Plasmid analysis of Salmonella enteritidis isolated from human gastroenteritis cases and from epidemiologically associated poultry flocks.

Plasmid analysis of Salmonella enteritidis isolates from human gastroenteritis cases and from two commercial egg-producing poultry flocks was performed to determine if the poultry flocks were the source of the human infections. The plasmid profile and restriction fragment pattern (fingerprint) of five S. enteritidis isolates from human cases matched those of nine isolates from internal organs of egg-laying hens in one flock which was the source of eggs consumed by the cases. Another commercial flock was epidemiologically associated as the source of eggs consumed by affected persons in four separate gastroenteritis outbreaks from which S. enteritidis isolates were available. Five S. enteritidis isolates from human cases in these four outbreaks had the same profile and fingerprint, and they all matched those of the 24 isolates from hens in this flock. These results provide further documentation of egg-borne transmission of S. enteritidis to humans.     

The identification of Salmonella enteritidis-infected poultry flocks associated with an outbreak of human salmonellosis.

In the summer of 1991 a human outbreak of Salmonella enteritidis infection occurred following a barbecue in which about 100 persons were involved. Eggs, supplied by one or more of 10 different layer farms, were the most probable source of the infection. To identify the S. enteritidis-positive flocks, an immunoassay was used to detect salmonella serogroup D-specific antibodies in the yolk of hens eggs. Antibody titres in the eggs from two layer farms, farm A and B, clearly exceeded the titres found in randomly collected eggs. Further investigation on farm A and B yielded high antibody titres in the eggs from flocks A1, A2 and B2, and low titres in the eggs from flock B1. S. enteritidis was isolated from the faecal samples of flocks A1, A2 and B2, whereas no salmonella was detected in the faecal samples of flock B1. The flocks present on both farms originated from the same breeder flock.     

Salmonella enteritidis phage type 4 from the contents of intact eggs: a study involving naturally infected hens

Two small flocks of egg-laying hens, naturally infected with Salmonella enteritidis, were housed in individual cages so that their eggs could be identified. During a longitudinal study where the contents of 1,119 eggs were examined, 11 were positive for S. enteritidis. One isolate was phage type (PT) 33 the others were PT4. The production of infected eggs was clustered though intermittent. The positive eggs, which were produced by 10 of the 35 hens, were all found to contain fewer than 10 salmonellas. Some birds were also apparently carrying S. hadar PT14 as this organism was isolated from the contents of six cracked eggs.     

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.     

The prevalence of Salmonella enteritidis and other Salmonella spp. among Canadian registered commercial layer flocks

A survey was conducted to estimate the prevalence of Salmonella enteritidis and other salmonellas among Canadian commercial egg producing flocks. Environmental (faecal and eggbelt) samples from 152 of 295 (52.9%) randomly selected flocks were contaminated with salmonellas. Thirty-five different salmonella serovars were isolated. Eggbelt samples were more often contaminated with salmonellas than faecal samples (25.7 v. 10.1%). The most prevalent serovars were S. heidelberg, S. infantis, S. hadar, and S. schwarzengrund; they were isolated from samples of 59/295 (20%), 18/295 (6.1%), 17/295 (5.8%), and 15/295 (5.1%) flocks, respectively. Feed samples of 21/295 (7.2%) flocks were contaminated with salmonellas. Salmonella enteritidis was isolated from the environmental samples of 8/295 (2.7%) flocks. Salmonella enteritidis phage type (PT) 8 was isolated from 5 flocks, PT 13a from 2 flocks, and PT 13 from 1 flock.     

Salmonella isolated from humans,animals and other sources in Canada, 1983-92.

