Human isolates of Listeria monocytogenes in Sweden during half a century (1958-2010).

Isolates of Listeria monocytogenes (n = 932) isolated in Sweden during 1958-2010 from human patients with invasive listeriosis were characterized by serotyping and pulsed-field gel electrophoresis (PFGE) (AscI). Of the 932 isolates, 183 different PFGE types were identified, of which 83 were each represented by only one isolate. In all, 483 serovar 1/2a isolates were distributed over 114 PFGE types; 90 serovar 1/2b isolates gave 32 PFGE types; 21 serovar 1/2c isolates gave nine PFGE types; three serovar 3b isolates gave one PFGE type; and, 335 serovar 4b isolates gave 31 PFGE types. During the 1980s in Sweden, several serovar 4b cases were associated with the consumption of European raw soft cheese. However, as cheese-production hygiene has improved, the number of 4b cases has decreased. Since 1996, serovar 1/2a has been the dominant L. monocytogenes serovar in human listeriosis in Sweden. Therefore, based on current serovars and PFGE types, an association between human cases of listeriosis and the consumption of vacuum-packed gravad and cold-smoked salmon is suggested.

Bovine subclinical mastitis caused by different types of coagulase-negative staphylococci.

Subclinical mastitis caused by intramammary infections (IMI) with coagulase-negative staphylococci (CNS) is common in dairy cows and may cause herd problems. Control of CNS mastitis is complicated by the fact that CNS contain a large number of different species. The aim of the study was to investigate the epidemiology of different CNS species in dairy herds with problems caused by subclinical CNS mastitis. In 11 herds, udder quarter samples were taken twice 1 mo apart, and CNS isolates were identified to the species level by biochemical methods. The ability of different CNS species to induce a persistent infection, and their associations with milk production, cow milk somatic cell count, lactation number, and month of lactation in cows with subclinical mastitis were studied. Persistent IMI were common in quarters infected with Staphylococcus chromogenes, Staphylococcus epidermidis, and Staphylococcus simulans. The results did not indicate differences between these CNS species in their association with daily milk production, cow milk somatic cell count, and month of lactation in cows with subclinical mastitis. In cows with subclinical mastitis, S. epidermidis IMI were mainly found in multiparous cows, whereas S. chromogenes IMI were mainly found in primiparous cows.

Subtyping of a frequent phagovar of Listeria monocytogenes in Sweden by use of restriction endonuclease analysis.

In Sweden, many Listeria monocytogenes strains belonging to serovar 4b and isolated during the last five years from different sources share the same phagovar--2389:2425:3274:2671:47:108:340. The object of the present study was to investigate if 31 L. monocytogenes serovar 4b strains belonging to this particular phagovar could be differentiated by use of a simple restriction endonuclease analysis (REA). Among the enzymes tested, Xho I was found to be the most useful, since this enzyme could divide the 31 strains into five groups. The profiles of all human clinical isolates were indistinguishable from each other, which indicates that these strains may represent a single clone. The food isolates and the strains of human origin did not share the same profile. This further characterization may be of epidemiological importance as this phagovar of L. monocytogenes has been associated with at least two outbreaks of human listeriosis in Europe.

Occurrence of Listeria monocytogenes in soft and semi-soft cheeses in retail outlets in Sweden.

Samples of 333 retail cheeses produced in or imported into Sweden were examined for the presence of Listeria monocytogenes. Listeria monocytogenes was isolated from 6% of the cheese samples. Cheeses made from raw milk were more frequently contaminated with L. monocytogenes (42%) than cheeses made from heat-treated milk (2%). The incidence of the organism in whole cheeses and pre-cut wedges was similar (6%). L. monocytogenes was only found in imported cheeses (18 from France, and one from Germany and Italy, respectively). The numbers of L. monocytogenes varied from < 1 x 10(2) to 1 x 10(5) cfu/g. All L. monocytogenes strains belonged to serogroup 1/2, except isolates from two samples that belonged to serogroup 4.

Campylobacter incidence on a chicken farm and the spread of Campylobacter during the slaughter process.

