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1.
Listeria monocytogenes is a foodborne pathogen of major concern for public health in industrialized countries. Listeria carriage by pigs at the herd level could be a primary source for carcass contamination. Forty-seven finishing pig facilities were involved in the present study designed to compare three environmental swabbing sites in order to detect Listeria spp. in piggeries. Swabs were taken from the pen walls, the perianal regions of the pigs and the trough/feeder of the piggery premises. Listeria contamination of wet or dry feed given to the pigs was also investigated. The capacity of the various sampling sites for Listeria spp. detection were compared with a maximum likelihood estimation method. Listeria spp. were recovered in 74% of the pens studied and L. monocytogenes was detected in 15% of pens. With a specificity of 99%, sensitivity estimates (and 95% CI) of the Listeria spp. detection method were 93.4% (72.7-98.7) for pen walls, 73.1% (54.9-85.9) for pigs and 66.6% (48.6-80.7) for the trough/feeder. Listeria spp. were isolated from 84% of wet feed samples and 5% of dry feed samples. Listeria monocytogenes was found in 13% of wet feed samples. The type of feeding (wet versus dry) was associated (P < 0.001) with Listeria spp. contamination of both the pen and the feed. The results of this study confirm that Listeria spp., including L. monocytogenes, are present in pig facilities. Pen wall swabbing appears to be an effective way to assess Listeria spp. status of finishing pigs. The type of feeding (wet versus dry) could play a role in pig contamination.  相似文献   

2.
Samples were taken from 100 camel sausages from the different retail markets in Aydin province in the south-west of Turkey and they were tested for the presence of Listeria spp by biochemical methods. Samples were enriched using Listeria Enrichment Broth and they were inoculated onto Listeria Selective Agar. Listeria monocytogenes was isolated from nine samples (9%), Listeria innocua from 14 samples (14%) and Listeria welshimeri from two samples(2%). A 701 bp fragment of listeriolysin O sequence for L. monocytogenes was amplified using specific primers by polymerase chain reaction (PCR) for confirmation of the identification. A random primer (OPA-11) was used in a random amplified polymorphic DNA (RAPD) assay. This detected five different band profiles amongst the L. monocytogenes isolates, indicating a relatively large amount of genetic heterogeneity amongst the nine isolates. The study has highlighted the need for improved strategies for food safety, in particular appropriate hygienic precautions to avoid contamination of sausage during the manufacturing process and appropriate preservation techniques during storage and transport, to prevent transmission of Listeria spp to consumers at home and abroad.  相似文献   

3.
A total of 1,705 fecal specimens or ileo-cecal contents of cattle, pigs, dogs, cats, chicken and rats were submitted for the isolation of Listeria monocytogenes by the use of the combination of Oxford-LPM agar plates after the cold enrichment in PBS at 4 degrees C for 4-6 weeks. Prevalence of L. monocytogenes was found to be 1.9% in cattle, 0.6% in pigs, 0.9% in dogs and 6.5% in rats. However, none of L. monocytogenes was isolated from chicken or cats. Among 26 isolates of L. monocytogenes, 13 strains (50%) were classified into types 1/2a (3 strains), 1/2b (5 strains) and 4b (5 strains) and were often associated with human listeriosis. The majority of the Listeria spp. other than L. monocytogenes isolated from these animals was found to be L. innocua.  相似文献   

4.
Inhibition of isolation of Listeria monocytogenes by bacteriocin-like substance (BLS)-producing Listeria innocua after enrichment culture was investigated. When 26 L. monocytogenes strains were examined in combination with eight L. innocua strains using the spot on lawn method, 52/208 (25.0%) combinations showed the growth inhibition of L. monocytogenes. When two Listeria species were cultured simultaneously in selective enrichment broth, inhibition of isolation of L. monocytogenes was observed in 12/52 of the combinations at 24h (23.1%), in 24/52 at 48h (46.2%) and in 30/52 (57.7%) after 7 days of incubation. The randomly amplified polymorphic DNA profiles showed no interstrain similarities between either strains of the BLS-producing L. innocua or the BLS-sensitive L. monocytogenes strains. Therefore inhibition by BLS-producing L. innocua of isolation of L. monocytogenes after enrichment culture is unlikely to be dependent upon a particular genetic profile.  相似文献   

