomethanes, and halophenoles. They are mainly derived from specific precursors such as halophenols, anisoles, and amino acids species during the disinfection or distribution networks. In addition, the change of disinfectant during chlor(am)ination was also one of the causes of disinfection odors. Due to the extremely low odor threshold concentrations (OTCs) of these odor substances, the effective sample pre-enrichment for instrument identification and quantification are essential. The control strategies of odor problems mainly include adsorption, chemical oxidation, and combined processes such as ozonation and biological activated carbon processes (O3/BAC) and ultraviolet-based advanced oxidation processes (UV-AOPs). Finally, the challenges and possible future research directions in this research field were discussed and proposed.

The application of UV-C laser in persulfate activation for micropollutant removal: Case study with iodinated X-ray contrast medias.

A novel light source UV-C laser was applied in persulfate (PS) activation to effectively remove iodinated X-ray contrast medias (ICMs) including iohexol (IOX), iopamidol (IPM) and diatrizoate (DTZ) in this study. Significant ICMs degradation was observed in UV-C laser/PS systems with pseudo first-order rate constants of 0.022-0.067 s-1. Sulfate radicals (SO4•-) were the main active species in the three ICMs degradation, and the steady-state concentrations ([SO4•-]ss) were 3.629 × 10-11 M (IOX), 1.702 × 10-11 M (IPM) and 1.148 × 10-11 M (DTZ), respectively. Under the high intensity of UV-C laser, the optimal reaction efficiency was achieved at pH = 7.0 with PS concentration of 1.0 mM, and the degradation efficiency for IOX reached 93.8% within only 40 s. Both bicarbonate and chloride ions could inhibit the three ICMs degradation and the inhibition rate increased with the increase of ions concentration. The kinetic models were established and the steady-state concentrations of radicals were calculated. Density functional theory (DFT) calculations combined with experiments were used to derive the reaction pathways for three ICMs. Cyclic voltammetry measurements detected a lower redox potential peak in IOX degradation, revealing the existence of electron shuttles under the UV-C laser irradiation to promote the redox reaction. This study is the first report of UV-C laser activation of persulfate. It is a new advanced oxidation process mediated by very effective photolysis and active species formation.

Iodinated trihalomethanes formation in iopamidol-contained water during ferrate/chlor(am)ination treatment.

Iopamidol is a commonly used iodinated X-ray contrast media in medical field, and its residue in water can react with disinfectants to form highly toxic iodinated disinfection by-products (I-DBPs). This study investigated the degradation of iopamidol and formation of DBPs, especially iodinated trihalomethanes (I-THMs), during ferrate (Fe(VI)) pre-oxidation and subsequent chlor(am)ination under raw water background. It was found that iopamidol degradation efficiency in raw water by Fe(VI) at pH 9 could reach about 80%, which was much higher than that at pH 5 and pH 7 (both about 25%). With Fe(VI) dose increasing, iopamidol removal efficiency increased obviously. During the iopamidol degradation by Fe(VI), IO3- was the dominant product among all the iodine species. After pre-treated by Fe(VI), yields of THM4 and I-THMs can be reduced in subsequent chlor(am)ination. Besides, pH was a crucial factor for Fe(VI) pre-oxidition controlling DBPs. With the pH increasing from 5 to 9, the yield of THM4 kept increasing in subsequent chlorination but showed the highest amount at pH 6 in subsequent chloramination. The yield of I-THMs increased first and then decreased with the increase of pH in both subsequent chlorination and chloramination. I-THM concentrations in chlorinated samples were lower than chloraminated ones under acidic conditions but became higher under neutral and alkaline conditions. The total CTI of THMs during Fe(VI)-chloramination was higher than that during Fe(VI)-chlorination under neutral condition, but sharply decreased under alkaline conditions. In summary, Fe(VI)-chloramination subsequent treatment under alkaline conditions should be an effective method for iopamidol removal and DBP control.

Formation of disinfection by-products in a UV-activated mixed chlorine/chloramine system.

In recent years, ultraviolet (UV) irradiation coupled with chlor(am)ination process is ubiquitous in secondary water supply systems in many cities of China. However, the disinfection by-products (DBPs) formation in a UV-activated mixed chlorine/chloramine system (MCCS) still remains unclear. In this study, the DBPs formation in a UV-activated MCCS was systematically investigated, considering influencing factors including the mass ratios of free chlorine to NH2Cl, UV irradiation, pH values, NOM types, Br- concentration and toxicity of the DBPs. Results indicated that DBPs formation decreased remarkably as mass ratio of free chlorine to NH2Cl changed from 5:0 to 0:5. The DBPs formation in humic acid (HA)-containing water was the highest, followed by those in fulvic acid (FA) and algal organic matter (AOM). Besides, better control of the DBP-related calculated toxicity can be achieved in acidic conditions regardless of the UV irradiation. Furthermore, in the presence of Br-, a significant reduction of DBPs formation could be achieved in a UV-activated MCCS. The findings also demonstrated that DBPs formation in real water can be effectively reduced at high UV fluence in a MCCS.

