Listeriosis Outbreaks Research Articles

A Nosocomial Outbreak of Invasive Listeriosis in An Italian Hospital: Epidemiological and Genomic Features.

Listeria monocytogenes (L. monocytogenes) is a widespread opportunistic pathogen that causes the listeriosis foodborne disease. This bacterium has become a common contaminant of handled food, and a relevant public health issue. Here we describe a nosocomial outbreak of listeriosis caused by an ST451 strain of L. monocytogenes involving three cancer and one immunocompromised patients hospitalized in different units from the same hospital during September and October 2020. The epidemiological investigation was conducted using traditional microbiological methodology combined with a whole genome sequencing approach. The source of contamination was identified in the kitchen hospital, where a meat slicer used to prepare patients’ meals was tested positive to the same sequence type (ST) of L. monocytogenes. This is the first report of an outbreak of listeriosis caused by ST451 in Italy.

Complete Genome Sequences of Three Clinical Listeria monocytogenes Sequence Type 8 Strains from Recent German Listeriosis Outbreaks.

We report here the closed genome sequences of three clinical Listeria monocytogenes strains of multilocus sequence typing (MLST) sequence type 8 (ST8). These strains are representatives of three separate listeriosis outbreak clusters (Alpha1, Pi4, and Sigma1) that affected Germany between 2012 and 2020.

Environmental dissemination of pathogenic Listeria monocytogenes in flowing surface waters in Switzerland

Listeria monocytogenes is an opportunistic pathogen that is widely distributed in the environment. The aquatic environment may represent a potential source for the transmission of L. monocytogenes to animals and the food chain. The present study assessed the occurrence of L. monocytogenes in 191 surface water samples from rivers, streams and inland canals throughout Switzerland. Twenty-five (13%) of the surface water samples contained L. monocytogenes. Whole genome sequence (WGS) data were used to characterize the 25 isolates. The isolates belonged to major lineages I and II, with the majority assigned to either serotype 1/2a (48%), or 4b (44%). The predominant CCs identified were the hypervirulent serotype 4b clones CC1 and CC4, and the serotype CC412; all three have been implicated in listeriosis outbreaks and sporadic cases of human and animal infection worldwide. Two (8%) of the isolates belonged to CC6 which is an emerging hypervirulent clone. All isolates contained intact genes associated with invasion and infection, including inlA/B and prfA. The four CC4 isolates all harbored Listeria pathogenicity island 4 (LIPI-4), which confers hypervirulence. The occurrence of L. monocytogenes in river ecosystems may contribute to the dissemination and introduction of clinically highly relevant strains to the food chain.

Prevalence of Listeria Species on Food Contact Surfaces in Washington State Apple Packinghouses.

The 2014 caramel apple listeriosis outbreak was traced back to cross-contamination between food contact surfaces (FCS) of equipment used for packing and fresh apples. For Washington state, the leading apple producer in the United States with 79% of its total production directed to the fresh market, managing the risk of apple contamination with Listeria monocytogenes within the packing environment is crucial. The objectives of this study were to determine the prevalence of Listeria spp. on FCS in Washington state apple packinghouses over two packing seasons and to identify those FCS types with the greatest likelihood to harbor Listeria spp. Five commercial apple packinghouses were visited quarterly over two consecutive year-long packing seasons. A range of 27 to 50 FCS were swabbed at each facility to detect Listeria spp. at two sample times, (i) postsanitation and (ii) in-process (3 h of packinghouse operation), following a modified protocol of the FDA's Bacteriological Analytical Manual method. Among 2,988 samples tested, 4.6% (n = 136) were positive for Listeria spp. Wax coating was the unit operation from which Listeria spp. were most frequently isolated. The FCS that showed the greatest prevalence of Listeria spp. were polishing brushes, stainless steel dividers and brushes under fans/blowers, and dryer rollers. The prevalence of Listeria spp. on FCS increased throughout apple storage time. The results of this study will aid apple packers in controlling for contamination and harborage of L. monocytogenes and improving cleaning and practices for sanitation of the FCS on which Listeria spp. are the most prevalent.IMPORTANCE Since 2014, fresh apples have been linked to outbreaks and recalls associated with postharvest cross-contamination with the foodborne pathogen L. monocytogenes These situations drive both public health burden and economic loss and underscore the need for continued scrutiny of packinghouse management to eliminate potential Listeria niches. This research assesses the prevalence of Listeria spp. on FCS in apple packinghouses and identifies those FCS most likely to harbor Listeria spp. Such findings are essential for the apple-packing industry striving to further understand and exhaustively mitigate the risk of contamination with L. monocytogenes to prevent future listeriosis outbreaks and recalls.

