Distribution of molecular subtypes within Salmonella enterica serotype Enteritidis phage type 4 and S. Typhimurium definitive phage type 104 in nine European countries, 2000-2004: results of an international multi-centre study

This study investigates the distribution of pulsed-field gel electrophoresis (PFGE) profiles within Salmonella enterica serotype Enteritidis phage type (PT) 4 and S. Typhimurium definitive phage type (DT) 104, from cases of human infection in nine European countries from 2000 to 2004. Isolates were subtyped using standardized methods and gel images submitted by each participating country to the coordinating centre (Health Protection Agency Centre for Infections, London, UK), where they were entered into a central database, developed within BioNumerics software, and designated using an agreed nomenclature. S. Enteritidis PT4 (n=3637) was differentiated into 38 different profiles. Simpson's index of diversity (D) of profiles ranged from 0.2 to 0.4. Profile SENTXB.0001 represented at least 80% of all profiles in each country. S. Typhimurium DT104 (n=1202) was differentiated into 28 different profile types. Simpson's D was at least 0.6 in all countries except in Austria and Italy. In both these countries over 74% of S. Typhimurium DT104 profiles were STYMXB.0013. Profile STYMXB.0061, was predominant in Denmark, Spain, Finland and England and Wales where it represented between 36% and 45% of profiles. Profile STYMXB.0001 represented nearly half of all profiles in Scotland and 23% in England and Wales. PFGE is proving useful for further discrimination within S. Enteritidis PT4 and S. Typhimurium DT104. Ascertainment of international outbreaks involving common serotypes and phage types may be increased by the timely pooling of PFGE profiles within a central database readily accessible to all participating countries.     

Relationship of pulsed-field profiles with key phage types of Salmonella enterica serotype Enteritidis in Europe: results of an international multi-centre study

Salmonella is one of the most common causes of foodborne infection in Europe with Salmonella enterica serovar Enteritidis (S. Enteritidis) being the most commonly identified serovar. The predominant phage type for S. Enteritidis is phage type (PT) 4, although PT 8 has increased in incidence. Within these phage types, pulsed-field gel electrophoresis (PFGE) provides a method of further subdivision. The international project, Salm-gene, was established in 2001 to develop a database of PFGE profiles within nine European countries and to establish criteria for real-time pattern recognition. It uses DNA fingerprints of salmonellas to investigate outbreaks and to evaluate trends and emerging issues of foodborne infection within Europe. The Salm-gene database contains details of about 11 700 S. Enteritidis isolates, demonstrating more than 65 unique PFGE profiles. The clonal nature of S. Enteritidis is evidenced by the high similarity and distribution of PFGE profiles. Over 56% (6603/11 716) of the submitted isolates of several different phage types were profile SENTXB.0001, although this profile is most closely associated with PT 4. The next most common profiles, SENTXB.0002 and SENTXB.0005, were closely associated with PT 8 and PT 21 respectively. Studies to investigate the relationship of profile types with outbreaks and possible vehicles of infection suggest that the incidence of PFGE profile SENTXB.0002, and thus PT 8, in some countries may be due to importation of foods or food production animals from Eastern Europe, where PT 8 is amongst the most frequently identified phage types. Collation of subtyping data, especially in the commonly recognized phage types, is necessary in order to evaluate trends and emerging issues in salmonella infection.     

Antimicrobial resistance and phage types of human and non-human Salmonella enterica isolates in Ireland, 1998-2003

Between 1998 and 2003, 5,161 isolates (3,182 human) of Salmonella enterica were received by the National Salmonella Reference Laboratory of Ireland. Serotyping, antimicrobial susceptibility testing and phage typing were performed by standard methods. The number of isolates of S. enterica serovar Typhimurium decreased from 579 (80%) in 1998 to 208 (19%) in 2003, while S. enterica serovar Enteritidis increased from 59 (8%) in 1998 to 219 (20%) in 2003. Definitive (DT) phage types 104 and DT104b accounted for a declining proportion of all Salmonella Typhimurium isolates (from n = 523 [90%] in 1998 to 126 [60%] in 2003). Numbers of Salmonella Enteritidis phage type 4 declined from 50 (85%) in 1998 to 59 (27%) in 2003. Twenty-eight isolates of typhoidal Salmonella were received with a history of recent travel in 17 cases. Resistance to multiple (four or more) antimicrobial agents was related to serotype and, where applicable, phage type, and was common in Salmonella Typhimurium. Salmonella Typhimurium predominated among isolates from cattle and pigs (n = 213 [58%]), while Salmonella Livingstone (n = 327) and S. Kentucky (n = 227) were predominant in isolates from poultry (total n = 554 [43%]). This paper discusses trends, and their implications, in Irish salmonella isolates since the establishment of the Reference Laboratory.     

