Enterococci from Appenzeller and Schabziger Raw Milk Cheese: Antibiotic Resistance, Virulence Factors, and Persistence of Particular Strains in the Products

2007 ◽  
Vol 70 (2) ◽  
pp. 450-455 ◽  
Author(s):  
S. P. TEMPLER ◽  
A. BAUMGARTNER
Keyword(s):  
Virulence Factors ◽  
Gel Electrophoresis ◽  
Raw Milk ◽  
Pulsed Field Gel Electrophoresis ◽  
Pulsed Field ◽  
Phenotypic Resistance ◽  
Genes Encoding ◽  
Resistance Patterns ◽  
Raw Milk Cheese ◽  
Selection Of

Enterococci are natural residents of human and animal intestinal tracts, and grow to high numbers in a variety of cheeses. The aim of this study was to determine the diversity of enterococci in two types of artisanal raw milk cheese (Schabziger and Appenzeller) and to investigate whether particular strains with triple resistance against chloramphenicol (Chl), tetracycline (Tet), and erythromycin (Ery) persist in the production system. Of 46 cheese samples, a total of 312 Enterococcus strains were isolated over a 5-month period on selective agar plates containing Chl, Tet, or, Ery. Enterococcus faecalis was the predominant species (80.7%), followed by Enterococcus faecium (5.1%), and Enterococcus durans (11.7%). According to the phenotypic resistance patterns, a selection of 150 strains was analyzed with PCR for the presence of genes encoding resistance to Ery (ereA, ereB, mphA, ermA, ermB, ermC, mrsA/mrsB, mefA/mefE), and Tet (tetM, tetL). Because virulence factors have been linked to the pathogenicity of enterococci, the strain selection was also tested for the presence of the following virulence factors: Agg, GelE, Cyl, Esp, EfaAfs, EfaAfm, Cpd, Cob, and Ccf. All tested strains contained at least two of the nine virulence genes taken into analysis. Pulsed-field gel electrophoresis patterns of the isolates showed a limited persistence of several strains over a period of 1 to 2 months in Schabziger, and more than 2 months in Appenzeller. Finally, the enterococcal flora in the two types of cheeses seems to be rather unrelated. Within 150 strains from 25 different cheese samples (11 Appenzeller and 14 Schabziger), 41 pulsed-field gel electrophoresis patterns could be identified, and only 1 of these was found in enterococci from both types of cheese.

Characteristics of Virulent Vibrio parahaemolyticus Isolated from Oregon and Washington

2007 ◽  
Vol 70 (4) ◽  
pp. 1011-1016 ◽  
Author(s):  
TSAI-HSIN CHIU ◽  
JINGYUN DUAN ◽  
YI-CHENG SU
Keyword(s):  
Pacific Northwest ◽  
Virulence Factors ◽  
Gel Electrophoresis ◽  
Vibrio Parahaemolyticus ◽  
Pulsed Field Gel Electrophoresis ◽  
Outbreak Strain ◽  
Pulsed Field ◽  
Clinical Strains ◽  
Virulent Strains ◽  
The Pacific

Thirty-four virulent strains of Vibrio parahaemolyticus containing tdh and/or trh genes isolated from Oregon and Washington coastal water were analyzed for O-group antigens and urease activity, and by pulsed-field gel electrophoresis. Six O serotypes (O1, O3, O4, O5, O10, and O11) were identified among the isolates, with the O5 group (19 isolates) being the most prevalent, followed by the O1 group (9 isolates). Nearly all (33 of 34) isolates were capable of producing urease, which reaffirmed the correlation between urease production and virulence factors of V. parahaemolyticus strains isolated from the Pacific Northwest. Pulsed-field gel electrophoresis analysis with NotI and SfiI digestions of the 34 V. parahaemolyticus isolates plus five clinical strains revealed 22 patterns (N1S1 to N20S22), with N1S1 (25.6%) being the most common, followed by N2S2 (10.3%). Nine Oregon isolates were grouped with a 1997 Oregon outbreak strain (027-1C1) with the same serotype (O5), virulence factors (tdh+ and trh+), and genotype (N1S1). Three Washington isolates were found to share the same serotype (O1), virulence factors (tdh+ and trh+), and genotype (N2S2) with a 1997 Washington outbreak strain (10293). The repetitive isolation of virulent strains of V. parahaemolyticus identical to clinical strains involved in previous outbreaks indicates potential hazards associated with oyster consumption. These data may be useful in risk assessment of V. parahaemolyticus infections associated with raw oyster consumption in Oregon and Washington.

