Context Of Food Safety Research Articles

Promoting social sustainability and transforming foodways through alternative food spaces: a case study of Beijing farmers’ market

ABSTRACT Organic farmers’ markets have rapidly expanded in major Chinese cities as a response to food safety concerns, with the concept of “organic” serving as the driving force behind this movement. This study examines the Beijing Farmers’ Market, the first organic farmers’ market in mainland China, to explore its role in reconfiguring foodways and promoting social sustainability. The development of the Beijing Farmers’ Market is driven by the changing values, attitudes, and ideologies of both producers and consumers, that set them apart from the mainstream food system. While this study is situated within the broader context of food safety concerns that contribute to the emergence of such markets, it primarily focuses on how the market fosters trust and proactive relationships between health-conscious consumers and new farmers committed to safe food production. These dynamic connections are shaped by shifting lifestyles among both producers and consumers. This research highlights how food serves as a socio-spatial space for reconnecting people with nature and cultivating food communities, engagement and resilience.

Quantitative microbial risk assessment of Salmonella in fresh chicken patties

Quantitative microbial risk assessment (QMRA) has witnessed rapid development within the context of food safety in recent years. As a means of contributing to these advancements, a QMRA for Salmonella spp. in fresh chicken patties for the general European Union (EU) population was developed. A two-dimensional (Second Order) Monte-Carlo simulation method was used for separating variability and uncertainty of model’s parameters. The stages of industrial processing, retail storage, domestic storage, and cooking in the domestic environment were considered in the exposure assessment. For hazard characterization, a dose-response model was developed by combining 8 published dose-response models using a Pert distribution for describing uncertainty. The QMRA model predicted a mean probability of illness of 1.19*10−4 (5.28*10−5 – 3.57*10−4 95 % C.I.), and a mean annual number of illnesses per 100,000 people of 2.13 (0.96 – 6.59 95 % C.I.). Moreover, sensitivity analysis was performed, and variability in cooking preferences was found to be the most influential model parameter (r = −0.39), followed by dose-response related variability (r = 0.22), and variability in the concentration of Salmonella spp. at the time of introduction at the processing facility (r = 0.11). Various mitigation strategy scenarios were tested, from which, “increasing the internal temperature of cooking” and “decreasing shelf life” were estimated to be the most effective in reducing the predicted risk of illness. Salmonella-related illnesses exhibit particularly high severity, making them some of the most prominent zoonotic diseases in the EU. Regular monitoring of this hazard in order to further highlight its related parameters and causes is a necessary procedure. This study not only provides an updated assessment of Salmonella spp. risk associated with chicken patties, but also facilitates the identification of crucial targets for scientific investigation and implementation of real-world intervention strategies.

The Occurrence of Freshwater Fish-Borne Zoonotic Helminths in Italy and Neighbouring Countries: A Systematic Review.

In recent years, the consumption of fish products has surged in European countries, being an essential part of a healthy diet. Despite representing a small part of EU production, freshwater fisheries hold considerable significance for lake-dwelling populations and tourists seeking traditional dishes. This increased fish consumption has brought to light potential health risks associated with fish-borne zoonotic helminths (FBZHs), now acknowledged as global food-borne parasites. Fish-borne zoonotic helminths belong to various taxonomic groups, including nematodes (Anisakidae), trematodes (Opisthorchiidae and Heterophyidae), and cestodes (Diphyllobothriidae). More than 50 species of FBZH are known to cause human infections, derived from eating raw or undercooked aquatic foods containing viable parasites. Despite increased attention, FBZHs remain relatively neglected compared to other food-borne pathogens due to factors like chronic disease progression and under-diagnosis. This systematic review concentrates on the prevalence of six freshwater FBZHs (Clinostomum complanatum, Contracaecum rudolphii, Dibothriocephalus latus, Eustrongylides excisus, Opisthorchis felineus, and Pseudamphistomum truncatum) in Italy and neighbouring countries. The study explores the expansion of these parasites, analysing their biological and epidemiological aspects, and the factors that influence their proliferation, such as the increased cormorant population and the lake eutrophication phenomena. In summary, this research highlights the necessity for further research, the development of spatial databases, and the establishment of a unified European policy to effectively manage these multifaceted health concerns. It strongly advocates adopting a One-Health approach to address the growing incidence of parasitic zoonoses within the context of food safety in EU countries.