5.
Listeria spp. isolated from cheese were tested for biochemical characteristics together with reference strains from culture collections. Microtitration plates were used for testing the fermentation patterns. The results were subjected to a numerical cluster analysis based on linkage maps. The variation of the group structures calculating the characteristics with and without the hemolysin reactions is demonstrated. The pathogenic species L. monocytogenes could only be separated from the avirulent species L. innocua by the hemolysin tests. Most of the cheese isolates were identified as L. innocua, some as L. monocytogenes and L. seeligeri. There is a need for an inexpensive commercial test kit to identify the serovars or virulence factors of Listeria spp. in the quality assessment of food. The present study sets up doubts for a sufficiently ensured separation of L. innocua from L. monocytogenes.  相似文献   

6.
Seasonal variation in the fecal shedding of Listeria spp. in dairy cattle was examined by collecting a total of 3,878 fecal samples during a period of two years. The prevalences of Listeria spp. and L. monocytogenes were higher during the indoor season (12.7% and 9.2%, respectively) than in samples collected from the animals on pasture (5.3% and 3.1%, respectively). The highest frequencies of Listeria spp. (19.4%) and L. monocytogenes (16.1%) were detected in December. Listeriae were isolated from at least one of the dairy cows from 45.8% of the 249 herds examined. 2.9% of the 314 milk samples collected from the farm bulk tanks on 80 dairy farms on four different occasions yielded L. monocytogenes. The seasonal occurrence of these bacteria in milk reflected the frequencies of Listeria in the fecal material but not those in the main roughage used; grass silage and pasture grass. Fecal material is considered to be a potential source of contamination of raw milk by L. monocytogenes. Investigation of the numbers of viable Listeria organisms in different animal fodders is considered essential in further epidemiological studies of these bacteria.  相似文献   

7.
Possible sources of exogenous contamination of raw milk by Listeria monocytogenes were examined on four dairy farms of different size and type of animal housing during morning milking. Feeds, including hay and concentrates, were found to be major sources of both pathogenic and nonpathogenic species of Listeria on the barns. L. innocua was the only species isolated from the grass silage, which was of good quality on all the farms. The numbers of Listeria were below 10(2)/g in all feed samples. Fecal shedding of listeriae was detected in 11.9% of the cows and the prevalences of L. monocytogenes among farms ranged from no detections to 8.7%. The number of Listeria isolations from constructions inside the barns and from the milking environment varied between the farms. Listeriae were detected almost everywhere on one of the farms whereas on another farm the only isolations were from feed passages and floors. 13.6% of the swab samples taken from the teats before washing and drying were Listeria positive, whereas no isolations were made after cleaning the udder. Good milking and barn hygiene is considered important for diminishing the risks of exogenous contamination of raw milk by listeriae.  相似文献   

8.
Qualitative and quantitative contamination of ready-to-eat food-stuffs with the pathogen Listeria monocytogenes was studied in 1586 samples collected from 103 supermarkets (n = 946) and 61 households (n = 640) in Vienna, Austria. Seventeen groups of ready-to-eat foods were classified into three risk categories for contamination (CP1-CP3). Three to four samples were randomly collected at the retail level from each CP. Regarding the households, the sampling procedure was started with food items of CP1, and if not available, was continued with sampling of food items of CP2 and finally of CP3. Additionally, 184 environmental samples (swabs from the kitchen area, dust samples from the vacuum cleaner) and faecal samples (household members and pet animals) were included. One-hundred and twenty-four (13.1%) and 45 (4.8%) samples out of 946 food samples collected from food retailers tested positive for Listeria spp. and L. monocytogenes, respectively, with five smoked fish samples exceeding the tolerated limit of 100 CFU/g food. Food-stuffs associated with the highest risk of contamination were twice as frequently contaminated with L. monocytogenes as food-stuffs associated with a medium risk of contamination. Products showing the highest contamination rate were fish and seafood (19.4%), followed by raw meat sausages (6.3%), soft cheese (5.5%) and cooked meat products/patés (4.5%). The overall contamination rate of foods collected at the household level was more than two times lower. Only 5.6% and 1.7% of 640 food-stuffs analysed tested positive for Listeria spp. and L. monocytogenes, respectively. However, CP1 foods were rarely collected. Pulsed-field gel electrophoresis (PFGE) typing of the collected L. monocytogenes isolates revealed a high degree of diversity between the isolates, with some exceptions. PFGE typing of isolates harvested from green-veined cheese revealed a match among strains, although the manufacturer seemed to be distinguishable. Typing of household strains revealed an epidemiological link within one family. In this case, food-stuffs and the kitchen environment were contaminated by an indistinguishable isolate. In addition, the same isolate was collected from a pooled faecal sample of the household members suggesting that consumption of even low contaminated food items (<100 CFU/g) results in Listeria shedding after the passage through the gut.  相似文献   