Retrospective Use of Whole-Genome Sequencing Expands the Multicountry Outbreak Cluster of Listeria monocytogenes ST1247.

Listeria monocytogenes sequence type 1247 clonal complex 8 caused a prolonged multicountry outbreak in five EU countries: Denmark, Estonia, Finland, France, and Sweden. A total of 22 disease cases were identified with onset of symptoms between July 2014 and February 2019. Five patients died due to, or with, the disease. The retrospective analysis of L. monocytogenes isolate VLTRLM2013 revealed the presence of an outbreak-related strain (cgMLST type L2-SL8-ST1247-CT4158) in ready-to-eat fish product more than a year prior to the first outbreak-related cases. Reference outbreak strain and VLTRLM2013 strain were compared using core genome and whole-genome multilocus sequence typing analyses. Genomic level differences of the persistent L. monocytogenes strains associated with a prolonged multicountry foodborne listeriosis outbreak are described. It was concluded that the persistent nature of the multicountry outbreak-related L. monocytogenes strain VLTRLM2013 together with stress island, virulence, and antibiotic resistance genes could potentially be the determining factors for the extensive and prolonged outbreak affecting five European Union countries. Our results support the systematic application of whole-genome sequencing in food and public health surveillance and further encourages its wide adoption.

Occurrence of Listeria monocytogenes in Counter-Sliced Turkey Meat Samples from Independent Delis in New York City

Research suggests that small, independent delicatessens are less likely to follow proper sanitation procedures, including slicer inspection, which could lead to a higher likelihood of these delis being a reservoir for Listeria monocytogenes growth and cross-contamination. This study was undertaken to determine the incidence of L. monocytogenes in counter-sliced turkey deli meat obtained from independent delis in an urban city. Deli meat, counter-sliced on site, was collected from 118 independent delis in New York City. The samples were analyzed for L. monocytogenes using the U.S. Department of Agriculture Microbiology Laboratory Guidebook methodology for isolation and confirmation. The selection criteria for delis included using the city's restaurant inspection and grading system. Two samples, from separate delis, were confirmed as positive for L. monocytogenes (1.69%). Analysis of the genomic sequences of one of the samples revealed a close match to a cluster of six clinical isolates, which were part of an ongoing multistate listeriosis outbreak spanning four states. The sequence of the second isolate matched a clinical isolate in a neighboring state. Both isolates were obtained from delis that did not have the top inspection grade. Although a snapshot of one urban area, this study is the first report on the current incidence of L. monocytogenes on counter-sliced deli meat from independent deli establishments. This study suggests that these delis can potentially serve as sources of L. monocytogenes contamination or contribute to downstream foodborne listeriosis. Information provided by city inspection and grading systems, in addition to the letter grade, may serve as a tool to identify delis with potential L. monocytogenes contamination issues and a basis for product and environmental sampling by public health authorities.

Clinical characteristics of patients with listeriosis.