Asparagine and dough quality: Gluten strength factors in hard red spring wheat

AbstractBackground and ObjectivesSafety aspects regarding free asparagine concentration in wheat arise because of its acrylamide formation potential in baked wheat products. Free asparagine concentration in wheat is affected by environment, genotype, and nitrogen and sulfur fertilization, which also affect wheat protein content and composition. Changing protein content and composition affects gluten strength, altering product quality. This study investigated how gluten strength was affected by environment, genotype, and fertilization treatments grown under commercially relevant conditions.FindingsEnvironment predominated in affecting dough extensibility and genotype primarily affected resistance to extension, with minimal effects from fertilization treatments. Differences in extensigraph parameters at two resting times segregated genotypes according to gluten strength.ConclusionsFactors affecting free asparagine accumulation in wheat also affect gluten strength. Genotype and environment require primary attention since they play an important role in both wheat safety and quality.Significance and NoveltyThe lack of studies investigating the impact of factors affecting free asparagine concentration on the gluten strength of wheat has implications for the global wheat market. From a novel perspective on dough extensibility and resistance to extension, the effects of environment, genotype, and fertilization on the gluten strength of commercially important wheat varieties are considered in a food safety context.

A socio-technical approach to food safety incident analysis using the AcciMap model in the hospitality sector

A theory-based systems approach, such as AcciMap accident analysis, has been widely used over the years in multiple safety critical sectors such as the nuclear, petrochemical, aviation and railway industries to provide a detailed understanding of complex systems and the chain of events contributing to accidents resulting from system failure. However, despite its advantages, the use of a systems approach in the food safety context has to date been limited. The purpose of this study was to investigate three established norovirus incidents using the AcciMap accident analysis approach to determine its efficacy at informing the design of food safety policies following a norovirus outbreak to prevent reoccurrence. This approach was found to be of value in analysing norovirus outbreaks. The findings of the AcciMap analysis reveal the norovirus outbreaks were not the outcome of a single causal incident, but a chain of events and interactions that involved governmental failure to control and enforce safety regulations and the impact on managerial and individual behaviours at a lower level in the system. The analysis identified the common contributory factors such as poor inspections, lack of regular monitoring of quality of water supply, inadequate management of wastewater and ineffective communication that led to each incident across the hierarchical levels within a socio-technical system. The value of using the AcciMap approach is that it does not constrain the analysis to individual components or particular types of incident allowing for a more holistic and interconnected risk assessment.

USE OF BACTERIOPHAGES AS A TARGET SPECIFIC THERAPY AGAINST FOOD-BORNE PATHOGENS IN FOOD INDUSTRY- A REVIEW

Within the context of food safety, the use of modern antimicrobial technologies to inactivate the food-borne pathogens is not infallible, as proven by the ceaseless increase in the food-borne illnesses. The extensive use of antibiotics to minimize the microbial contamination further results in the evolution of antibiotic-resistant bacterial pathogens. Moreover, some of the decontamination methods frequently used in the food industry cannot be directly applied to fresh fruits and vegetables, and RTE meals. Therefore, efforts are being made by researchers in developing a new approach so as to control the bacterial contamination. Consequently, bacteriophages have evolved as the safe, green, effective and natural alternative for treatment and complete destruction of pathogens in food industry. The review provides a general description about bacteriophages and has mainly focused on their potential use as antimicrobials during the postharvest processing of foods. Numerous research papers, review papers, book chapters and other relevant literature was used for writing this review paper.

Comparison of Statistical Methods for Identifying Risk Factors for Salmonella Contamination of Whole Chicken Carcasses

