Persistent Source Of Contamination Research Articles

Tracing of persistent Listeria monocytogenes contamination in ewe's milk farm.

Ewe's milk farm production is permanently associated with the risk of contamination by pathogenic bacteria, including Listeria monocytogenes. In the present study, the prevalence and diversity of L. monocytogenes strains repeatedly isolated from tank ewe's milk and the milking environment on a farm in Slovakia during a prolonged period were investigated to identify the source of potentially persistent contamination. A total of 140 samples along the milk production chain were collected during an 18-month period. From all these samples, 45 samples were found L. monocytogenes positive with 90.3% positivity of tank milk samples (28 positive samples from 31 analysed). Pulsed-field gel electrophoresis profiling resulted in strain discrimination into six profiles with one pulsotype (NS1) corresponding to MLST-ST14 being predominant. A total of 17 proportionally selected L. monocytogenes isolates, including 11 NS1/ST14 isolates, were subjected to whole genome sequencing. Resulted data were used to compare the genomes diversity and to confirm the persistent contamination when <10 allelic differences threshold in cgMLST analysis was applied. The source of persistent contamination was localized inside the milking apparatus, probably in shelters that were very difficult to clean. Despite great efforts, the ewe's milk contamination could not be eliminated during the reporting period.

Antibiotic susceptibility pattern of fish pathogens: A new approach of emerging the bacterial resistance through biofilm formation in in-vitro condition.

The ability of many bacteria to adhere on the host surfaces and forming biofilms has major implications in a wide variety of industries including the food industry, where biofilms may create a persistent source of contamination. In the same environmental condition, the multiple bacterial species can closely interact with each other and may easily enhance their drug resistance capability, which finally increases the multi-drug resistant (MDR) attribute of the species. The present study examined whether the mixed-species biofilm possesses any impact on the enhancement of the antibiotic resistance of the planktonic or single-cell bacterial isolates present in the fish samples. In this regard, Cyprinus rubrofuscus (Koi), Heteropneustes fossilis (Shing) and Mystus vittatus (Tengra) fishes were collected and subjected to form an in vitro biofilm by shaking condition into the wise bath. The drug-resistant pattern was determined by the Kirby Bauer technique. All the samples exhibited a huge array (up to 107 cfu/ml or g) of bacteria such as E. coli, Klebsiella spp., Vibrio spp., Salmonella spp., Proteus spp. and Staphylococcus spp. The isolates from both the bulk samples and their corresponding biofilms were subjected to antibiogram assay using antibiotics such as Ampicillin (10µg), Erythromycin (15μg), Streptomycin (STP 10μg), Oxacillin (10µg), Nalidixic acid (30µg). Before biofilm formation, few of the isolates were found to be sensitive and few were resistant against the antibiotics. But when the species were isolated from the biofilm the sensitive one acquired drug resistance and resistant strain unveiled more resistance towards the same antibiotics. The present study revealed extensive bacterial contamination in fish samples among those some were resistant against the supplied drugs. After the formation of multi-species biofilm, the isolates became more resistant against the same drugs that is alarming for consumers and major obstacles to maintain sustainable health.

Phenotypic and Genetic Determination of Biofilm Formation in Heat Resistant Escherichia coli Possessing the Locus of Heat Resistance.

Despite the effectiveness of thermal inactivation processes, Escherichia coli biofilms continue to be a persistent source of contamination in food processing environments. E. coli strains possessing the locus of heat resistance are a novel food safety threat and raises the question of whether these strains can also form biofilms. The objectives of this study were to determine biofilm formation in heat resistant E. coli isolates from clinical and environmental origins using an in-house, two-component apparatus and to characterize biofilm formation-associated genes in the isolates using whole genome sequencing. Optimal conditions for biofilm formation in each of the heat resistant isolates were determined by manipulating inoculum size, nutrient concentration, and temperature conditions. Biofilm formation in the heat resistant isolates was detected at temperatures of 24 °C and 37 °C but not at 4 °C. Furthermore, biofilm formation was observed in all environmental isolates but only one clinical isolate despite shared profiles in biofilm formation-associated genes encoded by the isolates from both sources. The circulation of heat resistant E. coli isolates with multi-stress tolerance capabilities in environments related to food processing signify that such strains may be a serious food safety and public health risk.

