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.     

Genomic diversity within phage types of Salmonella enterica ssp. enterica serotypes Enteritidis and Typhimurium

The diversity among 1354 strains of Salmonella enterica subsp. enterica, serotype Enteritidis (n = 847) and Typhimurium (n = 507) isolated in Finland in 1991-2002 (n = 608) and in 2003 (n = 746) were studied. The former strains were studied retrospectively by phage typing and pulsed-field gel electrophoresis (PFGE) harmonized in the European Salm-gene project. The latter strains were studied prospectively, and the results correlated to their antimicrobial susceptibility and association with travel to popular tourist destinations. During both periods, S. Enteritidis phage types (PTs) PT1 and PT4, and S. Typhimurium definite types (DTs) DT1 and DT104 were the major phenotypes. SENTXB.0001 was the dominating single PFGE type among S. Enteritidis strains (40% in 1991-2002; 57% in 2003), and accounted correspondingly for 23% and 63% of the PT1 strains, and 81% and 88% of the PT4 strains. No PFGE types dominated among the S. Typhimurium strains but a correlation was found between certain phage and PFGE types: among DT1 strains, STYMXB.0098 accounted for 66% (1991-2002) and 98% (2003) and among the DT104 strains STYMXB.0001 accounted for 84% and 97% in the two time periods, respectively. Of the S. Enteritidis strains isolated in 2003, 91% were associated with travel, most commonly to Spain, Greece, and Bulgaria. SENTXB.0001 was the major Salmonella PFGE type in these countries. In contrast, most (55%) S. Typhimurium strains were of domestic origin. While only 1.3% of the S. Enteritidis strains were multiresistant and 24% were resistant to nalidixic acid only, 30% of the S. Typhimurium strains were multiresistant. Among the multiresistant S. Typhimurium strains, R-type ACSSuT and PFGE type STYMXB.0001 of the DT104 complex dominated.     

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.     

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.     

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.     

Global monitoring of Salmonella serovar distribution from the World Health Organization Global Foodborne Infections Network Country Data Bank: results of quality assured laboratories from 2001 to 2007

Salmonella enterica is commonly acquired from contaminated food and is an important cause of illness worldwide. Interventions are needed to control Salmonella; subtyping Salmonella by serotyping is useful for targeting such interventions. We, therefore, analyzed the global distribution of the 15 most frequently identified serovars of Salmonella isolated from humans from 2001 to 2007 in laboratories from 37 countries that participated in World Health Organization Global Foodborne Infections Network and demonstrated serotyping proficiency in the Global Foodborne Infections Network External Quality Assurance System. In all regions throughout the study period, with the exception of the Oceania and North American regions, Salmonella serovars Enteritidis and Typhimurium ranked as the most common and second most common serovar, respectively. In the North American and Oceania (Australia and New Zealand) regions, Salmonella serovar Typhimurium was the most common serovar reported, and Salmonella serovar Enteritidis was the second most common serovar. During the study period, the proportion of Salmonella isolates reported from humans that were Salmonella serovar Enteritidis was 43.5% (range: 40.6% [2007] to 44.9% [2003]), and Salmonella serovar Typhimurium was 17.1% (range: 15% [2007] to 18.9% [2001]). Salmonella serovars Newport (mainly observed in Latin and North American and European countries), Infantis (dominating in all regions), Virchow (mainly observed in Asian, European, and Oceanic countries), Hadar (profound in European countries), and Agona (intense in Latin and North American and European countries) were also frequently isolated with an overall proportion of 3.5%, 1.8%, 1.5%, 1.5%, and 0.8%, respectively. There were large differences in the most commonly isolated serovars between regions, but lesser differences between countries within the same region. The results also highlight the complexity of the global epidemiology of Salmonella and the need and importance for improving monitoring data of those serovars of highest epidemiologic importance.     

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.     

International dissemination of antibiotic resistant strains of bacterial pathogens.

The increasing incidence of methicillin and multiple resistant Staphylococcus aureus (MRSA) in nosocomial infections is mainly associated with a wide, international dissemination of well defined clonal lineages (epidemic MRSA) which are clearly different from community acquired MRSA by molecular typing patterns and structure of the staphylococcal cassette chromosome containing the mecA gene. Although belonging to a definite subpopulation within the species Enterococcus faecium, hospital associated vancomycin resistant isolates also containing the esp gene have very likely evolved by acquisition of glycopeptide resistance gene clusters at different occasions and at different times by a susceptible already disseminated clonal lineage. There is obviously a continuous selection of new types of extended spectrum beta-lactamases in enterobacteriaceae and also horizontal spread of bla-genes. Intrahospital dissemination of particular strains has often been observed, however, an international dissemination until now has been described only for Salmonella enterica serovar Typhimurium producing the CTX-M-3 enzyme. Multiresistant isolates of Salmonella enterica serovar Typhimurium phage type DT104 harbour a multiresistant gene cluster with resistance genes from taxonomically more unrelated species (tetG, floR, bla(PSE1)). Although in vitro transduction has been demonstrated, this gene cluster has only rarely been reported from isolates exhibiting other phage patterns of the same serovar or from other serovars of S. enterica.     

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.     

