Severe Outbreak of Sorbitol-Fermenting Escherichia coli O157 via Unpasteurized Milk and Farm Visits, Finland 2012.

Shiga toxin-producing, sorbitol-fermenting Escherichia coli O157 (SF O157) has emerged as a cause of severe human illness. Despite frequent human findings, its transmission routes and reservoirs remain largely unknown. Foodborne transmission and reservoir in cattle have been suspected, but with limited supporting evidence. This study describes the outbreak of SF O157 that occurred in Finland in 2012. The outbreak originated from a recreational farm selling unpasteurized milk, as revealed by epidemiologic and microbiological investigations, and involved six hospitalized children and two asymptomatic adults with culture-confirmed infection. An identical strain of SF O157 was isolated from patients, cattle and the farm environment, and epidemiologic analysis suggested unpasteurized milk as the vehicle of transmission. This study reports the first milkborne outbreak of SF O157, provides supporting evidence of cattle as a reservoir and highlights the health risks related to the consumption of unpasteurized milk.

Variation in the hyphal growth rate and the virulence of two genotypes of the crayfish plague organism Aphanomyces astaci.

Crayfish plague, a devastating disease of freshwater crayfish, is caused by an oomycete organism, Aphanomyces astaci. Currently five genotypes of A. astaci are known, but variable features between the strains or genotypes have not been studied extensively. This study analysed 28 isolates of the As genotype and 25 isolates of the Ps1 genotype and reveals that the radial growth rate is significantly (P < 0.001) different between these two genotypes, although highly variable inside the genotype As. Two Ps1 genotype isolates and two As genotype isolates with different radial growth rates were tested in an infection trial. Clear differences were detected in the development of mortality in the test groups. The representatives of the Ps1 genotype caused total mortality within a short time span. The As genotype isolates were much less virulent. The slow-growing As isolate showed higher virulence than the As isolate with a high growth capacity. Although slow growth could be one survival strategy of the pathogen, several other mechanisms are involved in the pathogenicity and warrant further studies.

Hybrids of Shigatoxigenic and Enterotoxigenic Escherichia coli (STEC/ETEC) Among Human and Animal Isolates in Finland.

Diarrhoeagenic Escherichia coli (DEC) cause serious foodborne infections in humans. Total of 450 Shigatoxigenic E. coli (STEC) strains isolated from humans, animals and environment in Finland were examined by multiplex PCR targeting the virulence genes of various DEC pathogroups simultaneously. One per cent (3/291) of the human STEC and 14% (22/159) of the animal and environmental STEC had genes typically present in enterotoxigenic E. coli (ETEC). The strains possessed genes encoding both Shiga toxin 1 and/or 2 (stx1 and/or stx2 ) and ETEC-specific heat-stable (ST) enterotoxin Ia (estIa). The identified stx subtypes were stx1a, stx1c, stx2a, stx2d and stx2g. The three human STEC/ETEC strains were isolated from the patients with haemolytic uraemic syndrome and diarrhoea and from an asymptomatic carrier. The animal STEC/ETEC strains were isolated from cattle and moose. The human and animal STEC/ETEC strains belonged to 11 serotypes, of which O2:H27, O15:H16, O101:H-, O128:H8 and O141:H8 have previously been described to be associated with human disease. Identification of multiple virulence genes offers further information for assessing the virulence potential of STEC and other DEC. The emergence of novel hybrid pathogens should be taken into account in the patient care and epidemiological surveillance.

Characterization of Clostridium perfringens isolates from healthy turkeys and from turkeys with necrotic enteritis.

