Suitability of Repetitive-DNA-Sequence-Based PCR Fingerprinting for Characterizing Epidemic Isolates of Salmonella enterica Serovar Saintpaul

1998 ◽  
Vol 36 (6) ◽  
pp. 1549-1554 ◽  
Author(s):  
W. Beyer ◽  
F. M. Mukendi ◽  
P. Kimmig ◽  
R. Böhm

Three molecular typing methods, repetitive-sequence-based PCR (rep-PCR) fingerprinting, plasmid profiling, and arbitrarily primed PCR fingerprinting, were used to characterize isolates of Salmonella enterica serovar Saintpaul. Most of the isolates were obtained from epidemic human cases of food-borne salmonellosis, together with some from the food material suspected to be the source of infection, and a few were obtained from other cases apparently not related to the epidemic. All three methods adequately discriminated the epidemic strain from other strains of the serovar. In addition several isolates from human cases which are not identical to the epidemic strain were found. These isolates therefore must have been responsible for some sporadic infections, which were only temporally related to the epidemic. These strains showed a high degree of similarity to a strain isolated from a turkey. rep-PCR fingerprinting with REP-Dt primers and primer ERIC1R, applicable even to crude cell lysates, offers an attractive choice as a primary method for the discrimination of variousSalmonella serotypes as well as isolates within serotype Saintpaul.

Author(s):  
Reda M. Gaafar ◽  
Marwa M. Hamouda ◽  
Khalid A. El-Dougdoug ◽  
Sameh Fayez Fouad

Abstract Background Salmonella is considered to be the second largest source of infection in food-borne diseases. It is also considered one of the most important dangers particularly in the meat and dairy industry. Therefore, the main objective of our study was to determine the relationship between thermotolerance of a Salmonella serotype and the expression of DnaK and HtrA genes. Results In this study, expression of the two genes DnaK and HtrA was compared under four different temperatures 37 °C, 42 °C, 50 °C, and 55 °C in two serotypes of Salmonella enterica subsp. enterica. One of them was isolated from tahini product and identified as Salmonella enterica subsp. enterica serovar choleraesuis. This identified serotype was found to be more thermotolerant than the second serotype (Salmonella enterica subsp. enterica serovar typhimurium (ATCC 13311)), which was used as reference. This conclusion was based on D and Z values, which were used to compare thermoresistance ability of the two serotypes under four different temperatures 60 °C, 65 °C, 70 °C, and 75 °C. In addition, the results of qRT-PCR showed that after 43 °C (induction temperature), the relative expression (fold change) of DnaK and HtrA genes increased up to 5 and 47, respectively, comparing to their expression at 37 °C. Conclusions Thermotolerance of the identified S. choleraesuis serotype showed significantly high expression levels of DnaK and HtrA genes.


2018 ◽  
Vol 68 (3) ◽  
pp. 377 ◽  
Author(s):  
B. N. FASAEI ◽  
I. A. TAMAI

Salmonellosis is an important food-borne bacterial zoonotic disease that affects both people and animals. Contamination sources include direct or indirect animal contact. We determined and measured the risk of Salmonella infection from zoo animal to human contact by isolation of Salmonella serovars from zoo animals kept at Karaj zoo park, Iran. Salmonella was isolated from 21 (20%) of the 104 anal swap samples. From the 21 collected samples 4, 7 and 10 were originated from birds, mammals and reptiles, respectively. Serotyping detected nine different serotypes including Enteritidis (n=4), Seftenberg (n=1), Typhimurium (n=4), Virchow (3), Berkeley (n=1), Kingabwa (n=2), Newport (n=2), Marina (n=2) and Havana (n=1). All the isolates except one (serovar Marina subspecies Hautenae IV) were belonged to Salmonella enterica subspecies enterica I. Salmonella enterica serotypes Typhimurium and Enteritidis were the most commonly detected serotypes. All the isolates tested, were resistant to one or more antibiotics and five isolates including the monkeys’ and long-eared owls’ isolates were multiresistant. RAPD profiles of each isolate produced with two different primers were identical. These finding shows highly contamination of zoo animals from Salmonella especially from the multiresistant isolates. All of the animals in the zoo, pet owners, veterinarians, zookeepers and visitors are at high risk of salmonellosis with reptiles being the most important source of infection.


