Characteristics of microbial biofilm on wooden vats ('gerles') in PDO Salers cheese

The purpose of this study was to characterize microbial biofilms from 'gerles' (wooden vats for making PDO Salers cheese) and identify their role in milk inoculation and in preventing pathogen development. Gerles from ten farms producing PDO Salers cheese were subjected to microbial analysis during at least 4 periods spread over two years. They were distinguished by their levels of Lactobacillus (between 4.50 and 6.01 log CFU/cm(2)), Gram negative bacteria (between 1.45 and 4.56 log CFU/cm(2)), yeasts (between 2.91 and 5.57 log CFU/cm(2)), and moulds (between 1.72 and 4.52 log CFU/cm(2)). They were then classed into 4 groups according their microbial characteristics. These 4 groups were characterized by different milk inoculations (with either sour whey or starter culture, daily or not), and different washing procedures (with water or whey from cheese making). The farm gerles were not contaminated by Salmonella, Listeria monocytogenes or Staphylococcus aureus. Only one slight, punctual contamination was found on one gerle among the ten studied. Even when the milk was deliberately contaminated with L. monocytogenes and S. aureus in the 40 L experimental gerles, these pathogens were found neither on the gerle surfaces nor in the cheeses. Using 40 L experimental gerles it was shown that the microbial biofilms on the gerle surfaces formed in less than one week and then remained stable. They were mainly composed of a great diversity of lactic acid bacteria (Leuconostoc pseudomesenteroides, Lactococcus lactis, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus hilgardii,…), Gram positive catalase positive bacteria (Curtobacterium flaccumfaciens, Curtobacterium oceanosedimentum Citrococcus spp., Brachybacterium rhamnosum, Kocuria rhizophila, Arthrobacter spp.…) and yeast (Kluyveromyces lactis, Kluyveromyces marxianus). In less than 1 min, even in a 500 L farm gerle, the gerle's microbial biofilm can inoculate pasteurized milk with micro-organisms at levels superior to those in raw milk.     

Important vectors for Listeria monocytogenes transmission at farm dairies manufacturing fresh sheep and goat cheese from raw milk

The aim of this study was to determine the transmission routs of Listeria spp. in dairy farms manufacturing fresh cheese made from ovine and caprine raw milk and to evaluate the impact of Listeria monocytogenes mastitis on raw milk contamination. Overall, 5,799 samples, including 835 environmental samples, 230 milk and milk product samples, and 4,734 aseptic half-udder foremilk samples were collected from 53 dairy farms in the dairy intensive area of Lower Austria. Farms were selected for the study because raw milk was processed to cheese that was sold directly to consumers. A total of 153 samples were positive for Listeria spp., yielding an overall prevalence of 2.6%; L. monocytogenes was found in 0.9% of the samples. Bulk tank milk, cheese, and half-udder samples were negative for Listeria spp. Because none of the sheep and goats tested positive from udder samples, L. monocytogenes mastitis was excluded as a significant source of raw milk contamination. L. monocytogenes was detected at 30.2% of all inspected farms. Swab samples from working boots and fecal samples had a significantly higher overall prevalence (P < 0.001) of L. monocytogenes (15.7 and 13.0%, respectively) than did swab samples from the milk processing environment (7.9%). A significant correlation was found between the prevalence of L. monocytogenes in the animal and in the milk processing environment and the silage feeding practices. Isolation of L. monocytogenes was three to seven times more likely from farms where silage was fed to animals throughout the year than from farms where silage was not fed to the animals.     

Effect of pH and water activity on the growth limits of Listeria monocytogenes in a cheese matrix at two contamination levels

Listeria monocytogenes can proliferate at the beginning of cheesemaking as the conditions favor growth. The objective of this study was to establish the growth limits of L. monocytogenes in a cheese matrix, in case of potential contamination of the milk prior to cheese manufacture. A semisoft laboratory scale model cheese system was made at different initial pH and water activity (a(w)) levels with a mix of two strains of L. monocytogenes. A factorial design of five pH values (5.6 to 6.5), four a(w) values (0.938 to 0.96), and two L. monocytogenes inoculation levels (1 to 20 CFU/ml and 500 to 1,000 CFU/ml) was carried out. Each combination was evaluated in six independent replicates. In order to determine if there was a dominant strain, isolated colonies from the cheeses were analyzed by pulsed-field gel electrophoresis. The data relating to growth initiation were fitted to a logistic regression model. The a(w) of milk influenced the probability of growth initiation of L. monocytogenes at both low and high contamination levels. The pH, at the concentrations tested, had a lower effect on the probability of growth initiation. At pH 6.5 and a(w) of 0.99 for low contamination levels and pH 6.5 and a(w) of 0.97 for high contamination levels, increases in population of up to 4 and 2 log were observed at low and high contamination levels, respectively. This shows that if conditions are favorable for growth initiation at the early stages of the cheesemaking process, contamination of milk, even with low numbers, could lead to L. monocytogenes populations that exceed the European Union's microbiological limit of 100 CFU/g of cheese.     

