Canadian Food Inspection Agency Research Articles

Characterization of MLST-99 Salmonella Typhimurium and the monophasic variant I:4,[5],12:i:- isolated from Canadian Atlantic coast shellfish.

Salmonella enterica subsp. enterica Typhimurium and its monophasic variant I 1;4,[5],12:i:- (MVST) are responsible for thousands of reported cases of salmonellosis each year in Canada, and countries worldwide. We investigated S. Typhimurium and MVST isolates recovered from raw shellfish harvested in Atlantic Canada by the Canadian Food Inspection Agency (CFIA) over the past decade, to assess the potential impact of these isolates on human illness and to explore possible routes of shellfish contamination. Whole-genome sequence analysis was performed on 210 isolates of S. Typhimurium and MVST recovered from various food sources, including shellfish. The objective was to identify genetic markers linked to ST-99, a sequence type specifically associated with shellfish, which could explain their high prevalence in shellfish. We also investigated the genetic similarity amongst CFIA ST-99 isolates recovered in different years and geographical locations. Finally, the study aimed to enhance the molecular serotyping of ST-99 isolates, as they are serologically classified as MVST but are frequently misidentified as S. Typhimurium through sequence analysis. To ensure recovery of ST-99 from shellfish was not due to favourable growth kinetics, we measured the growth rates of these isolates relative to other Salmonella and determined that ST-99 did not have a faster growth rate and/or shorter lag phase than other Salmonella evaluated. The CFIA ST-99 isolates from shellfish were highly clonal, with up to 81 high-quality single nucleotide variants amongst isolates. ST-99 isolates both within the CFIA collection and those isolated globally carried numerous unique deletions, insertions and mutations in genes, including some considered important for virulence, such as gene deletions in the type VI secretion system. Interestingly, several of these genetic characteristics appear to be unique to North America. Most notably was a large genomic region showing a high prevalence in genomes from Canadian isolates compared to those from the USA. Although the functions of the majority of the proteins encoded within this region remain unknown, the genes umuC and umuD, known to be protective against UV light damage, were present. While this study did not specifically examine the effects of mutations and insertions, results indicate that these isolates may be adapted to survive in specific environments, such as ocean water, where wild birds and/or animals serve as the natural hosts. Our hypothesis is reinforced by a global phylogenetic analysis, which indicates that isolates obtained from North American shellfish and wild birds are infrequently connected to isolates from human sources. These findings suggest a distinct ecological niche for ST-99, potentially indicating their specialization and adaptation to non-human hosts and environments, such as oceanic habitats.

Draft genome sequences of 13 Salmonella enterica subsp. enterica isolates from chickens, cows, and Canadian Salmonella outbreaks.

Salmonella enterica is the etiological agent responsible for salmonellosis. Here, we report the draft whole genome sequences of 13 S. enterica subsp. enterica isolates from chickens and cows, as well as from previous Canadian Salmonella outbreaks investigated by the Canadian Food Inspection Agency.

The Development of a Risk Assessment Model for Inedible Rendering Plants in Canada: Identifying and Selecting Feed Safety-Related Factors.

The Canadian Food Inspection Agency (CFIA) is developing an establishment-based risk assessment model to categorize rendering plants that produce livestock feed ingredients (ERA-Renderer model) according to animal and human health risks (i.e., feed safety risks) and help in determining the allocation of inspection resources based on risk. The aim of the present study was to identify and select feed-safety-related factors and assessment criteria for inclusion in the ERA-Renderer model. First, a literature review was performed to identify evidence-based factors that impact the feed safety risk of livestock feed during its rendering processes. Secondly, a refinement process was applied to retain only those that met the inclusion conditions, such as data availability, lack of ambiguity, and measurability. Finally, an expert panel helped in selecting factors and assessment criteria based on their knowledge and experience in the rendering industry. A final list of 32 factors was developed, of which 4 pertained to the inherent risk of a rendering plant, 8 were related to risk mitigation strategies, and 20 referred to the regulatory compliance of a rendering plant. A total of 179 criteria were defined to assess factors based on practices in the Canadian rendering industry. The results of this study will be used in the next step of the model development to estimate the relative risks of the assessment criteria considering their impact on feed safety. Once implemented, the CFIA's ERA-Renderer model will provide an evidence-based, standardized, and transparent approach to help manage the feed safety risks in Canada's rendering sector.