Occurrence of phage types of non-typhoid Salmonella serovar in the Slovak Republic 2000-2003

Salmonellae of non-typhoidal serovars are the most important pathogens involved in foodborne diseases in humans all over the world. The incidence rates of two major Salmonella serovars, i.e. S. enterica serovar Enteritidis (S. Enteritidis) and S. enterica serovar Typhimurium (S. Typhimurium), in the Slovak Republic in 2000-2003 are given. Over the period studied, 829 S. Enteritidis strains and 258 S. Typhimurium strains isolated from patients with salmonellosis were investigated in the National Reference Centre for Salmonella Phage Typing. The S. Enteritidis strains were differentiated into 16 phage types, with phage type 8 being dominant since found in 73.6%, 53.8%, 62.8% and 45.6% of strains in 2000, 2001, 2002 and 2003, respectively. The following most frequent phage types were 4 and 13a. New phage types, i.e. 15, 5, 25 and 14b, were identified from salmonellosis outbreaks in 2003. The S. Typhimurium strains were also differentiated into 16 phage types with phage type DT104 strains being prevalent and showing an increase from 7.4% in 2000 to 44.6% in 2003; 54.2% of them were resistant and of R type ACSSut. The second most frequent phage type in 2000-2001 was 2b in 2003 DT41. The frequency of the other phage types was not epidemiologically significant.     

Multiresistant Salmonella enterica subspecies enterica serovar Typhimurium DT104 in the Czech republic 2000-2001

Salmonelloses belong to the most important alimentary diseases. From 1989 the main source of infection is S. Enteritidis (95% and more). The ratio of S. Typhimurium in the etiology declined gradually since 1994 from 3% in 1993 to 1.9% in 2001. The number of detected multiresistant strains STM DT104 is so far low in the Czech Republic. In 2000 laboratory tests confirmed 53 and in 2001 a total of 63 strains. Analysis of these strains revealed: 1. the proportion of multiresistant strains STM DT104 of the total number of isolated STM strains increased from 7% in 2000 to 11.5% in 2001, 2. the territory in the CR where these multiresistant strains were found expanded, 3. the relationship of proportions of the most frequently detected STM phagotypes in the human population changes: in 2000 this ratio was balanced, in 2001 the ratio of phagotype DT104 is roughly three times higher than of DT 141, 4. analysis of all STM strains from the environment and animals indicates that the frequency of STM DT104 is 73%. Almost 90% of these strains are multiresistant (resistant to one or several antimicrobial substances). The highest number was found in calves, piglets but also in poultry. With regard to the results of the submitted work it is important to concentrate attention on detection and further laboratory examination of STM strains in the human and animal population.     

Multidrug-resistant strains of Salmonella enterica Typhimurium, United States, 1997-1998

To evaluate multidrug-resistant strains of Salmonella enterica serotype Typhimurium, including definitive type 104 (DT104) in the United States, we reviewed data from the National Antimicrobial Resistance Monitoring System (NARMS). In 1997 to 1998, 703 (25%) of 2,767 serotyped Salmonella isolates received at NARMS were S. Typhimurium; antimicrobial susceptibility testing and phage typing were completed for 697. Fifty-eight percent (402) were resistant to > or = 1 antimicrobial agent. Three multidrug-resistant (> or = 5 drugs) strains accounted for (74%) 296 of all resistant isolates. Ceftriaxone resistance was present in 8 (3%), and nalidixic acid resistance in 4 (1%), of these multidrug-resistant strains. By phage typing, 259 (37%) of S. Typhimurium isolates were DT104, 209 (30%) were of undefined type and 103 (15%) were untypable. Fifty percent (202) of resistant (> or = 1 drug) isolates were DT104. Multidrug-resistant S. Typhimurium isolates, particularly DT104, account for a substantial proportion of S. Typhimurium isolates; ceftriaxone resistance is exhibited by some of these strains.     

