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1.
Huan Jing Ke Xue ; 42(4): 1939-1945, 2021 Apr 08.
Artículo en Chino | MEDLINE | ID: mdl-33742829

RESUMEN

In order to study the effects of chlorine dioxide (ClO2) disinfection on the super antibiotic resistance genes (SARGs), the final effluents before and after chlorine dioxide were sampled throughout one year in a wastewater treatment plant (WWTP). The bacteria and extracellular nucleic acid were collected using microporous membrane filtration and nucleic acid adsorption particles, respectively. A total of 9 SARGs was detected through a quantitative real-time polymerase chain reaction (qPCR). The results revealed that both intracellular and extracellular NDM-1, MCR-1, and MEC-A could be positively detected in the samples. Overall, ClO2 disinfection enhanced the relative abundance of the iSARGs (P<0.05), exhibiting a seasonal pattern, and increasing in the spring, summer, and autumn. In spring, it improved the most, up to twice the abundance. No SARGs were detected positive in the winter, either intracellularly or extracellularly. There was no significant variation in the concentrations of eSARGs before and after ClO2 disinfection. Therefore, ClO2 disinfection cannot effectively remove iSARGs and eSARGs in the final effluent from the WWTP.


Asunto(s)
Compuestos de Cloro , Desinfectantes , Purificación del Agua , Antibacterianos/farmacología , Cloro , Compuestos de Cloro/farmacología , Desinfectantes/farmacología , Desinfección , Farmacorresistencia Microbiana/genética , Óxidos/farmacología
2.
Sci Total Environ ; 743: 140641, 2020 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-32653709

RESUMEN

Human enteric virus occurrence in bathing beaches poses a potential health risk to swimmers. They may come from several sources, but the understanding of the seasonal contribution of contamination sources to virus occurrence is still lacking. Here, the surveillance of human enteric viruses at the First Bathing Beach in Qingdao was performed January-December 2018. The occurrence of Enteric viruses, assayed with quantitative polymerase chain reaction (qPCR), was analyzed at temporal and spatial levels to determine the viral contamination sources. The results showed that only Astroviruses (AstVs) and Adenoviruses (HAdVs) were found in the swimming area. Their occurrence correlated significantly with the sewage-polluted area, but HAdVs were only found in autumn and AstVs in spring. Meanwhile, enteric viruses in the swimming area showed significantly higher levels than the surrounding area, particularly AstVs in summer with the swimmer crowd. All these data imply that sewage discharge and swimmers co-contribute to the viral occurrence in a seasonal pattern, with the former being more focused in warm seasons (spring and autumn) and the latter in hot seasons (summer). These results indicate that sewage discharge and crowd swimmers, as unsafe swimming conditions, should be avoided to improve public health at the bathing beaches.


Asunto(s)
Microbiología del Agua , Agua , Playas , Monitoreo del Ambiente , Heces , Humanos , Estaciones del Año
3.
ISME J ; 14(7): 1847-1856, 2020 07.
Artículo en Inglés | MEDLINE | ID: mdl-32327733

RESUMEN

Chlorine disinfection to drinking water plays an important role in preventing and controlling waterborne disease outbreaks globally. Nevertheless, little is known about why it enriches the antibiotic resistance genes (ARGs) in bacteria after chlorination. Here, ARGs released from killed antibiotic-resistant bacteria (ARB), and culturable chlorine-injured bacteria produced in the chlorination process as the recipient, were investigated to determine their contribution to the horizontal transfer of ARGs during disinfection treatment. We discovered Escherichia coli, Salmonella aberdeen, Pseudomonas aeruginosa and Enterococcus faecalis showed diverse resistance to sodium hypochlorite, and transferable RP4 could be released from killed sensitive donor consistently. Meanwhile, the survival of chlorine-tolerant injured bacteria with enhanced cell membrane permeabilisation and a strong oxidative stress-response demonstrated that a physiologically competent cell could be transferred by RP4 with an improved transformation frequency of up to 550 times compared with the corresponding untreated bacteria. Furthermore, the water quality factors involving chemical oxygen demand (CODMn), ammonium nitrogen and metal ions (Ca2+ and K+) could significantly promote above transformation frequency of released RP4 into injured E. faecalis. Our findings demonstrated that the chlorination process promoted the horizontal transfer of plasmids by natural transformation, which resulted in the exchange of ARGs across bacterial genera and the emergence of new ARB, as well as the transfer of chlorine-injured opportunistic pathogen from non-ARB to ARB. Considering that the transfer elements were quite resistant to degradation through disinfection, this situation poses a potential risk to public health.