Prevalence and concentration of Listeria monocytogenes in sliced ready-to-eat meat products in the Hellenic retail market

The aim of this work was to estimate the prevalence and concentration of Listeria monocytogenes in packaged precut (slices or cubes) ready-to-eat (RTE) meat products available in the Hellenic retail market. Samples of these RTE meat products (n = 209) were taken from local supermarkets during a 3-month period and analyzed for the presence of L. monocytogenes with an automated enzymatic qualitative immunoassay followed by biochemical confirmation of positive results. The concentration of the pathogen in the positive samples was also determined. Seventeen samples (8.1%) were positive for L. monocytogenes. Eight (47.1%) of these 17 samples were from the same manufacturer; 36.4% of the products tested from this manufacturer were positive for L. monocytogenes. When bacon samples were not considered, the estimated prevalence of L. monocytogenes in sliced RTE meat products was much lower (3.1%). The L. monocytogenes populations in all positive samples were low, < or = 10 CFU/g. In 64.7% of the L. monocytogenes-positive samples, other Listeria species, including L. innocua and L. welshimeri, were also present at <10 to 690 CFU/g. These results indicate that L. monocytogenes is present in low numbers but is in a considerable proportion of the packaged precut RTE meat products that are sold in the Hellenic retail market. Cooked ham and bacon cut in cubes were the sample types most often contaminated with L. monocytogenes. The higher level of handling (e.g., cutting) associated with these products may further increase the risk of contamination with L. monocytogenes.     

Quantitative risk assessment of listeriosis due to consumption of raw milk

The objectives of this study were to estimate the risk of illness for raw milk consumers due to Listeria monocytogenes in raw milk sold by permitted dealers, and the risk for people on farms who consume raw milk. Three scenarios were evaluated for raw milk sold by dealers: raw milk purchased directly from bulk tanks, from on-farm stores, and from retail. To assess the effect of mandatory testing of raw milk by regulatory agencies, the number of listeriosis cases per year was compared where no raw milk testing was done, only a screening test to issue a permit was conducted, and routine testing was conducted and milk was recalled if it was L. monocytogenes positive. The median number of listeriosis cases associated with consumption of raw milk from bulk tanks, farm stores, and retail for an intermediate-age population was 6.6 × 10(-7), 3.8 × 10(-5), and 5.1 × 10(-5) cases per year, respective ly. In populations with high susceptibility, the estimated median number of cases per year was 2.7 × 10(-7) (perinatal, i.e., pregnant women and their fetuses or newborns) and 1.4 × 10(-6) (elderly) for milk purchased from bulk tanks, 1.5 × 10(-5 ) (perinatal) and 7.8 × 10(-5) (elderly) for milk from farm stores, and 2.1 × 10(-5) (perinatal) and 1.0 × 10(-4) (elderly) for milk from retail. For raw milk consumed on farms, the median number of listeriosis cases was 1.4 × 10(-7) cases per year. A greater risk of listeriosis was associated with consumption of raw milk obtained from retail and farm stores as compared with milk obtained from bulk tanks. This was likely due to additional time-temperature combination steps in the retail and farm store models, which increased the chances for growth of L. monocytogenes in raw milk. A close relationship between prevalence of L. monocytogenes in raw milk and the values of disease incidence was observed. Hence, a reduction in the number of cases per year in all populations was observed when a raw milk-testing program was in place, especially when routine testing and recalling of milk was conducted.     