scholarly journals Genome Sequence of Lactococcus garvieae IPLA 31405, a Bacteriocin-Producing, Tetracycline-Resistant Strain Isolated from a Raw-Milk Cheese

2012 ◽  
Vol 194 (18) ◽  
pp. 5118-5119 ◽  
Author(s):  
Ana Belén Flórez ◽  
Pilar Reimundo ◽  
Susana Delgado ◽  
Elena Fernández ◽  
Ángel Alegría ◽  
...  
Keyword(s):  
Virulence Factors ◽  
Genome Sequence ◽  
Draft Genome ◽  
Raw Milk ◽  
M Gene ◽  
Content Type ◽  
Lactococcus Garvieae ◽  
Genes Encoding ◽  
Raw Milk Cheese ◽  
Plasmid Replicons

ABSTRACTThis work describes the draft genome sequence ofLactococcus garvieaeIPLA 31405, isolated from a traditional Spanish cheese. The genome contains a lactose-galactose operon, a bacteriocin locus, two integrated phages, a transposon harboring an activetet(M) gene, and two theta-type plasmid replicons. Genes encoding virulence factors were not recorded.

scholarly journals A physical map of the human regulator of complement activation gene cluster linking the complement genes CR1, CR2, DAF, and C4BP.

1988 ◽  
Vol 167 (2) ◽  
pp. 664-669 ◽  
Author(s):  
J Rey-Campos ◽  
P Rubinstein ◽  
S Rodriguez de Cordoba
Keyword(s):  
Gene Cluster ◽  
Complement Activation ◽  
Gel Electrophoresis ◽  
Physical Map ◽  
Pulsed Field Gel Electrophoresis ◽  
Complement Components ◽  
Pulsed Field ◽  
Human Genes ◽  
Tight Linkage ◽  
Genes Encoding

We report the organization of the human genes encoding the complement components C4-binding protein (C4BP), C3b/C4b receptor (CR1), decay accelerating factor (DAF), and C3dg receptor (CR2) within the regulator of complement activation (RCA) gene cluster. Using pulsed field gel electrophoresis analysis these genes have been physically linked and aligned as CR1-CR2-DAF-C4BP in an 800-kb DNA segment. The very tight linkage between the CR1 and the C4BP loci, contrasted with the relative long DNA distance between these genes, suggests the existence of mechanisms interfering with recombination within the RCA gene cluster.

Staphylococcal Poisoning Foodborne Outbreak: Epidemiological Investigation and Strain Genotyping

2013 ◽  
Vol 76 (12) ◽  
pp. 2093-2098 ◽  
Author(s):  
S. GALLINA ◽  
D. M. BIANCHI ◽  
A. BELLIO ◽  
C. NOGAROL ◽  
G. MACORI ◽  
...  
Keyword(s):  
Nasal Mucosa ◽  
Biological Samples ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Food Handler ◽  
Staphylococcal Protein A ◽  
Spa Typing ◽  
Pulsed Field ◽  
Genes Encoding ◽  
Genetic Profiles

In June 2011, an outbreak of Staphylococcus aureus enterotoxin food poisoning gastroenteritis occurred in Turin, Italy, following a catered dinner party at a private home. Within a few hours, 26 of the 47 guests experienced gastrointestinal illness, and 9 were hospitalized. A retrospective cohort study using a standardized questionnaire was carried out, and the risk ratios for each food item were calculated. The analysis indicated consumption of seafood salad as the most probable cause of the outbreak (risk ratio = 11.72; 95% confidence interval, 1.75 to 78.54). Biological samples were collected from four of the hospitalized guests (stool and vomit), nasal mucosa swabs from three food handlers employed with the caterer, and available food residuals. All stool and vomit samples tested positive for enterotoxigenic S. aureus. As residues of the seafood salad were no longer available for sampling, suspected contamination could not be verified. However, no other food was found contaminated by S. aureus or its enterotoxins. All isolates from the biological samples were characterized at the genomic level by means of two multiplex PCR protocols to determine the presence of genes encoding staphylococcal enterotoxins, pulsed-field gel electrophoresis and staphylococcal protein A gene (spa) typing to describe their genetic profiles. All the isolates presented genes encoding SEA and SEI; the pulsed-field gel electrophoresis genetic profiles revealed the same pulsotype in the microorganism isolated from the hospitalized guests as in one of the isolates from a food handler's nasal mucosa, and the spa-typing analysis reported two closely related spa types (t701 and t267), implicating the food handler as the most likely outbreak source.