Identifying septic pollution exposure routes during a waterborne norovirus outbreak - A new application for human-associated microbial source tracking qPCR

In June 2017, the Pennsylvania Department of Health (PADOH) was notified of multiple norovirus outbreaks associated with 179 ill individuals who attended separate events held at an outdoor venue and campground over a month period. Epidemiologic investigations were unable to identify a single exposure route and therefore unable to determine whether there was a persistent contamination source to target for exposure mitigation. Norovirus was detected in a fresh recreational water designated swimming area and a drinking water well. A hydrogeological site evaluation suggested a nearby septic leach field as a potential contamination source via ground water infiltration. Geological characterization revealed a steep dip of the bedrock beneath the septic leach field toward the well, providing a viral transport pathway in a geologic medium not previously documented as high risk for viral ground water contamination. The human-associated microbial source tracking (MST) genetic marker, HF183, was used as a microbial tracer to demonstrate the hydrogeological connection between the malfunctioning septic system, drinking water well, and recreational water area. Based on environmental investigation findings, venue management and local public health officials implemented a series of outbreak prevention strategies including discontinuing the use of the contaminated well, issuing a permit for a new drinking water well, increasing portable toilet and handwashing station availability, and promoting proper hand hygiene. Despite the outbreaks at the venue and evidence of ground water contamination impacting nearby recreational water and the drinking water well, no new norovirus cases were reported during a large event one week after implementing prevention practices. This investigation highlights a new application for human-associated MST methods to trace hydrological connections between multiple fecal pollutant exposure routes in an outbreak scenario. In turn, pollutant source information can be used to develop effective intervention practices to mitigate exposure and prevent future outbreaks associated with human fecal contaminated waters.

Insights into Cronobacter sakazakii Biofilm Formation and Control Strategies in the Food Industry

Cronobacter sakazakii (C. sakazakii) is a foodborne opportunistic pathogen that can cause life-threatening invasive diseases, such as necrotizing enterocolitis, meningitis, and sepsis in infants. The potential risk of C. sakazakii contamination of powdered infant formula (PIF) is an issue that has attracted considerable attention from manufacturers, regulators, and consumers. C. sakazakii biofilms on the surfaces of equipment and in diverse food-production environments constitute a mode of cell growth that protects the pathogen from hostile environments, and are an important source of persistent contamination of food products. Bacterial biofilms are difficult to remove due to their resistant properties. Conventional cleaning and sterilizing procedures may be insufficient for biofilm control, and may lead to further biofilm development and dispersal. Consequently, novel control strategies are being developed, such as nanotechnology-based delivery systems, interspecies interactions, antimicrobial molecules of microbial origin, natural extracts, and phages. This review focuses on describing the mechanisms underlying the biofilm formation and behavior of C. sakazakii and discussing potential control strategies.

Population Dynamics of Thermotolerant Campylobacter in Broilers Reared on Reused Litter.

A study using sentinel broiler chickens was performed to address Campylobacter persistence in litter that was reused for successive flocks. Cloacal swabs, litter, drag swabs, darkling beetles, feed, and drinking water were weekly sampled and analyzed by standard microbiological procedures. Thermotolerant Campylobacter isolated strains were confirmed by polymerase chain reaction and subtyped by pulsed-field gel electrophoresis analysis. Campylobacter was not detected in samples collected immediately after downtime between broiler flocks. However, Campylobacter-positive samples were first detected at 21 d. After Campylobacter was initially isolated from the cloacal swabs, reused litter, drag swabs, or darkling beetles, these samples remained Campylobacter positive in the following weeks until the end of the rearing period. Campylobacter-positive cloacal swabs obtained from sentinel broilers ranged from 97.3% to 100% at 42 d. All isolated strains were identified as Campylobacter jejuni. Among the subtypes identified, an indistinguishable C. jejuni strain was predominant in sentinel broilers and was also detected in the other environmental samples analyzed, suggesting a common and persistent contamination source within the flocks. Sentinel broilers may have contributed to amplify the Campylobacter level, maintaining flock and broiler house contamination until the end of the production cycle.

Outbreak of Salmonella enterica serotype Poona in infants linked to persistent Salmonella contamination in an infant formula manufacturing facility, France, August 2018 to February 2019.

