The role of roof rats ( Rattus rattus) in the spread of Salmonella Enteritidis and S. Infantis contamination in layer farms in eastern Japan

The prevalence of Salmonella in four layer farms in eastern Japan was investigated between 2004 and 2006 to determine the role of roof rats (Rattus rattus) in the epizootology of Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis). Persistent S. Enteritidis and S. Infantis contamination of the environment and pooled egg samples were detected in three out of four layer farms. A total of 113 (13.3%) and 158 (18.6%) out of 851 rats examined were positive for S. Enteritidis and S. Infantis, respectively. By pulsed-field gel electrophoresis, only one indistinguishable pulsed-field pattern was yielded by S. Enteritidis strains from rats, eggs and environmental samples from each of the two contaminated layer farms. Although, a variety of pulsed-field patterns were generated by S. Enteritidis isolates from rats, eggs, and the environment of the other contaminated farms, there are, however, some S. Enteritidis strains that are closely related clones. These results suggest that roof rats are carriers of S. Enteritidis and S. Infantis and that persistent S. Enteritidis and S. Infantis infections in a rat population may play an important role in the spread and maintenance of these pathogens inside the layer premises.     

Vertical and horizontal transmission of salmonella within integrated broiler production system

Salmonella remains one of the leading causes of food-borne illness in the United States, and many key questions regarding the introduction and persistence in animal production systems still remain. In order to understand the ecology of Salmonella within an integrated commercial broiler production system, 289 Salmonella enterica were recovered from two integrated poultry farms during the production and processing of seven consecutive flocks. The variety and prevalence of Salmonella serotypes differed between farms. Overall, 15 serotypes were identified, with the most common being Typhimurium (55%), Montevideo (7.9%), Kentucky (9%), and Enteritidis (9.7%). Salmonella Typhimurium and Enteritidis isolates recovered from processed carcasses from Farm One were further characterized using pulsed-field gel electrophoresis (PFGE), and were shown to be indistinguishable from isolates recovered from the poultry house environment and mice trapped on this farm. Additionally, the same broiler S. Typhimurium and S. Enteritidis strains, identified by PFGE, were also isolated from samples taken at a company breeder farm, suggesting vertical transmission of these Salmonella serotypes in this poultry production system. Results indicate that management practices at the breeder level may have a profound effect on the transmission and persistence of salmonellae within an integrated production system, as well as on the potential contamination of poultry-derived products.     

Web-based surveillance and global Salmonella distribution, 2000-2002

Salmonellae are a common cause of foodborne disease worldwide. The World Health Organization (WHO) supports international foodborne disease surveillance through WHO Global Salm-Surv and other activities. WHO Global Salm-Surv members annually report the 15 most frequently isolated Salmonella serotypes to a Web-based country databank. We describe the global distribution of reported Salmonella serotypes from human and nonhuman sources from 2000 to 2002. Among human isolates, S. Enteritidis was the most common serotype, accounting for 65% of all isolates. Among nonhuman isolates, although no serotype predominated, Salmonella enterica serovar Typhimurium was reported most frequently. Several serotypes were reported from only 1 region of the world. The WHO Global Salm-Surv country databank is a valuable public health resource; it is a publicly accessible, Web-based tool that can be used by health professionals to explore hypotheses related to the sources and distribution of salmonellae worldwide.     

Salmonella enteritidis infections, United States, 1985-1999

Salmonella enterica serotype Enteritidis emerged as an important illness during the 1980s. Investigations showed that consumption of undercooked eggs was the major risk factor for disease, and a variety of prevention and control efforts were initiated during the 1990s. We describe sporadic infections and outbreaks of S. Enteritidis in the United States from 1985 through 1999 and discuss prevention and control efforts. After reaching a high of 3.9 per 100,000 population in 1995, S. Enteritidis infections declined to 1.98 per 100,000 in 1999. While the total number of outbreaks decreased by half, those in the western states tripled. Outbreaks of S. Enteritidis phage type 4 infections accounted for 49% of outbreaks in 1999. Outbreak-associated deaths in health facilities decreased from 14 in 1987 to 0 in 1999. Overall, rates of sporadic S. Enteritidis infection, outbreaks, and deaths have declined dramatically. For further reductions, control measures should continue to be applied along the entire farm-to-table continuum.     

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.     

