Listeria monocytogenes infection is most commonly recognized in ruminants, including cattle, sheep, and goats; but it is rarely diagnosed in poultry. This report describes an outbreak of L. monocytogenes in a backyard poultry flock. Also, it points out the importance of collaboration between veterinarians and public health departments and the possible implications of zoonotic diseases.
A 2014 multistate listeriosis outbreak was linked to consumption of caramel-coated apples, an unexpected and previously unreported vehicle for Listeria monocytogenes. This outbreak was unanticipated because both the pH of apples (<4.0) and the water activity of the caramel coating (<0.80) are too low to support Listeria growth. In this study, Granny Smith apples were inoculated with approximately 4 log10 CFU of L. monocytogenes (a cocktail of serotype 4b strains associated with the outbreak) on each apple's skin, stem, and calyx. Half of the apples had sticks inserted into the core, while the remaining apples were left intact. Apples were dipped into hot caramel and stored at either 7°C or 25°C for up to 11 or 28 days, respectively. Data revealed that apples with inserted sticks supported significantly more L. monocytogenes growth than apples without sticks under both storage conditions. Within 3 days at 25°C, L. monocytogenes populations increased >3 log10 in apples with sticks, whereas only a 1-log10 increase was observed even after 1 week for caramel-coated apples without sticks. When stored at 7°C, apples with sticks exhibited an approximately 1.5-log10 increase in L. monocytogenes levels at 28 days, whereas no growth was observed in apples without sticks. We infer that insertion of a stick into the apple accelerates the transfer of juice from the interior of the apple to its surface, creating a microenvironment at the apple-caramel interface where L. monocytogenes can rapidly grow to levels sufficient to cause disease when stored at room temperature.
The food-borne pathogen Listeria (L) monocytogenes is able to survive a variety of stress conditions leading to the colonization of different niches like the food processing environment. This study focuses on the hypervariable genetic hotspot lmo0443-lmo0449 haboring three inserts: the stress survival islet 1 (SSI-1), the single-gene insert LMOf2365_0481 and two homologous genes of the non-pathogenic species L. innocua: lin0464, a putative transcriptional regulator and lin0465, an intracellular PfpI protease. Our prevalence study revealed a different distribution of the inserts between human and food-associated isolates. The lin0464-lin0465 insert was predominantly found in food-associated strains of sequence type (ST) 121. Functional characterization of this insert showed that the putative PfpI protease Lin0465 is involved in alkaline and oxidative stress response, but not in acidic, gastric, heat, cold, osmotic and antibiotic stress. In parallel, deletion of lin0464 decreased the survival under alkaline and oxidative stress. The expression of both genes increased significantly under oxidative stress conditions independently of the alternative sigma factor σ(B) Furthermore, we showed that the expression of the protease lin0465 is regulated by the transcription factor lin0464 under stress conditions, suggesting that lin0464 and lin0465 form a functional unit.In conclusion, we identified a novel stress survival islet 2 (SSI-2), predominantly present in L. monocytogenes ST121 strains, beneficial for survival under alkaline and oxidative stress, potentially supporting adaptation and persistence of L. monocytogenes in food processing environments.IMPORTANCEListeria (L.) monocytogenes strains of ST121 are known to persist for months and even years in food processing environments, thereby increasing the risk of food contamination and listeriosis. However, the molecular mechanism underlying this remarkable niche-specific adaptation is still unknown. Here, we demonstrate that the genomic islet SSI-2, predominantly present in L. monocytogenes ST121 strains, is beneficial for survival under alkaline and oxidative stress conditions, which are routinely encountered in food processing environments. Our findings suggest that SSI-2 is part of a diverse set of molecular determinants contributing to niche-specific adaptation and persistence of L. monocytogenes ST121 strains in food processing environments.
Nisin is a natural preservative for many food products. This bacteriocin is mainly used in dairy and meat products. Nisin inhibits pathogenic food borne bacteria such as Listeria monocytogenes and many other Gram-positive food spoilage microorganisms. Nisin can be used alone or in combination with other preservatives or also with several physical treatments. This article reviews physicochemical and biological properties of nisin, the main factors affecting its antimicrobial effectiveness, and its food applications as an additive directly incorporated into food matrices.