Emergence of multidrug-resistant Salmonella enterica serotype Typhimurium phage-type DT104 among salmonellae causing enteritis in Israel

The relative frequency of salmonella strains isolated from hospitalized and non-hospitalized patients in Southern Israel changed during the period, 1994-6. Salmonella enterica serotype Typhimurium definitive phage-type 104 (DT104) appeared in Israel in 1994 and became the most prevalent strain in 1996. An outbreak of enteritis due to Salmonella enterica serotype Agona occurred in Israel, in October 1994 and lasted for 4 months. The relative frequency of Salmonella enterica serotype Enteritidis remained almost constant during these years, with seasonal fluctuations only. The importance of the increase in the prevalence of Typhimurium DT104 has been the epidemic spread of a multiresistant strain of R-type ACT (A, ampicillin; C, chloramphenicol; T, tetracycline) belonging to this phage-type. Since 1995 the frequency of Typhimurium DT104 isolates that possess, in addition to the above R-type, a chromosomally encoded resistance to the quinolone drug, nalidixic acid, increased tenfold. In 1996, 27% of the Typhimurium DT104 isolates were of R-type ACTN. S. Enteritidis exhibited over 95% susceptibility to at least eight of the most commonly used antibiotic drugs, and none of the isolates was resistant to quinolone or fluoroquinoline.     

Pheno-genotyping of Salmonella enterica serotype Enteritidis isolates identified in Sicily during a reemergence period

After an upward trend paralleling that occurring in most European countries, including Italy, since October 2002 Salmonella enterica serotype Enteritidis (S. Enteritidis) has again gained the first position among outbreak and sporadic human isolates of Salmonella in Sicily. Because phage typing of S. Enteritidis has many technical and epidemiological limitations and molecular methods have proved to be poorly discriminative for this organism, multiple typing, using phage typing together with pulsed field gel electrophoresis (PFGE) and plasmid profiling on a sample of fifty human and poultry isolates identified during the period October 2002 to May 2003 in Sicily, was chosen as the most valuable strategy to explore key features of this new epidemic wave. Although the limited number of strains imposes a cautious interpretation of the results, an apparently increasing phage type heterogeneity has emerged with rise in PT6 as the more striking event. While PFGE has confirmed the findings by other authors about the close genetic homogeneity between PT4 and PT6, plasmid profiling has provided discriminative patterns for PT6 strains. Combined phenotypic and genotypic profiles are necessary for epidemiological studies and public health investigations on S. enteritidis.     

Characterization of Salmonella enterica serotype typhimurium in the Czech Republic: phage types, antimicrobial and plasmid profiles

In this study a collection of 547 S. Typhimurium strains isolated in the years 2000 and 2001 both of the human and non-human origin were analysed. 21 different phage types were detected, the most frequent one was DT104 (46%) followed by DT141 (28%) and DT68 (3%). Resistance to one or more antimicrobial agents was found mainly in DT104 (77.4%). S. Typhimurium isolates resistant to 5 and more antimicrobial agents were found in three phagetypes DT104 (57%), DT120 and DT155. Plasmid profiling of DT104 isolates showed 10 different profiles. Pattern A found in 30.5% of tested strains was predominant and carried serovar specific plasmid and one additional small plasmid of approx. 2.5 kb.     

Epidemiological characteristics of Salmonella Typhimurium isolated from animals and feed in Poland

Fifty-seven Salmonella Typhimurium strains isolated from poultry, swine and animal feed in Poland during the years 1979-1998 and 2000-2002 were analysed with conventional and molecular techniques. Antimicrobial resistance as well as multiresistance was found, respectively, in 80.1% and 56.1% of the isolates and most frequently among isolates from 2000-2002. Of several phage types noted, DT104 was prevalent among poultry, swine and feed isolates. DT104, U302 and non-typable strains had a multiple resistant profile (ACSSuT) due to the presence of class I integrons. Pulse-field gel electrophoresis of XbaI and BlnI digest showed high genomic similarity between the strains and confirmed clonal spread of S. Typhimurium infections. Plasmid profiling allowed further differentiation of the strains. We have, therefore, confirmed the appearance of S. Typhimurium DT104 showing genome integrated integron-mediated antimicrobial resistance in Poland. These findings are significant for public and animal health risks and document the dissemination of DT104 epidemic strains into new geographical regions.     

