Comparative analysis ofShigella sonneibiotype g isolated from paediatric populations in Egypt, 1999–2005

2012 ◽  
Vol 141 (8) ◽  
pp. 1614-1624 ◽  
Author(s):  
J. DEJLI ◽  
R. A. NADA ◽  
A. MANSOUR ◽  
A. A. EL-MONIEM ◽  
M. O. WASFY ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Genetic Similarity ◽  
Susceptibility Testing ◽  
Biochemical Characterization ◽  
Multidrug Resistant ◽  
Pulsed Field Gel Electrophoresis ◽  
Antibiotic Susceptibility Testing ◽  
Severe Diarrhoea ◽  
Plasmid Profiling ◽  
High Degree

SUMMARYStrain characteristics of 51Shigella sonneiisolates obtained from children seeking medical care (MC) and 48 isolates recovered during a prospective diarrhoea birth cohort (BC) study were compared. Biochemical characterization and antibiotic susceptibility testing determined that allS. sonneiisolates were biotype g and multidrug-resistant. Plasmid profiling identified 15 closely related patterns andXbaI pulsed-field gel electrophoresis confirmed the high degree of genetic similarity between isolates. AllS.sonneiisolates harbouredipaHand class II integrase genes and 84·3 and 80% of the MC and BC isolates, respectively carried thesengene. Neither the class I integrase nor thesetgene was detected. Our results indicate thatS. sonneiisolates associated with severe diarrhoea were indistinguishable from those associated with mild diarrhoea. Additional genetic tests with greater discrimination might offer an opportunity to determine genetic differences within the globally disseminating biotype g clone.

Coagulase-negative staphylococci from non-mastitic bovine mammary gland: characterization ofStaphylococcus chromogenesandStaphylococcus haemolyticusby antibiotic susceptibility testing and pulsed-field gel electrophoresis

2011 ◽  
Vol 79 (2) ◽  
pp. 129-134 ◽  
Author(s):  
Mateja Pate ◽  
Irena Zdovc ◽  
Jana Avberšek ◽  
Matjaž Ocepek ◽  
Andrej Pengov ◽  
...  
Keyword(s):  
Mammary Gland ◽  
Dairy Cows ◽  
Gel Electrophoresis ◽  
Susceptibility Testing ◽  
Clinical Signs ◽  
Pulsed Field Gel Electrophoresis ◽  
Antibiotic Susceptibility Testing ◽  
Bovine Mammary Gland ◽  
Coagulase Negative Staphylococci ◽  
Milk Samples

During routine microbiological examination of milk samples from dairy cows without clinical signs of mastitis, quarter milk samples of 231 dairy cows from 12 herds were investigated for the presence of coagulase-negative staphylococci (CNS). The isolates were identified on the basis of colony morphology, Gram staining, catalase and coagulase test and the commercial kit, API Staph. CNS was detected in 29% (67/231) of the cows. A total of seven CNS species were identified with the most prevalent beingStaphylococcus (Staph.) chromogenes(30%) andStaph. haemolyticus(28·8%), followed byStaph. simulans(11·2%),Staph. xylosus(11·2%),Staph. epidermidis(7·5%),Staph. hyicus(6·3%) andStaph. sciuri(5%). The predominant species,Staph. chromogenesandStaph. haemolyticus, were further characterized by antibiotic susceptibility testing using the agar disc diffusion method (Kirby-Bauer) and by pulsed-field gel electrophoresis (PFGE). Considerable resistance to ampicillin and penicillin was observed in both species. Isolates with identical or highly similar PFGE profiles were detected at the herd level despite a marked heterogeneity seen for both species. On the basis of somatic cell count, absence of clinical signs of inflammation and heterogeneity of genotypes, we assume that CNS isolated in this study could not be considered as important causative agents of the bovine mammary gland inflammation.

scholarly journals Prevalence and molecular characterization ofSalmonella entericaisolates throughout an integrated broiler supply chain in China

