In this study, the performance of four alternative selective chromogenic B. cereus agar was compa... more In this study, the performance of four alternative selective chromogenic B. cereus agar was compared to the reference mannitol-yolk polymyxin (MYP) agar (ISO 7932) using inclusion and exclusion test strains (n = 110) and by analyzing naturally contaminated milk and other food samples (n = 64). Subsequently, the panC group affiliation and toxin gene profile of Bacillus cereus senso lato (s.l.) isolates were determined. Our results corroborate that the overall best performing media CHROMagar™ B. cereus (93.6% inclusivity; 82.7% exclusivity) and BACARA® (98.2% inclusivity, 62.7% exclusivity) are more sensitive and specific compared to Brilliance™ B. cereus, MYP and ChromoSelect Bacillus Agar. Both media allow unequivocal detection of B. cereus with low risks of misidentification. Media containing ß-D-glucosidase for the detection of presumptive B. cereus may form atypical colony morphologies resulting in a false negative evaluation of the sample. Naturally contaminated samples presente...
Listeria monocytogenes is an environmentally adapted saprophyte that can change into a human and ... more Listeria monocytogenes is an environmentally adapted saprophyte that can change into a human and animal bacterial pathogen with zoonotic potential through several regulatory systems. In this review, the focus is on the occurrence of Listeria sensu stricto and sensu lato in different ecological niches, the detection methods, and their analytical limitations. It also highlights the occurrence of L. monocytogenes genotypes in the environment (soil, water, and wildlife), reflects on the molecular determinants of L. monocytogenes for the saprophytic lifestyle and the potential for antibiotic resistance. In particular, the strain-specific properties with which some genotypes circulate in wastewater, surface water, soil, wildlife, and agricultural environments are of particular interest for the continuously updating risk analysis.
Nucleotide sequence-based methods focusing on the single-nucleotide polymorphisms (SNPs) of Liste... more Nucleotide sequence-based methods focusing on the single-nucleotide polymorphisms (SNPs) of Listeria monocytogenes and L. innocua housekeeping genes (multilocus sequence typing) and in the core genome (core genome MLST) facilitate the rapid and interlaboratory comparison in open accessible databases as provided by Institute Pasteur ( https://bigsdb.web.pasteur.fr/listeria/listeria.html ). Strains can be compared on a global level and help to track forward and trace backward pathogen contamination events in food processing facilities and in outbreak scenarios.
Most Austrian dairies and cheese manufacturers participated in a Listeria monitoring program, whi... more Most Austrian dairies and cheese manufacturers participated in a Listeria monitoring program, which was established after the first reports of dairy product-associated listeriosis outbreaks more than thirty years ago. Within the Listeria monitoring program, up to 800 mL of product-associated liquids such as cheese smear or brine are processed in a semi-quantitative approach to increase epidemiological sensitivity. A sampling strategy within cheese production, which detects environmental contamination before it results in problematic food contamination, has benefits for food safety management. The liquid-based sampling strategy was implemented by both industrial cheese makers and small-scale dairies located in the mountainous region of Western Austria. This report considers more than 12,000 Listeria spp. examinations of liquid-based samples in the 2009 to 2018 timeframe. Overall, the occurrence of L. monocytogenes in smear liquid samples was 1.29% and 1.55% (n = 5043 and n = 7194 tes...
The stress response of 11 strains of Listeria monocytogenes to oxidative stress was studied. The ... more The stress response of 11 strains of Listeria monocytogenes to oxidative stress was studied. The strains included ST1, ST5, ST7, ST6, ST9, ST87, ST199 and ST321 and were isolated from diverse food processing environments (a meat factory, a dairy plant and a seafood company) and sample types (floor, wall, drain, boxes, food products and water machine). Isolates were exposed to two oxidizing agents: 13.8 mM cumene hydroperoxide (CHP) and 100 mM hydrogen peroxide (H2O2) at 10 °C and 37 °C. Temperature affected the oxidative stress response as cells treated at 10 °C survived better than those treated at 37 °C. H2O2 at 37 °C was the condition tested resulting in poorest L. monocytogenes survival. Strains belonging to STs of Lineage I (ST5, ST6, ST87, ST1) were more resistant to oxidative stress than those of Lineage II (ST7, ST9, ST199 and ST321), with the exception of ST7 that showed tolerance to H2O2 at 10 °C. Isolates of each ST5 and ST9 from different food industry origins showed dif...
