scholarly journals Molecular Structure and Transferability of Tn1546-Like Elements in Enterococcus faecium Isolates from Clinical, Sewage, and Surface Water Samples in Iran

2008 ◽  
Vol 74 (5) ◽  
pp. 1350-1356 ◽  
Author(s):  
M. Talebi ◽  
M. R. Pourshafie ◽  
M. Katouli ◽  
R. Möllby
Keyword(s):  
Molecular Structure ◽  
Surface Water ◽  
Molecular Analysis ◽  
Enterococcus Faecium ◽  
Pcr Amplification ◽  
Molecular Size ◽  
Municipal Sewage ◽  
Clinical Samples ◽  
Polymorphism Analysis ◽  
Hospital Sewage

ABSTRACT The molecular structure and transferability of Tn1546 in 143 vancomycin-resistant Enterococcus faecium (VREF) isolates obtained from patients (n = 49), surface water (n = 28), and urban and hospital sewage (n = 66) in Tehran, Iran, were investigated. Molecular characterization of Tn1546 elements in vanA VREF was performed using a combination of restriction fragment length polymorphism analysis and DNA sequencing of the internal PCR fragments of vanA transposons. Long-PCR amplification showed that the molecular size of Tn1546 elements varied from 10.8 to 12.8 kb. The molecular analysis of Tn1546 showed that 45 isolates (31.5%) harbored a deletion/mutation upstream from nucleotide 170. No horizontal transfer of Tn1546 was observed following filter-mating conjugation with these isolates. Nevertheless, the rates of transferability for other isolates were 10−5 to 10−6 per donor. Insertion sequences IS1216V and IS1542 were present in 103 (72%) and 138 (96.5%) of the isolates, respectively. The molecular analysis of Tn1546 elements resulted in three genomic organizations. The genomic organization lineage 1 was dominated by the isolates from clinical samples (3.4%), lineage 2 was dominated mostly by sewage isolates (24.5%), and lineage 3 contained isolates obtained from all sources (72.1%). The genetic diversity determined using pulsed-field gel electrophoresis (PFGE) revealed a single E. faecium clone, designated 44, which was common to the samples obtained from clinical specimens and hospital and municipal sewage. Furthermore, the results suggest that lineage 3 Tn1546 was highly disseminated among our enterococcal isolates in different PFGE patterns.

scholarly journals Phenotypic and molecular characterization of HA-MRSA in Taif hospitals, Saudi Arabia

2015 ◽  
Vol 9 (03) ◽  
pp. 298-303 ◽  
Author(s):  
Emad M Eed ◽  
Mabrouk M Ghonaim ◽  
Yousry M Hussein ◽  
Taiser M Saber ◽  
Amany S Khalifa
Keyword(s):  
Saudi Arabia ◽  
Gene Polymorphism ◽  
Pcr Amplification ◽  
Clinical Samples ◽  
Molecular Characteristics ◽  
Polymorphism Analysis ◽  
Specific Sequence ◽  
Mrsa Strains ◽  
Hospital Acquired ◽  
Sccmec Types

Introduction: Methicillin-resistant S. aureus (MRSA) is one of the most important organisms causing hospital-acquired infections worldwide. Molecular analysis of MRSA strains from Taif, Saudi Arabia, had not been previously done. Phenotypic and molecular characteristics of MRSA isolated from Taif hospitals were investigated. Methodology: This study involved S. aureus strains isolated from different clinical samples from Taif hospitals. MRSA strains were identified and antimicrobial susceptibility profiles were determined. Multiplex polymerase chain reaction (PCR) was used to identify S. aureus-specific sequence, mecA genes, and type of staphylococcal cassette chromosome mec (SCCmec). MRSA strains were typed using coagulase gene polymorphism. Results: In total, 390 strains of S. aureus were isolated, and 58 MRSA strains – 40 hospital-acquired  MRSA (HA-MRSA) and 18 community-acquired MRSA (CA-MRSA) – were detected. HA-MRSA strains included three SCCmec types, while CA-MRSA strains included two SCCmec types. PCR amplification and restriction of the coagulase gene of the 58 MRSA isolates showed nine different patterns, while three strains were non-typable. HA-MRSA strains showed seven distinct restriction fragment length polymorphism (RFLP) patterns; the most frequent was pattern 2 (15 isolates), followed by patterns 1 and 4 (5 isolates each). CA-MRSA showed five RFLP patterns; the most frequent was pattern 3 (7 isolates) followed by pattern 8 (6 isolates). Conclusions: HA-MRSA strains were more common than CA-MRSA strains. SCCmec typing and coagulase gene polymorphism analysis may be useful methods for studying clonal relatedness of isolates and for discriminating between HA-MRSA and CA-MRSA.

scholarly journals Population-level surveillance of antibiotic resistance in Escherichia coli through sewage analysis