To investigate the clinical characteristics of patients with listeriosis and to provide a basis for diagnosis, treatment, prevention and control of hospital infection. A total of 10 inpatients, who suffered from the listeriosis in Xiangya Hospital, Central South University from January 2013 to June 2019, were retrospectively collected for this study. The characteristics of the patients' age, gander, basic information, case type, clinical manifestations, first consultation department, days of diagnosis, infection indicator, specimen type, results of drug sensitivity, treatment plan, hospital infection or not, outcome, follow-up data were analyzed. Two cases were pregnant women and other were non-pregnant adults among 10 patients with listeriosis. Among them, there were 3 cases with hospital acquired infection. The age of patient onset was 27-71 years old, and the time from onset to diagnosis was 5-36 days. Five cases had fever, and other 5 cases had not fever. There were headache, fatigue, local pain, and other specialized symptoms in the 10 patients.The white blood cell count,the neutrophil ratio, the inflammatory index C-reactive protein, the procalcitonin were all increased, and the erythrocyte sedimentation was accelerated in the 10 patients.All the patients were sensitive to ampicillin, penicillin G, meropenem, and compound sinomine. Listeriosis often affects the patients with low immunity, which often leads to misdiagnosis or missed diagnosis in clinic.So early prevention, early diagnosis, and early treatment can reduce mortality; it is important for departments of nosocomial infection management to manage patients' diet for avoiding outbreaks of listeriosis in hospital.

Effect of Zataria multiflora Boiss. essential oil, NaCl, acid, time, and temperature on the growth of Listeria monocytogenes strains in broth and minced rainbow trout.

The small outbreaks of listeriosis as one of the leading causes of food poisoning‐associated deaths occur more than previously reported. In current study, the growth ability of Listeria monocytogenes strains isolated from different sources of food and human origin was measured under salt stress (0.5%, 2.5%, 5%, 7.5%, and 10%) and acid environments (pH = 6.64 and 5.77) for 96 hr by using a Bioscreen C microbiology reader at 37°C. In further steps of this study, after analysis of constituents of Zataria multiflora Boiss. essential oil (ZMEO), the sensory evaluation of the treated fish meat with ZMEO was performed. Then, the fish isolate of L. monocytogenes was exposed to sensory acceptable and subminimum inhibitory concentrations (subMICs) of ZMEO in fish broth and minced fish meat during incubation at abuse (12°C), room (22°C), and optimum (37°C) temperatures for 48 hr. The MIC of NaCl against four strains of L. monocytogenes was 10% at 37°C. The maximum optical densities (ODs) and under curve areas (AUC) of growth patterns in higher pH value and lower contents of NaCl followed the order of 21C > 6F > 66C > 22C of L. monocytogenes strains, while the lag time was prolonged in the reverse order. The maximum OD, growth, and lag times of samples treated with higher contents of NaCl and lower pH value were affected in a different order. The organoleptic evaluation showed that the fish meat treated with less than 0.5% of ZMEO was sensory acceptable. The population of L. monocytogenes remained relatively constant at the inoculation level of 107 cfu/ml (or g) at 12°C in broth and minced fish mediums. The inhibitory antilisterial activity of essential oil as an extensive‐used plant for food and pharmacological applications is negligible due to possible adverse sensory and toxic effects at relevant high doses.

First Detection of Listeria monocytogenes in a Buffalo Aborted Foetus in Campania Region (Southern Italy).

Listeria monocytogenes (LM) is the causative agent of listeriosis in both animals and humans, representing one of the most severe food-borne diseases in humans. Out of 13 serotypes, only three (i.e., 1/2a, 1/2b, and 4b) are responsible for 95% of human outbreaks of listeriosis. Ruminants have been hypothesised to represent the main natural reservoir for this pathogen and to be involved in the transmission of Listeria to humans. During pregnancy, listeriosis in ruminants cause various reproductive disorders as well as abortion. However, little is known about abortion due to LM in water buffaloes (Bubalus bubalis). In this study, we report for the first time the detection of LM in a water buffalo foetus in the region of Campania, Italy. Complete necropsy was performed, and samples and swabs from the abomasum, kidneys, liver, lungs, and spleen were collected. Microbiological and molecular analyses were carried out to detect bacterial, viral, and protozoarian abortive pathogens. The results revealed the presence of LM in the liver, lungs, and abomasum, and no other agent was detected. Isolation was confirmed by biochemical and molecular tests. Molecular serotype characterisation was performed, and serogroup IVb was identified. In conclusion, because of the zoonotic implications of our findings, this report highlights the importance of including LM in the diagnostic panel in cases of bubaline abortion.