The genus Listeria now comprises up to now 21 recognized species and six subspecies, with L. mono... more The genus Listeria now comprises up to now 21 recognized species and six subspecies, with L. monocytogenes and L. innocua as the most prevalent sensu stricto associated species. Reports focusing on the challenges in Listeria detection and confirmation are available, especially from food-associated environmental samples. L. innocua is more prevalent in the food processing environment (FPE) than L. monocytogenes and has been shown to have a growth advantage in selective enrichment and agar media. Until now, the adaptive nature of L. innocua in FPEs has not been fully elucidated and potential persistence in the FPE has not been observed. Therefore, the aim of this study is to characterize L. innocua (n = 139) and L. monocytogenes (n = 81) isolated from FPEs and cheese products collected at five dairy processing facilities (A–E) at geno- and phenotypic levels. Biochemical profiling was conducted for all L. monocytogenes and the majority of L. innocua (n = 124) isolates and included a rh...
Staphylococcus aureus causing persistent, recurrent bovine intramammary infections are still a ma... more Staphylococcus aureus causing persistent, recurrent bovine intramammary infections are still a major challenge to dairy farming. Generally, one or a few clonal lineages are predominant in dairy herds, indicating animal-to-animal transfers and the existence of distinct pathotypic traits. The aim of this study was to determine if long term persistence and spreading of S. aureus are associated with specific phenotypic traits, including cellular invasion, cytotoxicity and biofilm formation. Mastitis isolates were collected over a 3-years period from a single dairy herd, resulting in two persistent subtypes, the high within-herd prevalent subtype ST9 (CC9)-methicillin-susceptible S. aureus (MSSA), designated HP/ST9, and the low within-herd prevalent subtype ST504 (CC705)-MSSA, designated LP/ST504. Characterization of the two different coexisting persistent subtypes showed that the following phenotypic traits are particularly associated with high within-herd prevalence: lack of capsular p...
ABSTRACTWe studied the colonization and distribution of Listeria monocytogenes in a heavily conta... more ABSTRACTWe studied the colonization and distribution of Listeria monocytogenes in a heavily contaminated poultry processing plant over a 1-year period. A total of 180 nonfood contact surfaces, 70 food contact surfaces, 29 personnel, and 40 food samples were analyzed. L. monocytogenes isolates were subtyped by PCR serotyping, pulsed-field gel electrophoresis (PFGE), and multilocus sequence typing. L. monocytogenes was detected in samples collected at every visit to the plant, and 43.8% (visit 4) to 65.6% (visit 7) of samples were positive, for an overall prevalence of 55.2%. The deboning area had the highest prevalence of positive samples (83.3%), and the processing area had the highest diversity of PFGE types. Ninety percent of the final products were positive for L. monocytogenes. Most of the isolates belonged to well-known persistent L. monocytogenes sequence types (ST9 and ST121). This study illustrates a well-established L. monocytogenes contamination problem in a poultry proces...
Unregulated international flow of foods poses a danger to human health, as it may be contaminated... more Unregulated international flow of foods poses a danger to human health, as it may be contaminated with pathogens. Recent studies have investigated neglected routes of pathogen transmission and reported the occurrence of Listeria monocytogenes in food illegally imported into the European Union (EU), either confiscated at four international airports or sold illegally on the Romanian black market. In this study we investigated the genotype diversity and the amino acid sequence variability of three main virulence factors of 57 L. monocytogenes isolates. These isolates were derived from 1474 food samples illegally imported into the EU and originated from 17 different countries. Multilocus sequence typing revealed 16 different sequence types (STs) indicating moderate genotype diversity. The most prevalent STs were ST2, ST9, and ST121. The pulsed-field gel electrophoresis (PFGE) analysis resulted in 34 unique pulsotypes. PFGE types assigned to the most prevalent STs (ST2, ST9, and ST121) w...
