Epidemiological studies on the occurrence of Listeria monocytogenes in the feces of dairy cattle

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.     

Isolation of pathogenic Listeria monocytogenes and detection of antibodies against phosphatidylinositol-specific phospholipase C in buffaloes

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.     

Prevalence of Listeria monocytogenes in intestinal contents of healthy animals in Japan.

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.     

A survey for Listeria species, motile aeromonads and Yersinia enterocolitica on bovine and ovine carcasses

One hundred ovine and 100 bovine carcasses in two abattoirs were sampled just after dressing for the presence of Yersinia enterocolitica, Listeria monocytogenes and motile aeromonads. Yersinia enterocolitica was not isolated and only two samples were positive for Listeria spp. In both cases, the Listeria species were not normally pathogenic to man. In contrast, motile aeromonads were isolated from 81% of ovine and 35% of bovine carcasses.     

Simultaneous occurrence of selected food-borne bacterial pathogens on bovine hides, carcasses and beef meat

The aim of this study was to determine the simultaneous occurence of Salmonella spp., L. monocytogenes, verotoxigenic E. coli (VTEC), and Campylobacter spp. in slaughtered cattle and in beef meat subjected for human consumption. A total of 406 bovine hides and 406 corresponding carcasses were used to collect the samples with a swab method after exsanguination and evisceration of animals, respectively. Furthermore, 362 beef meat samples were purchased in local retail shops over the same period of time as for the bovine samples. Food-borne bacterial pathogens were identified with standard ISO methods with some modification by the use of PCR for VTEC. The isolated bacteria were then molecularly speciated (Campylobacter), serotyped (L. monocytogenes) and characterized for the presence of several virulence marker genes (VTEC and Campylobacter). It was found that 49 hide (12.1%) and 3 (0.7%) carcass samples were contaminated with more than one bacterial pathogen tested. Most of the hides were positive for Campylobacter spp. and VTEC (27 samples) and Campylobacter spp. together with L. monocytogenes (12 samples). Eight bovine hides contained L. monocytogenes and VTEC while L. monocytogenes and Salmonella spp. were detected in one sample. Furthermore, 3 pathogens (Campylobacter spp., L. monocytogenes and VTEC) were simultaneously identified in one bovine hide tested. In case of bovine carcasses 2 samples contained Campylobacter spp. and VTEC whereas one carcass was positive for L. monocytogenes and VTEC. On the other hand, 10 out of 362 (2.8%) minced beef samples were contaminated with at least two pathogens tested. The majority of these samples were contaminated with L. monocytogenes and Salmonella spp. (6 samples). It was noticed that equal number of C. jejuni and C. coli were found, irrespective of the origin of the samples. Most of the strains possessed more than one pathogenic factor as identified by PCR. Molecular serotyping of L. monocytogenes revealed that the majority of the isolates (27 out of 31; 87.1%) belonged to 1/2a serogroup. It was found that most of the VTEC isolates possessed the Shiga toxin stx2 gene (12 strains) whereas only 2 strains were str1-positive. The eneterohemolysin and intimin markers were identified only in 7 and 2 isolates, respectively. PCR analysis revealed that 4 VTEC belonged to O91 serogroup, 2 strains were O145 and 1 isolate was identified as O113. None of the VTEC detected in the study was O157 serogroup.     

Listeria spp. contamination in piggeries: comparison of three sites of environmental swabbing for detection and risk factor hypothesis

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.     

Campylobacter spp. in Irish feedlot cattle: a longitudinal study involving pre-harvest and harvest phases of the food chain

The aim of this study was to investigate faecal shedding and transmission of Campylobacter spp. in cohorts of cattle within a feedlot, to assess subsequent contamination of carcasses with this pathogen and to identify risk factors associated with faecal shedding of Campylobacter spp. A cohort of 133 heifers housed in four adjacent pens was examined over a five and a half month period, from entering the feedlot to slaughter. A parallel investigation of individual rectal faecal samples and pen environmental samples were taken at monthly intervals from November to February. The entire outer and inner surfaces of a carcass side of each animal were swabbed immediately following slaughter. Campylobacter spp. were isolated from 322 (54%) of the 600 rectal faecal samples. Campylobacter jejuni and C. coli accounted for 69 and 29.7% of the isolate recovered, respectively. A total of 159 environmental samples were examined, of these Campylobacter spp. was isolated from 46 samples (29%). Campylobacter jejuni and C. coli accounted for 35 and 59% of these isolates, respectively. Campylobacter spp. was not isolated from any of the dressed carcasses. Logistic regression indicated prevalence of Campylobacter spp. faecal shedding within pens was positively correlated to the pen, the month of sampling and the Campylobacter spp. contamination status of the pen dividing bars and the water trough surface. Campylobacter spp. should be considered as a pathogen shed in the faeces of a substantial proportion of feedlot cattle. However, with good hygienic practice during harvest, a very low level of this pathogen can be achieved on dressed carcasses.     