4.
Ecotoxicol Environ Saf ; 195: 110461, 2020 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-32182530

RESUMEN

Antibiotic residues in the environment pose a great risk to global public health. They increase antibiotic resistance by enhancing plasmid conjugation among bacteria or mutations within bacterial genomes. However, little is known about whether the putative environmental levels of antibiotics are sufficient to influence plasmid-mediated transformability. In this study, we explored the effect of eight kinds of representative antibiotics and several other compounds on the plasmid transformability of competent Escherichia coli. Only levofloxacin (LEV) at the putative environmental levels was found to facilitate the frequency of PBR322-or RP4-plasmid-mediated transformation by up to 5.3-fold. Additionally, PBR322 transformation frequency could be further enhanced by copper ion or ammonia nitrogen but inhibited by humic acid. However, when competent E. coli was exposed to the minimal inhibitory concentrations (MIC) of the antibiotics, an enhanced plasmid-assimilation ability was observed and plasmid transformation frequency was increased by up to 98.6-fold for all the tested antibiotics. Furthermore, E. coli exhibited a preference for the uptake of plasmids harbouring the resistance genes to the antibiotics it had been exposed to. Among these antibiotics, cephalexin, tetracycline, and kanamycin induced the highest uptake of RP4. The putative environmental levels of LEV enhanced plasmid transformability regardless of the presence of corresponding antibiotic resistance gene (ARG) on the genetic elements, suggesting environmental LEV residues may facilitate dissemination of antibiotic resistance by any plasmid-mediated transformability, thereby posing a great risk to health.


Asunto(s)
Antibacterianos/farmacología , Farmacorresistencia Bacteriana/genética , Escherichia coli/efectos de los fármacos , Escherichia coli/genética , Levofloxacino/farmacología , Transformación Bacteriana/efectos de los fármacos , Plásmidos/efectos de los fármacos
5.
Zhongguo Ying Yong Sheng Li Xue Za Zhi ; 36(6): 668-672, 2020 Nov.
Artículo en Chino | MEDLINE | ID: mdl-33719279

RESUMEN

Objective: To establish a high performance liquid chromatography tandem mass spectrometry (HPLC / MS) method for the simultaneous determination of three antidepressant drugs in feces. Methods: Samples were pretreated with n-hexane isopropanol (95:5, v/v). Gradient elution was carried out with mixed liquid of ultrapure water and acetonitrile as mobile phase and separated by Agilent ZORBAX SB-C18 liquid chromatography column (2.1 mm×100 mm, 3.5 m). The samples were detected by electrospray ionization tandem mass spectrometry and quantified by internal standard method. Results: The recoveries of duloxetine, fluoxetine and escitalopram in fecal samples were 61.6% - 116.5%, with precision of 2.80% - 12.9% (n=5). The correlation coefficients (r) of linear equations were all greater than 0.995. The detection limits were 0.1, 1, and 0.001 µg/g, and the limits of quantification were 0.5, 2 and 0.005 µg/g, respectively. Conclusion: The method is simple and accurate to detect the contents of three antidepressants in feces, such as duloxetine, fluoxetine and escitalopram.


Asunto(s)
Citalopram , Espectrometría de Masas en Tándem , Antidepresivos , Cromatografía Líquida de Alta Presión , Heces
6.
Anal Bioanal Chem ; 411(26): 7039-7049, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31428817

RESUMEN

Antibiotic compounds in natural waters are normally present at low concentrations. In this paper, an easy and highly sensitive screening method using graphene oxide-functionalized magnetic composites (GO@NH2@Fe3O4) combined with matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS) was established for twelve quinolone antibiotics. GO@NH2@Fe3O4 composites were utilized as adsorbents for magnetic solid-phase extraction. This method combines the advantages of magnetic solid-phase extraction and MALDI-TOF MS, which allows for fast detection of quinolones at low concentrations. To improve absorption efficiency, the following parameters were individually optimized: sample acidity, extraction time, amount of adsorbent used, eluent used, and desorption time. Under the optimum conditions, the established method gave a low detection limit of 0.010 mg/L and allowed the high-throughput screening of twelve quinolone antibiotics (enoxacin, norfloxacin, ciprofloxacin, pefloxacin, fleroxacin, gatifloxacin, enrofloxacin, levofloxacin, sparfloxacin, danofloxacin, difloxacin, and lomefloxacin). The proposed method, having an easily prepared sorbent with a high affinity for quinolones and a convenient, high-throughput detection step, has been shown to have merit for the detection of antibiotics in water samples. Graphical abstract Schematic illustration of the (A) preparation of GO@NH2@Fe3O4 and (B) operating procedure for the MSPE and MALDI-TOF MS detection of QNs.


