The natural microflora of cold-smoked fish at the end of shelf-life are lactic acid bacteria (LAB). Some of these display a capacity to inhibit spoilage as well as several strains of pathogenic micro-organisms, e.g. Listeria monocytogenes which is isolated frequently from cold-smoked salmon (CSS). Eight batches of sliced vacuum-packed CSS from Norway, Scotland and Spain were collected at retail. Packs were stored at 5 degrees C and examined for chemical and microbiological characteristics, at purchase date and at expiration date. pH, water activity and salt content were similar to available data on lightly preserved fish products. There was a consistent pattern in the development of the microflora on CSS; the initial level of LAB was low on freshly produced CSS (10(2) cfu g(-1)); however, storage in vacuum packaging at refrigeration temperature was elective for LAB. At the end of the stated shelf-life these micro-organisms, represented mainly by Lactobacillus spp., attained ca.10(7) cfu g(-1) while Enterobacteriaceae counts were consistently lower (10(5) cfu g(-1)), which indicates the ability of LAB to grow and compete with few carbohydrates available and in the presence of moderate salt concentrations. L. monocytogenes was not found in any sample. Forty-one percent of LAB strains isolated exhibited inhibitory capacity against Listeria innocua, in a plate assay. A majority of the inhibitory effects were non-bacteriocinogenic, but nevertheless were very competitive cultures which may provide an additional hurdle for improved preservation by natural means.
Listeria monocytogenes, a psychrotrophic organism capable of growing at refrigeration temperatures, is of major concern in extended shelf life, refrigerated foods. Considering that as much as 80-90% of human listeriosis cases are linked to the ingestion of contaminated food, human cases are predominantly seen in high-risk individuals, including organ-transplant recipients, patients with AIDS and HIV-infected individuals, pregnant women, cancer patients, and the elderly. In 2001, the Canadian Listeriosis Reference Service (LRS) was created by the Bureau of Microbial Hazards (Health Canada) and the National Microbiology Laboratory (now part of the Public Health Agency of Canada). Major goals of the LRS include investigation of listeriosis cases and maintenance of a national collection of isolates. The LRS intends to create a comprehensive molecular epidemiological database of all isolates in Canada for use as a resource for outbreak investigations, research and other microbiological investigations. The PFGE profiles are being established and stored for clinical, food, environmental, and possibly animal strains of L. monocytogenes. The LRS pursues research activities for investigation and implementation of other molecular methods for characterizing L. monocytogenes isolates. Ribotyping, Multi-locus Sequence Typing (MLST), Variable Number of Tandem Repeats (VNTR), Multi-locus virulence sequence typing (MLVA), microarray- based technologies and sequence-based typing schemes, are being investigated on selected diversity sets. The LRS has also used PFGE typing for outbreak investigations. The molecular epidemiological data, timely coordination and exchange of information should help to reduce the incidence of listeriosis in Canada. In Canada, listeriosis is not a national notifiable disease, except for the province of Quebec, where it has been since 1999. The LRS, Canadian Public Health Laboratory Network, and federal epidemiologists are currently working on making human listeriosis notifiable throughout Canada.
Public health authorities place a high priority on investigating listeriosis outbreaks, and these epidemiological investigations remain challenging. Some approaches have been described in the literature to address these challenges. This review of listeriosis clusters and outbreaks investigated in the Province of Quebec (Quebec) highlights investigative approaches that contributed to identifying the source of these outbreaks.
The Laboratoire de Santé Publique du Québec (LSPQ) implemented pulsed-field gel electrophoresis (PFGE) molecular subtyping in 1997 to identify Listeria monocytogenes clusters among isolates from invasive listeriosis cases identified throughout Quebec. A cluster was defined as three cases or more with the same or similar PFGE profiles (=3 band difference) occurring over a 4-month period. An investigation was initiated if the epidemiologic indicators suggested a common source. Listeriosis data from LSPQ's database were reviewed to identify and describe clusters detected from 1997 to 2011, including those that led to an outbreak investigation. Epidemiological reports prepared following each outbreak were also reviewed.