[Degradation of RBK5 by High Crystallinity Mn-Fe LDH Catalyst Activating Peroxymonosulfate].

High crystallinity Mn-Fe LDH was synthesized by improved co-precipitation combined with the hydrothermal method and was utilized as a catalyst for peroxymonosulfate (PMS) activation to degrade reactive black 5. The high crystal purity and clear lamellar structure were characterized by X-ray diffraction (XRD), scanning electron microscope (SEM), energy dispersive X-ray spectrometer (EDS), Fourier transform infrared spectroscopy (FTIR), and X-ray photoelectron spectroscopy (XPS). The operating parameters such as Mn/Fe molar ratio, catalyst dosage, PMS concentration, and initial pH value on the absorption efficiency, catalytic degradation, and reaction kinetics of RBK5 were also investigated. The results demonstrated that high crystallinity Mn-Fe LDH has good adsorption capacity and high catalytic efficiency. The degradation efficiency of RBK5 (20 mg·L-1) could reach 86% within 90 min when the Mn/Fe molar ratio was 1, the catalyst dosage was 0.2 g·L-1, the PMS concentration was 1 mmol·L-1, and the initial pH value was 7.0. The reaction process follows pseudo-first-order reaction kinetics (R2>0.9). In addition, the quenching experiment indicated that SO4-·and·OH were the main active species that degraded RBK5 from the Mn-Fe LDH/PMS system. The XPS analysis of the catalyst before and after the reaction confirmed the synergistic effect between Mn and Fe. The charge balance between Mn(Ⅱ) and Fe(Ⅲ) on the LDH surface and CO32- in layers stabilized the structure, thus promoting the synergistic effect of Mn and Fe on the lamellar surface and improving the activation efficiency of PMS by Mn-Fe LDH. Three-dimensional fluorescence and the UV-Vis scanning spectral analysis were preliminarily discussed to understand the degradation process of RBK5.

[Degradation of RBk5 with Peroxymonosulfate Efficiently Activated by N-Doped Graphene].

The large loss of catalysts and secondary pollution problems are bottlenecks for the utilization of persulfate advanced oxidation processes. Thus, a modified Hummers method combined with a hydrothermal method was used to prepare N-doped graphene as a catalyst for peroxymonosulfate (PMS) activation. The produced sulfate radical (SO4-·) and hydroxyl radical (·OH) were able to degrade RBk5. N-doped graphene was characterized by Fourier transform infrared, X-ray photoelectron spectroscopy, Raman spectroscopy, and transmission electron microscopy. The influences of vital parameters (i. e., initial pH, catalyst dosage, and PMS dosage) on RBk5 removal were investigated systematically to examine the catalytic performance. The results showed that the N element doping can effectively improve the catalytic activity of graphene, and the activity is greatly affected by the N doping ratio. The initial pH of the wastewater had no significant effect on the degradation efficiency. Under the condition of 1.5 g·L-1 catalyst dosage and 0.3 g·L-1 PMS dosage, the removal rate of RBk5 dye reached 99% after 25 min of reaction. The reaction process accorded with first-order reaction kinetics. Radical quenching experiments were done and indicated that the degradation of RBk5 in N-doped graphene/PMS systems was a surface reaction, and SO4-· and ·OH were identified as the main radical species. The catalyst exhibited excellent stability over five successive degradation cycles.

Single nucleotide polymorphisms characteristics of the Yersinia Pestis in China / 中国地方病学杂志

Objective To study the single nucleotide polymorphisms (SNP) characteristics of Yersinia pestis strains from different natural foci in China.Methods Genome-wide comparison was done to find SNP sites by the Mummer program among 9 Yersinia pestis genome which was downloaded from NCBI.Then 13 genic fragments including 19 SNP sites were amplified by PCR and sequenced in 133 Yersinia pestis strains,and the results were cluster analyzed with the BioNumerics software.Results Three thousand seven hundred and eighty sequence variation sites were found by genome-wide comparison.Using the different combinations of SNP sites,UPGMA cluster analysis revealed obvious geographic regional and eco-aggregation characteristics of Yersinia pestis strains isolated from China.Conclusions As relatively stable genetic markers,SNP can better reflect the genome characteristics of Yersinia pestis in different plague natural foci of China.