Aflatoxin M1 and ochratoxin A in raw bulk milk from French dairy herds

Mycotoxins in milk are a public health concern and have to be regularly monitored. A survey on the presence of aflatoxin M₁ (AFM₁) and ochratoxin A (OTA) in raw bulk milk was conducted in 2003 in the northwest of France, the main French milk-producing basin. Randomly selected farms (n = 132) were characterized by a diet based on corn silage and containing a large proportion of on-farm produced cereals, feeding sources that are frequently contaminated by mycotoxins. Farms were surveyed twice in winter and in summer. At each sampling time, a trained surveyor completed a questionnaire recording farm management procedures and production traits. The AFM₁ was found in 3 out of 264 samples but at levels (26 ng/L or less) that are below the European legislation limit of 50 ng/L. Traces of AFM₁ (less than 8 ng/L) were also found in 6 other samples. The OTA was detected in 3 samples also at low levels, 5 to 8 ng/L. Farms that tested positive to the presence of mycotoxins, 12 in total including 6 farms that had traces of AFM₁, differed from negative farms by a more extensive use of total mixed rations, 58 vs. 27%. In addition, the positive farms tended to have lower milk yields. Although the incidence of milk contamination with AFM₁ and OTA at the farm level was low during the period studied, production and management data from the surveyed farms suggest a link between feeding management practices and mycotoxin contamination.     

Low incidence of foodborne pathogens of concern in raw milk utilized for farmstead cheese production

Overall milk quality and prevalence of four target pathogens in raw milk destined for farmstead cheesemaking was examined. Raw milk samples were collected weekly from June to September 2006 from 11 farmstead cheese operations manufacturing raw milk cheese from cow's, goat's, and sheep's milk. Samples were screened for Listeria monocytogenes, Staphylococcus aureus, Salmonella, and Escherichia coli O157:H7 both quantitatively (direct plating) and qualitatively (PCR). Overall, 96.8% of samples had standard plate counts of < 100,000 CFU/ml, 42.7% of which were < 1,000 CFU/ml. Although no federal standards exist for coliforms in raw milk, 61% of samples tested conformed to pasteurized milk standards under the U.S. Pasteurized Milk Ordinance (PMO) at < 10 CFU/ml. All cow and sheep milk samples and 93.8% of goat milk samples were within the limits dictated by the PMO for somatic cell counts. Of the 11 farms, 8 (73%) produced samples that were positive for S. aureus, which was detected in 34.6% (46 of 133) of milk samples. L. monocytogenes was isolated from three milk samples (2.3%), two of which were from the same farm. E. coli O157:H7 was recovered from one sample of goat's milk for an overall incidence of 0.75%. Salmonella was not recovered from any of the 133 samples. The findings of this study suggest that most raw milk intended for farmstead cheesemaking is of high microbiological quality with a low incidence of pathogens. These data will help inform risk assessments associated with the microbiological safety of farmstead cheeses, particularly those manufactured from raw milk.     

Prevalence and growth of Listeria on naturally contaminated smoked salmon over 28 days of storage at 4 degrees C

Only limited data are available on the growth characteristics of Listeria in naturally contaminated ready-to-eat foods. To evaluate Listeria contamination patterns and growth in smoked salmon, 72 smoked salmon product samples from two processing plants were tested for Listeria spp. and L. monocytogenes. Samples were divided into four approximately equal portions: one portion was tested on receipt, and the other three were vacuum sealed and stored at 4 degrees C for 7, 14, and 28 days. Listeria testing was performed using both an enrichment procedure and direct plating to enumerate Listeria in samples that contained >2 to 10 CFU/g. Five samples were positive for Listeria spp., including one sample that was positive for L. monocytogenes. Most samples yielded only sporadic positive results among the portions tested on days 0, 7, 14, and 28. Only one sample contained Listeria spp. in numbers above the detection limit for enumeration. For this sample, the portions tested on days 7 and 28 contained 46 and 52 CFU/g, respectively, whereas the portion tested on day 14 was negative. Overall, our data indicate that there is considerable heterogeneity in Listeria spp. distribution within a single positive smoked fish sample. Even with refrigerated storage for 28 days, none of the naturally contaminated samples reached Listeria spp. numbers >100 CFU/g, which indicates that Listeria growth was limited within a 4-week storage period. However, because of the apparent heterogeneity of Listeria distribution within samples, the interpretation of growth data collected on naturally contaminated samples is difficult.     