Evaluation of the protective efficiency of an autogenous Vibrio anguillarum vaccine in lumpfish (Cyclopterus lumpus) under controlled and field conditions in Atlantic Canada

Lumpfish (Cyclopterus lumpus) have become the predominant cleaner fish species used in North American salmon aquaculture. Vibrio anguillarum is a frequent pathogen of lumpfish in Atlantic Canada, and current vaccines against local isolates conferred low to moderate efficacy. This study evaluated the safety and efficiency of a V. anguillarum autogenous vaccine under controlled and field conditions. Two safety trials were conducted following the Canadian Food Inspection Agency (CFIA) regulations before field trial testing. The first safety trial was a common garden assay, and 250 PIT-tsgged lumpfish were used for five treatments per tank (PBS-negative control, Lab vaccine-positive control, autogenous intraperitoneal (IP) vaccine, autogenous dip vaccine, autogenous dip-IP boosted vaccine) in triplicates. Weight, gross pathology, and IgM titers were evaluated to determine the vaccine’s safety. After 10 weeks post-vaccination (wpv), lumpfish were bath-challenged with a lethal dose (1.24 x 106 CFU/mL) of V. anguillarum serotype O2. Survival rate, IgM titers, memory immune response, and cross-immune protection were evaluated. The second safety trial was conducted using a double vaccine dose in two groups, PBS and autogenous IP. The gross pathology score indicated a normal immune response without tissue damage, and no mortality nor fish health issues were observed in the immunized animals. The IP route of administration conferred the highest protection against the V. anguillarum challenge. The autogenous vaccine conferred long-term immunity but did not confer cross-protection against V. anguillarum serotype O1. To evaluate the efficiency of the autogenous vaccine under field conditions, 58,976 naïve lumpfish (8-10 g) were IP immunized with either the autogenous vaccine or the commercial Forte micro IV vaccine. After 7 wpv, the lumpfish were equally distributed into eight sea cages and co-cultivated with Atlantic salmon smolts (60,000 per cage). Lumpfish IgM titers, tissue bacterial loads, mortality, and sea lice count on salmon were monitored. The autogenous vaccine and Forte vaccine confer acceptable protection to the lumpfish in sea cages. No significant difference was observed between lumpfish vaccinated with autogenous and commercial vaccines under field conditions.

An integrated strategy involving high-throughput sequencing to characterize an unknown GM wheat event in Canada.

Glyphosate-resistant wheat plants were discovered in southern Alberta in 2017, representing an unauthorized GM release in Canada. The Canadian Food Inspection Agency undertook a series of experiments to characterize and identify this unknown GM wheat, as well as to develop and validate construct-specific and event-specific qPCR assays. Results of PCR-based assays and Sanger sequencing indicated the presence of CaMV 35S promoter (p35S), Rice Actin 1 intron (RactInt1), CP4-EPSPS gene and nopaline synthase terminator (tNOS) elements in the unknown GM wheat. Genome walking and bead capture strategies, combined with high-throughput sequencing, were used to identify the 5' and 3' wheat junctions and the subsequent mapping of the insert to chromosome 3B of the wheat genome. A probable transformation vector, pMON25497, was recognized, and further testing identified the unknown GM wheat as MON71200 event, one of two events obtained with the pMON25497 vector. The two construct-specific assays targeted the junctions of the RactInt1 and the CP4-EPSPS elements and the CP4-EPSPS and tNOS elements, while the event-specific assay was located at the 3' junction into the wheat genome. Both construct-specific and event-specific assays had limits of detection of 0.10% of MON71200 in a seed pool. As expected, the two construct-specific assays cross-reacted with other wheat and corn events containing the same elements in the same order. No cross-reactivity was observed for the event-specific assay. The integrated strategy employed in this study can serve as a model for other cases when facing similar challenges involving unknown GM events.