Staphylococcus aureus is one of the most important contagious mastitis pathogens in dairy cattle.... more Staphylococcus aureus is one of the most important contagious mastitis pathogens in dairy cattle. Due to its zoonotic potential, control of S. aureus is not only of great economic importance in the dairy industry but also a significant public health concern. The aim of this study was to decipher the potential of bovine udder associated S. aureus as reservoir for S. aureus contamination in dairy production and processing. From 18 farms, delivering their milk to an alpine dairy plant for the production of smeared semi-hard and hard cheese. one thousand hundred seventy six one thousand hundred seventy six quarter milk (QM) samples of all cows in lactation (n = 294) and representative samples form bulk tank milk (BTM) of all farms were surveyed for coagulase positive (CPS) and coagulase negative Staphylococci (CNS). Furthermore, samples from different steps of the cheese manufacturing process were tested for CPS and CNS. As revealed by chemometric-assisted FTIR spectroscopy and molecular subtyping (spa typing and multi locus sequence typing), dairy cattle represent indeed an important, yet underreported, entrance point of S. aureus into the dairy chain. Our data clearly show that certain S. aureus subtypes are present in primary production as well as in the cheese processing at the dairy plant. However, although a considerable diversity of S. aureus subtypes was observed in QM and BTM at the farms, only certain S. aureus subtypes were able to enter and persist in the cheese manufacturing at the dairy plant and could be isolated from cheese until day 14 of ripening. Farm strains belonging to the FTIR cluster B1 and B3, which show genetic characteristics (t2953, ST8, enterotoxin profile: sea/sed/sej) of the recently described S. aureus genotype B, most successfully contaminated the cheese production at the dairy plant. Thus, our study fosters the hypothesis that genotype B S. aureus represent a specific challenge in control of S. aureus in the dairy chain that requires effective clearance strategies and hygienic measures already in primary production to avoid a potential transfer of enterotoxic strains or enterotoxins into the dairy processing and the final retail product.
Campylobacteriosis is the leading zoonosis in the European Union with the majority of cases attri... more Campylobacteriosis is the leading zoonosis in the European Union with the majority of cases attributed to Campylobacter jejuni. Although the disease is usually selflimiting, some severe cases need to be treated with antibiotics, primarily macrolides and quinolones. However, the resistance to the latter is reaching alarming levels in most of the EU countries. To shed light on the expansion of antibiotic resistance in central Europe, we have investigated genetic similarity across 178 ciprofloxacin-resistant C. jejuni mostly isolated in Slovenia, Austria and Germany. We performed comparative genetic similarity analyses using allelic types of seven multilocus sequence typing housekeeping genes, and single nucleotide polymorphisms of a quinolone resistance determining region located within the DNA gyrase subunit A gene. This analysis revealed high genetic similarity of isolates from clonal complex ST-21 that carry gyrA allelic type 1 in all three of these central-European countries, suggesting these ciprofloxacin resistant isolates arose from a recent common ancestor and are spread clonally.
We investigated the pathogenicity, invasiveness, and genetic relatedness of 17 clinicalListeria m... more We investigated the pathogenicity, invasiveness, and genetic relatedness of 17 clinicalListeria monocytogenesstains isolated over a period of nine years (2006–2014). All isolates were phenotypically characterised and growth patterns were determined. The antimicrobial susceptibility ofL. monocytogenesisolates was determined in E-tests. Invasion assays were performed with epithelial HeLa cells. Finally,L. monocytogenesisolates were subtyped by PFGE and MLST. All isolates had similar phenotypic characteristics (β-haemolysis and lecithinase activity), and three types of growth curve were observed. Bacterial recovery rates after invasion assays ranged from 0.09% to 7.26% (1.62 ± 0.46). MLST identified 11 sequence types (STs), and 14 PFGE profiles were obtained, indicating a high degree of genetic diversity. Genetic studies unequivocally revealed the occurrence of one outbreak of listeriosis in humans that had not previously been reported. This outbreak occurred in October 2009 and affect...
Abstract: This chapter is focused on reported outbreaks of infectious disease linked to consumpti... more Abstract: This chapter is focused on reported outbreaks of infectious disease linked to consumption of contaminated milk and milk products. In many cases the pathogens are inactivated during the pasteurisation process, but recontamination after pasteurisation or process failures bring about the presence of these pathogens in pasteurised milk and milk products. Furthermore, an extendend summary of current European, US and international regulations for micriobiological requirements in milk and milk products is provided. Sections 4.3 and 4.4 comprise an overview on conventional microbiological methods and additional new developed techniques for the detection and enumeration of the most relevant milkborne pathogens.
The food-borne pathogen Listeria (L.) monocytogenes is often found in food production environment... more The food-borne pathogen Listeria (L.) monocytogenes is often found in food production environments. Thus, controlling the occurrence of L. monocytogenes in food production is a great challenge for food safety. Among a great diversity of L. monocytogenes strains from food production, particularly strains belonging to sequence type (ST)121 are prevalent. The molecular reasons for the abundance of ST121 strains are however currently unknown. We therefore determined the genome sequences of three L. monocytogenes ST121 strains: 6179 and 4423, which persisted for up to 8 years in food production plants in Ireland and Austria, and of the strain 3253 and compared them with available L. monocytogenes ST121 genomes. Our results show that the ST121 genomes are highly similar to each other and show a tremendously high degree of conservation among some of their prophages and particularly among their plasmids. This remarkably high level of conservation among prophages and plasmids suggests that s...