Differentiation of Salmonella enteritidis isolates by fluorescent amplified fragment length polymorphism

Salmonella Enteritidis is responsible for human gastroenteritis outbreaks worldwide, and the molecular characterization of isolates is an important tool for epidemiological studies. Fluorescent amplified fragment length polymorphism (FAFLP) analysis was performed on 31 Salmonella Enteritidis strains from South Brazil isolated from human, foods, swine, broiler carcasses, and other poultry-related samples to subtype isolates in comparison to pulsed-field gel electrophoresis (PFGE) analysis. Five strains of Salmonella Enteritidis from different geographical regions, Salmonella Enteritidis ATCC 13076, and four isolates of different Salmonella serovars were also tested. Among the 41 isolates tested, 96 polymorphic AFs and 40 distinct profiles were obtained, displaying a Simpson's index of diversity of 0.99; whereas the PFGE analysis presented 13 patterns and the resulting Simpson's index was 0.55. Nine FAFLP and seven PFGE clusters could be inferred based in Dice similarity coefficient. FAFLP clustering readily identified different serotypes of Salmonella but did not distinguish isolates epidemiologically nonrelated or distinct phage types. Therefore, these results indicate that FAFLP is a rapid method for epidemiological investigations of Salmonella outbreaks, presenting a high discriminatory power for subtyping of Salmonella Enteritidis.     

Laboratory-based surveillance of non-typhoidal Salmonella infections in Guangdong Province, China

Salmonella is one of the most common foodborne pathogens in humans. Laboratory-based surveillance for non-typhoidal Salmonella infection was conducted in Guangdong Province, China to improve understanding about the disease burden and detection of dispersed outbreaks. Salmonella isolated from patients with diarrhea were sent from 16 sentinel hospitals to local public health laboratories for confirmation, serotyping, antimicrobial susceptibility testing, and pulsed-field gel electrophoresis (PFGE). PFGE patterns were analyzed to identify clusters representing potential outbreaks. Between September 2009 and October 2010, 352 (4%) Salmonella isolates were obtained from 9167 stool specimens. Salmonella enterica serotype Typhimurium (45%) and Salmonella enterica serotype Enteritidis (13%) were the most common serotypes, and multidrug resistance was high, especially in Salmonella Typhimurium isolates. PFGE patterns of obtained Salmonella isolates were found to be diverse, but a unique PFGE pattern comprising 53 Salmonella Typhimurium isolates were found to occur almost exclusively in infants. Epidemiologic studies are ongoing to determine whether a common exposure is the source of the Salmonella Typhimurium strain frequently isolated from infants.     

Salmonella enteritidis in broiler chickens, United States, 2000-2005

US Department of Agriculture Food Safety and Inspection Service (FSIS) data on Salmonella enterica serotype Enteritidis in broiler chicken carcass rinses collected from 2000 through 2005 showed the annual number of isolates increased >4-fold and the proportion of establishments with Salmonella Enteritidis-positive rinses increased nearly 3-fold (test for trend, p<0.0001). The number of states with Salmonella Enteritidis in broiler rinses increased from 14 to 24. The predominant phage types (PT) were PT 13 and PT 8, 2 strains that a recent Foodborne Diseases Active Surveillance Network (FoodNet) case-control study associated with eating chicken. FSIS is directing more sampling resources toward plants with marginal Salmonella control to reduce prevalence in products including broilers. The policy targets establishments with common Salmonella serotypes of human illness, including Salmonella Enteritidis. Voluntary interventions should be implemented by industry.     

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.     

Emergence of Salmonella enteritidis outbreaks in broiler chickens in the Lebanon: epidemiological markers and competitive exclusion control