Survey of Food Product Labels to Assess Compliance With Labelling Provisions in Canada

Food product labels are a public health tool that guides consumers in making food choices. In December 2016, the Government of Canada amended the Food and Drug Regulations (i.e., nutrition labelling, list of ingredients, and food colour requirements). Health Canada and the Canadian Food Inspection Agency (CFIA) developed an implementation plan that focussed on education and compliance promotion of the new labelling provisions (Canadian Food Inspection Agency, 2022a). This progressive approach allows regulated parties to transition to the new changes gradually. As of December 15, 2021, the former regulations are no longer in effect, and the new labelling requirements apply. This study will survey food labels on products available in the marketplace, regarding the current labelling requirements, and determine whether there is a discrepancy in properly labelled food products between chain versus independent grocery stores of different ethnicity.

An Outbreak of Salmonella Typhimurium Infections Linked to Ready-To-Eat Tofu in Multiple Health Districts - Ontario, Canada, May-July 2021.

From May to mid-August 2021, the Ontario, Canada provincial public health agency, Public Health Ontario, in collaboration with local public health authorities and federal food safety partners, investigated a spatiotemporal cluster of 38 patients with Salmonella Typhimurium infections across multiple public health districts in Ontario. Five (13%) patients were hospitalized; no deaths were reported. The outbreak was linked to consumption of ready-to-eat seasoned tofu from one manufacturer that was distributed to multiple Ontario restaurants. Isolates from the seasoned tofu were within one or fewer allele differences to the outbreak strain by whole genome sequencing. Evidence from food safety investigations conducted by local public health authorities and the Canadian Food Inspection Agency (CFIA) revealed that unsanitary conditions could have led to cross-contamination of the tofu, and insufficient heating of the tofu at the production level likely resulted in failure to eliminate the pathogen. The CFIA issued a food recall for the tofu at hotel, restaurant, and institution levels. Tofu was identified as a novel outbreak-associated food vehicle for S. Typhimurium in this outbreak. Interventions that target the production level and all parts of the supply chain and include additional safeguarding steps that minimize microbial growth are important.

An Outbreak of Salmonella Typhimurium Infections Linked to Ready-To-Eat Tofu in Multiple Health Districts - Ontario, Canada, May-July 2021.

From May to mid-August 2021, the Ontario, Canada provincial public health agency, Public Health Ontario, in collaboration with local public health authorities and federal food safety partners, investigated a spatiotemporal cluster of 38 patients with Salmonella Typhimurium infections across multiple public health districts in Ontario. Five (13%) patients were hospitalized; no deaths were reported. The outbreak was linked to consumption of ready-to-eat seasoned tofu from one manufacturer that was distributed to multiple Ontario restaurants. Isolates from the seasoned tofu were within one or fewer allele differences to the outbreak strain by whole genome sequencing. Evidence from food safety investigations conducted by local public health authorities and the Canadian Food Inspection Agency (CFIA), revealed that unsanitary conditions could have led to cross-contamination of the tofu, and insufficient heating of the tofu at the production level likely resulted in failure to eliminate the pathogen. The CFIA issued a food recall for the tofu at hotel, restaurant, and institution levels. Tofu was identified as a novel outbreak-associated food vehicle for S. Typhimurium in this outbreak. Interventions that target the production level and all parts of the supply chain and include additional safeguarding steps that minimize microbial growth are important.

Distribution of West Nile virus cases in horses reveals different spatiotemporal patterns in eastern and western Canada.

West Nile virus (WNV) became notifiable in horses in 2003 in Canada and has been reported every year since. The objective of this study was to describe the spatiotemporal distribution of WNV in horses between 2003 and 2020 in Canada. The 848 symptomatic and laboratory-confirmed WNV cases in horses reported to the Canadian Food Inspection Agency between 2003 and 2020. Canada was divided into eastern and western regions for analysis. For each case, location and date of notification were captured. Triennial maps were made to describe the spatiotemporal distribution and expansion of reported cases. The association between year and latitude of cases was investigated with simple linear regressions, and space-time clusters were detected with a permutation scan test. Most of the western region showed an extended distribution of WNV cases from 2003 to 2005 and a high recurrence of cases at the census division level. In the eastern region, the expansion of cases was gradual, with new infected census divisions mostly contiguous to previous ones. There was no association between year and latitude of cases. Six spatiotemporal clusters were detected. This study confirmed the endemicity of WNV in parts of both regions with local peaks in risk varying in time. Prevention and control efforts should focus on previously infected areas based on the spatiotemporal regional distribution patterns. Incursions of WNV to new areas should also be anticipated. These findings could also contribute to enhancing monitoring and prevention of WNV infections in an integrated surveillance system.