A total of 89760 human and 22551 non-human isolates of salmonella were serotyped in Canada during the period 1983-92. There were 2180 reported outbreaks associated with 10065 cases during the 10-year period. The most common salmonella serovars isolated from human and non-human sources were S. typhimurium and S. hadar. The third and fourth most common serovars from human sources were S. enteritidis and S. heidelberg, respectively, and from non-human sources they were S. heidelberg and S. infantis. The number of S. typhimurium isolations from human and non-human sources showed a downward trend over the 10-year period. A total of 222 outbreaks of S. typhimurium associated with 1622 cases occurred. The S. hadar isolations from human and non-human sources reached a peak during the years 1987-90 and declined thereafter. The number of human isolates of S. enteritidis increased until 1985 and fluctuated at a level of 8.3-12.8% of all human isolates thereafter. Seventy-three outbreaks of S. enteritidis infection associated with 568 cases occurred. More than 50% of the S. enteritidis infections in humans were caused by phage type (PT) 8. During the review period, infections caused by PT4 were less common and were almost exclusively found in people who had travelled abroad. The annual isolation rates of S. heidelberg from human and non-human sources increased steadily during the period. Bacteriophage typing of serovars from outbreaks showed that contaminated food products of poultry and bovine origin were common sources of human infection. Salmonella typhi was identified as the cause of 43 small outbreaks affecting 116 persons.     

An in-depth investigation into a food poisoning outbreak

On and shortly after the 6th May 1990, 16 people were affected by food poisoning in an old peoples' residential home, of whom two died. The vehicle of infection was identified as a baked Alaska contaminated by Salmonella enteritidis phage type (PT) 8 and, at an early stage of the investigation, the source was attributed to a single infected egg. A separate investigation by the author, however, revealed that the baked Alaska meringue had been dispensed from an inadequately cleaned piping bag which had been recovered from the kitchen a month after the outbreak. A pure, profuse culture of S. enteritidis PT8 was isolated from it. At least one secondary case may have been attributable to food made with this bag. Ministry of Agriculture Investigations of the flocks suspected of producing the eggs used for the baked Alaska demonstrated an absence of S. enteritidis. On this basis, the author considered a more likely cause of the outbreak to be the piping bag, contaminated from source or sources unknown within the kitchen. Furthermore, the possibility of human carrier transmission cannot be wholly ruled out. The incident underlines the dangers of jumping to conclusions at the outset of food poisoning investigations and emphasises that hypotheses formulated on sources of contamination must be properly tested, the absence of which, in this instance, led the investigators to unwarranted conclusions as to the cause of the outbreak.     

Four linked outbreaks of Salmonella enteritidis phage type 4 infection--the continuing egg threat

Four outbreaks of Salmonella enteritidis phage type (PT) 4 occurred among guests at functions for which a single commercial caterer supplied food. Retrospective cohort studies were used to describe the epidemiology of three of these outbreaks and identify the vehicle(s) responsible. Of 172 guests at these three events, 47 fitted the clinical case definition for illness and 24 cases were confirmed to have S. enteritidis PT4 infection. Food containing raw egg was identified epidemiologically as the likely vehicle of infection in two of the three outbreaks (odds ratios (OR) and 95% confidence intervals 9.1 (2.2-39.9) and 6.9 (1.2-46.4)). Logistic regression analysis yielded OR = 10.7 (p = 0.0022) and OR = 9.3 (p = 0.015) for egg consumption in two of the outbreaks. These outbreaks highlighted the continuing need to remind the public and commercial caterers of the potential high risks of contracting salmonella from shell eggs. Education of caterers includes advice to obtain eggs and other products from reputable and identifiable suppliers.     

Investigation of an outbreak of Salmonella enteritidis gastroenteritis associated with consumption of eggs in a restaurant chain in Maryland

Salmonella enteritidis ser. enteritidis was isolated from patrons and employees of three restaurants in a restaurant chain in Maryland during August and September 1985. Isolates from all three restaurants had identical plasmid profiles; this profile was present in 13 of 40 randomly selected S. enteritidis isolates received by the Maryland state health department laboratory during a comparable time period. The outbreak in one restaurant resulted in at least 71 illnesses, with 17 persons known to have been hospitalized. Scrambled eggs served on a "breakfast bar" were implicated as the vehicle of transmission in this restaurant, with eggs a possible vehicle in another of the three restaurants. The data point out the risks associated with improper handling of eggs in food service establishments, provide further evidence for the observed association between S. enteritidis and eggs in the northeastern United States, and demonstrate the utility of plasmid analysis in investigation of outbreaks involving common Salmonella serotypes.     