Population Genomic Analysis of Listeria monocytogenes From Food Reveals Substrate-Specific Genome Variation.

Listeria monocytogenes is the major causative agent of the foodborne illness listeriosis. Listeriosis presents as flu-like symptoms in healthy individuals, and can be fatal for children, elderly, pregnant women, and immunocompromised individuals. Estimates suggest that L. monocytogenes results in ∼1,600 illnesses and ∼260 deaths annually in the United States. L. monocytogenes can survive and persist in a variety of harsh environments, including conditions encountered in production of fermented dairy products such as cheese. For instance, microbial growth is often limited in soft cheese fermentation because of harsh pH, water content, and salt concentrations. However, L. monocytogenes has caused a number of deadly listeriosis outbreaks through the contamination of cheese. The purpose of this study was to understand if genetically distinct populations of L. monocytogenes are associated with particular foods, including cheese and dairy. To address this goal, we analyzed the population genetic structure of 504 L. monocytogenes strains isolated from food with publicly available genome assemblies. We identified 10 genetically distinct populations spanning L. monocytogenes lineages 1, II, and III and serotypes 1/2a, 1/2b, 1/2c, 4b, and 4c. We observed an overrepresentation of isolates from specific populations with cheese (population 2), fruit/vegetable (population 2), seafood (populations 5, 8 and 9) and meat (population 10). We used the Large Scale Blast Score Ratio pipeline and Roary to identify genes unique to population 1 and population 2 in comparison with all other populations, and screened for the presence of antimicrobial resistance genes and virulence genes across all isolates. We identified > 40 genes that were present at high frequency in population 1 and population 2 and absent in most other isolates. Many of these genes encoded for transcription factors, and cell surface anchored proteins. Additionally, we found that the virulence genes aut and ami were entirely or partially deleted in population 2. These results indicate that some L. monocytogenes populations may exhibit associations with particular foods, including cheese, and that gene content may contribute to this pattern.

Closed Genome Sequences of Clinical Listeria monocytogenes PCR Serogroup IVb Isolates Associated with Two Recent Large Listeriosis Outbreaks in Germany.

Here, we report the closed genome sequences of two representative Listeria monocytogenes strains belonging to PCR serogroup IVb, which are related to two large outbreaks of human listeriosis that affected Germany in 2015 (Eta1) and 2018 to 2019 (Epsilon1a).

Development and Validation of a Quantitative PCR Method for Species Verification and Serogroup Determination of Listeria monocytogenes Isolates

Listeria monocytogenes (Lm) is one of the leading causes of death because of foodborne illness, affecting the elderly, pregnant women, neonates, and people who are immunocompromised. Serologically, Lm can be classified into 13 serotypes, although only 4 are typically linked with food contamination and illness. Since 2000, a shift in serotypes involved in listeriosis outbreaks has been observed, suggesting that tracking of serotypes could help identify emerging trends. A PCR method developed in 2004 allowed detection of the four major serotypes as molecular serogroups, corresponding to broad phylogenetic groups. In this study, a novel quantitative PCR (qPCR) method was developed that uses two multiplex qPCRs, one to confirm the Listeria genus and Lm species and the second for Lm molecular serogrouping. This method was compared with the U.S. Food and Drug Administration Bacteriological Analytical Manual (BAM) method for Lm and the seroagglutination method, using a 208-strain panel. Comparison of the genus and species qPCR assay with the BAM methods found an equal or slightly higher accuracy for the qPCR method (>98%), compared with the BAM protocol (>96%), when evaluated against independent characterization data. Molecular serogrouping using the qPCR method (96.6%) was more accurate than the seroagglutination assay (75.6%). The qPCR method identified Lm 4bV strains, which could not be resolved using seroagglutination. The qPCR could not identify lineage III and IV serotype 4b strains but did correctly identify 16 of 18 lineage III and IV strains. The qPCR method performed genus identification for the Listeria species Lm, L. innocua, L. welshimeri, L. ivanovii, and L. seeligeri. In addition, the method performed species identification for Lm and classified Lm into six molecular serogroups: 2A, 2B, 2C, 4B, NT, and 4bV. This method provided a rapid and accurate confirmation of Lm and serogroup determinations; furthermore, it could help identify otherwise unlinked strains by enabling whole genome sequencing analysis based on broad phylogeny, independent of other information.