Two Listeria monocytogenes outbreaks in a cancer centre: onsite food premises and their potential health risk to patients

BackgroundThis report describes two L. monocytogenes outbreak investigations that occurred in March and September of 2018 and that linked illness to a food premises located in an Ontario cancer centre. The cancer centre serves patients from across the province.MethodsIn Ontario, local public health agencies follow up with all reported laboratory-confirmed cases of listeriosis to identify possible sources of disease acquisition and to carry out investigations, including at suspected food premises. The Canadian Food Inspection Agency (CFIA) is notified of any Listeria-positive food product collected in relation to a case. The CFIA traces Listeria-positive product through the food distribution system to identify the contamination source and ensure the implicated manufacturing facility implements corrective measures.ResultsOutbreaks one and two each involved three outbreak-confirmed listeriosis cases. All six cases were considered genetically related by whole genome sequencing (WGS). In both outbreaks, outbreak-confirmed cases reported consuming meals at a food premises located in a cancer centre (food premises A) before illness onset. Various open deli meat samples and, in outbreak two, environmental swabs (primarily from the meat slicer) collected from food premises A were genetically related to the outbreak-confirmed cases. Food premises A closed as a result of the investigations.ConclusionsWhen procuring on-site food premises, healthcare facilities and institutions serving individuals with immuno-compromising conditions should consider the potential health risk of foods available to their patient population.

Production and certification of BOTS-1: bovine muscle-certified reference material for incurred veterinary drug residues.

A freeze-dried bovine muscle-certified reference material (CRM), known as BOTS-1 (DOI: https://doi.org/10.4224/crm.2018.bots-1 ), containing incurred residues of commonly used veterinary drugs was produced and certified for the mass fraction of eight veterinary drug residues. Value assignment was carried out using liquid chromatography tandem mass spectrometry (LC-MS/MS) methods in conjunction with isotope dilution and standard addition approaches involving stable isotope internal standards. Data from the National Research Council of Canada (NRC), Canadian Food Inspection Agency (CFIA), United States Department of Agriculture (USDA), and the Federal Office of Consumer Protection and Food Safety in Germany (BVL) were used for value assignment. Results for two drug residues were also obtained through an international inter-laboratory comparison CCQM-K141/P178 organized under the auspices of the International Bureau of Weights and Measures (BIPM). Quantitative NMR (1H-qNMR) was used to characterize primary standards of all veterinary drugs certified. The certified mass fractions of the veterinary drug residues were 490 ± 100µg/kg for chlorpromazine, 44 ± 4.4µg/kg for ciprofloxacin, 3.3 ± 1.4µg/kg for clenbuterol, 9.5 ± 0.8µg/kg for dexamethasone, 57 ± 4.8µg/kg for enrofloxacin, 3.0 ± 0.4µg/kg for meloxicam, 12.4 ± 1.2µg/kg for ractopamine, and 2290 ± 120µg/kg for sulfadiazine with expanded uncertainties quoted (95% confidence) which include the effects due to between-bottle inhomogeneity, instability during long-term storage and transportation, and characterization.