Methods in molecular biology (Clifton, N.J.), 2014
The Listeria monitoring program for Austrian cheese factories was established in 1988. The basic ... more The Listeria monitoring program for Austrian cheese factories was established in 1988. The basic idea is to control the introduction of L. monocytogenes into the food processing environment, preventing the pathogen from contaminating the food under processing. The Austrian Listeria monitoring program comprises four levels of investigation, dealing with routine monitoring of samples and consequences of finding a positive sample. Preventive quality control concepts attempt to detect a foodborne hazard along the food processing chain, prior to food delivery, retailing, and consumption. The implementation of a preventive food safety concept provokes a deepened insight by the manufacturers into problems concerning food safety. The development of preventive quality assurance strategies contributes to the national food safety status and protects public health.
Sampling approaches following the dairy chain, including microbiological hygiene status of critic... more Sampling approaches following the dairy chain, including microbiological hygiene status of critical processing steps and physicochemical parameters, contribute to our understanding of howStaphylococcus aureuscontamination risks can be minimised. Such a sampling approach was adopted in this study, together with rapid culture-independent quantification ofStaph. aureusto supplement standard microbiological methods. A regional cheese production chain, involving 18 farms, was sampled on two separate occasions. Overall, 51·4% of bulk milk samples were found to beStaph. aureuspositive, most of them (34·3%) at the limit of culture-based detection.Staph. aureuspositive samples >100 cfu/ml were recorded in 17·1% of bulk milk samples collected mainly during the sampling in November. A higher number ofStaph. aureuspositive bulk milk samples (94·3%) were detected after applying the culture-independent approach. A concentration effect ofStaph. aureuswas observed during curd processing.Staph. a...
The EU has issued several directives and regulations pertaining to the importation of animals and... more The EU has issued several directives and regulations pertaining to the importation of animals and products of animal origin (POAO) and veterinary controls on importation. Unfortunately, little information is available concerning associated risks and no attempts have been made to collect baseline data on the actual prevalence of zoonotic agents in POAO carried by travellers. To meet these challenges the EU recently introduced and financed a research project "PROMISE". Its main objectives were to assess the risks involved when foodborne pathogens are introduced to the EU via uncontrolled imports. With special permission of the Austrian health authorities, spot-checks were made of the luggage of 61,355 passengers from 240 flights from non-EU countries arriving at the Vienna International Airport (VIE airport). Over a period of eight months (August 2012 through March 2013) 1473 POAO items were confiscated. A total of 600 samples were suitable for Salmonella spp., Campylobacter spp., verotoxigenic Escherichia coli and Listeria monocytogenes prevalence analysis. Foodborne pathogens could be detected in 5% (30/600) of all samples. The highest prevalence was attributed to L. monocytogenes, at 2.5%, followed by VTEC and Salmonella spp. at 1.3% and 1.2%, respectively. Campylobacter spp. was not present in any of the 600 samples. Multi-locus sequence typing (MLST) of L. monocytogenes revealed that current sequence types (ST) corresponded to the worldwide most present clonal complexes 1, 2, 3, 5, 9, and 121. Generally, L. monocytogenes ST9 was the predominant allelic profile, which was mainly isolated from Turkish meat products.
SummaryThe aim of this study was to determine the prevalence of Listeria spp. and Listeria monocy... more SummaryThe aim of this study was to determine the prevalence of Listeria spp. and Listeria monocytogenes (L. monocytogenes) in urban public lavatories and on shoe soles of facility patrons in a European capital city. More than 91% of all municipal public lavatories in Vienna close to public hubs were included in this study. Overall, 373 swab samples of public lavatories and shoes of facility patrons were enriched, according to ISO 11290‐1. Listeria monocytogenes isolates were subtyped using pulsed‐field gel electrophoresis. A total of 24 samples were positive for Listeria spp., yielding an overall prevalence of 6.4% (24/373). Listeria monocytogenes was found in 2.1% (8/373) of all samples. Swabs from lavatories in parks, container lavatories and lavatories at markets had the highest prevalences of 20.7% (6/29), 20% (2/10) and 12.5% (1/8) Listeria spp., respectively. These detection rates were statistically significantly higher than those associated with lavatories in shopping centre...