Asunto(s)
Antibacterianos/análisis , Grafito/química , Quinolonas/análisis , Extracción en Fase Sólida/métodos , Contaminantes Químicos del Agua/análisis , Antibacterianos/aislamiento & purificación , Límite de Detección , Imanes/química , Modelos Moleculares , Quinolonas/aislamiento & purificación , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Agua/análisis , Contaminantes Químicos del Agua/aislamiento & purificación
7.
Biomed Environ Sci ; 32(5): 324-333, 2019 May.
Artículo en Inglés | MEDLINE | ID: mdl-31217049

RESUMEN

OBJECTIVE: To investigate the mechanisms underlying ozone-induced inactivation of poliovirus type 1 (PV1). METHODS: We used cell culture, long-overlapping RT-PCR, and spot hybridization assays to verify and accurately locate the sites of action of ozone that cause PV1 inactivation. We also employed recombinant viral genome RNA infection models to confirm our observations. RESULTS: Our results indicated that ozone inactivated PV1 primarily by disrupting the 5'-non-coding region (5'-NCR) of the PV1 genome. Further study revealed that ozone specifically damaged the 80-124 nucleotide (nt) region in the 5'-NCR. Recombinant viral genome RNA infection models confirmed that PV1 lacking this region was non-infectious. CONCLUSION: In this study, we not only elucidated the mechanisms by which ozone induces PV1 inactivation but also determined that the 80-124 nt region in the 5'-NCR is targeted by ozone to achieve this inactivation.


Asunto(s)
Genoma Viral/efectos de los fármacos , Oxidantes Fotoquímicos/farmacología , Ozono/farmacología , Poliovirus/efectos de los fármacos , Inactivación de Virus , Regiones no Traducidas 5' , Animales , Chlorocebus aethiops , Células Vero
8.
Water Res ; 156: 366-371, 2019 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-30928530

RESUMEN

Adaption to adverse environments plays an important role in bacterial survival and is receiving increasing globe attention now. Here, cultivable chlorine-injured Pseudomonas aeruginosa, produced on the chlorination process, was investigated about their resistance to antibiotics. Then, global transcriptional analyses, quantitative PCR (qPCR) validation and antioxidant enzymes measurement were performed to explore the underlying mechanisms. The results showed that chlorine injury enhanced antibiotic resistance in P. aeruginosa and cultivable chlorine-injured P. aeruginosa exposed to 4 mg/L sodium hypochlorite (half of the lethal dose) improved antibiotic resistance against ceftazidime, chloramphenicol and ampicillin by 1.4-5.6 fold. This increase in antibiotic resistance was not hereditable and over expression of the MexEF-OprN efflux pump resulting from oxidative stress contributed to it. These results demonstrate temporal physiological persistence to antibiotics in cultivable chlorine-injured pathogens, suggesting their survival from adverse environments with antibiotic exposure and thereby posing lasting hazards to human health.


Asunto(s)
Cloro , Pseudomonas aeruginosa , Antibacterianos , Cloranfenicol , Farmacorresistencia Microbiana , Humanos
9.
Mol Genet Genomics ; 294(5): 1123-1136, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31030276