Simplified Liquid Chromatography–Mass Spectrometry Methods for Gestagen Analysis in Animal Fat and Liver

Gestagens, a class of veterinary drugs also called progestogens, are synthetic hormones used to increase feed efficiency and rate of gain in heifers. The Canadian Food Inspection Agency analyzes progestogens melengestrol acetate (MGA), megestrol acetate, and chlormadinone acetate using liquid chromatography–mass spectrometry (LC–MS). Our conventional gestagen method for kidney fat has many time-consuming steps, including solid-phase extraction. A sample preparation procedure having fewer clean-up steps was developed for routine diagnostic analysis of kidney fat and provided similar results faster, and at lower cost. A confirmatory liver method for gestagens, developed using salt-assisted extraction, employed minimal clean-up steps that resulted in high chemical background at the desired lower limit of quantification (LLOQ). Differential ion mobility spectrometry, specifically high-field asymmetric waveform ion mobility spectrometry (FAIMS), was used to filter chemical background in the gas phase. The effect of the ionization probe position on FAIMS parameters, including sensitivity, is described. With LC-FAIMS-MS, chemical background for each gestagen was virtually eliminated, resulting in a quantitative liver method having the desired 0.6 ng/g LLOQ and estimated limits of detection (LODs) up to 140 times lower than LC-MS. Incurred MGA samples, analyzed using kidney fat and liver methods from the same animal, show levels within the quantitative ranges of both methods.

Attributing salmonellosis cases to foodborne, animal contact and waterborne routes using the microbial subtyping approach and exposure weights

Salmonella is a major cause of enteric disease in Canada. Cases of salmonellosis were attributed to retail meats, food animal manure contact, and surface water sources using a microbial subtyping approach coupled with adjustments for exposure. Results indicated that 64.7% of cases were attributed to chicken breast meat, followed by frozen raw breaded chicken products (12.9%), ground chicken (9.1%), water (3.0%), pork chops and sausage (1.3%), ground beef and veal (0.7%), turkey parts (0.5%), and molluscs (0.0%). The salmonellosis incidence rate in the FoodNet Canada sentinel sites fell by one third with a parallel drop of one third in the percent of cases attributed to chicken breast meat between 2015 and 2019. Decreases in the contribution of many of the top serovars to the percentage of cases attributed to chicken breast meat indicates some emerging success with broiler breeder chicken vaccination programs. In addition, preliminary prevalence results for frozen raw chicken products in late 2019 suggests the Canadian Food Inspection Agency intervention in 2019 requiring any Salmonella on these products to be below a detectable amount may be having an impact, though more data post intervention is needed to be more conclusive.

Baseline monitoring of contaminant concentrations in American lobster (Homarus americanus) tissues from coastal Northumberland Strait, Nova Scotia, Canada

A baseline survey was conducted in 2018 to characterize contaminants in American lobsters, Homarus americanus in the Northumberland Strait, Canada. Sampling included three age classes of lobsters at sites 4, 20, and 70 km from the Boat Harbour estuary, a historically contaminated site set to undergo remediation. Lobster tissues were measured for metal(loids), methylmercury, polycyclic aromatic hydrocarbons, and polychlorinated dibenzo-p-dioxins and polychlorinated dibenzo-p-furans. Contaminant concentrations were generally below the guidelines set by the Canadian Council of Ministers of the Environment and the Canadian Food Inspection Agency, except for arsenic which was elevated in all age classes from all sites (4.8–12.68 mg kg−1). Mercury and methylmercury (both ~0.04 mg kg−1) minimally exceeded one guideline in some age-classes and sites. There was also no consistent pattern of contaminant accumulation across either age classes or at particular sites. This study serves as a baseline for future monitoring following remediation of Boat Harbour.

Analysis and Discrimination of Canadian Honey Using Quantitative NMR and Multivariate Statistical Methods

To address the growing concern of honey adulteration in Canada and globally, a quantitative NMR method was developed to analyze 424 honey samples collected across Canada as part of two surveys in 2018 and 2019 led by the Canadian Food Inspection Agency. Based on a robust and reproducible methodology, NMR data were recorded in triplicate on a 700 MHz NMR spectrometer equipped with a cryoprobe, and the data analysis led to the identification and quantification of 33 compounds characteristic of the chemical composition of honey. The high proportion of Canadian honey in the library provided a unique opportunity to apply multivariate statistical methods including PCA, PLS-DA, and SIMCA in order to differentiate Canadian samples from the rest of the world. Through satisfactory model validation, both PLS-DA as a discriminant modeling technique and SIMCA as a class modeling method proved to be reliable at differentiating Canadian honey from a diverse set of honeys with various countries of origins and floral types. The replacement method of optimization was successfully applied for variable selection, and trigonelline, proline, and ethanol at a lower extent were identified as potential chemical markers for the discrimination of Canadian and non-Canadian honeys.

