scholarly journals Molecular characterization of Listeria monocytogenes isolated from animal products in a city of Northern Brazil

2013 ◽  
Vol 43 (8) ◽  
pp. 1443-1448 ◽  
Author(s):  
Lilyan Rosmery Luizaga de Monteiro ◽  
Albenones José de Mesquita ◽  
Maria Cláudia Dantas Porfirio Borges André ◽  
Juliana Lamaro Cardoso

Listeria monocytogenes, a foodborne pathogen causes listeriosis, a fatal disease in about 30% of cases that affects mainly immunocompromised persons. The aim of this research was to characterize L. monocytogenes pulsed-field gel electrophoresis (PFGE) types isolated from meat products collected at public markets in Araguaina city, TO. Sixty samples of raw ground beef and frescal sausage were analyzed during the second half of 2008. Five out of 30 samples (16.7%) of raw ground beef tested positive for L. monocytogenes, three of which were classified as serotype 1/2b and two as serotype 4b. Among the 30 samples of sausage collected, two strains of L. monocytogenes were isolated (6.7%), one of them belonging to serotype 1/2a and the other belonging to serotype 1/2b. The restriction enzymes used were ApaI and SmaI. Similarities among the strains were determined by Dice coefficient. The macro restriction profile obtained by using SmaI enzyme allowed the distribution of seven strains in two clusters, two pulsotypes and two subtypes. The result indicates that L. monocytogenes isolates, belonging to serotype 4b, 1/2a and 1/2b, are strongly correlated within the same serotype group, and in some cases among different serotypes, suggesting that they have a common source.

2017 ◽  
Vol 5 (49) ◽  
Author(s):  
Taylor W. Bailey ◽  
Naila C. do Nascimento ◽  
Arun K. Bhunia

ABSTRACT Listeria monocytogenes is an opportunistic invasive foodborne pathogen. Here, we performed whole-genome sequencing of L. monocytogenes strain F4244 (serotype 4b) using Illumina sequencing. The sequence showed 94.5% identity with strain F2365, serotype 4b, and 90.6% with EGD-e, serotype 1/2a.


2016 ◽  
Vol 4 (3) ◽  
Author(s):  
Aidan Casey ◽  
Olivia McAuliffe ◽  
Edward M. Fox ◽  
Dara Leong ◽  
Cormac G. M. Gahan ◽  
...  

Listeria monocytogenes is a foodborne pathogen and the causative agent of listeriosis among humans and animals. The draft genome sequences of L. monocytogenes serotype 4b strains 944 and 2993 and serotype 1/2c strains 198 and 2932 are reported here.


1995 ◽  
Vol 58 (4) ◽  
pp. 398-403 ◽  
Author(s):  
WERNER B. BARBOSA ◽  
JOHN N. SOFOS ◽  
GLENN R. SCHMIDT ◽  
GARY C. SMITH

Separate inocula of four strains of Listeria monocytogenes were prepared at 4°C and inoculated (3.58 to 4.67 log10 colony forming units [CFU]/g) in top round ground beef (< 4.0% fat) patties (78.8 ± 6.7 g) of normal (5.47 ± 0.03) or high (6.14 ± 0.08) pH, which were stored (4°C) vacuum-packaged for 56 days and analyzed for L. monocytogenes, total aerobic plate counts (APC) and pH. In normal-pH ground beef, strain N-7143 (serotype 3a), multiplied from 4.25 ± 0.71 log10 CFU/g at day of inoculation to 6.53 ± 0.34 log10 CFU/g at 35 days of storage (P < 0.05); a 2.3 log10 CFU/g increase. Populations of strain Na-19 (serotype 3b) increased 1.8 log10 CFU/g in 35 days of storage, while numbers of strain Na-16 (serotype 1/2a) did not change (P > 0.05) during the 56 days of storage. Strain Scott A (serotype 4b) decreased in numbers from 4.00 ± 1.21 log10 CFU/g at day-0 to 2.72 ± 0.98 log10 CFU/g at 56 days. Populations of strain Scott A were lower (P < 0.05) than other strains after 21 days of storage. In high-pH ground beef, populations of strains Na-19, N-7143 and Na-16 increased (P < 0.05) by 2.87, 2.64 and 2.24 log10 CFU/g, respectively, in 28 days. Populations of strain Scott A did not change significantly (P > 0.05), and they were lower (P< 0.05) than populations of other strains at 28 days. The results indicated that although growth of L. monocytogenes in vacuum-packaged, refrigerated ground beef was slow, it proceeded more rapidly in product of pH values above 6.0, and depended on strain of the pathogen tested.


