Subdivision of Salmonella enteritidis phage types by plasmid profile typing

Differentiation of Salmonella enteritidis by plasmid profile typing has been compared to differentiation by phage typing. Examination of the type strains of the 27 S. enteritidis phage types showed that only 11 profile patterns could be identified. Moreover, two profile patterns were found in 15 of the type strains, including those of the two most common phage types in Britain, types 4 and 8. On this basis, plasmid profile typing is not as sensitive as phage typing for the primary subdivision of S. enteritidis. When differentiation of 534 strains of the 27 phage types was attempted using plasmid profiles, variation in pattern suitable for epidemiological subdivision was found in 13 phage types and there were 9 profile patterns in strains of phage type 4. Plasmid profile typing can, therefore, be regarded as an effective adjunct to phage typing for the subdivision of S. enteritidis.     

Use of plasmid profile typing for surveillance of Salmonella enteritidis phage type 4 from humans, poultry and eggs.

Plasmids were found in 1022 of 1089 (94%) of drug-sensitive strains of Salmonella enteritidis phage type 4 from humans (sporadic and outbreak cases), poultry (chickens) and eggs in England and Wales in the 5-year period 1988-92 and 25 plasmid profile patterns were identified. Strains characterized by a single plasmid of 38 MDa predominated (= plasmid profile type SE 38), comprising over 90% of isolates from humans, 70% from poultry and 92% from eggs. Eleven profile types were identified in strains from humans, 21 in strains from poultry and 3 in strains from eggs. Eight of the 11 patterns identified in human isolates were found in strains from poultry and 2 in strains from eggs. In contrast 15 patterns seen in poultry were not found in strains from humans. Four percent of strains from humans and 13% from poultry did not carry the 38 MDa plasmid but all strains from eggs were found to carry this plasmid. The second most common profile type in strains isolated between 1981 and 1988 was not identified in strains isolated from 1988-92. It is concluded that plasmid profile typing is a useful method for rapid differentiation within phage type 4 of S. enteritidis but that methods which can discriminate within the predominant profile type, SE 38, are now required.     

Comparison of two Salmonella enteritidis phage typing schemes

The comparison of two phage typing schemes for Salmonella enteritidis was performed. A total of 517 strains were phage-typed according to the schemes of Lalko [27] and Ward et al. [21]. Strains were isolated from food-poisoning outbreaks and other common sources in Poland, between 1986–1995. Above 99% of all strains tested were differentiated to the definitive phage type using the Lalko collection of typing phages. Phage types 1 and 7 (PTs 1, 7) were the most isolated. The typing phages of Ward enabled to assign 56.5% of all strains (a total of 14 phage types were presented), 37.1% – reacted with phages without showing any of the designated phage types and 6.4% were untypable. Phage type 8 (PT8) predominated. The majority of Salmonella enteritidis strains from one phage type outbreaks of Lalko presented different types of lytic reactions with the Ward phages. Only the correspondence of Salmonella enteritidis PT7 of Lalko with PT8 of Ward et al. [21] was observed.     

Distribution of Salmonella enteritidis phage types in Canada

The distribution of Salmonella enteritidis phage types in Canada is described; 606 of 674 strains examined were of human origin. Typable strains of all sources, constituted 99.6% (671/674) of all strains examined, and were representative of 15 different phage types. Five phage types (8, 13, 4, 13a and 1) accounted for 92.4% of the total. Phage type 8 consistently showed the highest incidence in human (69.96%) and non-human (72.05%) sources and appeared to be the most common in North America. Phage type 4, the most prevalent in the UK, is infrequent in Canada (38/674). The distribution of phage types showed regional variation among infrequent phage types, whereas the common type, 8, was observed in different frequencies in all provinces. Examination of 29 outbreaks of S. enteritidis representing 254 isolates for humans revealed 5 different phage types, the highest number of outbreaks (11) were type 8. A study of these outbreaks and the animal-host-associations of the common phage types, 8 and 13, indicated that contaminated poultry appeared to be the most common source of human infection in Canada.     

Variations in biochemical phenotypes and phage types of Salmonella enteritidis in Germany 1980-92.