This study investigates the first emergence of Salmonella Enteritidis outbreaks among chickens in the Lebanon and identifies the epidemiological markers of selected recovered Enteritidis strains. In addition, the authors evaluate a competitive exclusion approach to control infection in broiler chickens by Enteritidis organisms which possess the prevalent identified markers. The basic procedure in this investigation involved recording signs and lesions in eleven broiler chicken flocks on eleven farms, and culturing livers, spleens, and caeca of ten randomly selected birds per flock for Salmonella isolation and serotyping. Furthermore, culturing for Salmonella and serotyping was attempted from the livers, spleens, caeca and oviduct swabs of ten hens in four broiler breeder flocks which provided hatching eggs for the broilers under study. The identification of epidemiological markers in recovered S. Enteritidis included the determination of drug-resistance patterns and plasmid profiling. The competitive exclusion was evaluated by spraying the microflora on day-old broilers in the hatchery, followed by a controlled oral challenge at three days of age, with 2.85 x 10(5) colony-forming units of S. Enteritidis organisms per bird. Exclusion was evaluated by culturing for S. Enteritidis in anal swabs, spleens, livers, and caeca of individual challenged birds treated with the microflora and in untreated challenged birds. A total of 112 invasive S. Enteritidis strains were recovered on eleven farms from individual organs of broiler chickens with typical signs and lesions of salmonellosis. The prevalent resistance to drugs in such strains was to furaltadone and gentamycin, a marker identified in 93 strains (83%), recovered from nine out of eleven farms. The same resistance pattern was present in S. Enteritidis strains recovered from breeders on one out of four farms. The prevalent plasmid profile in nine S. Enteritidis organisms selected randomly from a pool of 93 strains (one per each of the nine broiler farms) was 14.1 kilobases (kb) and approximately 50.0 kb, a typical pattern to that identified in S. Enteritidis organisms recovered from oviducts of breeders on one out of four breeder farms. The exclusion significantly reduced cumulative mortality in birds of up to 45 days of age by 3.93%, in comparison to that observed in untreated challenged birds (P < 0.05). At 45 days of age, exclusion resulted in a 15.6% reduction in the percentage infection rate by S. Enteritidis in spleens or livers and a 34.4% reduction in the percentage infection rate of the caeca (P < 0.05).     

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.     

Molecular characterization of Salmonella enterica serotype Enteritidis isolates from humans by antimicrobial resistance, virulence genes, and pulsed-field gel electrophoresis

Salmonella enterica serovar Enteritidis (S. Enteritidis) is a major serovar associated with human salmonellosis. A total of 425 clinical S. Enteritidis isolates of human origin were collected between June 2009 and September 2010 from North Carolina. The isolates were further characterized for antimicrobial susceptibility, antimicrobial resistance coding determinants, virulence genes, and fingerprint profiles to determine whether they were similar or different to the S. Enteritidis strain responsible for the human outbreak due to consumption of contaminated eggs. Ten different antimicrobial resistance phenotypes were observed with the highest frequency of resistance exhibited to ampicillin (n=10; 2.35%). The isolates were predominantly pansusceptible (n=409; 96.23%); however, seven isolates were multidrug resistant (MDR; i.e., resistant to three or more antimicrobials). Extended spectrum β-lactamase (ESBL) coding genes (bla(TEM) and bla(PSE)) were detected in the ampicillin-resistant isolates, whereas a single MDR isolate tested positive for class 1 integron (1 kb). The majority of the isolates (n=422; 99.3%) carried the invA, mgtC, stn, sopB, sopE1, and sefA virulence genes. However, 37 (8.7%) and 46 (10.82%) S. Enteritidis isolates tested negative for the plasmid encoded genes spvC and rck, respectively. Pulsed-field gel electrophoresis (PFGE) typing of 118 S. Enteritidis isolates by restriction enzymes XbaI and BlnI resulted in seven clusters, each with a discriminatory index (DI) of 0.715 and 0.785, respectively. The combination of XbaI-BlnI patterns generated a dendrogram with 14 clusters and a higher DI of 0.914. The PFGE profile of 80 isolates matched 100% with the S. Enteritidis strain that has been cited for the recent outbreak in the United States due to consumption of contaminated eggs. In conclusion, we identified a genotypic similar S. Enteritidis population in our study based on antimicrobial susceptibility, virulence gene, and PFGE fingerprint profiles.     

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.     