2016 ◽  
Vol 144 (14) ◽  
pp. 2989-2999 ◽  
Author(s):  
X. REN ◽  
M. LI ◽  
C. XU ◽  
K. CUI ◽  
Z. FENG ◽  
...  
Keyword(s):  
Supply Chain ◽  
Antimicrobial Susceptibility ◽  
Gel Electrophoresis ◽  
Susceptibility Testing ◽  
Guangdong Province ◽  
Multidrug Resistant ◽  
Pulsed Field Gel Electrophoresis ◽  
Production Chain ◽  
Similarity Threshold ◽  
Different Parts

SUMMARYA total of 1145 samples were collected from chicken breeder farms, hatcheries, broiler farms, a slaughterhouse and retail refrigerated chicken stores in an integrated broiler supply chain in Guangdong Province, China, in 2013. One-hundred and twoSalmonella entericastrains were isolated and subjected to serotyping, antimicrobial susceptibility testing, virulence profile determination and molecular subtyping by pulsed field gel electrophoresis (PFGE). The contamination rates in samples from breeder farms, hatcheries, broiler farms, the slaughterhouse and retail stores were 1·46%, 4·31%, 7·00%, 62·86% and 54·67%, respectively. The isolated strains ofS. entericabelonged to 10 serotypes; most of them wereS. Weltevreden (46·08%, 47/102) andS. Agona (18·63%, 19/102). Isolates were frequently resistant to streptomycin (38·2%), tetracycline (36·3%), sulfisoxazole (35·3%) and gentamicin (34·3%); 31·4% of isolates were multidrug resistant. The isolates were screened for 10 virulence factors. TheSalmonellapathogenicity island genesavrA, ssaQ, mgtC, siiD, andsopBand the fimbrial genebcfCwere present in 100% of the strains. PFGE genotyping of the 102S. entericaisolates yielded 24 PFGE types at an 85% similarity threshold. The PFGE patterns show that the genotypes ofS. entericain the production chain are very diverse, but some strains have 100% similarity in different parts of the production chain, which indicates that someS. entericapersist throughout the broiler supply chain.

Characterization of Shiga Toxin–Producing Escherichia coli O157 Isolates from Bovine Carcasses

2016 ◽  
Vol 79 (8) ◽  
pp. 1418-1423
Author(s):  
M. FONTCUBERTA ◽  
R. PLANELL ◽  
A. TORRENTS ◽  
S. SABATÉ ◽  
R. GONZALEZ ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Susceptibility ◽  
Molecular Typing ◽  
Gel Electrophoresis ◽  
Susceptibility Testing ◽  
Pulsed Field Gel Electrophoresis ◽  
Escherichia Coli O157 ◽  
Pulsed Field ◽  
E Coli ◽  
High Degree

ABSTRACT The main purpose of this study was to determine the prevalence of Escherichia coli O157 on bovine carcasses before and after chilling at a large slaughterhouse located in the city of Barcelona, Spain, to assess the effectiveness of dry chilling on reducing E. coli O157 contamination of carcasses. In addition, the study characterized the E. coli O157 strains isolated in terms of virulence factors, antibiotic susceptibility, and their genetic diversity. Individual bovine carcasses were sampled before (n = 300) and after (n = 300) chilling over an 8-month period. Positive samples for E. coli O157 were subjected to virulence screening by PCR (stx1, stx2, and eaeA genes and the fliCH7 gene), antimicrobial susceptibility testing, and molecular typing by pulsed-field gel electrophoresis. A total of 9.7% (29 of 300) of the nonrefrigerated carcasses examined and 2.3% (7 of 300) of the refrigerated carcasses were positive for E. coli O157. All the isolates were serotype O157:H7, 92% (33 of 36) carried the stx1, stx2, and eaeA genes, and 8% (3 of 36) carried the stx2 and eaeA genes. Antimicrobial susceptibility testing showed a high degree of resistance: 29 strains (81%) were resistant to at least 1 antimicrobial of the 12 antimicrobials tested; 69% (25 of 36) were resistant to 4 or more antimicrobials. Molecular typing by pulsed-field gel electrophoresis found a high diversity of genetic types, implying little cross-contamination in the slaughterhouse. This study confirms that E. coli O157:H7 is present on the carcasses slaughtered in Spain, although its prevalence is reduced by the dry chilling process used. The recovered isolates showed potential pathogenesis and a high degree of multidrug resistance, confirming the importance of bovine meat monitoring.