We describe a Salmonella Poona outbreak involving 31 infant cases in France. Following outbreak detection on 18 January 2019, consumption of rice-based infant formula manufactured at a facility in Spain was identified as the probable cause, leading to a recall on 24 January. Whole genome sequencing analysis linked present outbreak isolates to a 2010–11 S. Poona outbreak in Spain associated with formula manufactured in the same facility, indicating a persistent source of contamination.

Impact of the Isolation Source on the Biofilm Formation Characteristics of Bacillus cereus.

The human pathogen and food spoiler Bacillus cereus can form biofilms that act as a persistent source of contamination, which is of public health concern. This study aimed to understand how the source of isolation might affect the behavior of biofilm formation. Biofilm formation abilities of 56 strains of B. cereus isolated from different environments, including human food poisoning, farm, and food, were determined. Crystal violet assay results revealed significant (p < 0.05) differences in biofilm formation abilities among the strains isolated from different sources only at an early stage of incubation. However, strain origin showed no impact on later stage of biofilm formation. Next, correlation of the group of isolates on the basis of their biofilm-forming abilities with the number of sessile cells, sporulation, and extracellular polymeric substance (EPS) formation was determined. The number of sessile cells and spores in biofilms was greatly influenced by the groups of isolates that formed dense, moderate, and weak biofilms. The contribution of extracellular DNA and/or proteins to EPS formation was also positively correlated with biofilm formation abilities. Our results that the source of isolation had significant impact on biofilm formation might provide important information to develop strategies to control B. cereus biofilm formation.

Improving transfer functions to describe radiocesium wash-off fluxes for the Niida River by a Bayesian approach

This paper proposed methodological refinements of the generic transfer function approach to reconstruct radiocesium wash-off fluxes from contaminated catchments, by the integration of hydrological descriptors (passed volume of water, flow rate fluctuations and antecedent flow conditions). The approach was applied to the Niida River (Fukushima prefecture, Japan) for the period 03/2011-03/2015, for which daily flow rate (m3/s) and infrequent total radiocesium concentration (Bq/L) values were available from literature.Three models were defined, generic TF (Φ0), flow-corrected time variant (Φ1) and antecedent-flow corrected variant (Φ2). Calibration of these models’ parameters was performed with a Bayesian approach because it is particularly adapted to limited datasets and censored information, and it provides parameters distributions.The model selection showed strong evidence of model Φ2 (indicated by marginal likelihood), which integrates current and recent hydrology in its formulation, and lower prediction errors (indicated by RMSE and ME). Models Φ1 and Φ2 better described wash-off dynamics compared to model Φ0, due to the inclusion of one or several hydrological descriptors.From March 2011 to March 2015, model Φ2 estimated 137Cs export from Niida catchment between 0.32 and 0.67 TBq, with a median value of 0.49 TBq, which represents around 0.27% of the initial fallout and could represent a significant source-term to the Ocean compared to the direct release from Fukushima Dai-ichi Nuclear Power Plant (FDNPP). Moreover the remaining 99% of the initial radiocesium fallout within the catchment may constitute a persistent contamination source for wash-off.Although the proposed methodology brought improvements in the assessment of wash-off fluxes, it remains an empirical interpolation method with a limited predictive power, particularly for recent low activities. To improve predictions, modelling approaches require more observed data (particularly more activity values corresponding to more hydrological conditions), and the inclusion of more hydrological descriptors.

Outbreak of Salmonella Enteritidis linked to the consumption of frozen beefburgers received from a food bank and originating from Poland: northern France, December 2014 to April 2015.

A prolonged outbreak of Salmonella enterica serotype Enteritidis occurred in northern France between December 2014 and April 2015. Epidemiological investigations following the initial notification on 30 December 2014 of five cases of salmonellosis (two confirmed S. Enteritidis) in young children residing in the Somme department revealed that all cases frequented the same food bank A. Further epidemiological, microbiological and food trace-back investigations indicated frozen beefburgers as the source of the outbreak and the suspected lot originating from Poland was recalled on 22 January 2015. On 2 March 2015 a second notification of S. Enteritidis cases in the Somme reinitiated investigations that confirmed a link with food bank A and with consumption of frozen beefburgers from the same Polish producer. In the face of a possible persistent source of contamination, all frozen beefburgers distributed by food bank A and from the same origin were blocked on 3 March 2015. Microbiological analyses confirmed contamination by S. Enteritidis of frozen beefburgers from a second lot remaining in cases’ homes. A second recall was initiated on 6 March 2015 and all frozen beefburgers from the Polish producer remain blocked after analyses identified additional contaminated lots over several months of production.