Listeria monocytogenes is a major foodborne pathogen that causes life-threatening illnesses in humans. With emergence of antibiotic resistance in L. monocytogenes, there is considerable interest in testing the efficacy of alternative therapies for controlling listeriosis in humans. This study investigated the efficacy of three phytochemicals, namely trans-cinnamaldehyde (TC), carvacrol (CR), and thymol (TY) in reducing L. monocytogenes virulence in the recently established invertebrate model, Galleria mellonella. In addition, the effect of phytochemicals on the transcription of antimicrobial peptide genes in G. mellonella (responsible for host defense) was investigated using real-time quantitative polymerase chain reaction. G. mellonella larvae were inoculated with L. monocytogenes (10(5) CFU/larvae) either with or without the subinhibitory concentration (chemical concentration not inhibiting bacterial growth) of phytochemicals. The larvae were incubated at 37 °C for 5 days, and their mortality was scored at 24-h intervals. The transcriptional response of the defense genes was studied in inoculated and uninoculated larvae at 6 h post challenge. The experiments were repeated at least six times with replicates. All phytochemicals enhanced the survival rates of G. mellonella infected with lethal doses of L. monocytogenes (P < 0.05). CR and TC at 0.01 % concentration were found to be the most effective treatments, and increased larval survival rates by 80 % and 50 %, respectively, on day 5 (P < 0.05). The phytochemicals also upregulated the expression of antimicrobial peptide genes in G. mellonella larvae challenged with L. monocytogenes (P < 0.05). Results suggest that TC, CR, and TY could potentially be used to control listeriosis. Further investigation in an appropriate mammalian model is warranted.
Listeria monocytogenes possesses the highest number of leucine-rich repeat (LRR) containing proteins among all gram-positive bacteria; these LRR-containing molecules are known as the “internalin” family. To understand the functions of largely uncharacterized LRR-containing molecules, we constructed seven deletion mutants in L. monocytogenes H7858 strain targeting genes in this family and tested their virulence. Among the seven mutants, the ΔLMOh7858_0369 strain and the ΔLMOh7858_2546 strain showed significantly impaired invasiveness to HepG2 cells. We further tested the virulence of these two strains in the intravascular sepsis model using BALB/c mice. Interestingly, the ΔLMOh7858_0369 strain showed significant reduction in organ colonization, bacteremia, and invasion of the brain compared to the parental wild type strain. Host immune responses to listerial intravascular infection were measured at 24 h and 72 h post infection. Transcript levels of several proinflammatory cytokines and chemokines were significantly lower when induced by the ΔlmOh7858_0369 strain than when induced by wild type. These results suggest that the putative LRR-containing protein encoded by LMOh7858_0369 might be a novel virulence factor of the L. monocytogenes H7858 strain.
Controlling the food-borne pathogen Listeria (L.) monocytogenes is of great importance from a food safety perspective, and thus for human health. The consequences of failures in this regard have been exemplified by recent large listeriosis outbreaks in the USA and Europe. It is thus particularly notable that tolerance to quaternary ammonium compounds such as benzalkonium chloride (BC) has been observed in many L. monocytogenes strains. However, the molecular determinants and mechanisms of BC tolerance of L. monocytogenes are still largely unknown. Here we describe Tn6188, a novel transposon in L. monocytogenes conferring tolerance to BC. Tn6188 is related to Tn554 from Staphylococcus (S.) aureus and other Tn554-like transposons such as Tn558, Tn559 and Tn5406 found in various Firmicutes. Tn6188 comprises 5117 bp, is integrated chromosomally within the radC gene and consists of three transposase genes (tnpABC) as well as genes encoding a putative transcriptional regulator and QacH, a small multidrug resistance protein family (SMR) transporter putatively associated with export of BC that shows high amino acid identity to Smr/QacC from S. aureus and to EmrE from Escherichia coli. We screened 91 L. monocytogenes strains for the presence of Tn6188 by PCR and found Tn6188 in 10 of the analyzed strains. These isolates were from food and food processing environments and predominantly from serovar 1/2a. L. monocytogenes strains harboring Tn6188 had significantly higher BC minimum inhibitory concentrations (MICs) (28.5 ± 4.7 mg/l) than strains without Tn6188 (14 ± 3.2 mg/l). Using quantitative reverse transcriptase PCR we could show a significant increase in qacH expression in the presence of BC. QacH deletion mutants were generated in two L. monocytogenes strains and growth analysis revealed that ΔqacH strains had lower BC MICs than wildtype strains. In conclusion, our results provide evidence that Tn6188 is responsible for BC tolerance in various L. monocytogenes strains.