RESUMEN

Chromosome segment substitution lines (CSSLs) are ideal materials for identifying genetic effects. In this study, CSSL MBI7561 with excellent fiber quality that was selected from BC4F3:5 of CCRI45 (Gossypium hirsutum) × Hai1 (Gossypium barbadense) was used to construct 3 secondary segregating populations with 2 generations (BC5F2 and BC5F2:3). Eighty-one polymorphic markers related to 33 chromosome introgressive segments on 18 chromosomes were finally screened using 2292 SSR markers which covered the whole tetraploid cotton genome. A total of 129 quantitative trait loci (QTL) associated with fiber quality (103) and yield-related traits (26) were detected on 17 chromosomes, explaining 0.85-30.35% of the phenotypic variation; 39 were stable (30.2%), 53 were common (41.1%), 76 were new (58.9%), and 86 had favorable effects on the related traits. More QTL were distributed in the Dt subgenome than in the At subgenome. Twenty-five stable QTL clusters (with stable or common QTL) were detected on 22 chromosome introgressed segments. Finally, the 6 important chromosome introgressed segments (Seg-A02-1, Seg-A06-1, Seg-A07-2, Seg-A07-3, Seg-D07-3, and Seg-D06-2) were identified as candidate chromosome regions for fiber quality, which should be given more attention in future QTL fine mapping, gene cloning, and marker-assisted selection (MAS) breeding.


Asunto(s)
Cromosomas de las Plantas/genética , Gossypium/genética , Sitios de Carácter Cuantitativo/genética , Mapeo Cromosómico/métodos , Fibra de Algodón , Cruzamientos Genéticos , Genoma de Planta/genética , Fenotipo
10.
J Pathol ; 248(2): 155-163, 2019 06.
Artículo en Inglés | MEDLINE | ID: mdl-30680725

RESUMEN

The cellular mechanisms by which hepatitis C virus (HCV) replication might mediate cytopathic effects are controversial and not entirely clear. In this study, we found that blood-borne HCV (bbHCV) infection could lead to endoplasmic reticulum (ER)-stress and mitochondria-related/caspase-dependent apoptosis at the early stages of infection based on use of the highly efficient bbHCV cell culture model established previously. Sections of bbHCV-infected human fetal liver stem cells (hFLSCs) revealed convolution and nonlinear ER, cell vacuolization, swelling of mitochondria, and numerous double membrane vesicles (DMVs). The percentage of apoptotic hFLSCs infected by bbHCV reached 29.8% at 16 h postinfection, and the amount of cytochrome c increased remarkably in the cytosolic protein fraction. However, over time, apoptosis was inhibited due to the activation of NF-κB. The expression of NF-κB-p65, Bcl-xL, XIAP, and c-FLIPL in hFLSCs was increased significantly 24 h after in infection by bbHCV. The accelerated cell death cycles involving apoptosis, regeneration and repair by bbHCV infection might give rise to the development of cirrhosis, and ultimately to hepatocellular carcinogenesis. Copyright © 2019 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.


Asunto(s)
Apoptosis , Estrés del Retículo Endoplásmico , Células Madre Fetales/patología , Hepacivirus/crecimiento & desarrollo , Hepatitis C Crónica/virología , Hígado/patología , Replicación Viral , Proteínas Reguladoras de la Apoptosis/metabolismo , Línea Celular , Células Madre Fetales/metabolismo , Células Madre Fetales/virología , Hepatitis C Crónica/metabolismo , Hepatitis C Crónica/patología , Humanos , Hígado/metabolismo , Hígado/virología , Mitocondrias Hepáticas/metabolismo , Mitocondrias Hepáticas/patología , Mitocondrias Hepáticas/virología , Estrés Oxidativo , Transducción de Señal
11.
BMC Plant Biol ; 19(1): 19, 2019 Jan 11.
Artículo en Inglés | MEDLINE | ID: mdl-30634907

RESUMEN

BACKGROUND: Verticillium wilt (VW), also known as "cotton cancer," is one of the most destructive diseases in global cotton production that seriously impacts fiber yield and quality. Despite numerous attempts, little significant progress has been made in improving the VW resistance of upland cotton. The development of chromosome segment substitution lines (CSSLs) from Gossypium hirsutum × G. barbadense has emerged as a means of simultaneously developing new cotton varieties with high-yield, superior fiber, and resistance to VW. RESULTS: In this study, VW-resistant investigations were first conducted in an artificial greenhouse, a natural field, and diseased nursery conditions, resulting in the identification of one stably VW-resistant CSSL, MBI8255, and one VW-susceptible G. hirsutum, CCRI36, which were subsequently subjected to biochemical tests and transcriptome sequencing during V991 infection (0, 1, and 2 days after inoculation). Eighteen root samples with three replications were collected to perform multiple comparisons of enzyme activity and biochemical substance contents. The findings indicated that VW resistance was positively correlated with peroxidase and polyphenol oxidase activity, but negatively correlated with malondialdehyde content. Additionally, RNA sequencing was used for the same root samples, resulting in a total of 77,412 genes, of which 23,180 differentially expressed genes were identified from multiple comparisons between samples. After Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis on the expression profiles identified using Short Time-series Expression Miner, we found that the metabolic process in the biological process, as well as the pathways of phenylpropanoid biosynthesis and plant hormone signal transduction, participated significantly in the response to VW. Gene functional annotation and expression quantity analysis indicated the important roles of the phenylpropanoid metabolic pathway and oxidation-reduction process in response to VW, which also provided plenty of candidate genes related to plant resistance. CONCLUSIONS: This study concentrates on the preliminary response to V991 infection by comparing the VW-resistant CSSL and its VW-susceptible recurrent parent. Not only do our findings facilitate the culturing of new resistant varieties with high yield and superior performance, but they also broaden our understanding of the mechanisms of cotton resistance to VW.


