Search results for "microbial"
showing 10 items of 2041 documents
Comparison of different primer sets for use in Automated Ribosomal Intergenic Spacer Analysis of complex bacterial communities.
2004
ABSTRACT ITSF and ITSReub, constituting a new primer set designed for the amplification of the 16S-23S rRNA intergenic transcribed spacers, have been compared with primer sets consisting of 1406F and 23Sr (M. M. Fisher and E. W. Triplett, Appl. Environ. Microbiol. 65:4630-4636, 1999) and S-D-Bact-1522-b-S-20 and L-D-Bact-132-a-A-18 (L. Ranjard et al., Appl. Environ. Microbiol. 67:4479-4487, 2001), previously proposed for automated ribosomal intergenic spacer analysis (ARISA) of complex bacterial communities. An agricultural soil and a polluted soil, maize silage, goat milk, a small marble sample from the façade of the Certosa of Pavia (Pavia, Italy), and brine from a deep hypersaline anoxi…
Cumulative Effects of Short-Term Polymetal Contamination on Soil Bacterial Community Structure
2006
ABSTRACT In this study we evaluated the short-term effects of copper, cadmium, and mercury, added singly or in combination at different doses, on soil bacterial community structure using the bacterial automated ribosomal intergenic spacer analysis (B-ARISA) fingerprinting technique. Principal-component analysis of B-ARISA profiles allowed us to deduce the following order of impact: (Cu + Cd + Hg) >> Hg ≥ Cd > Cu. These results demonstrated that there was a cumulative effect of metal toxicity. Furthermore, the trend of modifications was consistent with the “hump-backed” relationships between biological diversity and disturbance described by Giller et al. (K. E. Giller, E. Witler, an…
High prevalence of integron-mediated resistance in clinical isolates of Salmonella enterica
2010
Salmonella enterica has become progressively resistant to antimicrobial agents worldwide as a result of genes carried on different classes of integrons. The aim of the current study was to investigate the molecular diversity of these integrons and their association with antimicrobial resistance in clinical S. enterica isolates from Tehran, Iran. Antimicrobial susceptibility testing was performed according to the Clinical and Laboratory Standards Institute. The presence of integrons was investigated by PCR using specific primers. Integrons were detected in 65 (47.1%) strains, with classes 1 and 2 being observed in 54 (39%) and 11 (8%) strains, respectively. Integron-positive isolates belonge…
Molecular relationship among Salmonella dublin isolates identified at the Center for Enterobacteriaceae of Palermo during the years 1971-85.
1987
SUMMARYA molecular epidemiological study was carried out on 60Salmonella dublinisolates identified at the Southern Italy Enterobacteriaceae Center between 1971 and 1985. These included 23 isolates from children with diarrhoea in Palermo obtained during 1984.All isolates from the outbreak of gastroenteritis in children were resistant to chloramphenicol and streptomycin and harboured two plasmids of 50 MDa and 3 MDa molecular weight, whereas the majority of the isolates identified before 1984 were susceptible to these antibiotics and carried only a 50 MDa molecular weight plasmid. FourS. dublinstrains successively identified from cattle (Palermo, Foggia, Portici) and from a child (Palermo) we…
pilF polymorphism-based real-time PCR to distinguish Vibrio vulnificus strains of human health relevance
2012
The Gram-negative bacterium Vibrio vulnificus is a common inhabitant of estuarine environments. Globally, V. vulnificus is a significant foodborne pathogen capable of causing necrotizing wound infections and primary septicemia, and is a leading cause of seafood-related mortality. Unfortunately, molecular methods for the detection and enumeration of pathogenic V. vulnificus are hampered by the genetically diverse nature of this pathogen, the range of different biotypes capable of infecting humans and aquatic animals, and the fact that V. vulnificus contains pathogenic as well as non-pathogenic variants. Here we report an alternative approach utilizing the development of a real-time PCR assay…
Development of a real-time PCR assay for detection and quantification of enterotoxigenic members of Bacillus cereus group in food samples
2009
A highly sensitive real-time PCR (qPCR) procedure, targeting the phosphatidylcholine-specific phospholipase C gene (pc-plc), was developed for specific detection and quantification of strains belonging to Bacillus cereus group. The target region was selected based on the enterotoxigenic profiles of 75 Bacillus strains. The inclusivity and exclusivity of the RTi-PCR assay were assessed with 59 isolates of the B. cereus group, 16 other Bacillus spp., and 4 non-Bacillus strains. The assay was also used to construct calibration curves for different food matrices, and it had a wide quantification range of 6 log units using both serial dilutions of purified DNA and calibrated cell suspensions of …
Ongoing spread of colistin-resistant Klebsiella pneumoniae in different wards of an acute general hospital, Italy, June to December 2011.
2012
We describe polyclonal spread of colistin-resistant Klebsiella pneumoniae in an acute general hospital in Italy. Between June and December 2011, 58 colistin-resistant K. pneumoniae isolates were recovered from 28 patients admitted to different wards, but mainly in the intensive care units. All isolates were tested for drug susceptibility and the presence of beta-lactamase (bla) genes. Clonality was investigated by repetitive extragenic palindromic (rep)-PCR and multilocus sequence typing (MLST). Fifty-two isolates had minimum inhibitory concentrations (MICs) for colistin of 6-128 mg/L, carried blaKPC3 and were attributed to sequence type ST258. The remaining six isolates were susceptible to…
Methicillin-resistant Staphylococcus pseudintermedius as causative agent of dairy cow mastitis.
2013
Staphylococcus pseudintermedius is a coagulase-positive specie similar to Staphylococcus intermedius , frequently associated with pyoderma, otitis and urinary tract infections of dogs and cats (van Duijkeren and others 2011). No information about bovine mastitis caused by S pseudintermedius is available in the literature. Antimicrobial resistance among S pseudintermedius strains is increasing: in the past, susceptibility to most antibiotics was common (Bond and Loeffler 2012), but in the last few years methicillin-resistant S pseudintermedius (MRSP) strains have emerged as a significant animal health problem in veterinary medicine (Schwarz and others 2008, van Duijkeren and others 2008, Wee…
PCR-based procedures for detection and quantification of Staphylococcus aureus and their application in food.
2006
Aims: To evaluate the specificity of nuc targeted primers for PCR detection of Staphylococcus aureus in different food matrices and to establish a RTQ-PCR procedure suitable for the routine detection and quantification of this pathogen in food. Methods and Results: Specificity of nuc targeted primers (Pri1–Pri2 and the newly designed RTQ-PCR primers) was tested on a total of 157 strains of genetically confirmed identity, including reference and food isolates. PCR detection on artificially inoculated beef samples by DNA extraction using a DNeasy Tissue Kit (Qiagen GmhH, Hilden, Germany) showed a sensitivity value around 103 CFU g−1. The two RTQ-PCR systems, incorporating SYBR-Green I and T…
Real-time quantitative PCR of Staphylococcus aureus and application in restaurant meals.
2006
Staphylococcus aureus is considered the second most common pathogen to cause outbreaks of food poisoning, exceeded only by Campylobacter. Consumption of foods containing this microorganism is often identified as the cause of illness. In this study, a rapid, reliable, and sensitive real-time quantitative PCR was developed and compared with conventional culture methods. Real-time quantitative PCR was carried out by purifying DNA extracts of S. aureus with a Staphylococcus sample preparation kit and quantifying it in the LightCycler system with hybridization probes. The assay was linear from a range of 10 to 10(6) S. aureus cells (r2 > 0.997). The PCR reaction presented an efficiency of >85%. …