Lytic characterization and application of listerial endolysins PlyP40 and PlyPSA in queso fresco

Queso fresco (QF) is a fresh Hispanic-style cheese that is commonly associated with the human foodborne pathogen Listeria monocytogenes and outbreaks of listeriosis in the United States. Endolysins, cell wall hydrolases derived from bacteriophages, are promising candidates for controlling bacterial pathogens in food systems. In this study, we characterized the lytic capabilities of 2 endolysins, PlyP40 and PlyPSA, under varying conditions (pH, temperature, salt concentration) and compared their activities with those of the previously described endolysin PlyP100. We showed that PlyP40 was effective, showing at least a 33% reduction in cellular debris, against a broader range of Listeria than PlyPSA, which showed little lytic activity toward Listeria strains not from serovar 4. Both endolysins were also capable of maintaining lytic activity to varying extents at refrigeration temperature. The effect of salt concentration and pH differed between PlyP40 and PlyPSA. Furthermore, we added the endolysins to QF and monitored their ability to control L. monocytogenes contamination over 28 d of cold storage. Both PlyP40 and PlyPSA were capable of lowering QF inoculum cell counts compared with the control; however, both were less effective than the previously characterized PlyP100. Further characterization of endolysins will continue to open opportunities to optimization and implementation in a variety of food matrices for controlling pathogen contamination.

Analysis of virulence genes and molecular typing of Listeria monocytogenes isolates from human, food, and livestock from 2008 to 2016 in Iran.

The frequency of Listeria monocytogenes isolates collected from a total of 1150 samples including food (n= 300), livestock (n= 50), and human clinical (n= 800) was evaluated during 2008-2016. Antimicrobial resistance patterns, virulence factors, and molecular characteristics of these isolates were analyzed using disk diffusion method, sequencing, serotyping, and pulsed-field gel electrophoresis (PFGE). The analysis of 44 L. monocytogenes isolates showed that 72.7% (32 of 44) of all the isolates belonged to Serotype 1/2c, and 15.9% (7 of 44) belonged to Serotype 3c. All 44 isolates were resistant to one or more antimicrobial agents with the most frequent resistance to penicillin (75%) and tetracycline (47.7%). Of the 44 L. monocytogenes strains, 100, 69.2, and 62.5% of livestock, human, and food strains were resistant to penicillin, respectively. Using pulsed-field gel electrophoresis (PFGE) technique, the isolates' genetic diversity was determined, and 28 PFGE patterns with 8 common (CT) and 20 single types (ST) were identified. This study highlights the high prevalence of Serotype 1/2c in clinical and livestock samples, while different serotypes were observed in food samples. The presence of rare serotypes such as 4c, belonging to the Lineage III, as well as 4e and 1/2c which are infrequent in Iran indicates that paying attention to uncommon serotypes, especially 1/2c, during the listeriosis outbreaks is necessary.

Molecular characterization and in vivo pathogenicity study of Listeria monocytogenes isolated from fresh and frozen local and imported fish in Jordan.