Asunto(s)
Cromosomas de las Plantas/genética , Gossypium/genética , Gossypium/microbiología , Transcriptoma/genética , Verticillium/patogenicidad , Regulación de la Expresión Génica de las Plantas/genética
12.
J Hazard Mater ; 365: 340-345, 2019 03 05.
Artículo en Inglés | MEDLINE | ID: mdl-30448547

RESUMEN

Antibiotic resistance genes (ARGs) have gained global attention due to their public health threat. Extracelluar ARGs (eARGs) can result in the dissemination of antibiotic resistance via free-living ARGs in natural environments, where they promote ARB transmission in drinking water distribution systems. However, eARG pollution in tap water has not been well researched. In this study, concentrations of eARGs and intracellular ARGs (iARGs) in tap water, sampled at Tianjin, China, were investigated for one year. Fourteen eARG types were found at the highest concentration of 1.3 × 105 gene copies (GC)/L. TetC was detected in 66.7% of samples, followed by sul1, sul2, and qnrA with the same detection frequency of 41.7%. Fifteen iARGs (including tetA, tetB, tetM, tetQ, tetX, sul1, sul2, sul3, ermB, blaTEM, and qnrA) were continuously detected in all collected tap water samples with sul1 and sul2 the most abundant. Additionally, both eARG and iARG concentrations in tap water presented a seasonal pattern with most abundant prevalence in summer. The concentration of observed intracellular sulfonamide resistance genes showed a significantly positive correlation with total nitrogen concentrations. This study suggested that eARG and iARG pollution of drinking water systems pose a potential risk to human public health.


Asunto(s)
Agua Potable/microbiología , Farmacorresistencia Microbiana/genética , Genes Bacterianos , Microbiología del Agua , China , Monitoreo del Ambiente
13.
J Clin Exp Hepatol ; 8(2): 195-204, 2018 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-29892184

RESUMEN

In the twenty-seven years since the discovery of hepatitis C virus (HCV) the majority of individuals exposed to HCV establish a persistent infection, which is a leading cause of chronic liver disease, cirrhosis and hepatocellular carcinoma. In developed nations, the cure rates of HCV infection could be over 90% with direct-acting antiviral (DAA) regimens, which has made the great progress in global eradication. However, the cost of these treatments is so expensive that the patients in developing nations, where the disease burden is the most severe, could not afford it, which highly restricted its access. Additionally, the largely asymptomatic nature of infection facilitates continued transmission in risk groups due to limited surveillance. Consequently a protective vaccine and likely emergence of drug-resistant viral variants call for further studies of HCV biology. In the current review, the development and the progress of preventive and therapeutic vaccines against the HCV have been reviewed in the context of peptide vaccines, recombinant protein vaccines, HCV-like particle, DNA vaccines and viral vectors expressing HCV genes.