Prevalence and Amounts of Salmonella Found on Raw California Inshell Pistachios

After harvest, pistachios are hulled with mechanical abrasion and then separated in a float tank containing water; the nuts that float (∼15%; floaters) and those that sink (∼85%; sinkers) are dried and stored separately. To determine the prevalence of Salmonella in pistachios, a total of 3,966 samples (1,032 floaters and 2,934 sinkers) were collected within 4 months of the 2010, 2011, and 2012 harvests from storage silos (12 samples from each silo, in most cases) and were stored at 4°C; 100-g subsamples were enriched for the presence of Salmonella. Twenty-one of the floater samples and 11 of the sinker samples were positive for Salmonella: 2.0% prevalence (95% confidence interval [CI], 1.3 to 3.1%) and 0.37% prevalence (95% CI, 0.21 to 0.67%), respectively, for a weighted average prevalence of 0.61%. Levels of Salmonella were determined for positive samples using a most-probable-number (MPN) method with multiple 50-g, three 5.6-g, and three 0.56-g subsamples. Geometric mean levels of Salmonella in floaters and sinkers were 0.66 MPN/100 g (0.14 to 5.3 MPN/100 g) and 0.18 MPN/100 g (0.10 to 0.62 MPN/100 g), respectively. Seven different serovars were identified among the isolates, with nine pulsed-field gel electrophoresis fingerprints; as many as four serovars were isolated from some samples. Salmonella serovars Montevideo (44%), Enteritidis (19%), Senftenberg (16%), Worthington (12%), and Liverpool (9.4%) were most commonly isolated from the initial 100-g samples. The prevalence and levels of Salmonella in pistachios are within those observed for other tree nuts, but the limited number of serovars isolated suggests a narrow and persistent contamination source.

209 Risk of Infection Transmission in Curvilinear Array Echoendoscopes: Results of a Prospective Reprocessing and Culture Registry

Outbreaks of multi-drug resistant organisms leading to occasional fatal infections have been attributed to transmission from reprocessed duodenoscopes. The complex design of the elevator mechanism is a recognized challenge for disinfection and has been implicated as the potential source of persistent contamination. Linear echoendoscopes also have an elevator mechanism, however there are no data or recommendations regarding the risk of persistent bacterial contamination of echoendoscopes.

Effects of Ca2+ and Mg2+ on the Biofilm Formation of Cronobacter Sakazakii Strains from Powdered Infant Formula

AbstractBiofilm of Cronobacter sakazakii on food contact surfaces is the important source of persistent contamination in powdered infant formula. This study aimed to assess the effects of Ca2+ and Mg2+ on biofilm formation. Results indicated that the biofilm‐forming ability was strain specific and divalent cation concentrations dependent. Most of C. sakazakii formed the highest amount of biomass at 1.50% MgCl2 (17 strains, 74%) and 0.25% CaCl2 (10 strains, 43.4%). Finally, confocal laser scanning microscope (CLSM) further indicated that effects of divalent cation were attributable to changing properties of extracellular matrix. The findings presented here provide new insights for control of biofilm formation in dairy products.Practical ApplicationsC. sakazakii is an important foodborne pathogen involved in infections of infant. Control of C. sakazakii contamination in powdered infant formula is an intractable problem in production of powdered infant formula. In this study, effects of Ca2+ and Mg2+ on the biofilm formation were performed. Furthermore, the changes of extracellular matrix in biofilm were revealed by CLSM. The results contribute to useful information for development of essential measures for precaution and control of biofilm of C. sakazakii.