Clostridium perfringens is an important bacterial pathogen, especially in poultry, where it can lead to both subclinical and clinical disease. The aim of this study was to present data on pathological findings at outbreaks of necrotic enteritis (NE) in turkey production in Finland during the period from 1998 to 2012. Furthermore, C. perfringens isolates from healthy and diseased turkeys were characterized and their genetic diversity was investigated using pulsed-field gel electrophoresis (PFGE). Isolates (n = 212) from birds with necrotic gut lesions and from healthy flocks of 30 commercial turkey farms were characterized for the presence of cpa, cpb, iA, etx, cpb2, and cpe and netB genes. A total of 93 C. perfringens isolates, including 55 from birds with necrotic gut lesions and 38 from healthy birds from 13 different farms, were analyzed with PFGE. All contract turkey farmers (n = 48) of a turkey company that produces 99% of domestic turkey meat in Finland were interviewed about background information, management at the farm, and stress factors related to NE outbreaks. Pulsed-field gel electrophoresis analysis with SmaI restriction enzyme resulted in 30 PFGE patterns among the 92 C. perfringens isolates of high diversity. Out of all isolates, 212 (100%) were α-toxin-positive and one isolate (0.5%) was both α- and ß2 toxin-positive. Fourteen isolates (6.6%) were necrotic enteritis toxin B (NetB) positive; all were recovered from turkeys with NE. In none of the isolates obtained from healthy turkeys was the netB toxin identified. In conclusion, a high diversity of C. perfringens isolates from turkeys with different health status was shown. All isolates produced α toxin, whereas only low percentages of isolates carried the netB toxin gene. The role of the netB toxin in NE in turkeys needs to be further investigated.

Distribution and epidemiology of genotypes of the crayfish plague agent Aphanomyces astaci from noble crayfish Astacus astacus in Finland.

The crayfish plague agent Aphanomyces astaci was isolated from 69 noble crayfish Astacus astacus samples in Finland between 1996 and 2006. All isolates were genotyped using randomly amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR). Altogether, 43 isolates belonged to the genotype group of Astacus strains (As), which is assumed to represent the genotype originally introduced into Europe around 1860 and into Finland in 1893. There were 26 crayfish plague isolates belonging to the group of Pacifastacus strain I (Ps1), which appeared in Europe after the stocking of the North American species signal crayfish Pacifastacus leniusculus. The geographical distribution of the 2 genotypes in Finland corresponded with the stocking strategies of signal crayfish. The majority of Ps1-strains (83%) were associated with a classical crayfish plague episode involving acute mortality, compared with only 33% of the As-strains. As-strains were found more often by searching for reasons for population declines or permanently weak populations, or through cage experiments in connection with reintroduction programmes. In some water bodies, isolations of the As-strains were made in successive years. This study shows that persistent crayfish plague infection is not uncommon in noble crayfish populations. The described epidemiological features suggest a difference in virulence between these 2 genotypes.

Molecular characterization of Salmonella enterica subsp. enterica serovar Typhimurium DT1 isolates.

Salmonella Typhimurium DT1 is endemic to Finland and has caused human outbreaks since the 1960s. Domestic DT1 isolates (n=235) from 1972 to 1999 from human cases, animals and other sources, as well as foreign DT1 isolates from human cases (n=20) were analysed by molecular methods. Pulsed-field gel electrophoresis (PFGE) yielded 38 XbaI profiles. Of these, XbaI profile 10 was seen in 49% (125/255) of the isolates. Twelve IS200 profiles were obtained; the most common IS200 profile D was seen in 64% (33/52) of the isolates. Two clusters were formed by compilation of the XhaI-, BlnI- and SpeI-PFGE and IS200 profiles and possession of the serovar-specific virulence plasmid. The major cluster contained eight IS200 profiles, including IS200 profile D and XhaI profile 10, and had no virulence plasmid, and can be regarded as typical of the endemic Typhimurium DT1 infection.

Discrimination between endemic and feedborne Salmonella Infantis infection in cattle by molecular typing.

Salmonella enterica serovar Infantis is endemic in Finnish cattle. Feed contaminated with S. Infantis was distributed to cattle farms in May 1995. Following increased sampling, S. Infantis was detected on 242 farms in 1995. Molecular typing was used to differentiate the farms that were infected by the feed-related Infantis from those infected by other endemic strains. Twenty-three isolates from feed in 1995 and 413 from cattle (72 from 19924, 324 from 1995, 17 from 1996-7) were analysed. The feed-related Infantis was clonally related to the endemic infection by the ribotype, IS200-type and XbaI-profile. The feed isolates had a distinctive plasmid that appeared in pulsed-field gel electrophoresis as a 60 kb band when cleaved with XbaI or linearized by S1-nuclease. This plasmid appeared in cattle only since the outbreak and seemed stable on the follow-up farms. In addition to contact farms, the feedborne strain was found on 19% of the farms infected with S. Infantis in 1995 but not having bought suspected feedstuffs, possibly as secondary infections.