14.
Sci Total Environ ; 635: 964-971, 2018 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-29710618

RESUMEN

Here we developed an integrated cell absorption process and quantitative (reverse transcription) polymerase chain reaction (ICAP-q(RT)PCR) assay to detect infectious viruses, which based on the detection of the viral nucleic acid (RNA or DNA) in the early stage of viral attachment and entry towards cells. The results showed that the poliovirus or adenovirus whose concentration was as low as 0.2 TCID50/mL could be detected by ICAP-q(RT)PCR after 4 h incubation. The ICAP-q(RT)PCR exhibited much higher sensitivity than the plaque assay. In parallel, it took shorter time to detect the viruses towards field samples compared with the integrated cell culture (ICC)-qPCR, but could still get the consistent detecting results with ICC-qPCR. This method is verified by detecting four different kinds of viruses including poliovirus, adenovirus, rotavirus, and astrovirus, which existed in the actual water samples. Among all the 24 Jinhe river samples, 50% (12/24) of river water samples were positive for poliovirus when detected by ICAP-q(RT)PCR, which was in accordance with the results detected by ICC-qPCR. However, 21% (5/24) and 68% (18/24) of the samples were detected to be positive for poliovirus by the plaque counting and the direct qPCR method, respectively. Compared with ICAP-q(PT)PCR and ICC-qPCR, the detecting results of qPCR or plaque assay displayed a marked expansion or decline, respectively, which lead to the evident deviations in the accuracy. The results demonstrated that our developed ICAP-q(RT)PCR method could dramatically reduce the test duration and quite improve the sensitivity towards infectious viruses. Therefore, the ICAP-q(RT)PCR method could be an effective and quantitative tool for detecting infectious viruses in water environments.


Asunto(s)
Bioensayo/métodos , Monitoreo del Ambiente/métodos , Agua Dulce/virología , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Rotavirus , Sensibilidad y Especificidad , Virus , Agua
15.
Water Res ; 136: 131-136, 2018 06 01.
Artículo en Inglés | MEDLINE | ID: mdl-29501757

RESUMEN

The emergence and spread of antibiotic resistance has posed a major threat to both human health and environmental ecosystem. Although the disinfection has been proved to be efficient to control the occurrence of pathogens, little effort is dedicated to revealing potential impacts of disinfection on transmission of antibiotic resistance genes (ARGs), particularly for free-living ARGs in final disinfected effluent of urban wastewater treatment plants (UWWTP). Here, we investigated the effects of chlorine disinfection on the occurrence and concentration of both extracellular ARGs (eARGs) and intracellular ARGs (iARGs) in a full-scale UWWTP over a year. We reported that the concentrations of both eARGs and iARGs would be increased by the disinfection with chlorine dioxide (ClO2). Specifically, chlorination preferentially increased the abundances of eARGs against macrolide (ermB), tetracycline (tetA, tetB and tetC), sulfonamide (sul1, sul2 and sul3), ß-lactam (ampC), aminoglycosides (aph(2')-Id), rifampicin (katG) and vancomycin (vanA) up to 3.8 folds. Similarly, the abundances of iARGs were also increased up to 7.8 folds after chlorination. In terms of correlation analyses, the abundance of Escherichia coli before chlorination showed a strong positive correlation with the total eARG concentration, while lower temperature and higher ammonium concentration were assumed to be associated with the concentration of iARGs. This study suggests the chlorine disinfection could increase the abundances of both iARGs and eARGs, thereby posing risk of the dissemination of antibiotic resistance in environments.


Asunto(s)
Cloro/farmacología , Desinfectantes/farmacología , Farmacorresistencia Bacteriana , Proteínas de Escherichia coli/genética , Escherichia coli/efectos de los fármacos , Escherichia coli/genética , Antibacterianos/farmacología , Cloro/análisis , Desinfectantes/análisis , Desinfección , Farmacorresistencia Microbiana , Escherichia coli/metabolismo , Proteínas de Escherichia coli/metabolismo , Halogenación , Tetraciclina/farmacología , Aguas Residuales/microbiología , Purificación del Agua
16.
BMC Genomics ; 18(1): 705, 2017 Sep 08.
Artículo en Inglés | MEDLINE | ID: mdl-28886694