The occurrence of Listeria in the production of processed meat, salami and mettwurst

In six Swiss meat-processing plants 206 samples of cured and air-dried beef (Bündnerfleisch), salami and Mettwurst were analyzed for the presence of Listeria spp. Samples were taken during the fabrication, fermentation and drying of the products. Out of 44.7% of all samples Listeria spp. could be detected. 6.8% turned out to be L. monocytogenes, 37.4% L. innocua and 0.5% L. seeligeri. Listeria spp. were found in all production stages of the tested foods. The concentration of L. monocytogenes was always less than or equal to 20 MPN/g. 86% of the isolated strains formed part of the serogroup 1/2 and 14% of the serogroup 4. Listeria spp. could only be found on the surface of Bündnerfleisch. Both, L. monocytogenes and L. innocua were able to survive the maturation process of salami, even when the initial concentration was very low. The ripening was more often survived by L. innocua than by L. monocytogenes. It appeared that Mettwurst had the highest contamination rate of Listeria spp. (94.4%), followed by salami (46.7%) and Bündnerfleisch (23.1%). The corresponding proportions for L. monocytogenes were 8.0% (salami), 5.8% (Bündnerfleisch) and 0% (Mettwurst). Listeria spp. positive samples were found in every examined plant, L. monocytogenes in five of therm. The Listeria spp. contamination rates moved from 10.0% to 86.2%, those of L. monocytogenes from 0% to 12.1%.     

Detection of pathogenic Listeria spp. in zoo animal faeces: use of immunomagnetic separation and a chromogenic isolation medium

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.     

Campylobacter spp. in Irish Feedlot Cattle: A Longitudinal Study Involving Pre‐harvest and Harvest Phases of the Food Chain

The aim of this study was to investigate faecal shedding and transmission of Campylobacter spp. in cohorts of cattle within a feedlot, to assess subsequent contamination of carcasses with this pathogen and to identify risk factors associated with faecal shedding of Campylobacter spp. A cohort of 133 heifers housed in four adjacent pens was examined over a five and a half month period, from entering the feedlot to slaughter. A parallel investigation of individual rectal faecal samples and pen environmental samples were taken at monthly intervals from November to February. The entire outer and inner surfaces of a carcass side of each animal were swabbed immediately following slaughter. Campylobacter spp. were isolated from 322 (54%) of the 600 rectal faecal samples. Campylobacter jejuni and C. coli accounted for 69 and 29.7% of the isolate recovered, respectively. A total of 159 environmental samples were examined, of these Campylobacter spp. was isolated from 46 samples (29%). Campylobacter jejuni and C. coli accounted for 35 and 59% of these isolates, respectively. Campylobacter spp. was not isolated from any of the dressed carcasses. Logistic regression indicated prevalence of Campylobacter spp. faecal shedding within pens was positively correlated to the pen, the month of sampling and the Campylobacter spp. contamination status of the pen dividing bars and the water trough surface. Campylobacter spp. should be considered as a pathogen shed in the faeces of a substantial proportion of feedlot cattle. However, with good hygienic practice during harvest, a very low level of this pathogen can be achieved on dressed carcasses.     

Isolation of Listeria monocytogenes from buffaloes with reproductive disorders and its confirmation by polymerase chain reaction

Listeria monocytogenes, a gram-positive, facultative intracellular pathogen was isolated from buffaloes with a history of reproductive disorders and polymerase chain reaction (PCR) analyses for the presence of virulence-associated genes were conducted. A total of 530 samples of faecal, nasal, vaginal swabs and blood samples from 135 buffaloes were screened. The prevalence of L. monocytogenes and other Listeria spp. was found to be 4.4 and 7.4%, respectively. All isolates were subjected to PCR for virulence-associated genes (prfA, plcA, hlyA, actA and iap) and to pathogenicity testing by the phosphatidylinositol phospholipase C (PI-PLC) assay and mice and chick-embryo inoculation. All L. monocytogenes isolates were hemolytic and positive for the hlyA gene. One L. monocytogenes isolate possessed all five virulence-associated genes and was also positive in the PI-PLC assay as well as in the in vivo pathogenicity tests. The remaining hemolytic L. monocytogenes isolates lacking the plcA gene and PI-PLC assay activity were, however, non-pathogenic via mice and chick-embryo inoculation tests, in spite of having the hlyA gene. The detection of multiple virulence-associated genes, in combination with in vitro pathogenicity tests, must be performed to identify pathogenic L. monocytogenes.     