Effects of culture conditions on the biofilm formation of Cronobacter sakazakii strains and distribution of genes involved in biofilm formation

Biofilm of Cronobacter sakazakii on the surfaces of equipment and processing environments is the important source of persistent contamination in food samples. Based on 828 tests (23 × 4 × 3 × 3), the biofilm-forming abilities of 23 C. sakazakii isolates were assessed under different pH values, temperatures, and culture time. Biofilm formed by C. sakazakii was evaluated in tryptone soy broth in 96-well plates using crystal violet staining and its quantification was counted using the optical density measurements (OD570 nm). Among the evaluated conditions, C. sakazakii formed highest amount of biofilm at pH 5.0 (9 strains, 39.1%), at 28 °C (23 strains, 100%) and for 48 h (14 strains, 60.9%). In addition, the biofilm under pH7-24 h-28 °C, pH5-24 h-28 °C, and pH7-48 h-28 °C was detected by confocal laser scanning microscopy for detailed analysis of biofilm. Finally, PCR methods were newly developed for detection of the genes involved in biofilm formation. The bcsC, bcsG, flgJ, bcsA, fliD, and flhE were present in 100%, 100%, 91.3%, 95.7%, 100%, and 34.8% of tested strains respectively. Results indicated that the properties of polysaccharides in biofilm formation of C. sakazakii were affected by microbial environments and the biofilm-forming ability was strain-specific. The findings presented here provide useful information for development of efficient procedures against biofilm formation.

Population Diversity of Campylobacter jejuni in Poultry and Its Dynamic of Contamination in Chicken Meat.

This study aimed to analyse the diversity of the Campylobacter jejuni population in broilers and to evaluate the major source of contamination in poultry meat. Eight rearing cycles over one year provided samples from three different broiler farms processed at the same slaughterhouse. A total of 707 C. jejuni were isolated from cloacal swabs before slaughter and from the breast skin of carcasses after slaughter and after chilling. All suspected Campylobacter colonies were identified with PCR assays and C. jejuni was genotyped by sequence analysis of the flaA short variable region (SVR) and by pulsed-field gel electrophoresis (PFGE) using SmaI enzyme. Phenotypic antibiotic resistance profiles were also assayed using minimal inhibitory concentration (MIC). The flocks carried many major C. jejuni clones possibly carrying over the rearing cycles, but cross contamination between farms may happen. Many isolates were resistant to fluoroquinolones, raising an issue of high public concern. Specific Campylobacter populations could be harboured within each poultry farm, with the ability to contaminate chickens during each new cycle. Thus, although biosecurity measures are applied, with a persistent source of contamination, they cannot be efficient. The role of the environment needs further investigation to better address strategies to control Campylobacter.

Quantification of Bacteria on Abiotic Surfaces by Laser Scanning Cytometry: An Automated Approach to Screen the Antifouling Properties of New Surface Coatings

Bacterial biofilms are a persistent source of contamination, and much effort has been invested in developing antifouling surfaces or coatings. A bottleneck in developing such coatings is often the time-consuming task of screening and evaluating a large number of surface materials. An automated high-throughput assay is therefore needed. In this study, we present a promising technique, laser scanning cytometry (LSC), for automated quantification of bacteria on surfaces. The method was evaluated by quantifying young Staphylococcus xylosus biofilms on glass surfaces using LSC and comparing the results with cell counts obtained by fluorescence microscopy. As an example of application, we quantified bacterial adhesion to seven different sol-gel-based coatings on stainless steel. The surface structure and hydrophobicity of the coatings were analyzed using atomic force microscopy and water contact angle measurements. Among the coatings tested, a significant reduction in adhesion of S. xylosus was observed only for one coating, which also had a unique surface microstructure. LSC was particularly sensitive for quantification at low cell densities, and the adhered bacteria could be quantified both as cell number and as area coverage. The method proved to be an excellent alternative to microscopy for fast and reproducible quantification of microbial colonization on abiotic surfaces.

Evaluation of rhamnolipid and surfactin to reduce the adhesion and remove biofilms of individual and mixed cultures of food pathogenic bacteria

