Petite colony formation by Listeria monocytogenes and Listeria species grown on esculin-containing agar

1990 ◽  
Vol 36 (10) ◽  
pp. 697-703 ◽  
Author(s):  
Gregory R. Siragusa ◽  
L. A. Elphingstone ◽  
P. L. Wiese ◽  
S. M. Haefner ◽  
M. G. Johnson

Several strains of Listeria species formed petite-sized colonies from parent stock cultures when grown on agar media containing 0.2 – 1% (w/v) esculin. This was observed in Listeria monocytogenes (7/22 strains), L. innocua (1/3), L. grayi (1/1), L. seeligeri (1/3), and L. welshimeri (1/1), but not in L. ivanovii (0/1) and L. murrayi (0/1). This phenomenon was only observed on agar media that contained esculin. All petite isolates had biotyping profiles identical to their larger, normal-sized counterpart isolates. Normal and petite-sized isolates from two L. monocytogenes strains, Scott A and V7, were pathogenic to immunosuppressed white mice. On media containing 0.5% (w/v) esculin + ferric iron, Listeria cultures produced colony diameters intermediate in size between those of normal and petite cultures. When pregrown in glucose broth, all petite isolates demonstrated visible β-glucosidase (esculinase) activity within 5 min, while the normal-sized isolates showed β-glucosidase activity only after at least 20–70 min. This evidence suggests that cells forming petite colonies are β-glucosidase constitutive variants within the parent population, while cells that form normal-sized colonies are inducible for β-glucosidase (esculinase) activity. A possible role for the esculin hydrolysis product, esculetin, in causing petite colony formation is discussed. Key words: Listeria species, petite colonies, esculinase, pathogenicity.


1992 ◽  
Vol 38 (2) ◽  
pp. 161-164 ◽  
Author(s):  
Pearl I. Peterkin ◽  
Mary-Ann Gardiner ◽  
Naeem Malik ◽  
Edmund S. Idziak

One hundred and twenty-two food, clinical, and veterinary strains of Listeria monocytogenes were examined for the presence of plasmids. Twenty-five (20%) contained plasmids, which varied from 1.3 to 66 MDa in size. Of 10 strainsof other Listeria species (L. innocua, L. ivanovii, L. welshimeri, L. seeligeri, L. grayi, and L. murrayi) examined, seven (70%) contained plasmids, varying from 38 to 53 MDa. No strains with multiple plasmids were found. Plasmids of identical size were isolated from related strains in some, although not all, cases. The presence of a plasmid in a strain was not related to phenotypic characters of known extrachromosomal inheritance. Key words: plasmid, Listeria spp., Listeria monocytogenes.



2007 ◽  
Vol 22 (8) ◽  
pp. 1664-1671 ◽  
Author(s):  
Padmapriya P. Banada ◽  
Songling Guo ◽  
Bulent Bayraktar ◽  
Euiwon Bae ◽  
Bartek Rajwa ◽  
...  


1987 ◽  
Vol 50 (3) ◽  
pp. 188-192 ◽  
Author(s):  
J. LOVETT ◽  
D. W. FRANCIS ◽  
J. M. HUNT

To determine the incidence of Listeria monocytogenes in raw milk, an isolation method was evaluated and used to analyze milk from three areas of the United States. The incidence varied by area from 0% in California to 7% in Massachusetts, with an overall incidence of 4.2%. The highest incidence found in any area during a single sampling period was 12% in Massachusetts in March 1985. During that same sampling, the incidence for all Listeria species was 26%. Of the 27 L. monocytogenes strains isolated during the survey, 25 were pathogenic in adult mice. One of three Listeria ivanovii isolated was pathogenic. No other isolates demonstrated pathogenicity.