Prevalence of Listeria species in camel sausages from retail markets in Aydin province in Turkey and RAPD analysis of Listeria monocytogenesisolates

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.     

Occurrence of Listeria spp. in captive antelope herds and their environment.

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.     

Survey on the Listeria contamination of ready-to-eat food products and household environments in Vienna, Austria

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.     

Comparison of direct colony count methods and the MPN-method for quantitative detection of Listeria in model and field conditions]

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.     

The occurrence of pathogenic Listeria monocytogenes and antibodies against listeriolysin-O in buffaloes

The occurrence of Listeria monocytogenes in meat and milk samples, and antilisteriolysin O (ALLO) 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. The pathogenicity of the isolates was tested by Christie, Atkins, Munch Petersen test and mouse inoculation test. Of 167 meat samples 2.4 and 10.17% were positive for L. monocytogenes and Listeria sp., respectively. Of the 64 milk samples 6.25 and 26.13% were positive for L. monocytogenes and Listeria sp., respectively. A total of 284 serum samples were tested by listeriolysin O (LLO)-based indirect enzyme-linked immunosorbent assay of which 25.35% were found to be seropositive. The culture positivity for L. monocytogenes and detection of ALLO did not show any agreement (kappa = 0.035). The prevalence of pathogenic L. monocytogenes in milk and meat and the occurrence of anti-LLO antibodies is of concern from the public health point of view.     

Incidence of Listeria monocytogenes in wild animals in Japan

Between 1991 and 1993, the intestinal contents and feces of wild animals in Japan were examined for the presence of Listeria. The wild animals examined included 623 mammals (11 species) and 996 birds (18 species). Listeria species were isolated from 38 (6.1%) of the 623 mammalian samples and 133 (13.4%) of 996 bird samples. The highest incidence of Listeria in the mammals was found in Japanese monkeys (20.0%) and that in birds was found in crows (43.2%). The incidence of Listeria in Japanese monkeys varied from 0 to 40.0% depending on the capture area. L. monocytogenes was isolated from II of these positive samples. Serovars 1/2a and 4b predominated in eight serotyped L. monocytogenes isolates.     

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.     

Evaluation of indirect and avidin-biotin enzyme linked immunosorbent assays for detection of anti-listeriolysin O antibodies in bovine milk samples

Listeria monocytogenes is a foodborne pathogen that causes a wide spectrum of diseases in humans and animals. Enzyme linked immunosorbent assays (ELISA) [indirect and avidin-biotin (A-B)] for detecting L. monocytogenes antibodies in bovine milk samples (n = 2060) were standardized and evaluated by comparison with bacteriological examination. The tests were standardized by checker board titration. Highly purified listeriolysin O (LLO) was used as an antigen. Receiver operating characteristic (ROC) analysis was performed to decide the cut-off values. The ROC analysis revealed the sensitivities of indirect and A-B ELISA as 100% and specificities as 97.1 and 99.9% respectively. Listeria monocytogenes was isolated from 105 (5.1%) milk samples collected from 52 farms. Anti-LLO IgG antibodies were detected from 137 and 112 milk samples when tested by indirect and A-B ELISA respectively. Of the 52 farms screened, 28 (53.8%) yielded one or more isolates of L. monocytogenes and 33 (63.5%) of the farms had one or more animals simultaneously positive by one or both the assays for anti-LLO antibodies.     

PCR detection of a putative N-acetylmuramidase gene from Listeria ivanovii facilitates its rapid identification

Listeria ivanovii is a Gram-positive bacterial pathogen that is capable of causing abortions and stillbirths in farm animals, particularly sheep and cattle. In terms of morphological, biochemical and molecular characteristics, L. ivanovii resembles other Listeria species such as L. monocytogenes, a pathogen of both man and animals. In this study, through comparative analysis of genomic DNA from the six Listeria species, a L. ivanovii specific clone (liv22-228) containing a 946 bp insert was isolated. This clone contained the 5' ends of two divergently transcribed L. ivanovii genes and an intergenic spacer region, similar in organization to homologous regions from the L. innocua and L. monocytogenes genomes. Regions of low homology in the clone were identified by comparing to the L. innocua and L. monocytogenes genomes, and oligonucleotide primers (liv22-228F and liv22-228R) were designed. These primers amplified a 463 bp band from genomic DNA of L. ivanovii strains only, but not from other Listeria species or common bacteria. Thus, PCR employing L. ivanovii specific primers (liv22-228F and liv22-228R) provides a useful and straightforward method for rapid and precise determination of L. ivanovii.