The evolution and distribution of phage ST160 within Salmonella enterica serotype Typhimurium

Salmonellosis is an internationally important disease of mammals and birds. Unique epidemics in New Zealand in the recent past include two Salmonella serovars: Salmonella enterica subsp. enterica serovar Typhimurium definitive type (DT) 160 (S. Typhimurium DT160) and S. Brandenburg. Although not a major threat internationally, in New Zealand S. Typhimurium DT160 has been the most common serovar isolated from humans, and continues to cause significant losses in wildlife. We have identified DNA differences between the first New Zealand isolate of S. Typhimurium DT160 and the genome-sequenced strain, S. Typhimurium LT2. All the differences could be accounted for in one cryptic phage ST64B, and one novel P22-like phage, ST160. The majority of the ST160 genome is almost identical to phage SE1 but has two regions not found in SE1 which are identical to the P22-like phage ST64T, suggesting that ST160 evolved from SE1 via two recombination events with ST64T. All of the New Zealand isolates of DT160 were identical indicating the clonal spread of this particular Salmonella. Some overseas isolates of S. Typhimurium DT160 differed from the New Zealand strain and contained SE1 phage rather than ST160. ST160 was also identified in New Zealand isolates of S. Typhimurium DT74 and S. Typhimurium RDNC-April06 and in S. Typhimurium DT160 isolates from the USA. The emergence of S. Typhimurium DT160 as a significant pathogen in New Zealand is postulated to have occurred due to the sensitivity of the Salmonella strains to the ST160 phage when S. Typhimurium DT160 first arrived.     

Use of AFLP and PFGE to discriminate between Salmonella enterica serovar Typhimurium DT126 isolates from separate food-related outbreaks in Australia

In 2001 Salmonella enterica serovar Typhimurium definitive phage-type (DT) 126 was isolated at higher frequency in Australia compared to other S. Typhimurium phage types and in comparison to previous years. Associated with this increase was the implication of this phage type in a number of food-related outbreaks. We compared fluorescent amplified fragment length polymorphism (FAFLP) to pulsed-field gel electrophoresis (PFGE), the current 'gold standard' for molecular typing of Salmonella for the discrimination between outbreak-associated isolates and epidemiologically unrelated DT126 strains. FAFLP showed a greater ability to discriminate between isolates than PFGE, with 16 groups of clusters or individual isolates with < 90% similarity to each other compared to three groups as determined by PFGE. Both methods were able to discriminate between isolates from two separate outbreaks in South Australia and isolates associated with an outbreak at a restaurant in New South Wales. The resolving power of both methods was not sufficient to separate all epidemiologically unrelated DT126 isolates from the outbreak isolates. We conclude that amplified fragment length polymorphism is a useful tool to assist in the discrimination of S. Typhimurium DT126 isolates.     

2011-2012年杭州市肠道沙门菌临床分离株的分子型别分析

目的研究2011-2012年杭州市肠道沙门菌临床分离株的型别,了解本地菌株分子流行病学特征。方法对66株肠道沙门菌临床分离株进行血清分型和多位点序列分型(MLST)。对其中主要血清型:鼠伤寒、甲型副伤寒、萨雷甲尼和肠炎沙门菌菌株进行脉冲场凝胶电泳(PFGE)分型。结果分布于21个血清型的66株沙门菌分成26个ST型别。发现一株纽波特沙门菌为新型ST1690。菌株血清型与MLST型别数据库中所对应的血清型符合率为100.00%。9株甲型副伤寒沙门菌的PFGE带型完全一致(P7型),与先前杭州流行菌株有差异(P1-P6型)。6株肠炎沙门菌分成4个PFGE型,型间最小相似性为92.70%。13株鼠伤寒沙门菌分为11个PFGE型,型间最小相似性为71.70%。7株萨雷甲尼沙门菌分成4个PFGE型别,型间最小相似性为91.00%。结论近年杭州腹泻病人中流行的肠道沙门菌菌株主要血清型为鼠伤寒、甲型副伤寒、萨雷甲尼和肠炎等。甲型副伤寒沙门菌菌株在杭州出现了新PFGE型别。MLST数据可以对沙门菌血清学鉴定提供一定的帮助。     

Molecular typing of Salmonella enterica serovars Enteritidis, Corvallis, Anatum and Typhimurium from food and human stool samples in Tunisia, 2001-2004

During the period from 2001 to 2004, a total of 72 isolates of Salmonella enterica serovars: Anatum (n=40), Enteritidis (n=18), Corvallis (n=8), and Typhimurium (n=6), of various origins (mainly food and diarrhoeagenic stool samples), were collected and further characterized by antibiotic resistance, plasmid analysis, and pulsed-field gel electrophoresis (PFGE). Forty-five isolates presented multidrug resistance to antibiotics. Among which one S. enterica serovar Anatum isolate was resistant to 11 antibiotics, and one S. enterica serovar Typhimurium DT104 isolate was resistant to eight antibiotics. Plasmid profiling identified eight plasmid profiles (with 1-5 plasmids) among the isolates, of which one plasmid profile (P01) was predominant. XbaI PFGE analysis revealed the presence of a predominant clone of the four studied Salmonella serovars circulating in Tunisia throughout the years 2001-2004.     