scholarly journals Monitoring and Characterization of Extended-Spectrum β-Lactamases in Escherichia coli Strains from Healthy and Sick Animals in Spain in 2003

2005 ◽  
Vol 49 (3) ◽  
pp. 1262-1264 ◽  
Author(s):  
Laura Briñas ◽  
Miguel Angel Moreno ◽  
Tirushet Teshager ◽  
Yolanda Sáenz ◽  
María Concepción Porrero ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Pulsed Field ◽  
Extended Spectrum ◽  
Genes Encoding ◽  
M 14

ABSTRACT Genes encoding CTX-M-14, CTX-M-9, CTX-M-1, CTX-M-32, SHV-12, TEM-52, or CMY-2 β-lactamases were detected in 21 Escherichia coli strains recovered during 2003 from sick animals (11 of 459 [2.4%] strains) and healthy animals (10 of 158 [6.3%] strains) in Spain. Twelve of these strains harbored bla CTX-M genes and showed unrelated pulsed-field gel electrophoresis patterns.

scholarly journals Detection and Pulsed-Field Gel Electrophoresis Typing of Listeria monocytogenes Isolates from Milk Vending Machines in Croatia

Beverages ◽  
2019 ◽  
Vol 5 (3) ◽  
pp. 46
Author(s):  
Nevijo Zdolec ◽  
Dean Jankuloski ◽  
Marta Kiš ◽  
Brigita Hengl ◽  
Nataša Mikulec
Keyword(s):  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Molecular Detection ◽  
Dna Amplification ◽  
Raw Milk ◽  
Pulsed Field Gel Electrophoresis ◽  
Effective Control ◽  
Pulsed Field ◽  
Milk Samples ◽  
Vending Machines

The aim of the present study was to evaluate the persistence of Listeria monocytogenes in raw milk from vending machines, based on culture and molecular detection of pathogen and Pulsed-Field Gel Electrophoresis typing. From December 2015 to January 2017, a total of 319 milk samples from 36 vending machines were examined for the presence and count of L. monocytogenes by reference methods ISO 11290:1 and ISO 11290:2. Molecular detection of pathogens was performed by loop-mediated isothermal DNA amplification (LAMP) coupled with bioluminescence (Molecular Detection Assay). L. monocytogenes was detected by MDA in 14 milk samples (4.38%) from four farms, compared to eight positive samples (2.5%) retrieved by a reference ISO method. Cultivable L. monocytogenes isolates were subjected to Pulsed-Field Gel Electrophoresis typing and pulsotypes were compared with those obtained during the previous survey in Croatia (2014‒2015). It was found that identical PFGE patterns of L. monocytogenes occur in milk samples of the same producer over a three-year period, indicating the persistence of pathogens in raw milk vending machines. The results obtained support the need for more effective control of milk in the entire food chain.

scholarly journals Serotype 19F Multiresistant Pneumococcal Clone Harboring Two Erythromycin Resistance Determinants [erm(B) and mef(A)] in South Africa

2001 ◽  
Vol 45 (5) ◽  
pp. 1595-1598 ◽  
Author(s):  
Lesley McGee ◽  
Keith P. Klugman ◽  
Avril Wasas ◽  
Thora Capper ◽  
Adrian Brink
Keyword(s):  
South Africa ◽  
Streptococcus Pneumoniae ◽  
Private Sector ◽  
Dna Fingerprinting ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Erythromycin Resistance ◽  
Pulsed Field ◽  
Resistance Determinants ◽  
Genes Encoding

ABSTRACT One hundred eighteen erythromycin-resistant Streptococcus pneumoniae (ERSP) strains (MICs of ≥0.5 μg/ml) from five laboratories serving the private sector in South Africa were analyzed for the genes encoding resistance to macrolides. Sixty-seven ERSP strains (56.8%) contained the erm(B) gene, and 15 isolates (12.7%) contained the mef(A) gene. Thirty-six isolates (30.5%) harbored both the erm(B) and mef(A) genes and were highly resistant to erythromycin and clindamycin. DNA fingerprinting by BOX-PCR and pulsed-field gel electrophoresis identified 83% of these strains as belonging to a single multiresistant serotype 19F clone.