2019 ◽  
Vol 24 (37) ◽  
Author(s):  
Marion Hutinel ◽  
Patricia Maria Catharina Huijbers ◽  
Jerker Fick ◽  
Christina Åhrén ◽  
Dan Göran Joakim Larsson ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Primary Care ◽  
Antibiotic Resistance ◽  
Municipal Sewage ◽  
Clinical Samples ◽  
E Coli ◽  
Clinical Resistance ◽  
Resistance Rates ◽  
Hospital Sewage ◽  
Resistance Data

Introduction The occurrence of antibiotic resistance in faecal bacteria in sewage is likely to reflect the current local clinical resistance situation. Aim This observational study investigated the relationship between Escherichia coli resistance rates in sewage and clinical samples representing the same human populations. Methods E. coli were isolated from eight hospital (n = 721 isolates) and six municipal (n = 531 isolates) sewage samples, over 1 year in Gothenburg, Sweden. An inexpensive broth screening method was validated against disk diffusion and applied to determine resistance against 11 antibiotics in sewage isolates. Resistance data on E. coli isolated from clinical samples from corresponding local hospital and primary care patients were collected during the same year and compared with those of the sewage isolates by linear regression. Results E. coli resistance rates derived from hospital sewage and hospital patients strongly correlated (r2 = 0.95 for urine and 0.89 for blood samples), as did resistance rates in E. coli from municipal sewage and primary care urine samples (r2 = 0.82). Resistance rates in hospital sewage isolates were close to those in hospital clinical isolates while resistance rates in municipal sewage isolates were about half of those measured in primary care isolates. Resistance rates in municipal sewage isolates were more stable between sampling occasions than those from hospital sewage. Conclusion Our findings provide support for development of a low-cost, sewage-based surveillance system for antibiotic resistance in E. coli, which could complement current monitoring systems and provide clinically relevant antibiotic resistance data for countries and regions where surveillance is lacking.

scholarly journals The role of wastewater treatment plants in surface water contamination by plastic pollutants

2018 ◽  
Vol 45 ◽  
pp. 00054 ◽  
Author(s):  
Bozena Mrowiec
Keyword(s):  
Wastewater Treatment ◽  
Surface Water ◽  
Water Contamination ◽  
Membrane Filtration ◽  
Municipal Wastewater ◽  
Wastewater Treatment Plants ◽  
Point Sources ◽  
Municipal Sewage ◽  
Municipal Wastewater Treatment ◽  
Surface Water Contamination

The aim of this paper was to review the literature data regarding the physico-chemical characteristic of plastic pollutants discharged with municipal sewage, the practical possibility of removing microplastic particles from wastewater during different treatment steps in WWTPs and the problem of surface water contamination within them. Microplastics (the size range of 1 nm to < 5 mm), have been recognized as an emerging threat, as well as an ecotoxicological and ecological risk for water ecosystems. Municipal wastewater treatment plants (WWTPs) are mentioned as the main point sources of microplastics in an aquatic environment. Microplastic particles can be effectively removed in the primary treatment zones via solids skimming and sludge settling processes. Different tertiary treatment processes such as: gravity sand filtration, discfilter, air flotation and membrane filtration provide substantial additional removal of microplastics, and the efficiency of wastewater treatment process can be at a removal level of 99.9%. Nevertheless, given the large volumes of effluent constantly discharged to receivers, even tertiary level WWTPs may constitute a considerable source of microplastics in the surface water.

Molecular analysis of the replication region of the pCIZ2 plasmid from the multiple bacteriocin producer strain Enterococcus faecium L50

Plasmid ◽  
2008 ◽  
Vol 60 (3) ◽  
pp. 181-189 ◽  
Author(s):  
R. Criado ◽  
J. Gutiérrez ◽  
A. Budin-Verneuil ◽  
P.E. Hernández ◽  
A. Hartke ◽  
...  
Keyword(s):  
Molecular Analysis ◽  
Enterococcus Faecium ◽  
Producer Strain ◽  
Bacteriocin Producer ◽  
Replication Region

scholarly journals Examination of Surface Water Along River-Rima Floodplain in Wamakko, Sokoto State, Nigeria.

2020 ◽  
Vol 2 (3) ◽  
Author(s):  
Abdulqadir Abubakar Usman ◽  
Murtala Abubakar Gada ◽  
Aminu Muhammad Bayawa ◽  
Ibrahim Mustapha Dankani ◽  
Saadu Umar Wali
Keyword(s):  
Drinking Water ◽  
Surface Water ◽  
Principal Component ◽  
Hierarchical Cluster ◽  
World Health ◽  
Coliform Bacteria ◽  
Municipal Sewage ◽  
Bacteria Growth ◽  
Anthropogenic Inputs ◽  
Health Organization