Comprehensive survey of pasteurized fluid milk produced in the United States reveals a low prevalence of Listeria monocytogenes

A comprehensive survey was undertaken to generate contemporary data on the prevalence of Listeria monocytogenes in pasteurized fluid milk produced in the United States. Samples (5,519) near the sell-by expiration date were purchased at retail outlets over a 5-week period and analyzed for presence of L. monocytogenes. Products consisted of whole milk, nonfat milk, and chocolate milk packaged in gallon, half gallon, quart, pint, and half-pint containers. Samples were collected from both large and small retail stores in urban and suburban locations in four FoodNet cities (Baltimore, Md., Atlanta, Ga., St. Paul/ Minneapolis, Minn., and San Francisco, Calif.). Samples were prescreened for L. monocytogenes by the AOAC-approved rapid Vitek immunodiagnostic assay system, enzyme-linked fluorescent assay method. Positive prescreening samples were cultured according to the Bacteriological Analytical Manual, enumerated for L. monocytogenes with a nine-tube most-probable-number (MPN) procedure, and confirmed by biochemical characterization. The frequency of isolation of L. monocytogenes in these products was 0% (0 of 1,897) in whole milk, 0.05% (1 of 1,846) in nonfat milk, 0% (0 of 1,669) in chocolate milk, and 0% (0 of 107) in other (reduced fat and low fat) milk samples. Overall, L. monocytogenes was confirmed in only 0.018% of pasteurized milk samples (1 of 5,519). Enumeration of the single confirmed positive nonfat milk sample revealed low-level contamination (<0.3 MPN/g), even when sampled 5 days past the expiration of the sell-by date. The results confirm the low frequency of contamination of pasteurized fluid milk products by L. monocytogenes for products sold in the United States and reaffirm the reduction of contamination frequency of fluid milk by L. monocytogenes when compared with earlier estimates from the U.S. Food and Drug Administration Dairy Safety Initiatives Program.     

Ecology of Listeria spp. in a fish farm and molecular typing of Listeria monocytogenes from fish farming and processing companies

This study focused on the ecology of Listeria monocytogenes in a fish farm by following the changes in its occurrence in different types of samples for a three year period. In addition, L. monocytogenes isolates from different seafood industry areas were compared with pulsed field gel electrophoresis (PFGE) typing to discover possible associations between primary production, further processing and final products. Weather conditions were found to have a strong influence on the probability of finding Listeria spp. in a fish farm environment. The number of samples contaminated with Listeria spp. was typically bigger after rainy periods. Brook and river waters as well as other runoff waters seemed to be the main contamination source at the farm studied. The farmed fish originally found to carry L. monocytogenes become gradually Listeria free. The time needed for the purification of the fish was several months. The sea bottom soil samples were the ones that preserved the L. monocytogenes contamination the longest time. It can be stated that the fish and fish farm equipment studied did not spread listeria contamination. On the contrary, they were found to suffer from listeria contamination coming from outside sources like the brook water. There was a wide range of different L. monocytogenes PFGE-pulsotypes (30) found at 15 Finnish fish farms and fish processing factories. L. monocytogenes isolates from the final products often belonged to the same pulsotypes as did the isolates from the processing environment as well as from the raw fish. This suggests that, in addition to the fish processing factory environment, the fish raw materials are important sources of L. monocytogenes contamination in final products.     

A cross-sectional study on the prevalence of Listeria monocytogenes and Salmonella in New York dairy herds

As part of our long-term objective of assessing risk for Listeria monocytogenes and Salmonella spp. in dairy herds, we carried out a cross-sectional study to determine the prevalence of the two organisms. The study population consisted of a sample of dairy herds enrolled in the Quality Milk Promotion Services at Cornell during the period of April 1998 to March 1999. The sample was stratified by geographical region to assure representation. Four hundred and four dairy farms were enrolled in the study. In-line milk filters were collected from each farm for bacteriological examination of L. monocytogenes and Salmonella spp. Four hypothesized risk factors were evaluated for their association with the likelihood of the presence of each of the two organisms using logistic regression analysis. Listeria monocytogenes was isolated from 51 (12.6%) of the milk filters. We found region-specific differences in the rate of farms with positive milk filters for this pathogen. Salmonella spp. were isolated from 6 (1.5%) milk filters. One isolate was confirmed as Salmonella enterica Serotype Typhimurium DT 104. There was no significant association between any of the hypothetical risk factors and the likelihood of Salmonella spp. isolation. Our study demonstrated that both L. monocytogenes and Salmonella spp. were prevalent in milk filters in New York dairy herds and that Salmonella was isolated at a significantly lower rate then L. monocytogenes.     