Author(s):  
Lisa Gorski ◽  
Samarpita Walker ◽  
Kelly F Romanolo ◽  
Sophia Kathariou

The foodborne pathogen Listeria monocytogenes lives as a saprophyte in nature and can adhere to and grows on surfaces as diverse as leaves, sediment, and stainless steel. To discern the mechanisms used by L. monocytogenes for attachment and growth on various surfaces, we studied interactions between the pathogen on lettuce and stainless steel. A panel of 24 strains (23 of Listeria monocytogenes and 1 L. innocua ) was screened for attachment and growth on lettuce at 4 o C and 25 o C and on stainless steel at 10 o C and 37 o C. Overnight growth of attached cells resulted in a 0 – 3 log increase on lettuce, depending on the strain and the temperature. Among the worst performing strains on lettuce were two from a large cantaloupe outbreak, indicating that factors important for interactions with cantaloupe may be different from those required on lettuce tissue. Strains that grew the best on lettuce belonged to serotypes 1/2a, 1/2b, and 4b and were from cheese, potatoes, and water/sediment near produce fields. Confocal microscopy of L. monocytogenes tagged with constitutively expressed green fluorescent protein indicated associations with the cut edges and veins of lettuce leaves. On stainless steel coupons, there was a 5 – 7 log increase at 10 o C after 7 d and a 4 – 7 log increase at 37 o C after 40 h. Statistically, surface growth on stainless steel was better for serotype 1/2a than for serotype 4b strains, even though certain serotype 4b strains grew well on the coupons. The latter included strains that originated from produce and water/sediment. Some strains were fit in both environments, while others showed variability between the two different surfaces. Further analysis of these strains should reveal molecular factors needed for adherence and surface growth of L. monocytogenes on different biotic and abiotic surfaces.


2015 ◽  
Vol 9 (09) ◽  
pp. 962-969 ◽  
Author(s):  
André Victor Barbosa ◽  
Aloysio de Mello Figueiredo Cerqueira ◽  
Leonardo Alves Rusak ◽  
Cristhiane Moura Falavina Dos Reis ◽  
Nilma Cintra Leal ◽  
...  

Introduction: Listeria monocytogenes is an important foodborne pathogen and the 4b serotype is responsible for many cases of human listeriosis reported in Brazil. Several listeriosis outbreaks worldwide have involved a small number of well-defined clonal groups, designated as epidemic clones (ECs). Methodology: We studied 71 strains of serotype 4b, including 25 isolates from human cases of listeriosis and 46 from meat-based foods, collected in Brazil between 1977 and 2010. The presence of ECs (I and II) markers and virulence genes (inlA, inlB, ilnC, inlJ and actA) were evaluated by PCR assay. The genetic relationship of ECs-positive strains was assessed by pulsed field gel electrophoresis. Results: ECI and ECII markers were found both in human and food strains, with 19.7% positive for the ECI marker and 40.8% for ECII. Most strains (97.2%) were positive for the virulence genes that were studied. Nevertheless, the actA gene amplicons showed two distinct sizes, with all ECI positive strains exhibiting a 105bp deletion. Pulsed field gel electrophoresis (PFGE) analysis allowed the recognition of highly related strains, particularly from two outbreaks of neonatal listeriosis in São Paulo State occurred in 1992 and 1997, both ECII-positive; and two ECI strains from a human case (1982) and from bovine meat (2009). Conclusions: The presence of ECs among clinical samples and beef isolates of serotype 4b from some regions of Brazil highlights the need for rigorous control of production procedures. Furthermore, the association of ECII with two nosocomial outbreaks suggests its ability to spread in these settings.


2014 ◽  
Vol 77 (2) ◽  
pp. 254-261 ◽  
Author(s):  
MARIOS MATARAGAS ◽  
ANNA GREPPI ◽  
KALLIOPI RANTSIOU ◽  
LUCA COCOLIN

A Listeria monocytogenes subgenomic array, targeting 54 genes involved in the adhesion, adaptation, intracellular life cycle, invasion, and regulation of the infection cycle was used to investigate the gene expression patterns of acid- and salt-stressed Listeria cells after exposure to conditions similar to those in gastric and pancreatic fluids. Three L. monocytogenes strains, one laboratory reference strain (EGDe) and two food isolates (wild strain 12 isolated from milk and wild strain 3 isolated from fermented sausage), were used during the studies. Differences in the expressed genes were observed between the gastric and pancreatic treatments and also between the serotypes. Increased transcripts were observed of the genes belonging to the adaptation and regulation group for serotype 4b (strain 12) and to the invasion and regulation group for serotype 1/2a (strain EGDe). Interestingly, no significantly differentially expressed genes were found for serotype 3c (strain 3) in most cases. The genes related to adaptation (serotype 1/2a) and to intracellular life cycle and invasion (serotype 4b) were down-regulated in order to cope with the hostile environment of the gastric and pancreatic fluids. These findings may provide experimental evidence for the dominance of serotypes 1/2a and 4b in clinical cases of listeriosis and for the sporadic occurrence of serotype 3c.