New Canadian and Provincial Records of Coleoptera Resulting from Annual Canadian Food Inspection Agency Surveillance for Detection of Non-Native, Potentially Invasive Forest Insects

Simple SummaryEarly detection of adventive and potentially invasive insects that may threaten the health of forests is essential for their successful management in Canada and the world at large. To that end, the Canadian Food Inspection Agency (CFIA) conducts annual surveys at sites with high risk of adventive species introductions (e.g., ports, industrial zones, disposal sites for solid wood packaging material) using semiochemical-baited traps and also by rearing insects from trunk sections collected from stressed trees. We report 31 new Canadian provincial records of beetle species detected in surveys conducted from 2011 to 2021, including 13 new records for Canada and 9 species adventive to North America. Nine of the new Canadian records were native North American species previously detected only south of the border. All but three species belong to the Curculionidae family (“snout beetles”) and most of these were in the subfamily Scolytinae (bark and ambrosia beetles). Rearing of insects from trunk sections of stressed trees accounted for two new species records whereas trapping accounted for the remainder. These surveys not only assist our efforts to manage forest insects by documenting new species introductions and apparent range expansions but also increase our knowledge of biodiversity.The arrival and establishment of adventive, invasive forest insects are a threat to the health, diversity, and productivity of forests in Canada and the world at large, and their early detection is essential for successful eradication and management. For that reason, the Canadian Food Inspection Agency (CFIA) conducts annual surveys at high risk sites such as international ports and freight terminals, industrial zones, and disposal sites for solid wood packaging material using two methods: (1) semiochemical-baited traps deployed in a total of about 63–80 sites per year in British Columbia (BC), Ontario (ON), Quebec (QC), New Brunswick (NB), Nova Scotia (NS), and Newfoundland and Labrador (NL); and (2) rearing of insects from bolts collected from stressed trees and incubated in modified shipping containers in four cities (Vancouver, Toronto, Montreal, and Halifax). We report 31 new Canadian provincial records of Coleoptera from surveys conducted in 2011–2021, including 13 new records for Canada and 9 species adventive to North America (indicated by †). Nine of the new Canadian records were native North American species previously detected only south of the border. All but three species belong to the Curculionidae family and most of these were in the subfamily Scolytinae. The records include: Xenomelanophila miranda (LeConte) (Canada, BC) (Buprestidae: Buprestinae); Neoclytus mucronatus mucronatus (Fabricius) (BC) (Cerambycidae: Cerambycinae); Amphicerus cornutus (Pallas) (Canada, BC) (Bostrichidae: Bostrichinae); Mecinus janthinus (Germar)† (ON) (Curculionidae: Curculioninae); Aulacobaris lepidii (Germar)† (Canada, ON); Buchananius striatus (LeConte) (ON) (Curculionidae: Baridinae); Cylindrocopturus binotatus LeConte (Canada, ON) (Curculionidae: Conoderinae); Himatium errans LeConte (ON); Phloeophagus canadensis Van Dyke (ON); Rhyncolus spretus Casey (Canada, BC); Stenomimus pallidus (Boheman) (Canada, ON); Tomolips quercicola (Boheman) (Canada, ON) (Curculionidae: Cossoninae); Strophosoma melanogrammum (Forster)† (NB) (Curculionidae: Entiminae); Conotrachelus aratus (Germar) (ON) (Curculionidae: Molytinae); Anisandrus maiche Stark† (Canada, ON, QC); Cnesinus strigicollis LeConte (Canada, ON); Cyclorhipidion pelliculosum (Eichhoff)† (Canada, ON, QC); Hylesinus fasciatus LeConte (QC); Hylesinus pruinosus Eichhoff (QC); Hypothenemus interstitialis (Hopkins) (Canada, ON); Lymantor alaskanus Wood (BC); Pityogenes bidentatus (Herbst)† (Canada, ON); Scolytus mali (Bechstein)† (BC); Scolytus schevyrewi Semenov† (QC); Trypodendron scabricollis (LeConte) (Canada, ON); Trypophloeus populi Hopkins (QC); Xylechinus americanus Blackman (NFLB); and Xylosandrus crassiusculus (Motschulsky)† (BC, QC) (Curculionidae: Scolytinae). We also provide additional data confirming the presence of the adventive Hylastes opacus Erichson† in NS. Rearing of insects from bolts accounted for two new records (H. pruinosus, R. spretus) and trapping accounted for the remainder. These surveys not only assist our efforts to manage forest insects by documenting new species introductions and apparent range expansions but also increase our knowledge of biodiversity.