Detection and enumeration of four foodborne pathogens in raw commingled silo milk in the United States

A nationwide survey was conducted to obtain qualitative and quantitative data on bacterial contamination of raw commingled silo milk intended for pasteurization. The levels of total aerobic bacteria, total coliforms, Enterobacteriaceae, Escherichia coli, and Staphylococcus aureus were determined using the TEMPO system. The prevalence rates and levels of presumptive Bacillus cereus, E. coli O157:H7, Listeria monocytogenes, and Salmonella spp. were determined in 214 samples. B. cereus was detected in 8.91% of samples, at 3.0 to 93 CFU/ml. E. coli O157:H7 was detected in 3.79 to 9.05% of samples, at <0.0055 to 1.1 CFU/ml, depending on the assay utilized. Salmonella spp. were recovered from 21.96 to 57.94% of samples, at <0.0055 to 60 CFU/ml. L. monocytogenes was detected in 50.00% of samples, at <0.0055 to 30 CFU/ml. The average log-transformed counts of total viable bacteria were slightly lower in samples containing no pathogens. No correlation was observed between the levels of organisms detected with the TEMPO system and the presence or levels of any pathogen except E. coli O157:H7. A higher average log-transformed count of total viable bacteria was observed in samples positive for this organism. The high prevalence rates of target pathogens may be attributed to a variety of factors, including detection methods, sample size, and commingling of the milk in the silo. The effects of commingling likely contributed to the high prevalence rates and low levels of target pathogens because of the inclusion of milk from multiple bulk tanks. The high prevalence rates also may be the result of analysis of larger sample volumes using more sensitive detection methods. These quantitative data could be utilized to perform more accurate risk assessments and to better estimate the appropriate level of protection for dairy products and processing technologies.     

SIMPLE oPSU BROTH-BAX® PCR-PICOGREEN® SYSTEM FOR RAPID DETECTION OF LISTERIA MONOCYTOGENES IN PASTEURIZED MILK AND HOT DOGS

Factors that significantly affect BAX® PCR detection of Listeria monocytogenes from optimized Penn State University (oPSU) broth were identified and optimized. Concentration of PCR product was significantly reduced by BAX™ protease and significantly increased by eliminating the lysis step and directly diluting oPSU broth prior to PCR. A simple oPSU broth-BAX® PCR-PicoGreen® (PSU-BAX-PicoGreen) system was developed and compared with current methods for detecting low levels of L. monocytogenes in commercial milk and hot dogs. All 30 milk samples inoculated with 10–20 CFU per mL L. monocytogenes were positive by FDA, BAX® and PSU-BAX-PicoGreen methods and all 42 uninoculated milk samples were negative by all of the above methods. All 30 hot dog samples inoculated with 10-20 CFU/g L. monocytogenes were positive by the USDA and PSU-BAX-PicoGreen methods, however, 2 hot dog samples gave indeterminate results with the standard BAX® method. All 42 uninoculated hot dog samples were negative by USDA, 9 were indeterminate by BAX® and 2 were positive by PSU-BAX-PicoGreen. The PSU-BAX-PicoGreen system may provide a simple and accurate method for rapidly screening pasteurized foods for both injured and noninjured L. monocytogenes.     

Identification and molecular characterization of Listeria monocytogenes isolated in raw milk in the region of Algiers (Algeria)

Listeria monocytogenes was isolated from raw milk, whey and curdled milk produced and collected in the region of Algiers and Blida between September 2003 and July 2004. Four out of 153 (2.61%) farm milk samples and 6 out of 80 (7.50%) tankers' samples tested positive for L. monocytogenes. All samples of whey and curdled milk (n=12) tested negative for L. monocytogenes, but 2 of 22 (9%) samples of whey were contaminated by L. innocua. L. monocytogenes isolates were grouped by a multiplex PCR assay; all isolates belonged to the PCR-group IVb, which corresponds to serovars 4b, 4d and 4e. L. monocytogenes isolates were characterized by Pulsed-Field Gel Electrophoresis (PFGE). The combination of AscI and ApaI macrorestriction patterns yielded five different pulsovars (I to V). The results indicate that raw milk, and raw milk products are potential sources of the L. monocytogenes and represent a potential risk for consumers.     