Background:Listeria monocytogenes (L. monocytogenes) is a serious zoonotic and food transmitted human pathogen causing meningitis and abortions. Several outbreaks of listeriosis have been associated with the consumption of ready-to-eat food products; dairy, meat, fish, and contaminated fruits and vegetables worldwide.Aim:This study was designed to detect and characterize L. monocytogenes isolated from local and imported fish in Jordan. Methods:A total of 170 fish (70 local and 100 imported), of which 140 fresh and 30 frozen samples were used in this study. Listeria monocytogenes was cultured and initially identified using conventional microbiological methods. For confirmation and serotyping of the L. monocytogenes isolates, PCR techniques were used. Using oral and intraperitoneal administration, mice were used to determine the pathogenicity and LD50 of the isolated L. monocytogenes. Results:A total of 72 Listeria spp. isolates were cultured from fish. Of those, 24 were positively identified as L. monocytogenes. Other strains of Listeria spp. were L. ivanovii (21), L. innocua (11), and L. grayi (16). Serotyping of the L. monocytogenes indicated that 14 isolates belonged to the 1/2b, 3b serotypes whereas 10 isolates belonged to the 4a and 4c serotypes. All isolates were virulent to mice with an LD50 dose ranging from 3 × 1010 CFU/ml to 3 × 107.5 CFU/ml. All the virulent isolates belonged to the serotype 1/2b. Histopathologically, dead mice showed multiple necrotic lesions in the liver and spleen.Conclusion:Results of this study showed the presence of potentially pathogenic L. monocytogenes in fresh and frozen, local, and imported fish in Jordan. Strict monitoring and quality control regulatory measures must be adopted to prevent future outbreaks of food poisoning associated with fish consumption.

Listeria monocytogenes isolates from Western Cape, South Africa exhibit resistance to multiple antibiotics and contradicts certain global resistance patterns.

Food-borne disease outbreaks are common and offer valuable insights into the causes, impacts, and mechanisms underlying food pathogens. This also serves as a good foundation to validate the performance of current best practice control methods, for example antibiotics, that are used in the fight against food pathogens. Listeriosis outbreaks, caused by Listeria monocytogenes, is no exception. In 2018, South Africa experienced the largest global listeriosis outbreak recorded to date. However, despite the scale of this outbreak, information on the bacterium and its resistance towards antibiotics is still severely lacking. Furthermore, until now it remained to be determined whether L. monocytogenes antibiotic resistance patterns in South Africa mirror resistance patterns elsewhere in the world. The aim of this study was therefore to evaluate the efficacy of antibiotics that are currently used against L. monocytogenes. Using the European Committee on Antimicrobial Susceptibility Testing (EUCAST) disc diffusion method, L. monocytogenes isolates (n = 177) from diverse origins in the Western Cape, South Africa (clinical, food, and environment) were tested for susceptibility against five different antibiotics, namely ampicillin, erythromycin, chloramphenicol, gentamicin, and tetracycline. Isolates were collected over a period of two years (2017–2019). All isolates were susceptible to ampicillin, the currently recommended antibiotic, while a large number of isolates were resistant to chloramphenicol, erythromycin, and tetracycline. Also, patterns of resistance observed here are different to patterns observed elsewhere. The findings of this study demonstrate that it is imperative to continuously monitor the efficacy of currently recommended antibiotics, since resistance patterns can quickly develop when such antibiotics are overutilized, and secondly, that it is crucial to assess local antibiotic resistance patterns in conjunction with global patterns, since the latter is not necessarily generalizable to local scales.

Listeriosis Caused by Persistence of Listeria monocytogenes Serotype 4b Sequence Type 6 in Cheese Production Environment

Abstract A nationwide outbreak of human listeriosis in Switzerland was traced to persisting environmental contamination of a cheese dairy with Listeria monocytogenes serotype 4b, sequence type 6, cluster type 7488. Whole-genome sequencing was used to match clinical isolates to a cheese sample and to samples from numerous sites within the production environment.