Norovirus GI and GII and hepatitis A virus in berries and pomegranate arils in Canada

Between 2016 and 2021, the Canadian Food Inspection Agency (CFIA) collected 4218 samples of fresh and frozen berries (blackberries, blueberries, raspberries, strawberries and mixed berries) and pomegranate arils at retail across 11 major cities in Canada and tested these samples for the presence of norovirus GI, norovirus GII and hepatitis A virus RNA. The purpose of this testing was to provide information on the prevalence of these viruses in berries and pomegranate arils on the Canadian marketplace. Of the 926 fresh fruit samples tested, norovirus GI RNA was detected in one raspberry sample and norovirus GII RNA was detected in one strawberry sample. Of the 3292 frozen fruit samples tested, norovirus GI RNA was detected in one blackberry sample, one raspberry sample and one strawberry sample, and norovirus GII RNA was detected in one blueberry sample, three raspberry samples, four strawberry samples, one pomegranate arils sample and one mixed berry sample. None of the fresh or frozen fruit samples tested positive for hepatitis A virus RNA. No statistically significant associations were observed between the prevalence of viral RNA in samples of fresh and frozen fruit, between the prevalence of viral RNA in samples of domestic and imported fruit or between the prevalence of viral RNA in samples of specific fruit types. Overall, the prevalence of norovirus GI and GII RNA together in fresh and frozen fruit samples in Canada was 0.36 %. The results of this study may be used to refine surveillance programs for norovirus and hepatitis A virus in fresh and frozen berries and pomegranate arils, e.g. by adapting the commodities tested and/or the numbers of planned samples to better target these hazards. This information may also be used to inform other Government of Canada approaches to better understand the controls associated norovirus and hepatitis A virus in fresh and frozen berries and pomegranate arils.

Sensing and Characterization of Agricultural Mycotoxins Using Electrochemistry

Mycotoxins are highly toxic and can be found as contaminants in products derived from cereals and grains.1 According to the Canadian Food Inspection Agency (CFIA), at least 25% of the grain produced each year worldwide present mycotoxin contamination.2 Among the mycotoxins of major concern in Canada is ochratoxin A (OTA),2 which is produced mainly by Aspergillus fungi species.3 Because of its severely negative health effects, the detection of this compound in grain and food samples is highly relevant and regulated by the legislation.2 OTA has been detected in food products and commodities through costly and time-consuming methods, such as chromatography.4 Existing electrochemical sensors are still based on systems that are too complex and expensive for industrial applications.5,6 Herein, we propose a rapid, reliable and cost-efficient sensing strategy for OTA detection, which is based on glassy carbon electrodes (GCE) modified with PtNPs.7 OTA is characterized for oxidation and reduction potentials, diffusion behaviour, and limits of detection and quantification. To this end, we aim to develop a practical and highly sensitive electroanalytical methodology that will enable us to detect OTA in grain samples in the future. References B. Scafuri et al., Scientific Reports, 7, 1–11 (2017).L. L. Charmley and H. L. Trenholm, RG-8 Regulatory Guidance: Contaminants in Feed (2017) https://inspection.canada.ca/animal-health/livestock-feeds/regulatory-guidance/rg-8/eng/1347383943203/1347384015909?chap=1.T. R. Bui-Klimke and F. Wu, Critical Reviews in Food Science and Nutrition, 55, 1860–1869 (2015).R. Chauhan, J. Singh, T. Sachdev, T. Basu, and B. D. Malhotra, Biosensors and Bioelectronics, 81, 532–545 (2016).A. Suea-Ngam, P. D. Howes, C. E. Stanley, and A. J. Demello, ACS Sensors, 4, 1560–1568 (2019).K. Kunene et al., Sensors and Actuators, B: Chemical, 305 (2020).E. Mazzotta et al., ACS Applied Nano Materials, 4, 7650–7662 (2021).