The Phene Plate system for typing Salmonella serotypes (PhP-S) is a simple automated typing method based on biochemical fingerprinting. It gives a quantitative value of the metabolism of various substrates by measuring the speed and intensity of each reaction. The ''biochemical fingerprint'' of each isolate is used to calculate similarities among the tested strains with a personal computer program. We used this system to examine a collection of 86 strains of Salmonella enteritidis isolated from human sporadic cases in Germany between 1980 and 1992. Twenty-three biochemical phenotypes (BPTs) consisting of 9 common (C) and 14 single (S) BPTs were identified. BPTs C2 and C4 containing 20 and 36 strains respectively accounted for 65% of the isolates. Strains of BPT C2 were found over a wide period of time whereas strains of BPT C4 were isolated during the period between 1988 and 1992. With phage typing, 11 discrete phage types (PTs) and 18 strains designated as non-specific type (NST) were identified. PTs 4 and 8 with 39 and 17 strains respectively were the dominant PTs. Strains of PT 8 were isolated over a wide period of time whereas all (except one) strains of PT 4 were isolated between 1988 and 1992. Combination of biochemical fingerprinting and phage typing divided the strains into 25 phenotypes (BPT:PTs). Whilst phenotype C2:8 was found over a number of different years, phenotype C4:4 was isolated only between 1988 and 1992. These findings indicate the presence of one persistent and one recently emerged phenotype among S. enteritidis strains in Germany.(ABSTRACT TRUNCATED AT 250 WORDS)     

Phage typing of Salmonella enteritidis isolated from clinical, food, and poultry samples in Chile]

Since 1994 an extensive epidemic of infections with Salmonella enteritidis (S. enteritidis) has affected Chile. In order to understand the diversity of infective sources, the possible origin of the epidemic, and the epidemiological relationships between clinical, food, and poultry isolates, we carried out phage typing of three groups of samples: 1) 310 S. enteritidis clinical samples collected between 1975 and 1996, 2) 47 food isolates obtained during S. enteritidis outbreaks, and 3) 27 strains isolated in surveillance studies of poultry-raising establishments. With the clinical samples, a total of 13 phage types were identified, 2 isolates could not be typed, and 1 was considered atypical. The phage types that were identified most frequently were 1 (56.8%) and 4 (31.3%), trailed by type 8 (4.8%) and type 28 (1.9%). Over time and in different regions of the country there were major changes in the distribution of the phage types. In the first years of collection the only phage types registered were 8 and 28, which disappeared around 1980 and then began reappearing sporadically in 1996. With the gradual S. enteritidis expansion that started in 1988, in the central and southern areas of the country phage type 4 began to appear; that type had not been found before in Chile. In 1991 in the northern area of the country phage type 1 began to predominate; it was another type that had not been reported before in Chile. In the food isolates the only phage types identified were 1 and 4, which were also the most common in the poultry isolates. Phage typing of S. enteritidis has proved to be useful in guiding the epidemiological analysis of the infections caused by this pathogen.     

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.     

A comparison of Salmonella enteritidis phage types from egg-associated outbreaks and implicated laying flocks.

Infections due to Salmonella enteritidis are increasing worldwide. In the United States, between 1985 and 1989, 78% of the S. enteritidis outbreaks in which a food vehicle was identified implicated a food containing raw or lightly cooked shell eggs. Under a US Department of Agriculture regulation published in 1990, eggs implicated in human food-borne S. enteritidis outbreaks were traced back to the source flock. The flock environment and the internal organs of a sample of hens were tested for S. enteritidis. We compared the S. enteritidis phage types of isolates from 18 human, egg-associated outbreaks and the 15 flocks implicated through traceback of these outbreaks. The predominant human outbreak phage type was recovered from the environment in 100% of implicated flocks and from the internal organs of hens in 88% of implicated flocks we tested. The results support the use of phage typing as a tool to identify flocks involved in human S. enteritidis outbreaks.     

Genetic relationships among strains of Salmonella enteritidis in a national epidemic in Switzerland.

A collection of Salmonella enteritidis strains isolated in Switzerland (1965-90) was characterized. The phage type and plasmid profile of isolates were compared with the copy number and insertion loci of the DNA insertion element IS200. Three clonal lines of S. enteritidis were identified by IS200 profile; the various phage types were subtypes reproducibly associated with one of these lines. All human and poultry isolates contained a 38 Mda plasmid which hybridized with a mouse virulence-associated gene probe. In S. enteritidis, the IS200 profile is a race-specific molecular marker of the chromosome, and may be particularly applicable for studying the epidemiology of less common serovars.     

Acquisition of a drug resistance plasmid converts Salmonella enteritidis phage type 4 to phage type 24

Salmonella enteritidis accounted for 55% of the 27,478 salmonellae isolated from humans in England and Wales during 1988. Within this serotype phage type 24 has increased from 24 isolations in 1987 to 201 in 1988. The high frequency of drug resistance in this phage type has been shown to be due to the presence of plasmids belonging to Inc N and coding for resistance to a range of antimicrobial drugs among which resistance to ampicillin, streptomycin, tetracycline (AST) and T predominate. These plasmids are phage-type determining and convert strains of phage type 4 to phage type 24.     

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.     