scholarly journals Phenotypic and Molecular Analysis of Penicillin-Nonsusceptible Streptococcus pneumoniae Isolates in Poland

2006 ◽  
Vol 51 (1) ◽  
pp. 40-47 ◽  
Author(s):  
Ewa Sadowy ◽  
Radosław Izdebski ◽  
Anna Skoczyńska ◽  
Paweł Grzesiowski ◽  
Marek Gniadkowski ◽  
...  
Keyword(s):  
Streptococcus Pneumoniae ◽  
Molecular Analysis ◽  
Gel Electrophoresis ◽  
Multilocus Sequence Typing ◽  
Susceptibility Testing ◽  
Bacterial Pathogen ◽  
Pulsed Field Gel Electrophoresis ◽  
Pulsed Field ◽  
Related Clone ◽  
Sequence Types

ABSTRACT β-Lactams are the drugs of choice for the treatment of infections caused by the important bacterial pathogen Streptococcus pneumoniae. The recent growth of resistance of this organism to penicillin observed worldwide is of the highest concern. In this study, using 887 surveillance pneumococcal isolates recovered in Poland from 1998 to 2002, we observed the increase in penicillin nonsusceptibility from 8.7% to 20.3%. All of the 109 penicillin-nonsusceptible S. pneumoniae (PNSP) isolates identified, together with 22 archival PNSP isolates from 1995 to 1997, were subsequently analyzed by susceptibility testing, serotyping, profiling of pbp genes, pulsed-field gel electrophoresis, and multilocus sequence typing (MLST). Four predominant serotypes, serotypes 6B, 9V, 14, and 23F, characterized 85.5% of the isolates. MLST revealed the presence of 34 sequence types, 15 of which were novel types. Representatives of seven multiresistant international clones (Spain23F-1, Spain6B-2, Spain9V-3, Taiwan23F-15, Poland23F-16, Poland6B-20, and Sweden15A-25) or their closely related variants comprised the majority of the study isolates. The spread of Spain9V-3 and its related clone of serotype 14/ST143 has remarkably contributed to the recent increase in penicillin resistance in pneumococci in the country.

scholarly journals Molecular Epidemiology of Salmonella Enterica Serovar Saintpaul Isolated from Animals, Food, and Humans in 12 European Countries

2012 ◽  
Vol 56 (4) ◽  
pp. 459-466 ◽  
Author(s):  
Dariusz Wasyl ◽  
Magdalena Zając ◽  
Derek J. Brown ◽  
Henry Kuronen ◽  
Kim Van Der Zwaluw ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Salmonella Enterica ◽  
Pulsed Field Gel Electrophoresis ◽  
European Countries ◽  
Animal Origin ◽  
Potential Threat ◽  
Food Of Animal Origin ◽  
Control Programmes ◽  
High Degree ◽  
Pfge Profiles