9.
Quantitative analyses of Listeria monocytogenes in imported ready-to-eat (RTE) foods sold at retail stores in Japan were performed. Of the 77 non-cooked meat products, 6 samples (7.8%) tested positive. The levels of contamination of 4 of the samples were below 100 colony-forming units (CFU)/g, which is the microbiological criterion for L. monocytogenes in RTE foods as determined by Codex. However, Listeria cells at levels of 100 and 400 CFU/g were detected in a salami sample and a raw ham sample, respectively. All of the 70 cheese samples and the 3 samples made from raw ham and cheese showed negative test results. These results suggest that imported RTE foods are potential sources of the causative agent of listeriosis.  相似文献   

10.
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.  相似文献   

11.
We attempted to isolate Listeria monocytogenes from skin, contents of large intestines and carcasses of cattle introduced to a slaughterhouse in order to identify source of contamination for this pathogen. Sixty skin samples, 60 samples of the contents of large intestines and 30 carcass samples were colleted in June, August and November 2003 for use in this study. Listeria spp. and L. monocytogenes were isolated from 30 (50%) and 3 (5%) of the cattle skin samples, respectively. However, no Listeria spp., including L. monocytogenes, were isolated from intestinal contents or carcasses. Seven isolates were obtained, of which five and two strains were serotypes 1/2a and 1/2b, respectively. Genetic analysis suggested that there was persistent inhabitation of the pathogen around the area investigated in this study.  相似文献   

12.
The selective L-PALCAMY differential enrichment broth, the Listeria enrichment broth of the International Dairy Federation, Oxford Listeria selective agar, and PALCAM Listeria selective agar were comparatively examined in the cultural isolation of Listeria spp. from ten ruminant brains. The L-PALCAMY medium proved to be superior to the IDF broth in both selectivity and productivity for Listeria spp. in the brain samples, which were also contaminated with other bacteria. The Oxford and PALCAM agars corresponded in their productivity for Listeria spp. The latter, however, was more selective than the Oxford agar. Bacterial counts of up to 1.2 x 10(9) CFU/g of brain stem sample were made from Listeria monocytogenes (L.m.), and up to 6.2 x 10(4) CFU/g from Listeria innocua. A total of 164 brains from ruminants showing CNS disturbances and/or pathoanatomical CNS alterations were examined using L-PALCAMY medium, and Oxford and PALCAM agar. L.m. could be isolated from 29 of the brains, and Listeria innocua from five. Cultural isolation of both Listeria spp. occurred in one brain. Of 27 brains containing L.m., which were also examined using cold enrichment, L.m. was isolated in 59.3% of the cases with direct culture, in 81.5% of the cases using selective warm enrichment, and in 77.8% of the cases by means of selective cold enrichment. Five cases each were identified solely by cold or warm enrichment, respectively. In investigations of further 69 ruminant brains the number of brains shown to contain L.m. could be increased from seven to 13 by means of selective cold enrichment for three months.  相似文献   

13.
Faecal samples, collected from 200 healthy animals in Antwerp Zoo, were examined for the presence of pathogenic Listeria spp. A two-stage standard isolation (ISO) method was combined with immunomagnetic separation (IMS). ALOA agar, a chromogenic isolation medium, differentiating Listeria spp. on the basis of beta-glucosidase and phosphatidylinositol-specific phospholipase C (PIPLC) activity, was compared with PALCAM agar. Confirmation of the isolates was based on conventional biochemical tests and a disc test, which detects a specific aminopeptidase produced by all Listeria spp. except Listeria monocytogenes. Listeria spp. were isolated from 42 (21.0%), L. monocytogenes from 14 (7.0%), and Listeria ivanovii from two (1.0%) faecal samples. The application of IMS after primary enrichment detected pathogenic Listeria spp. in 12 (6.0%) samples. The ISO method, combining primary and secondary enrichment, detected pathogenic Listeria spp. in 15 (7.5%) samples. The sensitivity of IMS compared to the ISO method was 73.3% and the specificity was 99.5%. ALOA agar was superior to PALCAM agar for isolation of Listeria spp. The disc test identified all L. monocytogenes isolates. IMS after primary enrichment was a suitable screening method, but secondary enrichment increased the number of positive samples.  相似文献   