The prevalence of Salmonella enteritidis and other Salmonella sp. among Canadian registered commercial chicken broiler flocks

A nation-wide survey was conducted to estimate the prevalence of Salmonella enteritidis and other salmonellas among Canadian commercial broiler flocks. Environmental (litter and/or water) samples from 226 of 294 (76.9%) randomly selected flocks were contaminated with salmonellas. Litter samples were more often contaminated with salmonellas than water samples (47.4 v. 12.3%). Fifty different salmonella serovars were isolated. The most prevalent serovars were S. hadar, S. infantis, and S. schwarzengrund; they were isolated from samples of 98/294 (33.3%), 26/294 (8.8%), and 21/294 (7.1%) flocks, respectively. Feed samples of 39/290 (13.4%) flocks were contaminated with salmonellas. Salmonella enteritidis was isolated from the environmental samples of 9/294 (3.1%) flocks. Salmonella enteritidis phage type (PT) 8 was isolated from seven flocks, and PT 13a from two flocks.     

A hospital outbreak of salmonella food poisoning due to inadequate deep-fat frying.

In an outbreak of plasmid-free Salmonella enteritidis phage type 4 (PT4) food poisoning at a hospital for mentally handicapped people in July 1990, 101 residents and 8 staff were affected and a cohort study implicated beef rissoles cooked by deep-fat frying as the vehicle of infection (relative risk 2.92, 95% confidence interval 1.73-4.93, P << 0.001). Replication of the cooking process demonstrated that the rissoles achieved core temperatures of only 48-60 degrees C despite external temperatures of 91-95 degrees C and an oil temperature of 142-154 degrees C. No residual food was available for microbiological testing but plasmid-containing S. enteritidis PT 4 was isolated in shell eggs from the hospital kitchen.     

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.     

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.     

Consecutive salmonella outbreaks traced to the same bakery.

Two consecutive community outbreaks of Salmonella enteritidis phage type 4 (PT4) traced to the same bakery occurred in Cardiff, Wales during August-September 1992. In the first outbreak, illness was associated with eating custard slices (odds ratio 23.8, 95% confidence interval 6.5-94.4, P < 0.0001), and in the second, with eating fresh cream cakes (odds ratio 15.8, 95% confidence interval 1.6-374, P = 0.004). Environmental investigations implicated cross-contamination during preparation of the cold-custard mix as the cause of the first outbreak, and inadequate cleaning and disinfection of nozzles used for piping cream in the second outbreak. S. enteritidis PT4 was isolated from fresh cream sponge cake retained by a case and from two fresh cream cakes and four environmental swabs obtained at the bakery. This incident illustrates the hazard of widespread environmental contamination with salmonella and the need for thorough environmental cleansing for any premises implicated in an outbreak of food poisoning.     

Two separate outbreaks of Salmonella enteritidis phage type 14b food poisoning linked to the consumption of the same type of frozen food

OBJECTIVES: To ascertain, using a combination of epidemiological, environmental and microbiological methods of investigation, a possible link between two outbreaks of salmonella food poisoning. METHODS: Case-control studies were carried out on the known at-risk populations. Environmental investigations took place in the food preparation areas used for the social functions and microbiological examinations were carried out on faecal specimens obtained from cases, environmental swabs, and food specimens when these were available. RESULTS: In both outbreaks, illness was associated with the consumption of sesame prawn toast (outbreak one P<0.004; outbreak two P<0.0001). Salmonella enteritidis phage type 14b was cultured from the faecal specimens of cases in both outbreaks and from a packet of sesame prawn toast used for the second outbreak function. Molecular typing methods indicated that the salmonella cultures obtained in both outbreaks were indistinguishable from each other and from cultures obtained from imported Spanish eggs in a previous survey. Imported Spanish eggs were used in the manufacture of the sesame prawn toast. CONCLUSIONS: Adequate cooking must take place of raw food products, which should be clearly labelled as such. Manufacturers should consider, when possible, the use of pasteurized egg in the preparation of food products.     

Numbers of Salmonella enteritidis in the contents of naturally contaminated hens' eggs.