Comparative genomic analysis of gene variations of two Chinese Yersinia pestis isolates from vaccine strain EV76 / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate genomic variations of two Chinese Yersinia pestis isolates that were isolated from different plague foci obtained from vaccine strain EV76 from the Yunnan province of China.</p><p><b>METHODS</b>A microarray containing 12 000 probes covering the entire genome of seven Yersinia pestis and two Yersinia pseudotuberculosis strains, was used. PCR assays were performed to confirm microarray results.</p><p><b>RESULTS</b>The gene variations detected included the absence of five genes related to the synthesis of betaine in both EV76 and another sequenced attenuated strain, KIM D27. Several genes related to phage-related membrane proteins were found to be absent in the Antiqua biovar Yunnan strain, 485, which was isolated from a rodent plague foci.</p><p><b>CONCLUSION</b>These findings provide initial insight into the distinct strains isolated from natural foci, within their genomic context, including Yunnan Y. pestis strains. This information will be used therefore to establish subsequent comparisons of these sequences with published complete genomes of other strains.</p>

Comparison of Yersinia pestis genomes from a new natural foci in China / 中国地方病学杂志

Objective Measurement and analysis of the complete genome sequences of Yersinia Pestis from a new plague natural foci and adjacent foci in China, to know the genetic relationship among the epidemic strain isolated in Yulong (D 106004) and Jianchuan strains (D 182038) and the Tibetan strain ( Z 176003 ). Methods Three complete genome sequences were sequenced using the whole-genome shotgun and Solexa method and comparative genomics analysis was done among the three sequences. Genome comparative analysis among the coding sequences was done by BLAST software, SNPs finding was done by the program, genome rearrangements were analyzed using MAUVE software. Results All of the genomes of Yersinia pestis strains D182038, D106004 and Z176003 consist of a single circular chromosome and three virulence plasmids, pMT1, pCD1 and pPCP1. They had similar characteristics in chromosome and plasmid features, and there were no significant difference in coming sequence (CDS) of the cluster of orthologous groups of proteins (COG) functional classification and the number of insertion sequence in the three strains (x2 =3.03, 0.257, all P ＞ 0.05). The comparative genomics results showed that the three bacteria had 2882 genes with 100％ homology, of 3636 genes predicted in D106004, 2994 were identical with D182038's and 3113 with Z176003's, and of which 240 had 90％ homology with D182038's and 200 with Z176003 's. Synonymous single nucleotide polymorphisms(sSNPs) were 59 and 68, and non-synonymous SNPs(nsSNPs) were 104 and 203 between strains D106004 and Z176003/D182038. There were 11 segments rearrangements between D106004 and Z176003, which was less than 16 segments rearrangements between D106004 and D182038. ConclusionsThe three strains are highly homologous, the Yulong strain has more similarity with Tibet strain than with Jianchuan strain, the strain from Yulong foci may be evolved from Tibet foci.

Study on the identification characteristics of rRNA genes on Yersinia pestis / 中华流行病学杂志

Objective To study the identification characteristics of rRNA genes on Yersinia (Y.)pestis.Methods By means of comparative genomics,we compared the rRNA genome sequences of nine completely sequenced strains of Y. pestis isolated from China and other countries by Clustal W software.we also compared the 2000 bp sequence adjacent to the rRNA genes,rRNA genes and 16S-23S rRNA spacer region respectively to determine the identification features of rRNA genes for Y. pestis.Results There were 6 rRNA gene clusters in the strains of D182038,D106004,Z176003 and CO92 respectively(6 copies strain).There were 7 rRNA gene clusters in the strains of 91001,KIM,Nepa1516,Antiqua and Pestoides F(7 copies strain).According to the 2000 bp sequence,13 types of rRNA gene clusters could classify the strains between the 6 copies and 7 copies.There were 4 types of tRNA gene among the 16S-23S rRNA spacer region that could classify the strains among the 6 copies and 7 copies strains respectively.The number of point mutation among the 23S rRNA gene was statistically different in some copies under ANOVA analysis(F=0.548,P=0.815>0.05 among the strains and F=5.228,P<0.01 among the copies).Conclusion The 2000 bp sequence adjacent to the rRNA genes,tRNA gene and 23S rRNA gene sequence could serve as the identification sign of rRNA genes for classifing the strains of Y. pestis.