The complexity of the food system makes analyzing microbiological data from food studies challenging because many of the assumptions (e.g., linear relationship between independent and dependent variable and independence of observations) associated with common analytical approaches (e.g., analysis of variance) are violated. Repeated sampling within an establishment introduces longitudinal correlation that must be accounted for during analyses. In this study, statistical methods for clustered or correlated data were used to determine how correlation impacts conclusions and to compare how assumptions associated with statistical methods impact the appropriateness of these methods within the context of food safety. Risk factor analyses for Salmonella contamination of whole chicken carcasses were conducted as a case study with regulatory data collected by the U.S. Department of Agriculture Food Safety and Inspection Service between May 2015 and December 2019 from 203 regulated establishments. Three models, generalized estimating equation, random effects, and logistic, were fit to Salmonella presence or absence data with establishment demographics and inspection history included as potential covariates. Beta parameter estimates and their standard errors and odds ratios and their 95% confidence intervals were compared across models. Conclusions drawn from the three models differed with respect to geographic region, whether the chicken establishment also slaughters turkeys, and establishment noncompliance with 9 CFR §417.4 (hazard analysis critical control point system validation, verification, and reassessment) in the 84 days leading up to sample collection. The results of this study reveal the need to consider clustering and correlation when analyzing food microbiological data, provide context for selecting a statistical method, and suggest that generalized estimating equation and random effects models are preferrable over logistic regression when analyzing correlated food data. These results support a renewed focus on statistical methodology in food safety.

Spirulina phenolic compounds: natural food additives with antimicrobial properties

Food safety is a scientific discipline to ensure consumers’ safety and prevent food-related harms, hazards, or risks along the entire food supply chain. Although fish and seafood products are the best means for securing food and nutrition in a population, they are also highly perishable, being vulnerable to pathogenic bacteria and fungi. This review thus aimed to provide updated scientific information on the role of the phenolic compounds of Spirulina as food additives, and their antimicrobial activities in fish and seafood products, in a food safety context. Recent applications of Spirulina phenolic compounds showed good results in contaminated fish and seafood products. Different studies have concluded that Spirulina phenolic compounds such as polyphenols, C-phycocyanin, γ-linolenic acid, fatty acids, and the combination of lauric and palmitoleic acids have antimicrobial activity in eliminating and/or controlling the growth of pathogenic bacteria such as Escherichia coli, Pseudomonas aeruginosa, and Bacillus subtilis, and fungi such as Aspergillus flavus and Aspergillus niger. Overall, Spirulina can be considered as an emerging natural food additive with antimicrobial activities against pathogenic fungi and bacteria.

USE OF BACTERIOPHAGES AS A TARGET SPECIFIC THERAPY AGAINST FOOD-BORNE PATHOGENS IN FOOD INDUSTRY- A REVIEW: Bacteriophage

Within the context of food safety, the use of modern antimicrobial technologies to inactivate the food-borne pathogens is not infallible, as proven by the ceaseless increase in the food-borne illnesses. The extensive use of antibiotics to minimize the microbial contamination further results in the evolution of antibiotic-resistant bacterial pathogens. Moreover, some of the decontamination methods frequently used in the food industry cannot be directly applied to fresh fruits and vegetables, and RTE meals. Therefore, efforts are being made by researchers in developing a new approach so as to control the bacterial contamination. Consequently, bacteriophages have evolved as the safe, green, effective and natural alternative for treatment and complete destruction of pathogens in food industry. The review provides a general description about bacteriophages and has mainly focused on their potential use as antimicrobials during the postharvest processing of foods. Numerous research papers, review papers, book chapters and other relevant literature was used for writing this review paper.

Consumers value healthy eating and environmental responsibility: how negative food contexts aid decision-making

Abstract The research aims to explore consumers’ intention toward traceable food in the dual context of the animal disease outbreak (i.e. African Swine Fever - ASF) and the current food safety issues. An extended Theory of Planned Behavior (TPB) model was used to predict purchase intention/attitude toward traceable pork. The study employed the structural equation modeling (SEM) to extract insights from the dataset of 230 students in Vietnam. The current detrimental context of food safety deemed promoting the consumption of traceable food, a risk-mitigating alternative. Healthy and environmentally responsible consumers held positive standpoints toward traceable pork as a solution to fulfill their responsibility with the environment and healthy eating lifestyle. The extended model was promising in explaining 52% of the variance of the purchase intention. The study confirmed both the positive impacts of the long-lasting unsafe food issues and the short-term animal disease outbreak on intention toward traceable food. To promote traceable food, industry food marketers should tailor their marketing and communication strategies to target healthy and responsible eaters.

Bovine Hemoglobin Enzymatic Hydrolysis by a New Eco-Efficient Process-Part II: Production of Bioactive Peptides.