Antibiotic susceptibility and genotypes of non-typhi Salmonella isolates from children in Kilifi on the Kenya coast

A 4-year retrospective study was undertaken at the Kilifi District Hospital on the coast of Kenya to determine the antibiotic-susceptibility patterns and genotypes of non-typhi Salmonella (NTS) isolates from children. Overall, during the period 1994-97, positive cultures were obtained from 543 (14%) of 3885 blood samples, 364 (30%) of 1210 stool samples and 143 (11%) of 1283 cerebrospinal fluid (CSF) samples. NTS were isolated from 151 (27.8%), 72 (19.8%), and 11 (7.7%) of these positive cultures, respectively. The total 234 NTS isolates were serotyped: the most frequent were Salmonella enterica serotype Enteritidis (41%) and S. enterica serotype Typhimurium (38%). Antibiotic sensitivity testing was done using ampicillin (amp), chloramphenicol (chl), gentamicin (cn), co-trimoxazole (s-t), cefuroxime (cxm), ciprofloxacin (cip), cefotaxime (ctx), amoxicillin-clavulanic acid 20 micrograms-10 micrograms (amc), and tobramycin (tob). Of the 234 isolates, 43 were sensitive to all antibiotics tested and 133 were multiple drug resistant (MDR). The most common resistance type seen was amp, cn, cxm, s-t, ctx, amc, tob (36/234). Our results indicate a high proportion of MDR amongst the isolates from Kilifi. We conclude that 2 major serotypes of salmonella, i.e., S. enterica serotype Typhimurium and S. enterica serotype Enteritidis, of micro-epidemic nature that have been previously unrecognized in Kilifi are responsible for infection in Kilifi district on the coast of Kenya and that over half (56.8%) of total NTS isolates are MDR.     

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.     

肠炎沙门菌食物中毒的病原学检测分析

目的 对一起食物中毒事件进行病原学分析和流行病学调查,探讨其中毒原因、传染源及传播途径。方法 通过现场流行病学调查对数据资料进行Fisher确切概率检验,对患者肛拭子、剩余食品、原材料等28份样品进行实时荧光PCR检测及分离培养;对分离菌株进行生化鉴定和血清分型,用改良K-B法进行药敏试验,应用脉冲场凝胶电泳（PFGE）分型技术进行同源性分析。 结果 从可疑食品蛋糕和6个患者中共分离到7株肠炎沙门菌,生物学性状、血清型和药敏试验结果一致, PFGE条带聚类分析显示所有菌株属同一克隆株。结论 这是一起肠炎沙门菌污染引起的食物中毒事件。食品卫生监督部门应加强网购食品卫生管理,减少食物中毒的发生。     

Representative Salmonella serovars isolated from poultry and poultry environments in Saudi Arabia

The authors describe the source and prevalence of pathogenic Salmonella serovars among poultry farms in Saudi Arabia. A total of 1,052 (4%) Salmonella isolates were recovered from 25,759 samples of poultry (broilers, layers, broiler breeders and layer breeders) and poultry environments (box liner, litter, drag swab, droppings, mice and feed) were examined bacteriologically between 1988 and 1997 at the Poultry Disease Laboratory at the National Agriculture and Water Research Center in Riyadh. Eleven Salmonella serogroups representing 38 different Salmonella serovars were identified by means of antigenic analysis. The majority of the 276 isolates (26.2%) of Salmonella typed, were recovered from liver, heart and intestines of the broilers and layers. The most prominent Salmonella serogroups isolated were as follows: serogroup C1 (392 isolates, 37.26%), B (289 isolates, 27.47%) and D1 (269 isolates, 25.69%). However, untypable and multiple serogroups were also encountered, the most frequent isolates serotyped belonged to groups C1 (97 isolates, 24.7%), D1 (86 isolates, 31.9%), and B (71 isolates, 24.6%). Salmonella Enteritidis (85 isolates, 98.8%), Salmonella Virchow (48 isolates, 57.8%), Salmonella Paratyphi B var. Java (41 isolates, 57.7%) and Salmonella Infantis (30 isolates, 20.6%) were distributed the most widely as all were encountered in poultry and in poultry environments. S. Enteritidis phage type 4 (30 isolates, 35.3%), was the phage type most frequently detected among group D1 phage types, while 39 (45.8%) of the isolates of S. Enteritidis could not be phage typed.     

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.     

Increasing ceftriaxone resistance in Salmonellae, Taiwan

In Taiwan, despite a substantial decline of Salmonella enterica serotype Choleraesuis infections, strains resistant to ciprofloxacin and ceftriaxone persist. A self-transferable bla(CMY-2)-harboring IncI1 plasmid was identified in S. enterica serotypes Choleraesuis, Typhimurium, Agona, and Enteritidis and contributed to the overall increase of ceftriaxone resistance in salmonellae.