Listeria monocytogenes (Listeria) causes the third highest number of foodborne illness deaths (an estimated 255) in the United States annually, after nontyphoidal Salmonella species and Toxoplasma gondii (1). Deli meats are a major source of listeriosis illnesses (2,3), and meats sliced and packaged at retail delis are the major source of listeriosis illnesses attributed to deli meat (4). Mechanical slicers pose cross-contamination risks in delis and are an important source of Listeria cross-contamination (5,6). Reducing Listeria contamination of sliced meats in delis will likely reduce Listeria illnesses and outbreaks (6). Good slicer cleaning practices can reduce this foodborne illness risk (7). CDC’s Environmental Health Specialists Network (EHS-Net) studied how often retail deli slicers were fully cleaned (disassembled, cleaned, and sanitized) at the Food and Drug Administration (FDA) Food Code-specified minimum frequency of every 4 hours and examined deli and staff characteristics related to slicer cleaning frequency (8). Interviews with staff members in 298 randomly-selected delis in six EHS-Net sites showed that approximately half of delis fully cleaned their slicers less often than FDA’s specified minimum frequency. Chain-owned delis and delis with more customers, more slicers, required manager food safety training, food safety-knowledgeable workers, written slicer-cleaning policies, and food safety-certified managers fully cleaned their slicers more frequently than did other types of delis, according to deli managers or workers. States and localities should require deli manager training and certification, as specified in the FDA Food Code. They should also consider encouraging or requiring delis to have written slicer-cleaning policies. Retail food industry leaders can also implement these prevention efforts to reduce risk in their establishments. Because independent and smaller delis had lower frequencies of slicer cleaning, prevention efforts should focus on these types of delis.
- Proceedings of the National Academy of Sciences of the United States of America
- Published over 1 year ago
Listeria monocytogenes is responsible for gastroenteritis in healthy individuals and for a severe invasive disease in immunocompromised patients. Among the three identified L. monocytogenes evolutionary lineages, lineage I strains are overrepresented in epidemic listeriosis outbreaks, but the mechanisms underlying the higher virulence potential of strains of this lineage remain elusive. Here, we demonstrate that Listeriolysin S (LLS), a virulence factor only present in a subset of lineage I strains, is a bacteriocin highly expressed in the intestine of orally infected mice that alters the host intestinal microbiota and promotes intestinal colonization by L. monocytogenes, as well as deeper organ infection. To our knowledge, these results therefore identify LLS as the first bacteriocin described in L. monocytogenes and associate modulation of host microbiota by L. monocytogenes epidemic strains to increased virulence.
The relationship between the number of ingested Listeria monocytogenes cells in food and the likelihood of developing listeriosis is not well understood. Data from an outbreak of listeriosis linked to milkshakes made from ice cream produced in 1 factory showed that contaminated products were distributed widely to the public without any reported cases, except for 4 cases of severe illness in persons who were highly susceptible. The ingestion of high doses of L. monocytogenes by these patients infected through milkshakes was unlikely if possible additional contamination associated with the preparation of the milkshake is ruled out. This outbreak illustrated that the vast majority of the population did not become ill after ingesting a low level of L. monocytogenes but raises the question of listeriosis cases in highly susceptible persons after distribution of low-level contaminated products that did not support the growth of this pathogen.