Bovine cruor, a slaughterhouse waste, was mainly composed of hemoglobin, a protein rich in antibacterial and antioxidant peptides after its hydrolysis. In the current context of food safety, such bioactive peptides derived from enzymatic hydrolysis of hemoglobin represent potential promising preservatives for the food sector. In this work, the hemoglobin hydrolysis to produce bioactive peptides was performed in a regulated pH medium without the use of chemical solvents and by an eco-efficient process: electrodialysis with bipolar membrane (EDBM). Bipolar/monopolar (anionic or cationic) configuration using the H+ and OH− generated by the bipolar membranes to regulate the pH was investigated. The aim of this study was to present and identify the bioactive peptides produced by EDBM in comparison with conventional hydrolysis and to identify their biological activity. The use of the EDBM for the enzymatic hydrolysis of hemoglobin has allowed for the production and identification of 17 bioactive peptides. Hydrolysates obtained by EDBM showed an excellent antimicrobial activity against six strains, antioxidant activity measured by four different tests and for the first time anti-fungal activities against five yeasts and mold strains. Consequently, this enzymatic hydrolysis carried out in regulated pH medium with bipolar membranes could provide bioactive peptides presenting antibacterial, antifungal and antioxidant interest.

Outbreaks of Escherichia coli O157:H7 Infections Linked to Romaine Lettuce in Canada from 2008 to 2018: An Analysis of Food Safety Context

Foodborne diseases are a major cause of illness in Canada. One of the main pathogens causing cases and outbreaks of foodborne illness in Canada is Escherichia coli O157:H7. From 2008 to 2018, 11 outbreaks of E. coli O157:H7 infection in Canada were linked to leafy greens, including 7 (63.6%) linked to romaine lettuce, 2 (18.2%) linked to iceberg lettuce, and 2 (18.2%) linked to other or unspecified types of leafy greens. The consumption of lettuce in Canada, the behavior of E. coli O157:H7 on lettuce leaves, and the production practices used for romaine and iceberg lettuce do not seem to explain why a higher number of outbreaks of E. coli O157:H7 infection were linked to romaine than to iceberg lettuce. However, the difference in the shape of iceberg and romaine lettuce heads could be an important factor. Among the seven outbreaks linked to romaine lettuce in Canada between 2008 and 2018, an eastern distribution of cases was observed. Cases from western provinces were reported only twice. The consumption of romaine and iceberg lettuce by the Canadian population does not seem to explain the eastern distribution of cases observed, but the commercial distribution, travel distances, and the storage practices used for lettuce may be important factors. In the past 10 years, the majority of the outbreaks of E. coli O157:H7 infection linked to romaine lettuce occurred during the spring (March to June) and fall (September to December). The timing of these outbreaks may be explained by the availability of lettuce in Canada, the growing region transition periods in the United States, and the seasonality in the prevalence of E. coli O157:H7. The consumption of romaine lettuce by the Canadian population does not explain the timing of the outbreaks observed.

Slaughterhouse By-Product Valorization: Hydrolysis Degree Modification for Higher Antimicrobial Recovery by Electroseparation

In the actual context of food safety and circular economy, the separation of the α137–141 antimicrobial peptide (TSKYR, 653 Da) coming from the hydrolysis of bovine hemoglobin, a non-valorized slaughterhouse by-product, and its reuse as a preservative agent for food products during storage would be of major interest. However, depending on the degree of hydrolysis (DH) of the bovine hemoglobin, the TSKYR peptide environment into the hydrolysate will be different and would impact its recovery yield by electrodialysis with ultrafiltration membranes (EDUF), a hybrid and eco-friendly technology. In this context, five DHs (3, 5, 10, 13 and 18%) were investigated to study the influence of the peptide populations on the α137–141 selective separation by EDUF. It appeared from these results that the most appropriate DH was of 5% since a lower population of peptides between 500 and 1000 Da was present and the subunits of hemoglobin were already digested. Hence, an enrichment factor of about 13 folds for the α137–141 was obtained in comparison with the initial hydrolysate. At our knowledge, it was the first time that the effect of the hydrolysis degree of a protein is demonstrated to impact preferentially the separation of a specific peptide by EDUF. Graphic

Inactivation of Single Strains of Listeria monocytogenes and Salmonella Typhimurium Planktonic Cells Biofilms With Plasma Activated Liquids.