Epidemiological analysis of Salmonella enterica Enteritidis isolates in Japan by phage-typing and pulsed-field gel electrophoresis.

Salmonella enterica serotype Enteritidis isolates of phage types (PTs) PT1, PT4, PT13a and PT22 derived from sporadic cases and outbreaks of food poisoning in Japan during 1994 and 1995 were analysed by pulsed-field gel electrophoresis (PFGE). While PT1 strains from 5 different outbreaks showed 14 PFGE patterns, 5 PFGE patterns were observed among PT4 isolates from 5 different outbreaks and 6 independent isolates from imported chicken. Interestingly, 8 out of 9 PT4 strains associated with foreign travel to Southeast Asia were indistinguishable in PFGE pattern from 5 independent isolates of imported chicken from England. Although both PT13a and PT22 were first reported in Japan in 1994, PT22 showed various PFGE patterns compared to PT13a which had the same pattern within an outbreak, unlike PT1. These results could indicate that multiple clonal lines of PT1 and PT22 had already spread while relatively fewer clonal lines of PT4 and PT13a might exist in Japan.     

Antimicrobial resistance of Salmonella enterica typhimurium DT104 isolates and investigation of strains with transferable apramycin resistance.

An examination of salmonella isolates collected by the Scottish Agricultural College Veterinary Services Division from April 1994 to May 1995 was conducted to determine the extent to which Salmonella enterica serotype Typhimurium phage type 104 (DT104) occurred and to investigate the antimicrobial resistance patterns of isolates. Typhimurium DT104 was the predominant salmonella and was isolated from nine species of animal. All isolates of this phage type possessed resistance to at least one antimicrobial and 98% of the isolates were resistant to multiple antimicrobials with R-type ACTSp the predominant resistance pattern. Various other resistance patterns were identified and transferable resistance to the veterinary aminoglycoside antimicrobial apramycin was demonstrated in three strains. A retrospective study for gentamicin resistance in isolates from the Scottish Salmonella Reference Laboratory collection revealed a human isolate of Typhimurium DT104 resistant to gentamicin but sensitive to apramycin and a bovine isolate with apramycin and gentamicin resistance.     

Salmonella enterica serotype Typhimurium DT 104 antibiotic resistance genomic island I in serotype paratyphi B

We have identified Salmonella genomic island I (SGI1) in an isolate of Salmonella enterica serotype Paratyphi B. This antibiotic-resistance gene cluster, which confers multidrug resistance, has been previously identified in S. enterica serotype Typhimurium phage types DT 104 and DT 120 and in S. enterica serotype Agona.     

An outbreak of human salmonellosis caused by ampicillin-resistant Salmonella enterica serovar Enteritidis PT13 in the Czech Republic

In summer 2004, an outbreak caused by Salmonella enterica serovar Enteritidis phage type 13 (S. Enteritidis PT13) was recorded in the Czech Republic. As well being a relatively rare phage type the strain was also ampicillin resistant. Outbreak (n=39) and pre-outbreak isolates (n=13) were characterized by pulsed-field gel electrophoresis (PFGE), beta-lactamase gene polymerase chain reaction and plasmid profile. The majority of outbreak isolates (n=37) were identical in XbaI PFGE profile, and two other outbreak isolates each differed from this profile by one or two fragments respectively. The pre-outbreak isolates were uniform in PFGE profile but distinct from the outbreak strain. Ampicillin resistance was confirmed to be encoded by the blaTEM gene located on the TnA transposon. This gene was readily transferable to a S. Enteritidis recipient strain and was associated with the transfer of a 200-kb plasmid. Our results indicate that all S. Enteritidis PT13 tested from 2004 belonged to a single outbreak strain which prior to 2004 had not been recognized in the Czech Republic.     

Multiply Resistant (MR) Salmonella enterica Serotype Typhimurium DT 12 and DT 120: A Case of MR DT 104 in Disguise?

Multiresistant Salmonella enterica serotype Typhimurium definitive phage type (DT) 12 and DT 120 are more closely related to DT 104 than to non-multiresistant strains of their respective phage types. Multiresistant DT 12 and DT 120 appear to have arisen due to changes in phage susceptibility of DT 104 rather than horizontal transfer of resistance genes.