In this study, the performance of four alternative selective chromogenic B. cereus agar was compa... more In this study, the performance of four alternative selective chromogenic B. cereus agar was compared to the reference mannitol-yolk polymyxin (MYP) agar (ISO 7932) using inclusion and exclusion test strains (n = 110) and by analyzing naturally contaminated milk and other food samples (n = 64). Subsequently, the panC group affiliation and toxin gene profile of Bacillus cereus senso lato (s.l.) isolates were determined. Our results corroborate that the overall best performing media CHROMagar™ B. cereus (93.6% inclusivity; 82.7% exclusivity) and BACARA® (98.2% inclusivity, 62.7% exclusivity) are more sensitive and specific compared to Brilliance™ B. cereus, MYP and ChromoSelect Bacillus Agar. Both media allow unequivocal detection of B. cereus with low risks of misidentification. Media containing ß-D-glucosidase for the detection of presumptive B. cereus may form atypical colony morphologies resulting in a false negative evaluation of the sample. Naturally contaminated samples presente...
Listeria monocytogenes is an environmentally adapted saprophyte that can change into a human and ... more Listeria monocytogenes is an environmentally adapted saprophyte that can change into a human and animal bacterial pathogen with zoonotic potential through several regulatory systems. In this review, the focus is on the occurrence of Listeria sensu stricto and sensu lato in different ecological niches, the detection methods, and their analytical limitations. It also highlights the occurrence of L. monocytogenes genotypes in the environment (soil, water, and wildlife), reflects on the molecular determinants of L. monocytogenes for the saprophytic lifestyle and the potential for antibiotic resistance. In particular, the strain-specific properties with which some genotypes circulate in wastewater, surface water, soil, wildlife, and agricultural environments are of particular interest for the continuously updating risk analysis.
Nucleotide sequence-based methods focusing on the single-nucleotide polymorphisms (SNPs) of Liste... more Nucleotide sequence-based methods focusing on the single-nucleotide polymorphisms (SNPs) of Listeria monocytogenes and L. innocua housekeeping genes (multilocus sequence typing) and in the core genome (core genome MLST) facilitate the rapid and interlaboratory comparison in open accessible databases as provided by Institute Pasteur ( https://bigsdb.web.pasteur.fr/listeria/listeria.html ). Strains can be compared on a global level and help to track forward and trace backward pathogen contamination events in food processing facilities and in outbreak scenarios.
Most Austrian dairies and cheese manufacturers participated in a Listeria monitoring program, whi... more Most Austrian dairies and cheese manufacturers participated in a Listeria monitoring program, which was established after the first reports of dairy product-associated listeriosis outbreaks more than thirty years ago. Within the Listeria monitoring program, up to 800 mL of product-associated liquids such as cheese smear or brine are processed in a semi-quantitative approach to increase epidemiological sensitivity. A sampling strategy within cheese production, which detects environmental contamination before it results in problematic food contamination, has benefits for food safety management. The liquid-based sampling strategy was implemented by both industrial cheese makers and small-scale dairies located in the mountainous region of Western Austria. This report considers more than 12,000 Listeria spp. examinations of liquid-based samples in the 2009 to 2018 timeframe. Overall, the occurrence of L. monocytogenes in smear liquid samples was 1.29% and 1.55% (n = 5043 and n = 7194 tes...
The stress response of 11 strains of Listeria monocytogenes to oxidative stress was studied. The ... more The stress response of 11 strains of Listeria monocytogenes to oxidative stress was studied. The strains included ST1, ST5, ST7, ST6, ST9, ST87, ST199 and ST321 and were isolated from diverse food processing environments (a meat factory, a dairy plant and a seafood company) and sample types (floor, wall, drain, boxes, food products and water machine). Isolates were exposed to two oxidizing agents: 13.8 mM cumene hydroperoxide (CHP) and 100 mM hydrogen peroxide (H2O2) at 10 °C and 37 °C. Temperature affected the oxidative stress response as cells treated at 10 °C survived better than those treated at 37 °C. H2O2 at 37 °C was the condition tested resulting in poorest L. monocytogenes survival. Strains belonging to STs of Lineage I (ST5, ST6, ST87, ST1) were more resistant to oxidative stress than those of Lineage II (ST7, ST9, ST199 and ST321), with the exception of ST7 that showed tolerance to H2O2 at 10 °C. Isolates of each ST5 and ST9 from different food industry origins showed dif...