Behaviour and adhesion capacity of Listeria monocytogenes on Mozzarella di Bufala Campana PDO cheese and in fluids involved in the production process

Listeria monocytogenes is a foodborne pathogen for humans. It can be found in ready-to-eat (RTE) products and its presence represents one of the biggest food safety concerns. Most of the cases of cheese contamination reported by the Rapid Alert System for Food and Feed (RASFF) in the last 10 years did not involve Mozzarella di Bufala Campana (MBC) Protected Designation of Origin (PDO) cheese. Suddenly, in the last years, RASFF has disposed of the withdrawn from the market of this product contaminated with L. monocytogenes on two different occasions (RASFF 2018/1198 and 2018/2459), and the Canadian Food Inspection Agency disposed one withdrawn in 2019. Following these events, this study aimed to evaluate the behaviour of L. monocytogenes in Mozzarella di Bufala Campana PDO cheese. First of all, the adhesion capacity of the pathogen to MBC cheese was studied through microbial challenge tests simulating the passage of MBC in hardening water and in covering liquid used for cheese storage during the production process. In addition, the ability of L. monocytogenes to grow in MBC cheese and covering liquid during shelf life, at refrigerated condition, was evaluated also this time employing microbial challenge tests. Results demonstrated that L. monocytogenes can adhere to the surface of the cheese and be released during the contact of contaminated cheese with other fluids. However, the presence of an abundant microbiota on the cheese exerted a protective role against this pathogen preventing the growth of L. monocytogenes. Indeed, the pathogens artificially contaminating the covering liquid at 2 log CFU/mL and 4 log CFU/mL remained at the inoculum level during 32 days of storage at 4 °C.

Assessing the extent and public health impact of bat predation by domestic animals using data from a rabies passive surveillance program.

Domestic animals can serve as consequential conveyors of zoonotic pathogens across wildlife-human interfaces. Still, there has been little study on how different domestic species and their behaviors influence the zoonotic risk to humans. In this study, we examined patterns of bat encounters with domestic animals that resulted in submission for testing at the rabies laboratories of the Canadian Food Inspection Agency (CFIA) during 2014-2020. Our goals were specifically to examine how the number of bats submitted and the number of rabies positive bats varied by the type of domestic animal exposure and whether domestic cats were indoor or free-roaming. The CFIA reported 6258 bat submissions for rabies testing, of which 41.5% and 8.7% had encounter histories with cats and dogs, respectively. A much smaller fraction of bat submissions (0.3%) had exposure to other domestic animals, and 49.5% had no domestic animal exposure. For the bat submissions related to cats, and where lifestyle was noted, 91.1% were associated with free-roaming cats and 8.9% with indoor cats. Model results indicated the probability of a rabies-positive bat was the highest with a history of dog association (20.2%), followed by bats with no animal exposure (16.7%), free-roaming cats (6.9%), cats with unspecified histories (6.0%) and the lowest probability associated with non-free-roaming (indoor) cats (3.8%). Although there was lower rabies prevalence in bats associated with cats compared to dogs, the 4.8 fold higher number of cat-bat interactions cumulatively leads to a greater overall rabies exposure risk to humans from any free-roaming outdoor cats. This study suggests that free-roaming owned cats may have an underappreciated role in cryptic rabies exposures in humans and as a significant predator of bats. Preventing free-roaming in cats is a cost-effective and underutilized public health recommendation for rabies prevention that also synergistically reduces the health burden of other feline-associated zoonotic diseases and promotes feline welfare and wildlife conservation.