To get a better understanding of the epidemiology of Campylobacter, a chicken farm was studied for 16 weeks with samplings in each flock weekly from input until the flock became colonized with Campylobacter or slaughtered. Samples were taken from fresh droppings and from drinkers during the rearing period, as well as from the environment in empty houses. The spread of Campylobacter during the slaughter process was also surveyed. No Campylobacter was found in samples from newly-hatched or one-week-old chickens or their drinkers. All flocks but one were colonized at two to five weeks of age. All Campylobacter isolates belonged to the same sero- and biotype; C. jejuni Penner 2. The spread of Campylobacter in the flock was rapid and usually all samples were positive once colonization had been proven. C. jejuni was isolated from flies in ante-rooms as well as from air in chicken units in houses with positive chicken flocks. Samples were taken at slaughter when some of the Campylobacter positive flocks from the farm were slaughtered. Campylobacter were isolated from all sampled equipment along the processing line, from the chicken transport crates to the chillers, as well as from the air.

Histamine formation by enterococci in goat cheese.

Cheeses were made of heat-treated goat milk inoculated with large numbers of a histamine-producing strain of Streptococcus faecium or a non-histamine-producing strain of S. faecalis. Every second week during ripening (91 days) the cheeses were sampled for histamine analyses and bacteriological analyses. The numbers of enterococci remained high throughout the whole period of investigation and the maximum amount of histamine detected was 8.2 micrograms/g in one of the cheeses. The enterococci seem to have no relevance from a histamine intoxication point of view in cheeses of this kind.

Lessons from an outbreak of listeriosis related to vacuum-packed gravad and cold-smoked fish.

The first lesson learned from this outbreak was that vacuum-packed rainbow trout is not only an excellent medium for the growth of Listeria monocytogenes, but may also cause human listeriosis. Another lesson is that one single fish processing plant may spread multiple clonal types of L. monocytogenes by selling contaminated products to consumers. Thus, when investigating fish-borne outbreaks of listeriosis one should identify and type several isolates of L. monocytogenes from each food and environmental sample, since multiple clonal types might be present. The outbreak described in this paper involved at least eight human cases, three clonal types of L. monocytogenes, and lasted for 11 months. During the outbreak investigation, L. monocytogenes was also isolated from another brand of rainbow trout found in the refrigerator of one of the patients. These latter isolates belonged to a clonal type not associated with the outbreak. However, this clonal type is of considerable interest since it has been associated with foodborne outbreaks of listeriosis in several countries, and is also the second most common clonal type among human clinical isolates of L. monocytogenes in Sweden. Besides the described outbreak, it is likely that vacuum-packed, cold-smoked and gravad rainbow trout have been involved in additional cases of foodborne listeriosis in Sweden.

Characterization of Listeria strains isolated from soft cheese.

Three soft cheeses were exposed to quantitative analysis for listeria and found to contain a large number of listeria. Thirty-five of the listeria strains isolated from the three cheeses were characterized by use of biochemical tests, serotyping, phagetyping and DNA restriction enzyme analysis. Seven isolates were identified as Listeria innocua and 28 as Listeria monocytogenes. Two to four different clones of L. monocytogenes could be identified from each cheese. In contrast, only one clone could be detected among the L. innocua isolates. From an epidemiological point of view the findings of different clones of L. monocytogenes in the same cheese emphasize the need for typing several listeria isolates from one and the same food sample. It is concluded that the best overview of the population of the listeria strains is obtained after direct plating of the sample followed by enumeration, isolation and extensive typing.

Experimental colonization of mice with Campylobacter jejuni.

The ability of one human and two chicken strains of Campylobacter jejuni to colonise and survive in three different strains of laboratory mice (NMRI, CBA and C57-Black) was studied. Mice were inoculated orally with Campylobacter jejuni and faeces samples were cultured at regular intervals during the following months. The length of colonisation of mice differed between mouse strains but also between Campylobacter strains. The mouse strain C57-Black was not colonised with C. jejuni to the same degree as the other mouse strains. It is concluded that mice can become colonised for prolonged periods and that they may act as reservoirs of Campylobacter for other species.

Comparison of a cold enrichment method and the IDF method for isolation of Listeria monocytogenes from animal autopsy material.

The method most often used in Sweden for isolation of Listeria monocytogenes from animal autopsy material is a cold enrichment method. This method is very slow. The International Dairy Federation (IDF) has recently presented a method for detection of L. monocytogenes in milk and milk products that is complete in one week. During a two year period 69 specimens from dead animals with suspected listeriosis were examined for L. monocytogenes in parallel analyses with both the cold enrichment method and the IDF method. Samples derived from different autopsy material representing a variety of animals. L. monocytogenes was isolated in 27.5% of the samples with the IDF method but only in 4.3% with the cold enrichment method. It is concluded that the IDF method was more sensitive than the cold enrichment method.

Occurrence of Listeria species in broilers pre- and post-chilling in chlorinated water at two slaughterhouses.