肠炎沙门氏菌脉冲场凝胶电泳分型研究

目的　为研究肠炎沙门氏菌菌株间的关系,特建立了分子流行病学研究方法脉冲场凝胶电泳分型方法,并对从食品中分离的肠炎沙门氏菌进行了分型研究。方法　1991～2 0 0 1年间,从河南、黑龙江等省份采集食品样品,分离出肠炎沙门氏菌,挑取单个菌落增菌,用溶菌酶、蛋白酶K和TE缓冲液依次提取菌体DNA后,用限制性内切酶XbaΙ消化胶块,然后在CHEFMapper仪上进行脉冲场凝胶电泳,电泳条件:0 . 5×TBE缓冲液、14℃、1%琼脂糖、脉冲时间5s～4 5s、2 6h、6V cm。应用λ噬菌体DNA作为脉冲场凝胶分子量标准。结果　从鸡肉、牛肉、熟肉及蔬菜中分离出30株肠炎沙门氏菌,其中18株来自河南省,绝大多数菌株(2 2株)分离于2 0 0 1年。对所得菌株进行脉冲场凝胶电泳后,结果显示30株肠炎沙门氏菌分为A、B两个型,两型图谱之间有8个条带的差异。A型菌株有19株,占全部菌株的6 3. 3% ,B型菌株有11株,占36 .7%。A型之内又可以分出2个亚型,其中A1型占78 .9% ,A2亚型与A1亚型相比的差别在于,A2型在5 0kb处有一条带缺失。在菌株分型与采集时间和采集地点之间没有相关性,但所有从鸡肉中分离到的菌株均为A1型,提示了某种有待进一步研究的流行病学趋势。结论　在研究肠炎沙门氏菌不同菌株之间的分子流行病学关系方面,脉冲场凝胶电泳是一种有     

Salmonella enteritidis phage types in Germany

In order to monitor the epidemiological situation ofS. enteritidis in Germany, in 1990–91 1138 isolates from more than 180 locations in West Germany were phage typed. 1124 strains (98.8%) from all sources were typeable, belonging to 21 different phage types (PT). PT4 strains were isolated most frequently (70.8%). In addition, PT7, 25, 34 and 8 were of epidemiological relevance with incidences of 7.2 to 4.5%. The comparison of data shows that in Germany, like in other parts of Europe, PT4 predominates. This phage type is, however, infrequent in North America, where PT8 has the highest incidence.     

Salmonellosis in Indonesia: phage type distribution of Salmonella paratyphi A.

The distribution of phage types among 221 human strains of Salmonella paratyphi A in Indonesia was studied. Approximately 50% were phage type 5, a rare type elsewhere in the world. Most other isolates were the cosmopolitan phage type 1. The ratio of phage type 1-5 was compared for two cities on Java, Jakarta and Yogyakarta. The ratios were significantly different, phage type 5 predominating in Jakarta. The ratio of phage types among 10 S. paratyphi A strains isolated from Jakarta river water was similar to that found in Jakarta patients.     

Foodborne outbreaks caused by Salmonella in Italy, 1991-4.

This report summarizes studies on 1699 foodborne outbreaks, in Italy, reported to the Istituto Superior di Sanità (ISS) (the National Institute of Health of Italy, Rome) during the period 1991-4. The most frequently reported foodborne outbreaks were caused by salmonellae (81%), in particular by Salmonella enteritidis and non-serotyped group D salmonella (34% and 33% of the total salmonella outbreaks, respectively). A vehicle was implicated in 69% of the salmonella outbreaks; eggs were implicated in 77% of the outbreaks for which a vehicle was identified or suspected. Salmonella strains isolated in 54 outbreaks were studied for phenotypic and genotypic characteristics. The isolates belonged to S. enteritidis (50 outbreaks), S. typhimurium (three outbreaks) and S. hadar (one outbreak). In the S. enteritidis outbreaks, phage type 4 was most frequently isolated (64.8%), followed by phage type 1 (14.8%). The virulence plasmid of 38 megadaltons was found in many different phage types of S. enteritidis.     

Genome fingerprinting of Salmonella typhi by pulsed-field gel electrophoresis for subtyping common phage types.

The genomic DNA of 39 strains of Salmonella typhi isolated from local residents and patients who had visited countries in the Asian region was analysed for restriction fragment length polymorphisms (RFLP). Pulsed-field gel electrophoretic (PFGE) analysis of Xba I- and Spe I-generated genomic restriction fragments established 22 PFGE types whereas phage typing differentiated the 39 isolates into 9 distinct phage types. This study showed that PFGE is more discriminatory than phage typing as it is capable of subtyping S. typhi strains of the same phage types. Genetic relatedness among the isolates was determined. Seven major clusters were identified at SABs of > 0.80 and the remaining 13 isolates were distributed into minor clusters which were related at SABs of less than 0.80. In conclusion, PFGE analysis in conjunction with distance matrix analysis served as a useful tool for delineating common S. typhi phage types of diverse origins from different geographical locales and separated in time.     