Biofilms represent a great concern for food industry, since they can be a source of persistent contamination leading to food spoilage and to the transmission of diseases. To avoid the adhesion of bacteria and the formation of biofilms, an alternative is the pre-conditioning of surfaces using biosurfactants, microbial compounds that can modify the physicochemical properties of surfaces changing bacterial interactions and consequently adhesion. Different concentrations of the biosurfactants, surfactin from Bacillus subtilis and rhamnolipids from Pseudomonas aeruginosa, were evaluated to reduce the adhesion and to disrupt biofilms of food-borne pathogenic bacteria. Individual cultures and mixed cultures of Staphylococcus aureus, Listeria monocytogenes and Salmonella Enteritidis were studied using polystyrene as the model surface. The pre-conditioning with surfactin 0.25% reduced by 42.0% the adhesion of L. monocytogenes and S. Enteritidis, whereas the treatment using rhamnolipids 1.0% reduced by 57.8% adhesion of L. monocytogenes and by 67.8% adhesion of S. aureus to polystyrene.Biosurfactants were less effective to avoid adhesion of mixed cultures of the bacteria when compared with individual cultures. After 2 h contact with surfactin at 0.1% concentration, the pre-formed biofilms of S. aureus were reduced by 63.7%, L. monocytogenesby 95.9%, S. Enteritidis by 35.5% and the mixed culture biofilm by 58.5%. The rhamnolipids at 0.25% concentration removed 58.5% the biofilm of S. aureus, 26.5% of L. monocytogenes, 23.0% of S. Enteritidis and 24.0% the mixed culture after 2 h contact. In general, the increase in concentration of biosurfactants and in the time of contact decreased biofilm removal percentage. These results suggest that surfactin and rhamnolipids can be explored to control the attachment and to disrupt biofilms of individual and mixed cultures of the food-borne pathogens.

Biofilm formation and the food industry, a focus on the bacterial outer surface

The ability of many bacteria to adhere to surfaces and to form biofilms has major implications in a variety of industries including the food industry, where biofilms create a persistent source of contamination. The formation of a biofilm is determined not only by the nature of the attachment surface, but also by the characteristics of the bacterial cell and by environmental factors. This review focuses on the features of the bacterial cell surface such as flagella, surface appendages and polysaccharides that play a role in this process, in particular for bacteria linked to food-processing environments. In addition, some aspects of the attachment surface, biofilm control and eradication will be highlighted.

Reaerosolization of Fluidized Spores in Ventilation Systems

This project examined dry, fluidized spore reaerosolization in a heating, ventilating, and air conditioning duct system. Experiments using spores of Bacillus atrophaeus, a nonpathogenic surrogate for Bacillus anthracis, were conducted to delineate the extent of spore reaerosolization behavior under normal indoor airflow conditions. Short-term (five air-volume exchanges), long-term (up to 21,000 air-volume exchanges), and cycled (on-off) reaerosolization tests were conducted using two common duct materials. Spores were released into the test apparatus in turbulent airflow (Reynolds number, 26,000). After the initial pulse of spores (approximately 10(10) to 10(11) viable spores) was released, high-efficiency particulate air filters were added to the air intake. Airflow was again used to perturb the spores that had previously deposited onto the duct. Resuspension rates on both steel and plastic duct materials were between 10(-3) and 10(-5) per second, which decreased to 10 times less than initial rates within 30 min. Pulsed flow caused an initial spike in spore resuspension concentration that rapidly decreased. The resuspension rates were greater than those predicted by resuspension models for contamination in the environment, a result attributed to surface roughness differences. There was no difference between spore reaerosolization from metal and that from plastic duct surfaces over 5 hours of constant airflow. The spores that deposited onto the duct remained a persistent source of contamination over a period of several hours.

THE SPECIATION OF ARSENIC IN IRON OXIDES IN MINE WASTES FROM THE GIANT GOLD MINE, N.W.T.: APPLICATION OF SYNCHROTRON MICRO-XRD AND MICRO-XANES AT THE GRAIN SCALE

Understanding the solid-phase speciation of arsenic in soils and sediments is important in evaluations of the potential mobility of arsenic and of its bio-availability in the environment. This is especially true in mine-infl uenced environments, where arsenic commonly is present at concentrations two and three orders of magnitude above quality criteria for soils and sediments. Arsenicbearing particulates dispersed through hydraulic transport or aerosol emissions can represent a persistent source of contamination in sediments and soils adjacent to past mining and metallurgical operations. The stability and mobility of arsenic associated with these phases depend on the chemical form and oxidation state of the arsenic and the interaction with post-depositional geochemical conditions. The Giant mine in Yellowknife, Northwest Territories, roasted arsenic-bearing gold ore from 1949 to 1999. The roasting process decomposed arsenic-bearing sulfi des (pyrite and arsenopyrite) to produce a calcine containing fi ne (generally <50 � m) arsenic-bearing iron oxides. We have applied synchrotron As K-edge micro X-ray Absorption Near-Edge Structure