The genus Listeria now comprises up to now 21 recognized species and six subspecies, with L. mono... more The genus Listeria now comprises up to now 21 recognized species and six subspecies, with L. monocytogenes and L. innocua as the most prevalent sensu stricto associated species. Reports focusing on the challenges in Listeria detection and confirmation are available, especially from food-associated environmental samples. L. innocua is more prevalent in the food processing environment (FPE) than L. monocytogenes and has been shown to have a growth advantage in selective enrichment and agar media. Until now, the adaptive nature of L. innocua in FPEs has not been fully elucidated and potential persistence in the FPE has not been observed. Therefore, the aim of this study is to characterize L. innocua (n = 139) and L. monocytogenes (n = 81) isolated from FPEs and cheese products collected at five dairy processing facilities (A–E) at geno- and phenotypic levels. Biochemical profiling was conducted for all L. monocytogenes and the majority of L. innocua (n = 124) isolates and included a rh...
Staphylococcus aureus causing persistent, recurrent bovine intramammary infections are still a ma... more Staphylococcus aureus causing persistent, recurrent bovine intramammary infections are still a major challenge to dairy farming. Generally, one or a few clonal lineages are predominant in dairy herds, indicating animal-to-animal transfers and the existence of distinct pathotypic traits. The aim of this study was to determine if long term persistence and spreading of S. aureus are associated with specific phenotypic traits, including cellular invasion, cytotoxicity and biofilm formation. Mastitis isolates were collected over a 3-years period from a single dairy herd, resulting in two persistent subtypes, the high within-herd prevalent subtype ST9 (CC9)-methicillin-susceptible S. aureus (MSSA), designated HP/ST9, and the low within-herd prevalent subtype ST504 (CC705)-MSSA, designated LP/ST504. Characterization of the two different coexisting persistent subtypes showed that the following phenotypic traits are particularly associated with high within-herd prevalence: lack of capsular p...
ABSTRACTWe studied the colonization and distribution of Listeria monocytogenes in a heavily conta... more ABSTRACTWe studied the colonization and distribution of Listeria monocytogenes in a heavily contaminated poultry processing plant over a 1-year period. A total of 180 nonfood contact surfaces, 70 food contact surfaces, 29 personnel, and 40 food samples were analyzed. L. monocytogenes isolates were subtyped by PCR serotyping, pulsed-field gel electrophoresis (PFGE), and multilocus sequence typing. L. monocytogenes was detected in samples collected at every visit to the plant, and 43.8% (visit 4) to 65.6% (visit 7) of samples were positive, for an overall prevalence of 55.2%. The deboning area had the highest prevalence of positive samples (83.3%), and the processing area had the highest diversity of PFGE types. Ninety percent of the final products were positive for L. monocytogenes. Most of the isolates belonged to well-known persistent L. monocytogenes sequence types (ST9 and ST121). This study illustrates a well-established L. monocytogenes contamination problem in a poultry proces...
Unregulated international flow of foods poses a danger to human health, as it may be contaminated... more Unregulated international flow of foods poses a danger to human health, as it may be contaminated with pathogens. Recent studies have investigated neglected routes of pathogen transmission and reported the occurrence of Listeria monocytogenes in food illegally imported into the European Union (EU), either confiscated at four international airports or sold illegally on the Romanian black market. In this study we investigated the genotype diversity and the amino acid sequence variability of three main virulence factors of 57 L. monocytogenes isolates. These isolates were derived from 1474 food samples illegally imported into the EU and originated from 17 different countries. Multilocus sequence typing revealed 16 different sequence types (STs) indicating moderate genotype diversity. The most prevalent STs were ST2, ST9, and ST121. The pulsed-field gel electrophoresis (PFGE) analysis resulted in 34 unique pulsotypes. PFGE types assigned to the most prevalent STs (ST2, ST9, and ST121) w...