Over 5700 hens eggs from 15 flocks naturally infected with Salmonella enteritidis were examined individually for the presence of the organism in either egg contents or on shells. Thirty-two eggs (0.6%) were positive in the contents. In the majority, levels of contamination were low. Three eggs, however, were found to contain many thousands of cells. In eggs where it was possible to identify the site of contamination, the albumen was more frequently positive than the yolk. Storage at room temperature had no significant effect on the prevalence of salmonella-positive eggs but those held for more than 21 days were more likely (P less than 0.01) to be heavily contaminated. In batches of eggs where both shells and contents were examined, 1.1% were positive on the former site and 0.9% in the latter.     

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.     

Epizootic-epidemiologic aspects of the Salmonella infection chain in poultry production

Infections with S. typhimurium and S. enteritidis develop in poultry flocks as persistent flock enzootics, often without any clinical manifestation. Infected parenteral hen flocks are the source of vertical, so-called pseudo-trans-ovarian infection chains. These chains may induce horizontally self-maintaining cycles of the infection in the flocks. Environmental Salmonella may enter such cycles. Each of these cycles is able to transmit Salmonella to poultry products so entailing human health hazards permanently. The intensification in modern poultry production causes the permanent presence of Salmonella in the flocks. Hitherto all known control measures (biological, chemoprophylactic, physical) didn't result in a remarkable success. Therefore, the only alternative is the consequent implementation of hygiene regimes in all stages of production and during processing and marketing of poultry products in order to dilute Salmonella as much as possible.     

Egg-related Salmonella enteritidis, Italy, 1991

In recent years, Salmonella enteritidis has become an increasingly important public health problem in Italy. In some parts of the country, the fraction of total human salmonella isolates accounted for by S. enteritidis has risen from 3-4% in the mid-1980s to more than 30% in 1990. Between 1990 and 1991, the number of reported S. enteritidis outbreaks increased more than sixfold. The 33 outbreaks reported in 1991 occurred in seven contiguous regions in northern and central Italy and were clustered in time between June and October; in the majority, products containing raw or undercooked shell eggs were implicated. Five of the egg-related outbreaks that occurred within a 30 kilometre radius over a 7-week period were investigated in detail. A phage type 1 strain containing a 38·9 MDa plasmid appeared responsible for three of the outbreaks, while in the remaining two a phage type 4 strain, also with a 38·9 MDa plasmid was isolated. Efforts are being made to enhance epidemiological surveillance and laboratory evaluation, and the use of pasteurized eggs has been recommended for high-risk populations.     

Monitoring the incidence and causes of diseases potentially transmitted by food in Australia: annual report of the Ozfoodnet Network, 2006

In 2006, OzFoodNet sites reported 24,598 notifications of seven diseases or conditions that are commonly transmitted by food, representing an increase of 2.5% over the mean of the previous 5 years. The most frequently notified aetiological agents were Campylobacter (15,492 notifications) and Salmonella (8,331 notifications). Salmonella notifications increased in 2006 by 5.2% when compared to historical reports. The most common Salmonella serotype notified in Australia during 2006 was Salmonella Typhimurium, as in previous years. S. Saintpaul was the second most common serotype notified during 2006 as a result of a large multistate outbreak associated with rockmelons. During 2006, OzFoodNet sites reported 1,544 outbreaks of gastrointestinal illness including those transmitted by contaminated food. In total, these outbreaks affected 34,916 people and resulted in 769 people being admitted to hospital and 27 deaths. Food was suspected or confirmed as the primary cause for 115 of these outbreaks and affected 1,522 persons, hospitalised 146 persons but did not result in any deaths. S. Typhimurium was the most common aetiological agent in foodborne outbreaks and restaurants were the most common setting for foods implicated in foodborne outbreaks. Sixteen outbreaks were related to eggs during 2006 compared to five outbreaks in 2005. Eighty-one per cent (13/16) of these egg-associated outbreaks were due to various phage types of S. Typhimurium. Fresh fruits and vegetables, categorised as fresh produce were responsible for four outbreaks, all due to salmonellosis. Public health laboratories provided complete serotype and phage type information on more than 97% of all Salmonella notifications during 2006. Completeness of reporting for Salmonella was equivalent to 2005 and was essential to identifying and investigating outbreaks. This report demonstrates OzFoodNet's ability to detect and investigate the burden and causes of foodborne disease in Australia. OzFoodNet efforts assist agencies to develop food safety policy and prevent foodborne illness.