Recent research has proven the ability of cold atmospheric plasma (CAP) for assuring food safety. A more flexible and transportable alternative is the use of plasma activated liquids (PAL), which are also known to have antimicrobial properties. However, within the context of food safety, little is known on its potential regarding decontamination. This research therefore focusses on identifying the impact of (i) the microbial species and its cell type (planktonic cells or biofilms), (ii) the CAP settings (i.e., gas composition and generation time) and (iii) PAL related factors (treatment time and PAL age) on the technologies efficacy. Cell densities were monitored using the plate counting technique for which the results were analyzed by means of predictive inactivation models. Moreover, the pH and the concentrations of long-lived species (i.e., hydrogen peroxide, nitrite, and nitrate) were measured to characterize the PAL solutions. The results indicated that although the type of pathogen impacted the efficacy of the treatment, mainly the cell mode had an important effect. The presence of oxygen in the operating gas ensured the generation of PAL solutions with a higher antimicrobial activity. Moreover, to ensure a good microbial inactivation, PAL generation times needed to be sufficiently long. Both CAP related factors resulted in a higher amount of long-lived species, enhancing the inactivation. For 30 min. PAL generation using O2, this resulted in log reductions up to 3.9 for biofilms or 5.8 for planktonic cells. However, loss of the PAL activity for stored solutions, together with the frequent appearance of a tailing phase in the inactivation kinetics, hinted at the importance of the short-lived species generated. Different factors, related to (i) the pathogen and its cell mode, (ii) the CAP settings and (iii) PAL related factors, proved to impact the antimicrobial efficacy of the solutions and should be considered with respect to future applications of the PAL technology.

A genomic data mining pipeline for 15 species of the genus Olea.

In the big data era, conventional bioinformatics seems to fail in managing the full extent of the available genomic information. The current study is focused on olive tree species and the collection and analysis of genetic and genomic data, which are fragmented in various depositories. Extra virgin olive oil is classified as a medical food, due to nutraceutical benefits and its protective properties against cancer, cardiovascular diseases, age-related diseases, neurodegenerative disorders, and many other diseases. Extensive studies have reported the benefits of olive oil on human health. However, available data at the nucleotide sequence level are highly unstructured. Towards this aim, we describe an in-silico approach that combines methods from data mining and machine learning pipelines to ontology classification and semantic annotation. Fusing and analysing all available olive tree data is a step of uttermost importance in classifying and characterising the various cultivars, towards a comprehensive approach under the context of food safety and public health.

Comparison between random forest and gradient boosting machine methods for predicting Listeria spp. prevalence in the environment of pastured poultry farms.

Foodborne pathogens such as Listeria spp. contain the ability to survive and multiply in poultry farming environments, which provides a route of contamination for poultry processing environments and final poultry products. An understanding of the effect of meteorological variables on the prevalence of Listeria spp. in the farming environment is lacking. Soil and feces samples were collected from 11 pastured poultry farms from 2014 to 2017. Random forest (RF) and gradient boosting machine (GBM) predictive models were generated to describe and predict Listeria spp. prevalence in feces and soil samples based on meteorological factors at the farming location. This study attempted to demonstrate the use of GBM models in a food safety context and compare their use to RF models. Both feces models performed very well, with area under the curve (AUC) values of 0.905 and 0.855 for the RF and GBM models, respectively. The soil GBM model outperformed the RF model with AUCs of 0.873 and 0.700, respectively. The developed models can be used to predict the prevalence of Listeria spp. in pastured poultry farm environments and should be of great use to poultry farmers, producers, and risk managers.

Using Farm Practice Variables as Predictors of Listeria spp. Prevalence in Pastured Poultry Farms

Predictive models offer food scientists, farmers, and processors tools to help identify variables that lead to an increase in the food safety risk of a product. Foodborne pathogens, such as Listeria spp., pose a major problem for the pastured poultry industry. Currently, there is a lack of understanding of what farm practices lead to higher prevalence of Listeria spp. This study constructed random forest (RF) models to predict the prevalence of Listeria spp. in pastured poultry farming environments and the final broiler product based on major farm practices and variables. Feces, soil, and whole carcass rinse samples were collected from 11 farms in the southeastern United States and evaluated for Listeria spp. presence. The preharvest sample RF model identified the time of year and age of the broiler flock at time of sampling as factors of increased probability of Listeria spp. presence in feces and soil samples. The final product RF model identified brood feed and the presence of chlorine in processing rinse water as the two most important variables associated with an increased likelihood of Listeria spp. presence. Both the preharvest RF model and final sample RF model performed well on a held-out test set, with area under the receiver operating characteristic curve values of 0.867 and 0.887, respectively. The presented models showed the usefulness of RF models in a food safety context. Both RF models will help pastured poultry farmers and processors guide control strategies to manage Listeria contamination in pastured poultry farms and products.