Eleven clusters were identified in the province by LSPQ between 1997 and 2011. Outbreak investigations were initiated for six clusters, four of which involved more than 10 cases. Factors that contributed to identifying the source for three of these outbreaks highlighted the value of (1) making all stakeholders (food safety and inspection services, public health authorities, and laboratories) aware of any ongoing investigation and sharing relevant information even if the source is not yet identified; (2) promptly collecting food samples identified and considered as possible vehicles of infection identified during the interview of a Listeria case; (3) collecting food items and/or environmental samples in locations reported in common by cases in the same cluster.
Multiple approaches should be considered when investigating L. monocytogenes clusters. Networks to facilitate continuous exchange of human and food data between public health and food safety partners should be encouraged.
Since 1986, 68% of the Listeria monocytogenes isolates from human cases of invasive listeriosis in Sweden are available for retrospective studies. The aim of the present study was to characterize 601 human invasive isolates of L. monocytogenes in Sweden from 1986 to 2007 by using serotyping and pulsed-field gel electrophoresis. Since 1996, serovar 4b was permanently reduced to the second or third most common serovar in human cases in Sweden. During the latter period, 2000-2007, only 13% belonged to serovar 4b and 71% to 1/2a. The dendrogram, based on pulsovars, reveals two clusters with different serovars. Cluster 1 exhibits serovars 4b and 1/2b, whereas cluster 2 consists of serovar 1/2a. Serovar 1/2a seems to be more heterogeneous than serovar 4b.
Minimally processed spinach has been recently associated with outbreaks of foodborne illnesses. This study investigated the effect of commercial minimal processing of spinach on the coliform and Escherichia coli counts and the prevalence of E. coli O157:H7, Salmonella, Shigella spp., and Listeria monocytogenes on two types of spinach before and after minimal processing. A total of 1,356 spinach samples (baby spinach, n = 574; savoy spinach, n = 782) were collected daily in two processing plants over a period of 14 months. Raw spinach originated from nine farms in the United States and three farms in Canada. Overall, the proportion of samples positive for coliforms increased from 53% before minimal processing to 79% after minimal processing (P 0.1) was observed. E. coli O157:H7 and Shigella spp. were not isolated from any of the samples. Salmonella and L. monocytogenes were isolated from 0.4 and 0.7% of samples, respectively. Results demonstrate that commercial minimal processing of spinach based on monitored chlorine washing and drying may not decrease microbial load on spinach leaves as expected. Further research is needed to identify the most appropriate measures to control food safety risk under commercial minimal processing of fresh vegetables.
Australian Food Safety Centre of Excellence, Tasmanian Institute of Agricultural Research, School of Agricultural Science, University of Tasmania, Private Bag 54, Hobart 7001, Tasmania, Australia. firstname.lastname@example.org
Two commercially available organic acid salts, potassium lactate (PURASAL HiPure P) and a potassium lactate-sodium diacetate blend (PURASAL Opti. Form PD 4), were assessed as potential inhibitors of Listeria monocytogenes growth in modified atmosphere packaged (MAP) sliced ham in challenge studies. The influence of the initial inoculation level of L. monocytogenes (10(1) or 10(3) CFU g(-1)) and storage temperature (4 or 8 degrees C) was also examined. The addition of either organic acid salt to MAP sliced ham strongly inhibited the growth of L. monocytogenes during the normal shelf life of the product under ideal refrigeration conditions (4 degrees C) and even under abusive temperature conditions (i.e., 8 degrees C). During the challenge studies and in the absence of either organic acid salt, L. monocytogenes numbers increased by 1000-fold after 20 days at 8 degrees C and 10-fold after 42 days at 4 degrees C. Both organic acid salt treatments were found to be listeriostatic rather than listericidal. The addition of either organic acid salt to the MAP ham also reduced the growth of indigenous microflora, i.e., aerobic microflora and lactic acid bacteria. The influence of these compounds on the risk of listeriosis in relation to product shelf life is discussed.