Abstract The molecular epidemiological relationship among isolates of Salmonella enterica serovar (S.) Saintpaul, which was identified in animals, food, and humans in several EU countries, was investigated. Pulsed-field gel electrophoresis (PFGE) revealed a high degree of genetic diversity (82 XbaI PFGE profiles with 42.6% similarity) among 159 S. Saintpaul isolates from animals (n=91), food of animal origin (n=29), and humans (n=36) in 12 European countries during 2005 to 2009. Most frequent profiles (n=12) comprised almost 50% of the tested isolates. Profiles obtained in isolates from a single source within a particular geographical region or particular period of time were indistinguishable or closely related. Turkeys were confirmed as the major reservoir for S. Saintpaul. Indistinguishable PFGE profiles were identified in up to 19 isolates from turkey breeding and fattening flocks and food over the study period. Other animals, including food and pets, may also contribute to S. Saintpaul spread. International trade of animals and food, as well as travelling contributes to the spread of a specific clone to different geographical areas. Although control programmes in breeding turkey flocks, together with improved biosecurity, may interrupt the major transmission routes, it was concluded that S. Saintpaul will continue to represent a potential threat to human health.

scholarly journals Tigecycline susceptibility of multidrug-resistant Acinetobacter baumannii from intensive care units in the western Balkans

2020 ◽  
Vol 67 (3) ◽  
pp. 176-181
Author(s):  
Ina Gajic ◽  
Lazar Ranin ◽  
Dusan Kekic ◽  
Natasa Opavski ◽  
Aleksandra Smitran ◽  
...  
Keyword(s):  
Acinetobacter Baumannii ◽  
Susceptibility Testing ◽  
Multidrug Resistant ◽  
Inappropriate Therapy ◽  
Antibiotic Susceptibility Testing ◽  
Broth Microdilution ◽  
Western Balkans ◽  
Colistin Resistance ◽  
European Committee ◽  
Carbapenem Resistant

AbstractTigecycline can be effective to treat infections of carbapenem-resistant Acinetobacter baumannii (CRAB) however, no interpretive criteria have been approved so far. The objectives of this study were to evaluate the proportion of CRAB isolates and to compare gradient test with a broth microdilution (BMD) method for tigecycline susceptibility testing of A. baumannii.This study included 349 multidrug-resistant (MDR) Acinetobacter spp. collected from Serbia, Montenegro, Bosnia and Herzegovina in 2016 and 2017. Antibiotic susceptibility testing was performed by disk diffusion, VITEK2, gradient, ComASP Colistin. Tigecycline susceptibilities were interpreted according to breakpoints of European Committee on Antimicrobial Susceptibility Testing (EUCAST) and Food and Drug Administration (FDA).Majority of the tested isolates were CRAB (92.8%). Tigecycline MIC50/MIC90 values were 4/8 μg/mL by BMD and 0.5/4 μg/mL by gradient test. Essential agreement for BMD and gradient test amounted to 65.1%. With EUCAST breakpoints, categorical agreement (CA) was achieved in 38% isolates. Major discordance (MD-false susceptibility/resistance) and minor discordance (mD-false categorization involving intermediate results) were observed in 10% and 57% A. baumannii, respectively. With FDA breakpoints, CA, MD and mD were observed in 44%, 16% and 47% isolates, respectively. Colistin resistance was 2.1%.The study highlights a high proportion of CRAB and several discordances between BMD and gradient test which may lead to inappropriate therapy.

scholarly journals The Aquatic Environment as a Reservoir ofVibrio choleraeO1 in Hydrographic Basins of the State of Pernambuco, Brazil

2013 ◽  
Vol 2013 ◽  
pp. 1-5
Author(s):  
Carina Lucena Mendes-Marques ◽  
Vladimir da Mota Silveira Filho ◽  
Ana Paula Rocha da Costa ◽  
Mariana de Lira Nunes ◽  
Sandoval Vieira da Silva Filho ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Genetic Similarity ◽  
Virulence Genes ◽  
Intergenic Spacer ◽  
Pulsed Field Gel Electrophoresis ◽  
N Gene ◽  
Intergenic Spacer Region ◽  
Quiescent Period ◽  
Pathogenic Genes ◽  
Culture Negative