Mapping of genes encoding glycoside hydrolases on the chromosome of Cellulomonas fimi

2001 ◽  
Vol 47 (12) ◽  
pp. 1063-1067 ◽  
Author(s):  
Dominik Stoll
Keyword(s):  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Glycoside Hydrolases ◽  
Circular Chromosome ◽  
Cellulomonas Fimi ◽  
Pulsed Field ◽  
A Genome ◽  
Genome Map ◽  
Genes Encoding ◽  
Single Circular Chromosome

Cellulomonas fimi genomic DNA was digested with HpaI, MunI, HindIII, and NsiI, producing fragments ranging in size from 20 to 1400 kbp that were resolved by pulsed field gel electrophoresis. Genetic and physical linkages were determined by Southern blotting and were used to construct a genome map. Cellulomonas fimi has a single circular chromosome of approx. 4000 kbp. Except for two closely linked genes, cbh6A and cel5A, the genes known to encode glycoside hydrolases are scattered widely on the chromosome.Key words: Cellulomonas fimi, genome map, pulsed field gel electrophoresis, glycoside hydrolases.

scholarly journals Characterization of Recurrent and Sporadic Listeria monocytogenes Isolates from Raw Milk and Nondairy Foods by Pulsed-Field Gel Electrophoresis, Monocin Typing, Plasmid Profiling, and Cadmium and Antibiotic Resistance Determination

2001 ◽  
Vol 67 (2) ◽  
pp. 840-847 ◽  
Author(s):  
J. Harvey ◽  
A. Gilmour
Keyword(s):  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Raw Milk ◽  
Pulsed Field Gel Electrophoresis ◽  
Enzyme Electrophoresis ◽  
Genetic Changes ◽  
Pulsed Field ◽  
Time Period ◽  
Plasmid Profiling

ABSTRACT Following previous surveys to assess the incidence ofListeria monocytogenes in raw milk and nondairy foods processed in Northern Ireland, isolates were characterized as recurrent or sporadic on the basis of multilocus enzyme electrophoresis (MEE) analysis and restriction fragment length polymorphism typing. In the present study, 45 representative recurrent and sporadic electrophoretic types (ETs) previously identified by MEE were subjected to pulsed-field gel electrophoresis (PFGE) of genomic DNA macrorestriction fragments, monocin typing, plasmid profiling, and an examination of resistance to cadmium and nine different antibiotics. Although PFGE proved to be capable of subdividing a number of recurrent and sporadic ETs, the grouping of strains arrived at by PFGE and MEE were in broad agreement, and previous conclusions regarding the designation of L. monocytogenes strains as recurrent or sporadic remained unaltered. It is considered that PFGE was able to detect minor genetic changes in recurrent ETs which occurred during the time period in which food surveys were carried out. Production of type E monocin (Types A to E were found among the 45 strains), plasmid carriage, and resistance to cadmium occurred more frequently in recurrent than in sporadic strains and may be important with regard to the ability of L. monocytogenes to persist in food and food-processing environments. Only 2 of 45 strains showed resistance to any of the nine antibiotics tested: two sporadic strains were resistant to tetracycline (MIC, 64 μg ml−1).

Arcobacter Species and Their Pulsed-Field Gel Electrophoresis Genotypes in Finnish Raw Milk during Summer 2011

2013 ◽  
Vol 76 (9) ◽  
pp. 1630-1632 ◽  
Author(s):  
JOANA REVEZ ◽  
MARIANNE HUUSKONEN ◽  
MARJO RUUSUNEN ◽  
MIIA LINDSTRÖM ◽  
MARJA-LIISA HÄNNINEN
Keyword(s):  
Gel Electrophoresis ◽  
Raw Milk ◽  
Positive Sample ◽  
Pulsed Field Gel Electrophoresis ◽  
Selective Detection ◽  
Potential Hazard ◽  
Pulsed Field ◽  
Milk Samples ◽  
Unpasteurized Milk ◽  
Arcobacter Butzleri

The aim of this study was to investigate the occurrence of Arcobacter species in raw milk in Finland. A total of 177 raw milk samples, each from a separate farm, were examined from June to August 2011. Arcobacter species were isolated using an enrichment and selective detection procedure. Overall, 26 (15%) of the 177 samples yielded Arcobacter spp. Samples from 25 farms were positive for Arcobacter butzleri and from 1 farm for Arcobacter cryaerophilus. Moreover, both Arcobacter butzleri and A. cryaerophilus were recovered from 1 positive sample. To evaluate a possible genetic variability, one strain of A. butzleri from each farm and the A. cryaerophilus sample were analyzed by pulsed-field gel electrophoresis. Genotyping revealed that Arcobacter spp. populations are heterogeneous, and no dominant clone has spread in the investigated samples. Our study is the first report on the isolation of both A. butzleri and A. cryaerophilus in raw milk in Finland. Based on our findings, the presence of Arcobacter species in raw milk may pose a potential hazard for human health, in particular for consumers who prefer drinking unpasteurized milk.

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