Molecular epidemiology of Salmonella enteritidis phage type 1b and 6a isolates in Portugal

Salmonella enterica serotype Enteritidis is an important serovar comprising 76 % of Salmonella isolates in Portugal in 2001. For better understand the epidemiology of salmonellosis, a total of 47 isolates of S. Enteritidis phage type (PT) 1b and 6a were analysed by pulsed-field gel electrophoresis (PFGE) and genomic DNA was subjected to macro restriction with XbaI. For PT 1b isolates, only three different patterns were observed, and PT6a showed a total of 10 digestion patterns. Curiously, the main pattern among PT1b isolates seams quite similar to main pattern of PT6a isolates, but when the two patterns were analysed with Bionumerics, we observed that they exhibited some differences. It was concluded that, in 2001, there was one predominant pattern for PT1b and PT6a and, possibly, we were in presence of clonal strains that exists all over the country.     

Salmonellosis in pigmy hogs (Sus salvanius)--a critically endangered species of mammal

The pigmy hog (Sus salvanius) is the smallest and the rarest wild suid in the world. This species is on the verge of extinction and the World Conservation Union has rated it among the most endangered of all mammals. This paper reports the investigation into an outbreak of salmonellosis among captive pigmy hogs at the Research and Breeding Centre of the pigmy hog conservation programme, Guwahati, Assam, India. Of 75 pigmy hogs (38 males and 37 females) maintained at the Centre, seven (9.3%) died within five days. The causative organism associated with the outbreak was identified as Salmonella Typhimurium (syn. S. enterica serovar Typhimurium). All the isolates of S. Typhimurium belonged to phage type DT193. The isolates harboured multiple plasmids. Five isolates harboured four (65.0 MDa, 4.2 MDa, 3.0 MDa, 1.3 MDa), while two isolates carried three plasmids (65.0 MDa, 4.2 MDa, 3.0 MDa). All strains showed resistance to amikacin, ampicillin, streptomycin and sulfamerazin; five strains were resistant to oxytetracycline and trimethoprim. All the strains were sensitive to chloramphenicol, ciprofloxacin, enrofloxacin and gentamicin. All seven isolates of S. Typhimurium were found to harbour stn, sopB and pefA genes. However, none of them was found to carry sefC and sopE genes.     

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.     

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.     

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。     

The virulence for gnotobiotic pigs of live attenuated vaccine strains of Salmonella enterica serovars Typhimurium and Enteritidis

Gnotobiotic pigs were inoculated orally with wild-type or defined aroA or cya crp vaccine strains of Salmonella enterica serovars Typhimurium and S. Enteritidis. The parental strains were virulent, inducing severe diarrhoea, dehydration and systemic disease. The cya crp mutant was also highly virulent whereas the aroA strains were attenuated.     

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.     

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.     

Drastic decrease of Salmonella Enteritidis isolated from humans in Belgium in 2005, shift in phage types and influence on foodborne outbreaks

In Belgium, non-typhoidal salmonellosis and campylobacteriosis are the two most frequently reported foodborne illnesses. During 2005, a 71% decrease of Salmonella Enteritidis infections compared with the average annual number cases in the period 2000-2004 was recorded by the Belgian National Reference Centre for Salmonella and Shigella. After the peak of 1999, the total number of salmonellosis cases decreased gradually, with the exception of 2003 when an increase was again recorded due to the rise of isolates belonging to the serotype Enteritidis. PT4, the predominant phage type of serotype Enteriditis over recent years (except in 2003), became the second most prevalent phage type in 2005 after PT21. We present in this paper the epidemiology (incidence and trends) of human salmonellosis in Belgium and assess the role of the vaccination programme in layer flocks on the decline of the incidence of human salmonellosis and foodborne outbreaks due to S. Enteritidis.     

Risk factors for Salmonella Enteritidis and Typhimurium (DT104 and non-DT104) infections in The Netherlands: predominant roles for raw eggs in Enteritidis and sandboxes in Typhimurium infections

Since 1996 Salmonella Typhimurium DT104 salmonellosis has increased in The Netherlands. This prompted a case-control study of risk factors for salmonellosis to inform transmission routes for this phage type. Cases were laboratory-confirmed patients with a Salmonella infection and controls were selected from population registries by frequency matching for age, sex, degree of urbanization and season. Cases and controls received a questionnaire on risk factors. Of the 1171 cases, 573 (49%) responded: 245 S. Enteritidis and 232 S. Typhimurium cases (both DT104 and non-DT104), of which 58 were DT104. Of the 10250 controls, 3409 (33%) responded. Use of H2 antagonists [odds ratio (OR) 4.4, 95% CI 1.6-12.2] and proton pump inhibitors (OR 4.2, 95% CI 2.2-7.9), consumption of raw eggs (OR 3.1, 95% CI 1.3-7.4) and products containing raw eggs (OR 1.8, 95% CI 1.1-3.0) were associated with endemic S. Enteritidis infection. Risk factors for endemic S. Typhimurium infection were use of proton pump inhibitors (OR 8.3, 95% CI 4.3-15.9), occupational exposure to raw meat (OR 3.0, 95% CI 1.1-7.9), playing in a sandbox (for children aged 4-12 years) (OR 2.4, 95% CI 1.6-3.7), consumption of undercooked meat (OR 2.2, 95% CI 1.1-4.1) and use of antibiotics (OR 1.9, 95% CI 1.0-3.4). Use of proton pump inhibitors (OR 11.2, 95% CI 3.9-31.9) and playing in a sandbox (OR 4.4, 95% CI 1.8-10.7) were the only risk factors for S. Typhimurium DT104 salmonellosis. This study confirms known risk factors for salmonellosis. However, playing in a sandbox was a predominant new risk factor for S. Typhimurium salmonellosis in children [population attributable risk (PAR) 14%], and especially for S. Typhimurium DT104 (PAR 32%).