Linking food safety training with whistle-blowing

PurposeThe purpose of this paper is to develop a validated food safety whistle-blowing scale and examine how training influences food handlers’ whistle-blowing, the mediating roles of job satisfaction and food safety self-efficacy and the moderating roles of organization type and gender.Design/methodology/approachA sample of 734 food handlers from the food service industry and 306 food handlers from the food processing industry were recruited. A two-step psychometric process was conducted to validate the scale, and a moderated mediation model was used to examine the mechanisms through which food safety training influences whistle-blowing.FindingsThe results showed that job satisfaction and self-efficacy fully mediated the relationship between training and whistle-blowing. Organization type moderated the first step of the two indirect paths and gender moderated the second step, thereby supporting the mediated-moderated model.Practical implicationsThe results indicate the influence of food safety training programs on whistle-blowing behaviors and suggest other methods of enhancing employee whistle-blowing through human resource management.Originality/valueA validated scale is lacking to measure whistle-blowing in the food safety context, and little research has examined the influence of human resource practices on whistle-blowing. This study provides meaningful insights for researchers by developing and validating food safety the whistle-blowing scale, connecting training with whistle-blowing, and provides useful information for practitioners by offering the methods of enhancing whistleblowing in both the food processing and food service industry.

Untargeted food contaminant detection using UHPLC-HRMS combined with multivariate analysis: Feasibility study on tea.

Powerful data pretreatment strategies inspired from the field of metabolomics were adapted to chemical food safety context to enable samples discrimination by multivariate methods based on low abundance ions. A highly automated workflow was produced. The open-source XCMS package was used and efficient data filtration strategies were set up. Data were treated using Independent Components Analysis, and data mining strategies developed to automatically detect and annotate ions of low abundance by coupling blind data exploration strategies with a broad scale database approach. Our method was efficient in discriminating tea samples based on their contamination levels (even at 10 µg.kg-1) and detecting unexpected impurities in the spiking mix. Several "tracer" contaminants were considered, covering a broad range of physicochemical properties and structural diversity with overall 66% detected and annotated blindly. The methodology was successfully applied to a data set exhibiting only 3 "tracer" contaminants (at 50 µg.kg-1) and more product diversity.

Quantifying the Responses of Three Bacillus cereus Strains in Isothermal Conditions and During Spray Drying of Different Carrier Agents.

Spray drying is a widely used method for producing milk powder. This process is not aimed to cause microbial inactivation, thus sporeforming bacteria may be abundant in the microbiota of milk powder. The first aim of this study was to determine the inactivation kinetics parameters in capillary tubes of three Bacillus cereus strains (436, B63, 540) in three menstrua (whole milk, phosphate buffer, and talc suspension) at 90, 100, and 110°C. D-values for B. cereus in the three menstrua were not significantly different at the highest tested temperature (p > 0.05). Thus, talc was chosen as a carrier agent to allow the recovery of B. cereus from spray dried materials given its low interference on inactivation kinetics. B. cereus spores were also inoculated in whole milk and skim milk following spray drying at 95, 105, and 110°C (outlet temperature). After the spray drying runs, B. cereus spores were counted and the number of decimal reductions (γ) calculated. A correlation between the small diameter of the particles with the survival of spores of three B. cereus strains was found, and B. cereus 436 presented consistently the lowest γ no matter temperature and a carrier agent. The highest γ was found when talc powder was used, which suggest that this carrier agent does not protect B. cereus spores during spray drying. Spray drying of milk can lead to up to 4 γ (strain 540) of B. cereus spores but depending on the strain less than one γ (strain 436) could be observed. This study contributes to the knowledge on the microbiology of low water activity foods by providing novel findings regarding the fate of three B. cereus strains to different spray drying conditions. Acknowledging the variability of inactivation of B. cereus during spray drying is key in the current context of food safety in which the quantification of effects of unit operations must be known for the validation of processes and development of more robust formulations.