RESUMEN

BACKGROUND: How to develop new cotton varieties possessing high yield traits of Upland cotton and superior fiber quality traits of Sea Island cotton remains a key task for cotton breeders and researchers. While multiple attempts bring in little significant progresses, the development of Chromosome Segment Substitution Lines (CSSLs) from Gossypium barbadense in G. hirsutum background provided ideal materials for aforementioned breeding purposes in upland cotton improvement. Based on the excellent fiber performance and relatively clear chromosome substitution segments information identified by Simple Sequence Repeat (SSR) markers, two CSSLs, MBI9915 and MBI9749, together with the recurrent parent CCRI36 were chosen to conduct transcriptome sequencing during the development stages of fiber elongation and Secondary Cell Wall (SCW) synthesis (from 10DPA and 28DPA), aiming at revealing the mechanism of fiber development and the potential contribution of chromosome substitution segments from Sea Island cotton to fiber development of Upland cotton. RESULTS: In total, 15 RNA-seq libraries were constructed and sequenced separately, generating 705.433 million clean reads with mean GC content of 45.13% and average Q30 of 90.26%. Through multiple comparisons between libraries, 1801 differentially expressed genes (DEGs) were identified, of which the 902 up-regulated DEGs were mainly involved in cell wall organization and response to oxidative stress and auxin, while the 898 down-regulated ones participated in translation, regulation of transcription, DNA-templated and cytoplasmic translation based on GO annotation and KEGG enrichment analysis. Subsequently, STEM software was performed to explicate the temporal expression pattern of DEGs. Two peroxidases and four flavonoid pathway-related genes were identified in the "oxidation-reduction process", which could play a role in fiber development and quality formation. Finally, the reliability of RNA-seq data was validated by quantitative real-time PCR of randomly selected 20 genes. CONCLUSIONS: The present report focuses on the similarities and differences of transcriptome profiles between the two CSSLs and the recurrent parent CCRI36 and provides novel insights into the molecular mechanism of fiber development, and into further exploration of the feasible contribution of G. barbadense substitution segments to fiber quality formation, which will lay solid foundation for simultaneously improving fiber yield and quality of upland cotton through CSSLs.


Asunto(s)
Cromosomas de las Plantas/genética , Fibra de Algodón , Perfilación de la Expresión Génica , Gossypium/crecimiento & desarrollo , Gossypium/genética , Hibridación Genética , Pared Celular/metabolismo , Gossypium/citología , Fenotipo , Reproducibilidad de los Resultados
17.
Mol Ecol ; 26(19): 5318-5333, 2017 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-28742284

RESUMEN

The aqueous environment is one of many reservoirs of antibiotic resistance genes (ARGs). Fish, as important aquatic animals which possess ideal intestinal niches for bacteria to grow and multiply, may ingest antibiotic resistance bacteria from aqueous environment. The fish gut would be a suitable environment for conjugal gene transfer including those encoding antibiotic resistance. However, little is known in relation to the impact of ingested ARGs or antibiotic resistance bacteria (ARB) on gut microbiota. Here, we applied the cultivation method, qPCR, nuclear molecular genetic marker and 16S rDNA amplicon sequencing technologies to develop a plasmid-mediated ARG transfer model of zebrafish. Furthermore, we aimed to investigate the dissemination of ARGs in microbial communities of zebrafish guts after donors carrying self-transferring plasmids that encode ARGs were introduced in aquaria. On average, 15% of faecal bacteria obtained ARGs through RP4-mediated conjugal transfer. The hindgut was the most important intestinal region supporting ARG dissemination, with concentrations of donor and transconjugant cells almost 25 times higher than those of other intestinal segments. Furthermore, in the hindgut where conjugal transfer occurred most actively, there was remarkable upregulation of the mRNA expression of the RP4 plasmid regulatory genes, trbBp and trfAp. Exogenous bacteria seem to alter bacterial communities by increasing Escherichia and Bacteroides species, while decreasing Aeromonas compared with control groups. We identified the composition of transconjugants and abundance of both cultivable and uncultivable bacteria (the latter accounted for 90.4%-97.2% of total transconjugants). Our study suggests that aquatic animal guts contribute to the spread of ARGs in water environments.


Asunto(s)
Bacterias/clasificación , Farmacorresistencia Bacteriana/genética , Microbioma Gastrointestinal , Transferencia de Gen Horizontal , Genes Bacterianos , Pez Cebra/microbiología , Animales , Conjugación Genética , Heces/microbiología , Intestinos/microbiología , Plásmidos/genética , ARN Ribosómico 16S/genética , Agua
18.
J Microbiol Methods ; 139: 143-149, 2017 08.
Artículo en Inglés | MEDLINE | ID: mdl-28577899