2006 ◽  
Vol 72 (10) ◽  
pp. 6632-6637 ◽  
Author(s):  
J. M. Adaska ◽  
A. J. Silva ◽  
A. C. B. Berge ◽  
W. M. Sischo

ABSTRACT Fifty-six human and 24 adult dairy cattle isolates of Salmonella enterica serovar Typhimurium from a single county in California were compared using ribotyping, insertion sequence typing (IS200), pulsed-field gel electrophoresis, plasmid typing, phage typing, and antimicrobial resistance testing. The majority of the isolates fell into one of two groups which were phage types DT104 and DT193. Combining the information from all typing methods, a total of 45 different “clusters” were defined, with 35 of those including only a single isolate. The library of isolates had a high degree of variability, but antibiotic resistance and plasmid typing each defined single clusters in which human or bovine isolates predominated (χ2, P < 0.05).


2022 ◽  
Vol 12 ◽  
Author(s):  
Xenia Vázquez ◽  
Vanesa García ◽  
Javier Fernández ◽  
Margarita Bances ◽  
María de Toro ◽  
...  

Colistin is a last-resort antibiotic in fighting severe infections caused by multidrug resistant Gram negative pathogens in hospitals. Zoonotic bacteria acquire colistin resistance in animal reservoirs and mediate its spread along the food chain. This is the case of non-typhoid serovars of Salmonella enterica. Colistin-resistant S. enterica in foods represents a threat to human health. Here, we assessed the prevalence of colistin-resistance in food-borne isolates of S. enterica (2014–2019; Asturias, Spain), and established the genetic basis and transferability of this resistance. Five out of 231 isolates tested (2.2%) were resistant to colistin. Four of them, belonging to the European monophasic ST34 clone of S. Typhimurium, were characterized in the present study. They were collected from pork or pork and beef meat-derived products, either in 2015 (three isolates) or 2019 (one isolate). Molecular typing with XbaI-PFGE and plasmid profiling revealed distinct patterns for each isolate, even though two of the 2015 isolates derived from the same sample. The MICs of colistin ranged from 8 to 16 mg/L. All isolates carried the mcr-1.1 gene located on conjugative plasmids of the incompatibility groups IncX4 (2015 isolates) or IncHI2 (2019 isolate). Apart from colistin resistance, the four isolates carried chromosomal genes conferring resistance to ampicillin, streptomycin, sulfonamides and tetracycline [blaTEM–1, strA-strB, sul2, and tet(B)] and heavy metals, including copper and silver (silESRCFBAGP and pcoGE1ABCDRSE2), arsenic (arsRSD2A2BCA1D1) ± mercury (merEDACPTR), which are characteristically associated with the European ST34 monophasic clone. The 2019 isolate was also resistant to other antibiotics, comprising third generation cephalosporins and cephamycins. The latter phenotype was conferred by the blaCMY–2 gene located on an IncI1-I(α)-ST2 plasmid. Results in the present study identified meat-derived products as a reservoir of a highly successful clone harboring transferable plasmids which confer resistance to colistin and other clinically important antibiotics. An important reduction in the number of food-borne S. enterica detected during the period of the study, together with the low frequency of colistin resistance, underlines the success of One Health initiatives, such as those implemented at the UE, to control zoonotic bacteria along the food chain and to halt the spread of antimicrobial resistance.


2008 ◽  
Vol 190 (24) ◽  
pp. 8155-8162 ◽  
Author(s):  
Fiona J. Cooke ◽  
Derek J. Brown ◽  
Maria Fookes ◽  
Derek Pickard ◽  
Alasdair Ivens ◽  
...  