Listeriosis Caused by Persistence of Listeria monocytogenes Serotype 4b Sequence Type 6 in Cheese Production Environment.

A nationwide outbreak of human listeriosis in Switzerland was traced to persisting environmental contamination of a cheese dairy with Listeria monocytogenes serotype 4b, sequence type 6, cluster type 7488. Whole-genome sequencing was used to match clinical isolates to a cheese sample and to samples from numerous sites within the production environment.

Prevalence of Listeria monocytogens in Farm Milk and Some Dairy Products with Comparing its Sensitivity to Different Antibiotics and H2O2 Loaded on Silver Nanoparticles in Dakahlia Governorate, Egypt.

L. monocytogens is one of the major food-borne pathogens which can affect humans and a wide variety of animal species. It usually invades Milk and its products at several steps in the food chain, Listeriosis outbreaks remain an important problem globally. High resistance of L. monocytogens to the most of antibiotics due to misusing of it in both veterinary and human field, it necessitates searching about alternatives. Drug laden nanoparticles possibly will overwhelm the restrictions of conservative antibiotic actions related with poisonousness, unsuitable distribution, or enzymatic dilapidation. The aim of this research is to detect prevalence of this bacteria in farm milk and some milk products and to mark the result of H2O2 loaded on silver nanoparticles on Multidrug resistance strains of L. monocytogenes. Therefore, One hundred samples (25 farm bulk tank, 25 plain yoghurt and 50 fresh soft cheese (Talaga)) were haphazardly composed from dairy farms and different shops in different areas of Mansoura city, Dakahlia governorate. Egypt. The isolated strains of L. monocytogenes were evaluated phenotypically and genotypically for detection of virulence factors (invasive linked protein (iap), haemolysin (hylA) and actin polymerization protein (actA)) that were carried out by using multiplex PCR. Out of 100 samples 7 listeria monocytogenes isolates from Fresh soft cheese (Talaga) samples (14%). These 7 isolates had iap gene, while 6 isolates had hly gene and 5 isolates had actA gene. Also the three types of virulence genes were in four isolates at which a higher multiple antibiotic resistance between L.monocytogenes isolates. Assessment of antibacterial action of silver nanoparticles and hydrogen peroxide Antibacterial doings was assessed by broth micro-dilution test and our results revealed that H2O2/AgNPs with 6.25-3.13 µg/mL MIC values of the original concentrations 13 µg/mL MIC values of the original concentrations highly effective against L. monocytogens.

A Cross-Protective Vaccine Against 4b and 1/2b Listeria monocytogenes.

Listeria monocytogenes (Lm) is a foodborne zoonotic pathogen that causes listeriosis with a mortality rate of 20–30%. Serovar 4b and 1/2b isolates account for most of listeriosis outbreaks, however, no listeriosis vaccine is available for either prophylactic or therapeutic use. Here, we developed a triple-virulence-genes deletion vaccine strain, and evaluated its safety, immunogenicity, and cross-protective efficiency. The virulence of NTSNΔactA/plcB/orfX was reduced 794-folds compared with the parental strain. Additionally, it was completely eliminated in mice at day 7 post infection and no obvious pathological changes were observed in the organs of mice after prime-boost immunization for 23 days. These results proved that the safety of the Lm vaccine strain remarkably increased. More importantly, the NTSNΔactA/plcB/orfX strain stimulated higher anti-Listeriolysin O (LLO) antibodies, induced significantly higher expression of IFN-γ, TNF-α, IL-17, and IL-6 than the control group, and afforded 100% protection against serovar 4b and 1/2b challenges. Taken together, our research demonstrates that the triple-genes-deletion vaccine has high safety, can elicit strong Th1 type immune response, and affords efficient cross-protection against two serovar Lm strains. It is a promising vaccine for prevention of listeriosis.