Altogether 323 pooled samples of neck skins from 1615 broilers from 2 processing plants (A and B) were examined for the presence of Listeria species. The broilers were sampled pre-chilling-after leaving the final rinser but before entering the chiller with chlorinated water-and post-chilling-immediately upon leaving the chiller. Free available chlorine in the chilling water varied from 2 to 15 ppm in plant A and was about 10 ppm in plant B. Listeria monocytogenes was only isolated from broilers in plant A sampled post-chilling (58% of 62 samples). L. innocua was isolated from 19% and 39% of broilers sampled pre-chilling in plants A and B, respectively. Post-chilling, L. innocua was isolated from 3% and 6% of samples from plants A and B, respectively.

Prevalence of Listeria monocytogenes and other Listeria spp.in smoked and 'gravad' fish.

Altogether 150 samples of vacuum-packed fish were examined for the presence of Listeria species. Listeria monocytogenes were isolated from 12 of 58 'gravad' fish samples, 3 of 26 cold-smoked and one of 66 hot-smoked fish samples. Ten of these 16 positive samples harboured more than 100 L. monocytogenes cfu/g. The highest level (132,000) was found in a sample of hot-smoked rainbow trout (Oncorhynchus mykiss). Serogroup 1/2 was most frequently found, followed by 4 and 3. One sample of gravad rainbow trout harboured more than one serogroup of L. monocytogenes. L. innocua and L. seeligeri were isolated from 12 and 1 samples, respectively.

Occurrence of Listeria species in an abattoir for cattle and pigs in Bosnia and Hercegovina.

Altogether 496 samples of meat, lymph nodes, process water and swabs from different places in the abattoir were examined for the presence of Listeria spp. L. monocytogenes was isolated from 31 (6%) and other Listeria spp. from 65 (13%) samples L. monocytogenes was isolated from 2 of 10 beef meat samples, 4 of 50 pig meat samples and 1 of 21 lymph nodes of pigs. No Listeria bacteria were isolated from lymph nodes of cattle. The highest percentage of Listeria was recovered from the unclean sections (cattle 22% and pigs 27%) and the highest frequency was observed during the winter months.

Listeria monocytogenes in faeces from clinically healthy dairy cows in Sweden.

Faecal samples from 102 clinically healthy dairy cows, representing 34 farms in the Swedish province of Uppsala, were analysed for the presence of Listeria spp. using an enrichment procedure. Listeria monocytogenes was isolated from six (6%) and L. innocua from 2 (2%) cows. From each of the 6 samples positive for L. monocytogenes, 5 isolates were further characterised by restriction enzyme analysis using the 3 enzymes Apa I, Sma I, and Asc I, followed by pulsed-field gel electrophoresis. Three of the L. monocytogenes positive cows lived at the same farm, and they all harboured the same clonal type. One of these 3 cows also harboured a further clonal type of L. monocytogenes. The fact that one of the cows harboured 2 different clonal types of L. monocytogenes is important from an epidemiological point of view when routes of infection are to be investigated.

Isolation of Listeria monocytogenes from goat cheese associated with a case of listeriosis in goat.

Listeria monocytogenes was isolated from the brain of a goat, which was euthanized due to listeriosis. A few weeks later a similar subtype of L. monocytogenes was isolated from an on-farm manufactured fresh cheese which did not contain any milk from the goat which had suffered from listeriosis. A similar subtype was also found on 1 of the shelves in the refrigerator where cheeses were stored. Prior to the onset of listeriosis, 1 fresh cheese had been made of milk from the actual goat, which may have excreted L. monocytogenes in her milk. Thus, the cheese made of this milk may have contaminated the shelves in the refrigerator which then has served as a Listeria reservoir for new cheeses during several weeks.

Listeria monocytogenes subtypes associated with mortality among fallow deer (Dama dama).

Different subtypes of Listeria monocytogenes were isolated from various animal and environmental samples during an episode of increased mortality on a fallow deer (Dama dama) farm. During a 4-wk period, six fallow deer died, including four does, one fawn, and one adult buck. Prior to death, one of the does had exhibited central nervous system signs characteristic of listeriosis. Postmortem examination of the six deer showed no histologic changes typical of listeriosis, although inflammatory changes were present in several organs. Different subtypes of L. monocytogenes were isolated from brain samples from six deer, from fodder and soil from the deer feeding area, and from faces of some healthy animals on the farm. Listeria monocytogenes, which was frequently isolated in the environment of the farm, was considered the probable major cause of mortality in these fallow deer.