Staphylococcus aureus is one of the most important contagious mastitis pathogens in dairy cattle.... more Staphylococcus aureus is one of the most important contagious mastitis pathogens in dairy cattle. Due to its zoonotic potential, control of S. aureus is not only of great economic importance in the dairy industry but also a significant public health concern. The aim of this study was to decipher the potential of bovine udder associated S. aureus as reservoir for S. aureus contamination in dairy production and processing. From 18 farms, delivering their milk to an alpine dairy plant for the production of smeared semi-hard and hard cheese. one thousand hundred seventy six one thousand hundred seventy six quarter milk (QM) samples of all cows in lactation (n = 294) and representative samples form bulk tank milk (BTM) of all farms were surveyed for coagulase positive (CPS) and coagulase negative Staphylococci (CNS). Furthermore, samples from different steps of the cheese manufacturing process were tested for CPS and CNS. As revealed by chemometric-assisted FTIR spectroscopy and molecular subtyping (spa typing and multi locus sequence typing), dairy cattle represent indeed an important, yet underreported, entrance point of S. aureus into the dairy chain. Our data clearly show that certain S. aureus subtypes are present in primary production as well as in the cheese processing at the dairy plant. However, although a considerable diversity of S. aureus subtypes was observed in QM and BTM at the farms, only certain S. aureus subtypes were able to enter and persist in the cheese manufacturing at the dairy plant and could be isolated from cheese until day 14 of ripening. Farm strains belonging to the FTIR cluster B1 and B3, which show genetic characteristics (t2953, ST8, enterotoxin profile: sea/sed/sej) of the recently described S. aureus genotype B, most successfully contaminated the cheese production at the dairy plant. Thus, our study fosters the hypothesis that genotype B S. aureus represent a specific challenge in control of S. aureus in the dairy chain that requires effective clearance strategies and hygienic measures already in primary production to avoid a potential transfer of enterotoxic strains or enterotoxins into the dairy processing and the final retail product.
Campylobacteriosis is the leading zoonosis in the European Union with the majority of cases attri... more Campylobacteriosis is the leading zoonosis in the European Union with the majority of cases attributed to Campylobacter jejuni. Although the disease is usually selflimiting, some severe cases need to be treated with antibiotics, primarily macrolides and quinolones. However, the resistance to the latter is reaching alarming levels in most of the EU countries. To shed light on the expansion of antibiotic resistance in central Europe, we have investigated genetic similarity across 178 ciprofloxacin-resistant C. jejuni mostly isolated in Slovenia, Austria and Germany. We performed comparative genetic similarity analyses using allelic types of seven multilocus sequence typing housekeeping genes, and single nucleotide polymorphisms of a quinolone resistance determining region located within the DNA gyrase subunit A gene. This analysis revealed high genetic similarity of isolates from clonal complex ST-21 that carry gyrA allelic type 1 in all three of these central-European countries, suggesting these ciprofloxacin resistant isolates arose from a recent common ancestor and are spread clonally.
We investigated the pathogenicity, invasiveness, and genetic relatedness of 17 clinicalListeria m... more We investigated the pathogenicity, invasiveness, and genetic relatedness of 17 clinicalListeria monocytogenesstains isolated over a period of nine years (2006–2014). All isolates were phenotypically characterised and growth patterns were determined. The antimicrobial susceptibility ofL. monocytogenesisolates was determined in E-tests. Invasion assays were performed with epithelial HeLa cells. Finally,L. monocytogenesisolates were subtyped by PFGE and MLST. All isolates had similar phenotypic characteristics (β-haemolysis and lecithinase activity), and three types of growth curve were observed. Bacterial recovery rates after invasion assays ranged from 0.09% to 7.26% (1.62 ± 0.46). MLST identified 11 sequence types (STs), and 14 PFGE profiles were obtained, indicating a high degree of genetic diversity. Genetic studies unequivocally revealed the occurrence of one outbreak of listeriosis in humans that had not previously been reported. This outbreak occurred in October 2009 and affect...
Abstract: This chapter is focused on reported outbreaks of infectious disease linked to consumpti... more Abstract: This chapter is focused on reported outbreaks of infectious disease linked to consumption of contaminated milk and milk products. In many cases the pathogens are inactivated during the pasteurisation process, but recontamination after pasteurisation or process failures bring about the presence of these pathogens in pasteurised milk and milk products. Furthermore, an extendend summary of current European, US and international regulations for micriobiological requirements in milk and milk products is provided. Sections 4.3 and 4.4 comprise an overview on conventional microbiological methods and additional new developed techniques for the detection and enumeration of the most relevant milkborne pathogens.
The food-borne pathogen Listeria (L.) monocytogenes is often found in food production environment... more The food-borne pathogen Listeria (L.) monocytogenes is often found in food production environments. Thus, controlling the occurrence of L. monocytogenes in food production is a great challenge for food safety. Among a great diversity of L. monocytogenes strains from food production, particularly strains belonging to sequence type (ST)121 are prevalent. The molecular reasons for the abundance of ST121 strains are however currently unknown. We therefore determined the genome sequences of three L. monocytogenes ST121 strains: 6179 and 4423, which persisted for up to 8 years in food production plants in Ireland and Austria, and of the strain 3253 and compared them with available L. monocytogenes ST121 genomes. Our results show that the ST121 genomes are highly similar to each other and show a tremendously high degree of conservation among some of their prophages and particularly among their plasmids. This remarkably high level of conservation among prophages and plasmids suggests that s...