Author(s):  
Rachel K Streufert ◽  
Susanne E Keller ◽  
Joelle K Salazar

Growth on solid media as sessile cells is believed to increase the desiccation tolerance of Salmonella enterica . However, the reasons behind increased resistance have not been well explored. In addition, the same effect has not been examined for other foodborne pathogens such as pathogenic Escherichia coli or Listeria monocytogenes . The purpose of this research was two-fold: first, to determine the role of oxygenation during growth on the desiccation resistance of S. enterica , E. coli , and L. monocytogenes , and second, to determine the effect of sessile versus planktonic growth on the desiccation resistance of these pathogens. Three different serotypes each of Salmonella , E. coli , and L. monocytogenes were cultured in trypticase soy broth with 0.6% yeast extract (TSBYE), with (aerobic) shaking or on TSBYE with agar (TSAYE) under either aerobic or anaerobic conditions and harvested in stationary phase. After adding cell suspensions to cellulose filter disks, pathogen survival was determined by enumeration at 0 and after drying for 24 h. Results showed statistical differences in harvested initial populations prior to drying (0 h). For Salmonella , a correlation was found between high initial population and greater survival on desiccation (p = 0.05). In addition, statistical differences (p ≤ 0.05) between survival based on growth type were identified. However, differences found were not the same for the three pathogens, or between their serotypes. In general, Salmonella and E. coli desiccation resistance followed the pattern of aerobic agar media ≥ liquid media ≥ anaerobic agar media. For L. monocytogenes serotypes, resistance to desiccation was not statistically different based on mode of growth. These results indicate growth on solid media under aerobic conditions is not always necessary for optimal desiccation survival but may be beneficial when the desiccation resistance of the test serotype is unknown.



1993 ◽  
Vol 56 (3) ◽  
pp. 256-259 ◽  
Author(s):  
A. EL MARRAKCHI ◽  
A. HAMAMA ◽  
F. EL OTHMANI

Examination of 227 samples of milk and dairy products for Listeria monocytogenes showed that raw milk and some Moroccan traditionally made dairy products such as Iben and raib (fermented milks) and jben (fresh cheese) were contaminated with this pathogen. L. monocytogenes was the only Listeria species isolated except in one case in which it was associated with Listeria innocua. Pasteurized milk, fresh cream, and fresh and ripened cheeses (industrially made) were free from L. monocytogenes.



1997 ◽  
Vol 80 (4) ◽  
pp. 775-790 ◽  
Author(s):  
Philip T Feldsine ◽  
Andrew H Lienau ◽  
Robin L Forgey ◽  
Roger D Calhoon ◽  
S Al-Hasani ◽  
...  

Abstract Six foods representing a variety of food products were analyzed by the Assurance Listeria polyclonal enzyme immunoassay (EIA) and by either the Bacteriological Analytical Manual or the U.S. Department of Agriculture culture method for detecting Listeria monocytogenes and related Listeria species. Samples of each food type, at each inoculation level, were analyzed simultaneously by both methods. A total of 19 laboratories representing federal government agencies and private industry in the United States and Canada participated. Food types were inoculated with Listeria species including L. monocytogenes, with the exception of 3 lots of green beans, which were naturally contaminated. During this study, 1764 samples and controls were analyzed and confirmed, of which 492 were positive and 947 were negative by both methods. There were 159 samples that were positive by culture method but negative by the EIA and 188 that were negative by culture method but positive by EIA. Twenty-two samples were negative by EIA and by culture method but confirmed positive when Assurance selective enrichment broths were subcultured to selective agar. The Assurance polyclonal EIA for detecting L. monocytogenes and related Listeria species in foods has been adopted first action by AOAC INTERNATIONAL.



1990 ◽  
Vol 53 (8) ◽  
pp. 642-647 ◽  
Author(s):  
CURTT M. PERRY ◽  
CATHERINE W. DONNELLY

Silage samples representing approximately 10% of Vermont's dairy farms were tested for the presence of Listeria species. Listeria innocua was isolated from 15.3% of the silage samples, while Listeria monocytogenes was isolated from 2.9% of the examined samples. As silage pH increased, the incidence of Listeria increased concomitantly. Seventy-eight mesophilic lactic acid bacteria, indigenous to silage, were screened for specific and nonspecific antagonism against four L. monocytogenes indicator strains. Most of the silage isolates demonstrated nonspecific inhibition via lactic acid production against the L. monocytogenes indicator strains. None of the indigenous silage isolates tested in this survey demonstrated specific antagonism via production of bacteriocinogenic compounds.