14.
Brain samples of 849 wild ruminants (654 roe deer, 189 red deer and 6 chamois) from Bavaria were examined for the occurrence of encephalopathies caused by bacteria, using cultural, serological and genetic methods. In addition, 87 brain samples were investigated histologically for clarification of the pathogenetic relevance of specific microorganisms. Using conventional bacteriological methods, 464 different bacteria were isolated. 229 of them could be differentiated to the genus level and 235 to the species level. Totally, 35 different bacteria species were isolated, most frequently Micrococcus spp., Bacillus spp. and E. coli. Listeria spp. were detected in 43 brain samples (37 from roe deer, 5 from red deer and 1 from chamois). Sixteen strains were identified as L. innocua, 14 as L. monocytogenes, 9 as L. seeligeri and 4 as L. grayi. Serological investigations of L. monocytogenes showed that 9 strains belong to serotype 1/2a and five to 4b. Analysis of the geographical distribution of the Listeria findings indicate a statistically significant (p<0.011) regional aggregation in Unterfranken (prevalence for roe deer: 12.2%, versus 4.5% in Oberbayern-Schwaben, 6.1% in Niederbayern-Oberpfalz and 0% in Oberfranken-Mittelfranken). The histological investigation (HE staining) of 87 tissue samples contaminated with encephalitis relevant bacteria showed inflammation of different severity (mild meningitis and choroiditis (n = 26) to moderate (meningo)encephalitis (n = 13)) in 41 cases.  相似文献   

15.
In order to compare the plate count method for quantitating Listeria, as published in the "Official Collection of Testing Methods" in section 35 LMBG (L. 00.00-22), to an MPN-method for Listeria based on the same mediums, these two detection methods for Listeria were tested in three sets of experiments and a routine sample status evaluation. A pure broth culture of L. monocytogenes, artificially with L. monocytogenes contaminated ground meat, artificially contaminated and cold stored ground meat as well as 77 ground beef samples from Berlin retail food stores were used in the four trials. The detection limit of the MPN-method is about 66% lower than the plate count method allowing detection of a clearly greater number of Listeria-positive samples from naturally contaminated ground meat. The MPN-method yielded more Listeria spp.-positive samples (rel. 43%) and more L. monocytogenes-positive samples (rel. 21%) versus the colony count method based on the results from the field trial using ground beef samples from retail food stores in Berlin. Nevertheless the standardized colony count method is preferred over the MPN-method for routine use because of its slightly higher productivity and much smaller variation in the results. However, the MPN-method is preferable for epidemiological studies because of the significance of the lower detection level. The random sampling evaluation of ground beef from retail stores indicated that 39% of the samples were Listeria spp.-positive and 31% were L. monocytogenes-positive when using the colony count method. A total of 56% of the meat samples were found to be Listeria spp.-positive and 38% L. monocytogenes-positive when the MPN-method was used. Population levels ranged from 10 to 580 cfu/g (Listeria spp.-positive samples) and from 10 to 270 cfu/g (L. monocytogenes-positive samples) for the colony count method. The MPN-method yielded population levels of 3.6 to 930 MPN/g for Listeria spp.-positive samples and 3.6 to 150 MPN/g for L. monocytogenes-positive samples. L. monocytogenes strains isolated using the colony count method belonged to the following serovars: 1/2a (46%), 1/2b (13%), 1/2c (33%), 3b (4%) and 4c (4%). A similar serovar isolation pattern was found for L. monocytogenes-positive MPN-tubes. The most common serotype was 1/2a (43%), followed by 1/2c (32%) and 1/2b (14%). The serotypes 3c, 4b and 4c were all isolated 4% of the time.  相似文献   

16.
Environmental samples and samples of partially processed fish from a cold-smoked salmon processing and packing plant, and product samples purchased from retail outlets, were examined for the presence of Listeria monocytogenes and other Listeria spp., motile aeromonads and Yersinia enterocolitica. Listeria spp. were not isolated from raw fish but were a contaminant of processed and partially processed fish. Listeria spp. were also detected in 18.8% of surfaces in contact with fish. Unlike Listeria spp., motile aeromonads were isolated from the raw fish but not from the finished product. They were more frequently isolated from environmental sources than Listeria spp. Yersinia enterocolitica was not isolated from any of the samples tested. Little evidence was found to show a coincidence of motile aeromonads and Listeria spp., since only one subset of samples showed such a link. It is concluded that contamination by Listeria spp. was from environmental sources at the processing plant at, or beyond, the slicing stage. Reducing the number of wet areas and special cleaning and sanitation considerations for a contaminated site (the freezer seal) are suggested as ways of reducing contamination of the product.  相似文献   