Methods in molecular biology (Clifton, N.J.), 2014
The Listeria monitoring program for Austrian cheese factories was established in 1988. The basic ... more The Listeria monitoring program for Austrian cheese factories was established in 1988. The basic idea is to control the introduction of L. monocytogenes into the food processing environment, preventing the pathogen from contaminating the food under processing. The Austrian Listeria monitoring program comprises four levels of investigation, dealing with routine monitoring of samples and consequences of finding a positive sample. Preventive quality control concepts attempt to detect a foodborne hazard along the food processing chain, prior to food delivery, retailing, and consumption. The implementation of a preventive food safety concept provokes a deepened insight by the manufacturers into problems concerning food safety. The development of preventive quality assurance strategies contributes to the national food safety status and protects public health.
Sampling approaches following the dairy chain, including microbiological hygiene status of critic... more Sampling approaches following the dairy chain, including microbiological hygiene status of critical processing steps and physicochemical parameters, contribute to our understanding of howStaphylococcus aureuscontamination risks can be minimised. Such a sampling approach was adopted in this study, together with rapid culture-independent quantification ofStaph. aureusto supplement standard microbiological methods. A regional cheese production chain, involving 18 farms, was sampled on two separate occasions. Overall, 51·4% of bulk milk samples were found to beStaph. aureuspositive, most of them (34·3%) at the limit of culture-based detection.Staph. aureuspositive samples >100 cfu/ml were recorded in 17·1% of bulk milk samples collected mainly during the sampling in November. A higher number ofStaph. aureuspositive bulk milk samples (94·3%) were detected after applying the culture-independent approach. A concentration effect ofStaph. aureuswas observed during curd processing.Staph. a...
The EU has issued several directives and regulations pertaining to the importation of animals and... more The EU has issued several directives and regulations pertaining to the importation of animals and products of animal origin (POAO) and veterinary controls on importation. Unfortunately, little information is available concerning associated risks and no attempts have been made to collect baseline data on the actual prevalence of zoonotic agents in POAO carried by travellers. To meet these challenges the EU recently introduced and financed a research project "PROMISE". Its main objectives were to assess the risks involved when foodborne pathogens are introduced to the EU via uncontrolled imports. With special permission of the Austrian health authorities, spot-checks were made of the luggage of 61,355 passengers from 240 flights from non-EU countries arriving at the Vienna International Airport (VIE airport). Over a period of eight months (August 2012 through March 2013) 1473 POAO items were confiscated. A total of 600 samples were suitable for Salmonella spp., Campylobacter spp., verotoxigenic Escherichia coli and Listeria monocytogenes prevalence analysis. Foodborne pathogens could be detected in 5% (30/600) of all samples. The highest prevalence was attributed to L. monocytogenes, at 2.5%, followed by VTEC and Salmonella spp. at 1.3% and 1.2%, respectively. Campylobacter spp. was not present in any of the 600 samples. Multi-locus sequence typing (MLST) of L. monocytogenes revealed that current sequence types (ST) corresponded to the worldwide most present clonal complexes 1, 2, 3, 5, 9, and 121. Generally, L. monocytogenes ST9 was the predominant allelic profile, which was mainly isolated from Turkish meat products.
SummaryThe aim of this study was to determine the prevalence of Listeria spp. and Listeria monocy... more SummaryThe aim of this study was to determine the prevalence of Listeria spp. and Listeria monocytogenes (L. monocytogenes) in urban public lavatories and on shoe soles of facility patrons in a European capital city. More than 91% of all municipal public lavatories in Vienna close to public hubs were included in this study. Overall, 373 swab samples of public lavatories and shoes of facility patrons were enriched, according to ISO 11290‐1. Listeria monocytogenes isolates were subtyped using pulsed‐field gel electrophoresis. A total of 24 samples were positive for Listeria spp., yielding an overall prevalence of 6.4% (24/373). Listeria monocytogenes was found in 2.1% (8/373) of all samples. Swabs from lavatories in parks, container lavatories and lavatories at markets had the highest prevalences of 20.7% (6/29), 20% (2/10) and 12.5% (1/8) Listeria spp., respectively. These detection rates were statistically significantly higher than those associated with lavatories in shopping centre...
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Papers by Beatrix Stessl