2002 ◽  
Vol 184 (15) ◽  
pp. 4177-4186 ◽  
Author(s):  
Peter Lauer ◽  
Man Yin Nora Chow ◽  
Martin J. Loessner ◽  
Daniel A. Portnoy ◽  
Richard Calendar

ABSTRACT Two site-specific shuttle integration vectors were developed with two different chromosomal bacteriophage integration sites to facilitate strain construction in Listeria monocytogenes. The first vector, pPL1, utilizes the listeriophage U153 integrase and attachment site within the comK gene for chromosomal insertion. pPL1 contains a useful polylinker, can be directly conjugated from Escherichia coli into L. monocytogenes, forms stable, single-copy integrants at a frequency of ∼10−4 per donor cell, and can be used in the L. monocytogenes 1/2 and 4b serogroups. Methods for curing endogenous prophages from the comK attachment site in 10403S-derived strains were developed. pPL1 was used to introduce the hly and actA genes at comK-attBB′ in deletion strains derived from 10403S and SLCC-5764. These strains were tested for second-site complementation in hemolysin assays, plaquing assays, and cell extract motility assays. Unlike plasmid-complemented strains, integrated pPL1-complemented strains were fully virulent in the mouse 50% lethal dose assay. Additionally, the PSA phage attachment site on the L. monocytogenes chromosome was characterized, and pPL1 was modified to integrate at this site. The listeriophage PSA integrates in the 3′ end of an arginine tRNA gene. There are 17 bp of DNA identity between the bacterial and phage attachment sites. The PSA prophage DNA sequence reconstitutes a complete tRNAArg gene. The modified vector, pPL2, was integration proficient at the same frequency as pPL1 in common laboratory serotype 1/2 strains as well as serotype 4b strains.


2010 ◽  
Vol 76 (16) ◽  
pp. 5577-5584 ◽  
Author(s):  
Suleyman Yildirim ◽  
Driss Elhanafi ◽  
Wen Lin ◽  
Anthony D. Hitchins ◽  
Robin M. Siletzky ◽  
...  

ABSTRACTListeria monocytogenesis a food-borne pathogen with a clonal population structure and apparently limited gene flow between strains of different lineages. Strains of epidemic clone I (ECI) have been responsible for numerous outbreaks and invariably have DNA that is resistant to digestion by Sau3AI, suggesting methylation of cytosine at GATC sites. A putative restriction-modification (RM) gene cassette has been identified in the genome of the ECI strain F2365 and all other tested ECI strains but is absent from other strains of the same serotype (4b). Homologous RM cassettes have not been reported amongL. monocytogenesisolates of other serotypes. Furthermore, conclusive evidence for the involvement of this RM cassette in the Sau3AI resistance phenotype of ECI strains has been lacking. In this study, we describe a highly conserved RM cassette in certain strains of serotypes 1/2a and 4a that have Sau3AI-resistant DNA. In these strains the RM cassette was in the same genomic location as in the ECI reference strain F2365. The cassette included a gene encoding a putative recombinase, suggesting insertion via site-specific recombination. Deletion of the RM cassette in the ECI strain F2365 and the serotype 1/2a strain A7 rendered the DNA of both strains susceptible to Sau3AI digestion, providing conclusive evidence that the cassette includes a gene required for methylation of cytosine at GATC sites in both strains. The findings suggest that, in addition to its presence in ECI strains, this RM cassette and the accompanying genomic DNA methylation is also encountered among selected strains of other lineages.


2008 ◽  
Vol 75 (2) ◽  
pp. 366-373 ◽  
Author(s):  
Janet R. Donaldson ◽  
Bindu Nanduri ◽  
Shane C. Burgess ◽  
Mark L. Lawrence

ABSTRACT Listeria monocytogenes is a gram-positive, food-borne pathogen that causes disease in both humans and animals. There are three major genetic lineages of L. monocytogenes and 13 serovars. To further our understanding of the differences that exist between different genetic lineages/serovars of L. monocytogenes, we analyzed the global protein expression of the serotype 1/2a strain EGD and the serotype 4b strain F2365 during early-stationary-phase growth at 37°C. Using multidimensional protein identification technology with electrospray ionization tandem mass spectrometry, we identified 1,754 proteins from EGD and 1,427 proteins from F2365, of which 1,077 were common to both. Analysis of proteins that had significantly altered expression between strains revealed potential biological differences between these two L. monocytogenes strains. In particular, the strains differed in expression of proteins involved in cell wall physiology and flagellar biosynthesis, as well as DNA repair proteins and stress response proteins.


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