ABSTRACT Salmonella enterica serovar Typhimurium definitive phage type 104 (DT104) has caused significant morbidity and mortality in humans and animals for almost three decades. We completed the full DNA sequence of one DT104 strain, NCTC13348, and showed that significant differences between the genome of this isolate and the genome of the previously sequenced strain Salmonella serovar Typhimurium LT2 are due to integrated prophage elements and Salmonella genomic island 1 encoding antibiotic resistance genes. Thirteen isolates of Salmonella serovar Typhimurium DT104 with different pulsed-field gel electrophoresis (PFGE) profiles were analyzed by using multilocus sequence typing (MLST), plasmid profiling, hybridization to a pan-Salmonella DNA microarray, and prophage-based multiplex PCR. All the isolates belonged to a single MLST type, sequence type ST19. Microarray data demonstrated that the gene contents of the 13 DT104 isolates were remarkably conserved. The PFGE DNA fragment size differences in these isolates could be explained to a great extent by differences in the prophage and plasmid contents. Thus, here the nature of variation in different Salmonella serovar Typhimurium DT104 isolates is further defined at the gene and whole-genome levels, illustrating how this phage type evolves over time.


2004 ◽  
Vol 70 (7) ◽  
pp. 4030-4034 ◽  
Author(s):  
Jaime Martinez-Urtaza ◽  
Ernesto Liebana ◽  
Lourdes Garcia-Migura ◽  
Pelayo Perez-Piñeiro ◽  
Montserrat Saco

ABSTRACT Twenty-three Salmonella enterica serovar Typhimurium isolates from marine environments were characterized by phage typing, pulsed-field gel electrophoresis (PFGE) analysis, plasmid analysis, and antibiotic resistance, and the distribution of the different types in the coastal waters were subsequently analyzed. Five phage types were identified among the isolates (PT41, PT135, PT99, DT104, and DT193). PT135 isolates were exclusively detected during the winter months from 1998 to 2000, whereas DT104 and PT41 isolates were detected exclusively in the summer months from 2000 to 2002. XbaI PFGE analysis revealed 9 PFGE types, and plasmid profiling identified 8 plasmid types (with 1 to 6 plasmids) among the isolates. Only three isolates presented multidrug resistance to antibiotics. Two DT104 isolates were resistant to 8 and 7 antibiotics (profiles ACCeFNaSSuT and ACeFNeSSuT), whereas a PT193 isolate presented resistance to 6 antibiotics (profile ACFSSu). In addition, four PT41 isolates were resistant to a single antibiotic. The detection of multidrug-resistant phage types DT104 and DT193 in shellfish emphasizes the importance of monitoring the presence of Salmonella in routine surveillance of live bivalve molluscs.


1998 ◽  
Vol 36 (10) ◽  
pp. 2957-2963 ◽  
Author(s):  
Mary Ellen Rafferty ◽  
Aldona L. Baltch ◽  
Raymond P. Smith ◽  
Lawrence H. Bopp ◽  
Carol Rheal ◽  
...  

During an outbreak of diarrhea in a general hospital in 1992, 166Clostridium difficile isolates from 102 patients were typed by restriction enzyme analysis (REA), arbitrarily primed PCR (AP-PCR), and protein profile analysis (PP) techniques. A total of 18 types and 5 subtypes were identified by REA, 32 types were identified by AP-PCR, and 9 types were identified by PP. Analysis of the data indicated the presence of a predominant strain among 76, 75, and 84% of the isolates by REA, AP-PCR, and PP, respectively. Subsequently, 45C. difficile isolates which had been collected in 1990 from 33 patients in the same hospital following a significant increase in the number of cases of diarrhea caused by C. difficile were studied by REA, AP-PCR, and PP typing techniques. Thirteen types and one subtype were identified by REA, 12 types were identified by AP-PCR, and 5 types were identified by PP. As with the isolates from 1992, a dominant strain was identified. This strain was represented by 53, 64, and 70% of the total number of isolates when the strains were typed by REA, AP-PCR, and PP, respectively. Every isolate (210 of 211) from both 1990 and 1992 that was available for typing was typeable by all three methods. Furthermore, the same dominant strain was identified in both 1990 and 1992 by each method. This study demonstrates that each of the three typing methods can be useful in epidemiologic investigations of C. difficileoutbreaks and that one strain can be dominant in an institution over a number of years.


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