广东省2007年度非伤寒沙门菌监测及病原学特征分析

目的 了解广东省腹泻病患者中非伤寒沙门菌感染情况和菌株的血清型别、分布、耐药性变化.方法 对纳入研究的腹泻病患者进行非伤寒沙门菌检测,对分离到的菌株进行血清分型、药物敏感性试验和脉冲场凝胶电泳(PFGE)分型.结果 2007年度共检测1128份腹泻粪便标本,分离到71株沙门菌,阳性检出率为6.29%;共分得29种血清型,肠炎和鼠伤寒沙门菌居多;大多数沙门菌对常用的头孢类和喹诺酮类抗生素敏感,鼠伤寒沙门菌的耐药率普遍较肠炎和斯坦利沙门菌高;除肠炎沙门菌外,其余的血清型没有同一PFGE型别的菌株;用Xba Ⅰ酶切17株肠炎沙门菌可分为PFGE-XbaⅠ 1～8型,其中PFGE-XbaⅠ 4型为优势型别.用Sfi Ⅰ和Not Ⅰ酶对12株肠炎沙门菌进行再分型,综合用Xba Ⅰ/Sfi Ⅰ/Not Ⅰ三种酶的结果 发现仍有三组菌的PFGE图谱是完全一致的.结论 2007年度广东省非伤寒沙门菌的感染多数为散发病例,头孢类和喹诺酮类抗生素是治疗非伤寒沙门菌感染的首选药物.     

Integrated surveillance and potential sources of Salmonella enteritidis in human cases in Canada from 2003 to 2009

Salmonella enteritidis has emerged as the most prevalent cause of human salmonellosis in Canada. Recent trends of S. enteritidis subtypes and their potential sources were described by integrating Salmonella data from several Canadian surveillance and monitoring programmes. A threefold increase in S. enteritidis cases from 2003 to 2009 was identified to be primarily associated with phage types 13, 8 and 13a. Other common phage types (4, 1, 6a) showed winter seasonality and were more likely to be associated with cases linked to international travel. Conversely, phage types 13, 8 and 13a had summer seasonal peaks and were associated with cases of domestically acquired infections. During agri-food surveillance, S. enteritidis was detected in various commodities, most frequently in chicken (with PT13, PT8 and PT13a predominating). Antimicrobial resistance was low in human and non-human isolates. Continued integrated surveillance and collaborative prevention and control efforts are required to mitigate future illness.     

Molecular characterization of plasmids in Salmonella enteritidis phage types.

Plasmids in selected type strains of 26 of the Salmonella enteritidis phage types have been characterized by restriction enzyme fingerprinting and by DNA-DNA hybridization with oligonucleotide probes for Salmonella plasmid virulence (Spv) genes. With one exception, the fingerprints of the 38 MDa plasmids studied were homogeneous but there was heterogeneity in the fingerprints of 59 MDa plasmids found in 4 of the type strains. However all 38 MDa and 59 MDa plasmids were related as was a 45 MDa plasmid identified in the type strain of phage type 19. A 3.5 kb fragment homologous to SpvC was conserved in Hind III digests of all 38 MDa and 59 MDa plasmids, and in the related 45 MDa plasmid. In contrast a 65 MDa plasmid found in the type strain of phage type 10 was not related to these three plasmid molecular weight groups and did not carry the SpvC gene.     

Application of physico-chemical typing methods for the epidemiological analysis of Salmonella enteritidis strains of phage type 25/17.

Eighty-nine Salmonella enteritidis phage type 25/17 strains isolated from a localized outbreak in the German state Nordrhein-Westfalen (outbreak NWI) could not be further differentiated by biochemotyping and plasmid pattern analysis. They were submitted to a complex typing system consisting of modern physico-chemical analytical procedures. In lipopolysaccharide pattern analysis the strains proved to be homogeneous. In multilocus enzyme electrophoresis, outer membrane and whole cell protein pattern (WCPP) analysis, and Fourier-transform infrared (FT-IR) spectroscopy (increasing extent of differentiation in the given order) strains deviating from each basal pattern were found. The extent of correspondence in these deviations was satisfactory. Forty-six strains of the same sero- and phage type, however, obtained from different outbreaks, were additionally typed. The results obtained with them indicate that the data of the first group were not restricted to strains from outbreak NWI, but of general validity. It was found that both WCPP and FT-IR represent valuable methods for the sub-grouping of bacteria.