Assessment of comparative genomic hybridization experiment by an in situ synthesized CombiMatrix microarray with Yersinia pestis vaccine strain EV76 DNA / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>The quality of microarray data influences the accuracy of comparative genomic analyses to a large extent. To ensure that the results obtained by using an in situ synthesized microarray are accurate, data quality is to be assessed by evaluating the melting temperature (Tm) of probes, probability of false synthesis rates, and fragmentation of labeled targets.</p><p><b>METHODS</b>DNA from the Yersinia pestis vaccine strain EV76 was used for microarray analyses. Microarray results were confirmed by PCR. Statistical and bioinformatics methods were employed to perform microarray data analyses and evaluation.</p><p><b>RESULTS</b>Correlation coefficients of the three datasets were above 0.95 after two-time stripping and hybridization with a labeled DNA with the size of fragmentation being 200 bp - 2 kb, which showed that the hybridization results were highly reproducible. Correlation coefficients were lower with the values ranging from 0.87 to 0.92 between the datasets generated from hybridization with different sizes of the labeled DNA fragment. For the relationship between Tm and signal intensity, there was a different distribution of Tm in the lowest 300 or 3,000 probes with a range of 70 °C-72 °C and the highest 300 or 3,000 probes with a range of 72 °C-74 °C.</p><p><b>CONCLUSION</b>The results of this study suggest that the initial microarray design may affect the accuracy of final analyses and that the probe Tm and the size of the labeled fragment may be the two factors of the greatest importance.</p>

Clone expression of the caf1 gene of Yersinia pestis and immunological evaluation on recombinant F1 antigen / 中华流行病学杂志

Objective To get recombinant F1 antigen (rF1) and to construct the detection dipstick of plague antibody. Methods The cafl gene removing the signal peptide coding sequence was cloned into plasmid pET32a ( +) by double-digested sites of BamHI and Not I. Recombinant plasmid caf1-pET32a(+) was transformed into BL21 (DE3) and the rFl was expressed. Expression products were purified by affinity chromatography. Dual detection dipstick of plague antibody was constructed with purified rF1 and natural F1, and evaluated with 528 human serum samples of Zhejiang province. Results The fusion protein rF1 of 35.5 KD was expressed by BL21 strains containing caf1-pET32a( + ). The sensitivity of rF1 showed equivalent to or higher than the natural Fl antigen in detecting plague antibody. It seemed that there was a better consistency of 97.9% (k= 0.466) when 528 human sera was detected by rF1 and natural F1. Conclusion We successfully extracted the rF1 with good immunological activity that might be used to detecting Yersinia pestis.

16S rRNA gene clone library analysis of bacterial communities of the tick with infection of 4 species of pathogens / 中国地方病学杂志

Objective To develop the method of 16S rRNA gene clone library for tick bacterial flora analysis, and to analyze the detection effective of pathogens in tick and capacity of bacterial flora diversity. Methods Primers were designed according to the specific gene of Borrelia burgdorferi, Bartonella henselae, Anaplasma phagocytophilum, Ehrlichia chaffeensis and templates were choosen by positive PCR result to amplify the DNA extracted from the ticks. One set of primers targeting 16S rRNA gene conserved region were chosen to amplify certain fragments, DNA extraction, PCR reaction, cloning and sequencing. Nucleotide sequences were compared with GenBank database. Calculated Coverage values of clone library and Shannon-Wiener diversity index. Results Sixteen defined genus-or species-bacteria were detected in 103 valid sequences. Eight species were edge type (Clone No. > 5). Three kinds of pathogens were identified (Borrelia burgdorferi, Bartonella henselae and Rickettsia sp). Three kinds of pathogens were not edge type(Clone No. < 5). Coverage value was 96.11%, and Shannon-Wiener index was 2.40. Analysis results of cloning sequence showed that tick-parasitic bacteria mainly were α and γ deformation mycetes which accounted for 56.25% (9/16). Conclusions The 16S rRNA gene sequences technology could make relative quantitative of bacterial flora, and detect many kinds of pathogens in tick. It's a good method for detection of pathogens and bacterial flora analysis.

Locus-specific analysis of IS100 insertion within Yersinia pestis genome / 中国地方病学杂志

Objective To study the genotyping distribution of the Yersinia pestis(Y.pestis)strains by characterizing the diversity of the insertion sequence IS100 within the Y.pestis genome.Methods Derived fromthe known sequence of oriental strain CO92,5 pairs of locus-specific primers originating from both sides of the adjacent region of IS100 copies were designed,and two other complementary primers inside the IS100 sequence were designed to correspond with the outer primers.Then,91 Y.pestis strains and l pseudotubebculosis strain were tested by the specific PCR method using the primers described above and the PCR products were conformed by the sequence analysis,then further analysis WaS performed after the IS100 status was marked on the map of the plague focus type of china.Results The 91 Y.pestis strains had different IS100 status in their genome on tested loci.some possessed IS100 insertion,some didn't,and others changed their genome constitution.The IS100 possession on the 5 loci also suggested a distribution of regionality.Conclusion The analysis of some IS100 insertion element loci reveals that the IS100 genotyping distribution is consistent with the plague focus of type of China.And IS100genotyping pattern of the Y.pestis stains well reflects its genome constitution and the high flowability in its natural evolution.