Riitta Maijala

An outbreak of Listeria monocytogenes serotype 3a infections from butter in Finland.

In February 1999, an outbreak of listeriosis caused by Listeria monocytogenes serotype 3a occurred in Finland. All isolates were identical. The outbreak strain was first isolated in 1997 in dairy butter. This dairy began delivery to a tertiary care hospital (TCH) in June 1998. From June 1998 to April 1999, 25 case patients were identified (20 with sepsis, 4 with meningitis, and 1 with abscess; 6 patients died). Patients with the outbreak strain were more likely to have been admitted to the TCH than were patients with other strains of L. monocytogenes (60% vs. 8%; odds ratio, 17.3; 95% confidence interval, 2.8-136.8). Case patients admitted to the TCH had been hospitalized longer before cultures tested positive than had matched controls (median, 31 vs. 10 days; P=.008). An investigation found the outbreak strain in packaged butter served at the TCH and at the source dairy. Recall of the product ended the outbreak.

Contaminant lactic acid bacteria of dry sausages produce histamine and tyramine

Exogenous biogenic amines present a food poisoning hazard in fermented foods especially with additional risk factors, e.g. amine oxidase inhibiting drugs, alcohol and gastrointestinal diseases. Forty-two lactic acid bacteria (LAB) strains were isolated from seven dry sausages during ripening (0 day, 21 day and 49 day). Their ability to produce histamine and tyramine was studied by HPLC detection from broth cultures after 2 days incubation. The tyramine and histamine concentrations in sausages increased during fermentation. 10 of 42 LAB strains produced 402-1087 ppm tyramine. Four of these 10 contaminant LABs were also histamine-positive (725-1083 ppm). Most of the amine positive strains were found in sausages at the end of ripening and with highest amine levels. On the basis of these results the contaminant LABs play an important role in tyramine and histamine formation during the ripening of dry sausages. Therefore it could be possible to decrease the levels of amines formed by limiting the initial level and growth of these contaminant bacteria.

Exposure of Listeria monocytogenes within an epidemic caused by butter in Finland

Data on the levels of bacteria and the amounts of food consumed in food-borne outbreaks provides an excellent opportunity to study the effects of exposure to Listeria monocytogenes. Between June 1998 and April 1999, an outbreak caused by L. monocytogenes serotype 3a in butter occurred in Finland. The majority of the cases were immunocompromised and hospitalized at the Helsinki University Central Hospital (HUCH), where 7-g butter packages produced by a dairy plant were used as the only butter brand. The butter had also been sold to 10 other central hospitals as well as to the retail market. Based on the data on hospital stay, butter consumption and the qualitative and quantitative analyses of L. monocytogenes in butter, the attack rates and exposure were estimated. Incubation studies on the naturally contaminated small butter packages showed that the levels found in the packages at the time of detection of the outbreak could reliably be used for these estimations. However, the levels of L. monocytogenes in 500-g packages increased. The attack rate among HUCH patients varied from 70 to 117 cases per 1000 patients at risk, depending on which estimate of the contamination level of butter (100-60%) was used. The highest single dose (7.7 x 10(4) CFU in one meal) could have been sufficient to cause the listeriosis cases at HUCH. However, this data also supports another hypothesis, according to which these listeriosis cases were caused by a prolonged daily consumption of contaminated butter during the hospital stay. The estimated daily dose, based on the hospital kitchen data or the highest detected level in a wholesale sample (11,000 CFU/g), would have varied from 1.4 x 10(1) to 2.2 x 10(3) CFU/day or from 2.2 x 10(4) to 3.1 x 10(5) CFU/day, respectively. The choice of the hypothesis has a crucial impact on the interpretation of this data for the dose-response estimations as well as for the discussion on Food Safety Objectives. Due to the susceptibility of hospital patients, special care must be taken in order to avoid even low levels of L. monocytogenes in food served.