RESUMEN

Bacteriophages are widely used to the treatment of drug-resistant bacteria and the improvement of food safety through bacterial lysis. However, the limited investigations on bacteriophage restrict their further application. In this study, a novel and highly efficient method was developed for isolating bacteriophage from water based on the electropositive silica gel particles (ESPs) method. To optimize the ESPs method, we evaluated the eluent type, flow rate, pH, temperature, and inoculation concentration of bacteriophage using bacteriophage f2. The quantitative detection reported that the recovery of the ESPs method reached over 90%. The qualitative detection demonstrated that the ESPs method effectively isolated 70% of extremely low-concentration bacteriophage (100 PFU/100L). Based on the host bacteria composed of 33 standard strains and 10 isolated strains, the bacteriophages in 18 water samples collected from the three sites in the Tianjin Haihe River Basin were isolated by the ESPs and traditional methods. Results showed that the ESPs method was significantly superior to the traditional method. The ESPs method isolated 32 strains of bacteriophage, whereas the traditional method isolated 15 strains. The sample isolation efficiency and bacteriophage isolation efficiency of the ESPs method were 3.28 and 2.13 times higher than those of the traditional method. The developed ESPs method was characterized by high isolation efficiency, efficient handling of large water sample size and low requirement on water quality.


Asunto(s)
Bacteriófagos/aislamiento & purificación , Gel de Sílice , Virología/métodos , Microbiología del Agua , Adsorción , Bacterias/virología , Técnicas Electroquímicas/métodos , Filtración/métodos , Concentración de Iones de Hidrógeno , Ríos/virología , Temperatura , Calidad del Agua
19.
Artículo en Inglés | MEDLINE | ID: mdl-28555063

RESUMEN

Antibiotic failure is occurring worldwide. In a routine surveillance study on antibioticresistance genes (ARGs) in natural water bodies, we noted the detection of colistin-resistance gene mcr-1, previously identified in Escherichia coli and Klebsiella pneumoniae isolates from human beings and animals in several countries. The mcr-1 gene might be present in water environments, because aquatic ecosystems are recognized as reservoirs for antibiotic resistant bacteria (ARB) and ARGs. In this study, a qPCR assay was developed to monitor and quantify the mcr-1 gene in the Haihe River, China. The results showed that all 18 samples collected from different locations over 6 months along the Haihe River were positive for the mcr-1 gene, and the highest level of mcr-1 reached 3.81 × 105 gene copies (GC) per liter of water. This is the first study to quantify mcr-1 in a natural water system by qPCR. Our findings highlight the potential for this antibiotic resistance determinant to spread extensively, suggesting a significant health and ecological impact.


Asunto(s)
Farmacorresistencia Bacteriana/genética , Proteínas de Escherichia coli/genética , Genes Bacterianos , Ríos , Antibacterianos , China , Colistina , Monitoreo del Ambiente , Escherichia coli/aislamiento & purificación
20.
Med Hypotheses ; 102: 112-122, 2017 May.
Artículo en Inglés | MEDLINE | ID: mdl-28478815

RESUMEN

Although many hypotheses for tumorigenesis have been proposed, none can explain the occurrence and development of tumors comprehensively until now. We put forward a new hypothesis: tumors arise from the excessive repair of damaged stem cells. There are stem cells in all tissues and organs, and the stem cells have perfect damage repair mechanisms, including damage repair systems and repair-inhibiting systems. Tumors arise from the excessive repair of damaged stem cells, i.e., carcinogens induce stem cell damage, leading to overexpression of damage repair systems, and simultaneous inactivation of repair-inhibiting systems through genetic or non-genetic mechanisms, finally forming tumors. The outcome (forming clinically significant tumors or death) and development (tumor recurrence, metastasis or spontaneous healing) of the tumor cells depends on whether the injury and the excessive repair persists, whether immune surveillance function is normal and the tumor microenvironment is appropriate. This hypothesis not only addresses the issues of where tumor cells arise from, how tumors form and where they go, but also provides a reasonable explanation for many unresolved issues in tumor occurrence, development, metastasis or healing. In addition, this hypothesis could guide the early diagnosis, reasonable treatment and effective prevention of tumors.


Asunto(s)
Carcinogénesis/efectos de los fármacos , Carcinogénesis/patología , Carcinógenos/administración & dosificación , Neoplasias/patología , Neoplasias/fisiopatología , Células Madre Neoplásicas/efectos de los fármacos , Células Madre Neoplásicas/fisiología , Animales , Medicina Basada en la Evidencia , Humanos , Modelos Biológicos , Células Madre Neoplásicas/patología
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