1988 ◽  
Vol 51 (11) ◽  
pp. 840-841 ◽  
Author(s):  
MICHAEL B. LIEWEN ◽  
MARK W. PLAUTZ

Raw milk samples were obtained from bulk storage tanks of individual dairy farms in eastern Nebraska during February and July of 1986. One hundred different farms were tested during each period. One-tenth ml of each sample was plated directly onto McBride's Listeria Agar (MLA) and 30 ml was subjected to a four-week cold enrichment procedure. Suspect colonies from MLA were subjected to biochemical tests to confirm identity. Nine percent of all raw milk samples examined were determined to be positive for Listeria species after the cold enrichment procedure. Four percent contained L. monocytogenes and five percent contained L. innocua. Six percent and two percent of samples were found to contain L. monocytogenes in February and July respectively.



2002 ◽  
Vol 65 (5) ◽  
pp. 780-785 ◽  
Author(s):  
IRENE V. WESLEY ◽  
KAREN M. HARMON ◽  
JAMES S. DICKSON ◽  
ANN RAMOS SCHWARTZ

A multiplex polymerase chain reaction was developed to simultaneously identify Listeria monocytogenes and species of the genus Listeria. Two sets of primers were used, with the first amplifying a 938-bp region of the 16S rRNA gene that is highly conserved in all Listeria species and the second amplifying a 174-bp region of the listeriolysin (hlyA) gene of L. monocytogenes. Thus, isolates of Listeria spp. yield a single 938-bp product, whereas L. monocytogenes isolates yield both the 938-bp product and a 174-bp product. The specificity of the assay was verified with all six Listeria species and 11 serotypes of L. monocytogenes, as well as nonrelated bacteria. The multiplex PCR assay was used to determine the incidence of Listeria spp., especially L. monocytogenes, in mechanically separated turkey samples (n = 150 samples). L. monocytogenes strains were selected by using the University of Vermont two-step enrichment protocol and plating to selective Palcam agar. The multiplex PCR assay was used for verification of presumptive Listeria colonies. Approximately 38% of mechanically separated turkey samples (57 of 150) yielded L. monocytogenes; an additional 18% of these samples (27 of 150) harbored other Listeria spp. Fifty-one percent (29 of 57) of the L. monocytogenes isolates were of serogroup 1, 44% (25 of 57) were of serogroup 4, and 2% (1 of 57) were assigned to serogroups other than 1 and 4.



2008 ◽  
Vol 71 (4) ◽  
pp. 781-789 ◽  
Author(s):  
WON-BO SHIM ◽  
JIN-GIL CHOI ◽  
JI-YOUNG KIM ◽  
ZHENG-YOU YANG ◽  
KYU-HO LEE ◽  
...  

An enzyme-linked immunosorbent assay (ELISA), immunochromatography (ICG) strip test, and immunomagnetic bead separation (IMBS) system based on a monoclonal antibody were individually developed for the detection and isolation of Listeria monocytogenes in meat samples. The three methods showed a strong reaction with Listeria species and a weak reaction with Staphylococcus aureus. To increase the rapidity of L. monocytogenes detection, combinations of the ELISA and ICG strip test with the IMBS system (ELISA-IMBS and ICG-IMBS) were investigated. In comparative analyses of artificially inoculated meat and samples of processed meat, the ELISA and ICG strip test required 24 h of enrichment time to detect the inoculated meat samples with ≥ 1 × 102 CFU/10 g, whereas the ELISA-IMBS and ICG-IMBS required only 14 h of enrichment. Analyses of naturally contaminated meat samples (30 pork samples, 20 beef samples, 26 chicken samples, 20 fish samples, and 20 processed meat samples) performed by ELISA-IMBS, ICG-IMBS, and API kit produced similar results. The ELISA-IMBS and ICG-IMBS provide a more rapid assay than the individual ELISA and the ICG strip test and are appropriate for rapid and qualitative detection of L. monocytogenes (or Listeria species) in meat samples. With the ICG-IMBS, L. monocytogenes could be detected in meat samples within 15 h and the method has potential as a rapid, cost-effective on-site screening tool for the detection of L. monocytogenes in food samples and agricultural products at a minimum detection level of ~100 CFU/10 g.



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