The Effect of GDL-induced pH Decrease on the Formation of Biogenic Amines in Meat

The effect of pH on the formation of biogenic amines has mainly been studied in broths in which pH has been fixed before incubation. However, in the fermentation of dry sausage, pH quite rapidly decreases from the initial value to a certain level. In this study glucono-delta-lactone (GDL) was used to decrease pH in meat, Six minced meat samples were each divided into three portions (A-C): 0% (A), 0.5% (B), or 1.0% (C) of GDL was added and the samples were incubated at 20-22°C for 7 d. The amounts of biogenic amines (histamine, tyramine, putrescine, cadaverine, phenylethylamine, tryptamine, spermine, and spermidine) as well as pH, water activity, and the bacterial counts of lactic acid bacteria, fecal streptococci, coliforms, and total plate count were measured. Addition of GDL resulted in a significant decrease in pH and in the levels of histamine and putrescine as well as in the levels of fecal streptococci, coliforms, and total plate counts. Of 87 fecal streptococci, seven Enterococcus faecalis strains produced tyramine. All the coliforms and related strains isolated from violet red bile agar produced tyramine, putrescine, and cadaverine on agar plates. However, the proportion of histamine-positive strains of these strains, especially Hafnia alvei , increased from 0 to 57% during the incubation. The rate and level of pH decrease clearly affected amine formation in meat, indicating that the levels of, e.g., histamine produced could be decreased by optimizing the pH decrease during fermentation. Addition of GDL facilitates study of the effect of pH decrease without interactions between the starter culture and contaminant flora.

Contamination of Carcasses with Human Pathogenic Yersinia enterocolitica 4/O:3 Originates from Pigs Infected on Farms

Pigs are considered as a major reservoir of human pathogenic Yersinia enterocolitica and a source of human yersiniosis. However, the transmission route of Y. enterocolitica from farm to pork is still unclear. The transmission of pathogenic Y. enterocolitica from pigs to carcasses and pluck sets was investigated by collecting samples from 364 individual ear-tagged pigs on the farm and at the slaughterhouse. In addition, isolated strains were analyzed, using pulsed-field gel electrophoresis. Isolation of similar genotypes of pathogenic Y. enterocolitica 4/O:3 in animals on the farm and at the slaughterhouse and in carcasses shows that carcass contamination originates from the strains a pig carries during the fattening period. Direct contamination from the carrier pig to its subsequent pluck set is also the primary contamination route for pluck sets, but cross-contamination appears to have a larger impact on pluck set contamination than on carcasses. In this study, the within-farm prevalence of pathogenic Y. enterocolitica varied from 0% to 100%, indicating specific farm factors affect the prevalence of Y. enterocolitica in pigs. The association of farm factors with the high prevalence of pathogenic Y. enterocolitica on farms was studied for the first time, using correlation and two-level logistic regression analyses. Specific farm factors, i.e. drinking from a nipple, absence of coarse feed or bedding for slaughter pigs, and no access of pest animals to pig house, were associated with a high prevalence of pathogenic Y. enterocolitica 4/O:3.

Transmission of Yersinia pseudotuberculosis in the Pork Production Chain from Farm to Slaughterhouse

ABSTRACT The transmission of Yersinia pseudotuberculosis in the pork production chain was followed from farm to slaughterhouse by studying the same 364 pigs from different production systems at farm and slaughterhouse levels. In all, 1,785 samples were collected, and the isolated Y. pseudotuberculosis strains were analyzed by pulsed-field gel electrophoresis. The results of microbial sampling were combined with data from an on-farm observation and questionnaire study to elucidate the associations between farm factors and the prevalence of Y. pseudotuberculosis. Following the same pigs in the production chain from farm to slaughterhouse, we were able to show similar Y. pseudotuberculosis genotypes in live animals, pluck sets (containing tongue, tonsils, esophagus, trachea, heart, lungs, diaphragm, liver, and kidneys), and carcasses and to conclude that Y. pseudotuberculosis contamination originates from the farms, is transported to slaughterhouses with pigs, and transfers to pluck sets and carcasses in the slaughter process. The study also showed that the high prevalence of Y. pseudotuberculosis in live pigs predisposes carcasses and pluck sets to contamination. When production types and capacities were compared, the prevalence of Y. pseudotuberculosis was higher in organic production than in conventional production and on conventional farms with high rather than low production capacity. We were also able to associate specific farm factors with the prevalence of Y. pseudotuberculosis by using a questionnaire and on-farm observations. On farms, contact with pest animals and the outside environment and a rise in the number of pigs on the farm appear to increase the prevalence of Y. pseudotuberculosis.