After the worldwide cholera epidemic in 1993, permanent environmental monitoring of hydrographic basins was established in Pernambuco, Brazil, where cholera is endemic. After a quiescent period, 4rfbN (serogroup O1) positive water samples that were culture negative were detected by multiplex single-tube nested PCR (MSTNPCR); 2 of these were alsoctxA (cholera toxin) positive. From May to June 2012, 30V. choleraeO1 isolates were obtained by culturing samples. These isolates were analyzed for the presence of virulence genes by PCR, intergenic spacer region 16S-23S PCR (ISR-PCR), and pulsed field gel electrophoresis (PFGE). The isolates were positive for therfbN gene and negative for the assessed pathogenic genes and were classified into 2 groups by ISR and the same profile by PFGE. Close genetic similarity was observed between them (2012) and environmental strains from 2004 to 2005, indicating the permanence of endemicV. choleraeO1 in the region.

scholarly journals Sensitivity of Listeria monocytogenes to Sanitizers Used in the Meat Processing Industry

2002 ◽  
Vol 68 (12) ◽  
pp. 6405-6409 ◽  
Author(s):  
Nadya Romanova ◽  
Stacy Favrin ◽  
Mansel W. Griffiths
Keyword(s):  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Efflux Pump ◽  
Pulsed Field Gel Electrophoresis ◽  
Meat Processing ◽  
Processing Industry ◽  
Automated Ribotyping ◽  
Plasmid Profiling ◽  
The Relationship ◽  
Ammonium Compounds

ABSTRACT Nineteen Listeria monocytogenes strains were characterized by automated ribotyping, pulsed-field gel electrophoresis, and plasmid profiling to determine the relationship between genotype and sanitizer resistance. Isolates within a ribogroup had a consistent sensitivity or resistance phenotype except for ribogroup C isolates. All isolates with resistance phenotypes harbored two plasmids. The sensitivity of L. monocytogenes strains to quaternary ammonium compounds (QACs) was correlated with sensitivity to sanitizers and antibiotics with other modes of action. All isolates tested contained the mdrL gene, which encodes an efflux pump that confers resistance to QACs and is both chromosome and plasmid borne.

scholarly journals Optimization of Pulsed-Field Gel Electrophoresis for Legionella pneumophila Subtyping

2010 ◽  
Vol 76 (5) ◽  
pp. 1334-1340 ◽  
Author(s):  
Haijian Zhou ◽  
Hongyu Ren ◽  
Bingqing Zhu ◽  
Biao Kan ◽  
Jianguo Xu ◽  
...  
Keyword(s):  
Legionella Pneumophila ◽  
Gel Electrophoresis ◽  
Water System ◽  
Epidemiological Data ◽  
Pulsed Field Gel Electrophoresis ◽  
Discriminatory Power ◽  
Similarity Coefficients ◽  
Pulsed Field ◽  
Software Analysis ◽  
High Degree

ABSTRACT A total of 32 strains of Legionella pneumophila were used to optimize pulsed-field gel electrophoresis (PFGE) for subtyping of L. pneumophila. Twenty-six isolates of L. pneumophila with various origins and 11 isolates from five different water systems were used as the panels. For optimization of electrophoretic parameters (EPs) of SfiI PFGE, 26 isolates were analyzed with SfiI digestion, using four EPs yielding the same D value. The EP of a switch time of 5 to 50 s for 21 h had the smallest similarity coefficients and was declared the optimal EP for SfiI PFGE of L. pneumophila. By software analysis and pilot study, AscI was chosen as another PFGE enzyme. AscI PFGE could cluster the isolates from each water system into the same or very similar patterns and had a high degree of typing concordance with other molecular methods. In evaluating the discriminatory power of AscI with the panel of 26 isolates, AscI PFGE gave one single pattern and a D value of 100%. AscI PFGE had a high discriminatory power and a high degree of consistency with epidemiological data and other molecular typing methods for L. pneumophila subtyping, and hence, AscI could be used as a restriction enzyme in PFGE subtyping of L. pneumophila.

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