This study examined the hydrochemistry of surface water along the River-Rima floodplain area. Five sampling locations were purposively selected, and, in each point, three samples were taken from surface water (river). The sampling was repeated after 20 days. Thus, a total of 30 samples were collected. Water samples obtained were subjected to laboratory tests. Results revealed that BOD, TDS, Mg2+, and Fe3+ are above the World Health Organization (WHO) and Standard Organization of Nigeria (SON) reference guidelines for drinking water quality. Isolates detected from the coliform bacteriological analysis include Enterobacter aerogene, Escherichia coli, and Citrobacter freundii with most of the samples showing coliform bacteria growth above the SON standard for drinking water. Hence, the water in the River-Rima floodplain of the Wamakko area is of low quality and unsafe for drinking. Results of principal component analysis (PCA) revealed external influences such as pollutant wash off and rock weathering as controls on hydrochemistry of surface water. There is some indication of anthropogenic inputs (Cl-, NO3-, and PO42-) based on hierarchical cluster analysis. Elements including Cl-, NO3-, and PO42- are increasingly added into surface water from human activities, mainly agriculture, and municipal sewage.

scholarly journals Improved Genotyping Vaccine and Wild-Type Poliovirus Strains by Restriction Fragment Length Polymorphism Analysis: Clinical Diagnostic Implications

2000 ◽  
Vol 38 (12) ◽  
pp. 4337-4342 ◽  
Author(s):  
Amalia Georgopoulou ◽  
Panayotis Markoulatos ◽  
Niki Spyrou ◽  
Nicholas C. Vamvakopoulos
Keyword(s):  
Restriction Fragment Length Polymorphism ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Clinical Samples ◽  
Polymorphism Analysis ◽  
Rt Pcr ◽  
Restriction Fragment Length

The combination of preventive vaccination and diagnostic typing of viral isolates from patients with clinical poliomyelitis constitutes our main protective shield against polioviruses. The restriction fragment length polymorphism (RFLP) adaptation of the reverse transcriptase (RT)-PCR methodology has advanced diagnostic genotyping of polioviruses, although further improvements are definitely needed. We report here on an improved RFLP procedure for the genotyping of polioviruses. A highly conserved segment within the 5′ noncoding region of polioviruses was selected for RT-PCR amplification by the UC53-UG52 primer pair with the hope that it would be most resistant to the inescapable genetic alteration-drift experienced by the other segments of the viral genome. Complete inter- and intratypic genotyping of polioviruses by the present RFLP method was accomplished with a minimum set of four restriction endonucleases (HaeIII, DdeI, NcoI, andAvaI). To compensate for potential genetic drift within the recognition sites of HaeIII, DdeI, orNcoI in atypical clinical samples, the RFLP patterns generated with HpaII and StyI as replacements were analyzed. The specificity of the method was also successfully assessed by RFLP analysis of 55 reference nonpoliovirus enterovirus controls. The concerted implementation of these conditional protocols for diagnostic inter- and intratypic genotyping of polioviruses was evaluated with 21 clinical samples with absolute success.

Genome Analysis of the Enterococcus faecium Entfac.YE Prophage

2022 ◽  
Author(s):  
Yara Elahi ◽  
Ramin Mazaheri Nezhad Fard ◽  
Arash Seifi ◽  
Saeideh Mahfouzi ◽  
Ali Akbar Saboor Yaraghi
Keyword(s):  
Genome Analysis ◽  
Enterococcus Faecium ◽  
Clinical Sample ◽  
Bacterial Species ◽  
Bacterial Genome ◽  
Rapid Evolution ◽  
Housekeeping Genes ◽  
Water Sources ◽  
Clinical Samples ◽  
Transfer Resistance

Background: Bacteriophages are viruses that infect bacteria. Bacteriophages are widely distributed in various environments. The prevalence of bacteriophages in water sources, especially wastewaters, is naturally high. These viruses affect evolution of most bacterial species. Bacteriophages are able to integrate their genomes into the chromosomes of their hosts as prophages and hence transfer resistance genes to the bacterial genomes. Enterococci are commensal bacteria that show high resistance to common antibiotics. For example, prevalence of vancomycin-resistant enterococci has increased within the last decades. Methods: Enterococcal isolates were isolated from clinical samples and morphological, phenotypical, biochemical, and molecular methods were used to identify and confirm their identity. Bacteriophages extracted from water sources were then applied to isolated Enterococcus faecium (E. faecium). In the next step, the bacterial genome was completely sequenced and the existing prophage genome in the bacterial genome was analyzed. Results: In this study, E. faecium EntfacYE was isolated from a clinical sample. The EntfacYE genome was analyzed and 88 prophage genes were identified. The prophage content included four housekeeping genes, 29 genes in the group of genes related to replication and regulation, 25 genes in the group of genes related to structure and packaging, and four genes belonging to the group of genes associated with lysis. Moreover, 26 genes were identified with unknown functions. Conclusion: In conclusion, genome analysis of prophages can lead to a better understanding of their roles in the rapid evolution of bacteria.

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