Listeria monocytogenes contamination in pork can originate from farms.

The presence of Listeria monocytogenes in the pork production chain was followed from farm to slaughterhouse by examining the farm and slaughterhouse levels in the same 364 pigs, and finally by analyzing the cut meats from the same pig lots. Both organic and conventional farms were included in the study. Altogether, 1,962 samples were collected, and the 424 L. monocytogenes isolates were analyzed by pulsed-field gel electrophoresis. The results from microbial analyses were combined with data from an on-farm observation and a questionnaire to clarify the associations between farm factors and prevalence of L. monocytogenes. The prevalence of L. monocytogenes was 11, 1, 1, 24, 5, 1, and 4% in feed and litter, rectal swabs, intestinal contents, tonsils, pluck sets (including lungs, heart, liver, and kidney), carcasses, and meat cuts, respectively. The prevalence was significantly higher in organic than in conventional pig production at the farm and slaughterhouse level, but not in meat cuts. Similar L. monocytogenes genotypes were recovered in different steps of the production chain in pigs originating from the same farm. Specific farm management factors, i.e., large group size, contact with pet and pest animals, manure treatment, use of coarse feed, access to outdoor area, hygiene practices, and drinking from the trough, influenced the presence of L. monocytogenes in pigs. L. monocytogenes was present in the production chain, and transmission of the pathogen was possible throughout the chain, from the farm to pork. Good farm-level practices can therefore be utilized to reduce the prevalence of this pathogen.

Trapping the food safety performance of a small or medium-sized food company using a risk-based model. The HYGRAM® system

The requirements of implementing Hazard Analysis and Critical Control Points (HACCP) principles in food production are increasing. A practical risk quantification model, HYGRAM, was developed for small and medium-sized enterprises to meet this challenge. The model makes the user familiar with the HACCP principles by software-assisted guidance through the procedure, connecting special microbiological hazards, good hygiene practice, and other prerequisite programs to HACCP. HYGRAM is a tool to analyze and quantify risks of different processes, and to compare them. It is developed to relieve enterprises with limited resources in confirming the food safety of their production.

Histamine and Tyramine Production by a Lactobacillus Strain Subjected to External pH Decrease

The preservative effect of fermentation is based on the pH decrease during processing. However, most studies conceming the influence of pH on the formation of biogenic amines have been made in broths with different initial pH values. A histamine- and tyramine-positive Lactobacillus strain isolated from dry sausage was added at an initial level of 3.9- to 4.4-logl0 CFU/ml and incubated on a shaker at room temperature (20.0°C) for 6 days. The pH was decreased in histidine- or tyrosine-fortified MRS broth by adding glucono-delta-lactone (GDL) or lactic acid during the incubation. The external acidification decreased the growth and the production of histamine and tyramine by the strain. GDL was more effective as a preventative than lactic acid. According to these results, a rapid pH decrease resulting in decreased growth of amine-positive lactic acid bacteria in the beginning of fermentation may be a means of preventing the formation of high levels of amines in foods.

The temporal, PFGE and resistance pattern associations suggest that poultry products are only a minor source of human infections in Western Finland

In order to compare human and retail poultry meat thermophilic Campylobacter isolates originating in a regional area in Western Finland, minimum inhibitory concentration (MICs) for six antimicrobials (96 isolates) and pulsed-field gel electrophoresis (PFGE) (102 isolates) were analysed. Campylobacter spp. were detected in 10.5% out of 305 fresh poultry products studied; 29 (90.5%) isolates were identified as Campylobacter jejuni. Among the 70 human isolates, 66 (94.3%) isolates were identified as C. jejuni. Only one C. jejuni domestic poultry isolate showed resistance (ampicillin), whereas domestic human C. jejuni isolates were more commonly resistant to ciprofloxacin, nalidixic acid, ampicillin and tetracycline. The resistance in foreign human isolates was significantly more common than among domestic isolates. PFGE analysis with KpnI restriction enzyme resulted in 59 different PFGE types among the poultry and human isolates. Three types were detected first in poultry meat and thereafter during the following month in domestic human samples, whereas the other conjoint types were detected only after many months. This study suggests that poultry products play only a minor role in human campylobacteriosis in the study area and that the resistance found in domestic human isolates is not likely related to retail poultry meat products.