Microbiological profile of greenhouses in a farm producing hydroponic tomatoes

Produce, including tomatoes, has been implicated in several outbreaks of foodborne illness. A number of the sources of contamination for produce grown in open fields are known. However, as an alternative agricultural system, hydroponic greenhouses are reasonably expected to reduce some of these sources. The objective of the present study was to determine the microbiological profile of tomatoes grown in greenhouses at a Mexican hydroponic farm with a high technological level and sanitary agricultural practices (SAPs) in place. Tomatoes and other materials associated with the farm were analyzed for the presence of Salmonella enterica and populations of Escherichia coli, coliforms, and Enterobacteriaceae. Tomatoes showed median levels of 0.8 log CFU per tomato for Enterobacteriaceae, < 0.5 log CFU per tomato for coliforms, and 0.5 most probable number per tomato for E. coli. Despite the physical barriers that the facilities provide and the implemented SAPs, we found that 2.8% of tomatoes were contaminated with Salmonella and 0.7% with E. coli. Other Salmonella-positive materials were puddles, soil, cleaning cloths, and sponges. Samples from the nursery and greenhouses were positive for E. coli, whereas Salmonella was found only in the latter. Although hydroponic greenhouses provide physical barriers against some sources of enteric bacterial contamination, these results show that sporadic evidence of fecal contamination and the presence of Salmonella can occur at the studied greenhouse farm.     

Efficiency of electrolyzed oxidizing water on reducing Listeria monocytogenes contamination on seafood processing gloves

Food processing gloves are typically used to prevent cross-contamination during food preparation. However, gloves can be contaminated with microorganisms and become a source of contamination. This study investigated the survival of Listeria monocytogenes on gloves and determined the efficacy of electrolyzed oxidizing (EO) water for reducing L. monocytogenes contamination on seafood processing gloves. Three types of reusable gloves (natural rubber latex, natural latex, and nitrile) and two types of disposable gloves (latex and nitrile) were cut into small pieces (4 x 4 cm(2)) and inoculated with 5-strain L. monocytogenes cocktail (5.1 x 10(7) CFU/cm(2)) with and without shrimp meat residue attached to surfaces. L. monocytogenes did not survive well on clean reusable gloves and its populations decreased rapidly to non-detectable levels within 30 min at room temperature. However, high levels of Listeria cells were recovered from clean disposable gloves after 30 min of inoculation. Presence of shrimp meat residue on gloves enhanced the survival of L. monocytogenes. Cells of L. monocytogenes were detected on both reusable and disposal gloves even after 2 h at room temperature. Soaking inoculated gloves in EO water at room temperature for 5 min completely eliminated L. monocytogenes on clean gloves (>4.46 log CFU/cm(2) reductions) and significantly (p<0.05) reduced the contamination on soil-containing gloves when compared with tap water treatment. EO water could be used as a sanitizer to reduce L. monocytogenes contamination on gloves and reduce the possibility of transferring L. monocytogenes from gloves to RTE seafoods.     

Evaluation of an enumeration method for Listeria monocytogenes at low contamination levels in cold-smoked salmon

For the enumeration of Listeria monocytogenes in cold-smoked salmon, a sensitive enumeration method, based on membrane filtration followed by transfer of the filter on a selective medium has been recently developed (Gnanou Besse et al., 2004, A contribution to the improvement of L. monocytogenes enumeration in cold-smoked salmon. International Journal of Food Microbiology, 91, 119-127). The aim of the study was to assess the performance of this enumeration method through an inter-laboratory study, using cold-smoked salmon artificially contaminated at 2 different levels (approximately 0.6 and 1.6 log10 CFU g(-1)). A reproducibility standard deviation of 0.23 log10 CFU g(-1)and 0.15 log10 CFU g(-1) was obtained for the method respectively at the lower level and the higher level. Under certain conditions, the uncertainty of measurement can be derived from the method reproducibility standard deviation and was calculated to be 0.46 log10 CFU g(-1) for the lower contamination level and 0.30 log10 CFU g(-1) for the higher contamination level. These values can be considered as satisfactory for such low contamination levels.     