Rapid methods still rely on a prior (shortened) enrichment step before application. Quantitative information is a prerequisite for understanding the resuscitation kinetics of the growth during the enrichment step. In this study various basal and newly introduced selective enrichment broths were evaluated. First, growth parameters (lambda, mu(max)) of both healthy and sub-lethally injured cells were determined. Next, a selection of enrichment broths was compared for their capacity to support detection within 24h of low numbers of Listeria monocytogenes in artificially and naturally contaminated food samples. Detection was performed either by phage protein-based capture (Listeria Capture kit, Profos, Regensburg, Germany) combined with plating on chromogenic medium or by fluorescence in situ hybridization (FISH) using the VIT-Listeria kit (Vermicon, Munich, Germany). Kinetics of resuscitation and growth of L. monocytogenes in various enrichment broths showed that for detection of low numbers of sub-lethally injured L. monocytogenes cells at least an overnight enrichment was needed. A selective enrichment broth was needed to enable proliferation of L. monocytogenes within the indigenous bacterial flora present in foods. However, combination of an appropriate enrichment condition with advanced detection techniques may enable a 24h detection of L. monocytogenes.
Among 504 clinical lineage II isolates of Listeria monocytogenes isolated during 1958-2010 in Sweden, 119 pulsed-field gel electrophoresis (PFGE) types (AscI) have been identified based on the number and distribution of all banding patterns in each DNA profile. In this study, these types were further divided into PFGE groups based on the configuration of small bands with sizes 145.5?kb.
This 13-month survey was conducted to estimate the prevalence and counts of foodborne pathogenic bacteria and indicator bacteria on swine carcasses in Sweden. A total of 541 swine carcasses were sampled by swabbing prechill at the 10 largest slaughterhouses in Sweden. Pathogenic Yersinia enterocolitica was detected by PCR in 16% of the samples. The probability of finding Y. enterocolitica increased with increasing counts of Escherichia coli. No samples were positive for Salmonella. The prevalences of Campylobacter, Listeria monocytogenes, and verocytotoxin-producing E. coli were low (1, 2, and 1%, respectively). None of the verocytotoxin-positive enrichments, as determined by a reverse passive latex agglutination assay, tested positive for the virulence genes eaeA or hlyA by PCR. Coagulase-positive staphylococci, E. coli, and Enterobacteriaceae were recovered from 30, 57, and 87% of the samples, respectively, usually at low levels (95th percentiles, 0.79, 1.09, and 1.30 log CFU/cm2, respectively). The mean log level of Enterobacteriaceae was 0.35 log CFU/cm2 higher than that of E. coli on carcasses positive for both bacteria. The mean log level of aerobic microorganisms was 3.48 log CFU/cm2, and the 95th percentile was 4.51 log CFU/cm2. These data may be useful for risk assessment purposes and can serve as a basis for risk management actions, such as the use of E. coli as an alternative indicator organism for process hygiene control.
In British Columbia (BC), Canada, food processing facilities licensed under provincial authority are not required to sample for Listeria monocytogenes in food products or processing environments. In 2009, we conducted a survey of dairy, fish, and meat facilities under BC authority to estimate the prevalence of Listeria spp. and L. monocytogenes in ready-to-eat (RTE) foods and production environments. From August to October, 250 RTE food samples and 258 swabs from the food processing environments of 43 facilities were collected. Standard culture methods were applied to both food samples and swabs. Of swabs collected from all 258 environmental surfaces, 15% were positive for Listeria spp. Significantly (P, 0.001) more fish facilities than dairy and meat facilities had food contact surfaces contaminated with Listeria spp. L. monocytogenes was found in RTE foods from fish facilities alone (5 of 12); in all five of the fish facilities with contaminated product, one or more environmental swabs were also positive for L. monocytogenes. The results suggest that while control of L. monocytogenes in BC-inspected dairy and meat facilities is effective in limiting food contamination, there is a need for provincial inspectors to initiate improved monitoring and management of contamination by L. monocytogenes in RTE fish processing facilities.