17.
A culture technique employing cold enrichment at 4 degrees C followed by selective enrichment and plating at higher temperatures (30 degrees C) was used to isolate Listeria monocytogenes from faecal samples. The samples were held at 4 degrees C for 15 weeks and cultured weekly to assess the sensitivity of the culture after cold storage for different lengths of time. No media, Listeria selective enrichment broth (LSEB), nutrient broth (NB) and saline were used as cold storage medium. Cold storage increased the frequency of Listeria positive samples. The sensitivity of the culture for Listeria spp. and L. monocytogenes was 72 and 94%, and 56 and 61% after third and seventh week of cold storage, respectively. When the results of third and seventh week of cold storage were combined, the sensitivity was 100% for Listeria spp. and 94% for L. monocytogenes. LSEB and NB as storage medium increased Listeria positive samples after the first week of cold storage but did not maintain the increase thereafter while saline had an adverse effect on the growth of the bacteria. However, samples held in no media in a pilot study involving monthly sampling of a herd revealed better results. Detection limit of the culture media was also investigated. The lowest concentration detected by culture media was 3.17 organisms/ml. This was seven organisms/g for known Listeria positive sample. The faecal samples spiked with 10-fold dilutions of L. monocytogenes and held at 4 degrees C revealed that the sample spiked with 3.17 x 10-1 cfu/ml organisms resulted in growth after the second week of cold storage. The results suggest that the culture technique employing cold enrichment followed by selective enrichment and plating is more sensitive, the storage of faecal samples in no media when compared with the samples in storage medium, LSEB, NB and saline, during cold enrichment is a better application and culture of faeces, immediately after collection, at third and seventh week of cold enrichment produce more satisfactory results.  相似文献   

18.
The isolation of pathogenic Listeria spp. in bacteriological samples, and anti-phosphatidylinositol-specific phospholipase C (anti-PIPLC) antibodies in sera of buffaloes were studied. Isolation of the pathogen was attempted from the samples by selective enrichment in University of Vermont Medium and plating onto Dominguez-Rodriguez isolation agar. Pathogenicity of the isolates was tested by Christie, Atkins, Munch Petersen test and mice incoulation test. Listeria spp. and L. monocytogenes were isolated from 8.8 and 2.4%, and 4.8 and 1.6% of 125 each meat and blood samples, respectively. Out of the 125 samples each of feacal, nasal and vaginal swabs from buffaloes 8 and 4%, 13.6 and 2.4%, and 6.4 and 2.4% were positive for Listeria spp. and L. monocytogenes, respectively. L. ivanovii was confirmed from 0.8% vaginal sample. A total of 125 serum samples were tested by phosphatidylinositol-specific phospholipase C (PIPLC) based indirect ELISA of which 4.0% turned out to be seropositive.  相似文献   

19.
Two juvenile scimitar-horned oryx (Oryx dammah) at the Wild Animal Park Planckendael died from acute septicemia caused by Listeria monocytogenes serovar 4b. Subsequently, Listeria spp. were isolated from the feces, food, and environment of seven antelope species and examined using a two-stage enrichment procedure in Fraser Broth, followed by isolation on PALCAM agar. A total of 40/170 samples (23.5%) was positive for Listeria spp. No organisms were cultured in 83/170 samples (48.8%), and 47 samples (27.6%) were overgrown with Bacillus spp. Nonpathogenic Listeria spp. were isolated from 16/70 fecal samples, 22/40 soil samples, and 2/60 feed samples. Listeria monocytogenes serovar 1/2b was isolated from two soil samples collected in the enclosure of the scimitar-horned oryx.  相似文献   

20.
The objective of this study was to identify potential risk factors for Listeria monocytogenes contamination in French poultry production. Eighty-four flocks of layer hens kept in cages and 142 broiler flocks were included in this study. For each production type, a questionnaire was submitted to farmers and fecal samples were taken to assess the L. monocytogenes status of the flocks during a single visit to the farm. Two logistic regression models (specific to each production) were used to assess the association between management practices and the risk of L. monocytogenes contamination of the flock. The prevalence of L. monocytogenes-positive flocks was 30.9% (95% CI: 21.0; 40.9) and 31.7% (95% CI: 24.0; 39.4) for cage-layers and broiler flocks, respectively. For layer flocks, the risk of L. monocytogenes contamination was increased when pets were present on the production site. When droppings were evacuated by conveyor belt with deep pit storage, the risk of L. monocytogenes contamination decreased significantly. Feed meal was found to be associated with a higher risk of L. monocytogenes contamination than feed crumb. For broiler flocks, the risk of L. monocytogenes contamination was increased when farmers did not respect the principle of two areas (clean and dirty) at the poultry house entrance. A first disinfection by thermal fogging and the absence of pest control of the poultry house before the arrival of the next flock was found to increase the risk of contamination. When litter was not protected during storage and when farm staff also took care of other broiler chicken houses, the risk of L. monocytogenes contamination increased significantly. In the case of the watering system, nipples with cups were found to decrease the risk of contamination.  相似文献   

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