Dairy farm reservoir of Listeria monocytogenes sporadic and epidemic strains

Identifying the reservoirs of a pathogen is vital for control of sporadic disease and epidemics. Listeria monocytogenes is a zoonotic foodborne pathogen that is responsible for 28% of food-related deaths in the United States annually, as well as a major cause of massive product recalls worldwide. To examine the role of the dairy farm as a potential source or reservoir for L. monocytogenes subtypes shown to cause human listeriosis, we compared the pulsed-field gel electrophoresis (PFGE) restriction enzyme digestion profiles of L. monocytogenes dairy farm-associated strains (milk, environmental, and bovine) to human sporadic and epidemic disease strains. Twenty-three percent of human sporadic strains had PFGE patterns identical to that of farm isolate(s). Additionally, three farm environmental strains and one human sporadic strain had a PFGE pattern identical to a strain of L. monocytogenes responsible for the 1985 California epidemic. These data indicate that this epidemic strain continues to cause sporadic human illness and has a potential dairy farm as a reservoir.     

Occurrence of Listeria monocytogenes in freshwater fish farms and fish-smoking plants

Three Swiss fish farms, farming rainbow trout (Oncorhynchus mykiss), and their affiliated smoking plants were analyzed for the presence of Listeria spp. 590 samples were collected from the farming environment (raceway water, sludge), faecal content and skin of the fish, fish during processing, and the processing environment.Listeria spp. were found at prevalences of 2·3% in plant A, 31·6% in plant B (mainly L. monocytogenes), and 13·8% in plant C (mainly L. innocua). This high contamination rate in plant B may be explained by the following facts: (i) farm B uses river water flowing through agricultural land; (ii) plant B rears fish in earth ponds instead of concrete ponds or raceways; (iii) fish from farm B had not been denied feed prior to slaughter; and (iv) total lack of regular mechanical and chemical cleaning in the fish farm B and processing plant B.In all three plants samples taken after smoking but before packaging did not contain Listeria spp., although in plant B and C the raw fish was contaminated. Hygienic defaults during packaging can lead to contaminated ready-to-eat products, detected in plant B (L. monocytogenes) and plant C (L. innocua) with one sample each. To minimize a possible health hazard to the consumer, it is of great importance to prevent postprocessing contamination of smoked fish.Finally, means of preventing Listeria contamination during farming, slaughtering, processing and storage are suggested.     

Occurrence and typing of Listeria monocytogenes strains in retail vacuum-packed fish products and in a production plant.

One hundred and ten samples of ready-to-eat, vacuum-packed, smoked and cold-salted fish products were collected from retail outlets in southern Finland during 1996 for examination of the occurrence and level of Listeria monocytogenes. The samples originated from 12 producers. Positive samples with levels exceeding 100 CFU/g were encountered mainly in one of the producers (no. 8). Therefore, 200 samples from the plant and the products of this producer were studied during August-September 1996 and May-September 1997, as well as 55 samples from the six fish farms providing raw material fish to this plant, during September 1997-January 1998. The isolates were characterised by serotyping and pulsed-field gel electrophoresis (PFGE). L. monocytogenes was isolated in 20% (22/110) of the samples from the retail market, originating from 6 producers. Ten of these positive samples contained L. monocytogenes at > 100 CFU/g (maximum 1.37 X 10(4) CFU/g). Seventeen percent (5/30) of cold-smoked and 50% (16/32) of cold-salted rainbow trout samples were contaminated. Only one hot-smoked fish product (2%) was found to be positive by enrichment. Nineteen (86%) of the strains isolated from the retail samples belonged to serovar 1/2a and three (14%) to serovar 4b. In further studies the production line of plant no. 8 was found to be contaminated. All of isolates from up until autumn, 1997 both the products and the production plant were serovar 1/2a; thereafter one strain of 4b and one of 1/2 (H-antigen untypeable) were isolated from the plant. The samples from raw material fish were all negative for L. monocytogenes. The samples from retail market fell into seven PFGE types. Five and nine PFGE types, respectively, were found from the products and the plant of producer no. 8. PFGE type A was detected from the retail products of four producers and was also dominant among the isolates from production plant no. 8. PFGE type A was the only one found repeatedly from skinning, salting and slicing units as well as from products throughout the whole period. PFGE proved to be a powerful tool for studying contamination points and routes in the production plant. The measures based on hazard analysis critical control points (HACCP) program resulted in L. monocytogenes negative samples at production plant no. 8 from the beginning of January 1998.