Monitoring Changes in the Antimicrobial-Resistance Gene Set (ARG) of Raw Milk and Dairy Products in a Cattle Farm, from Production to Consumption.

Raw milk and dairy products can serve as potential vectors for transmissible bacterial, viral and protozoal diseases, alongside harboring antimicrobial-resistance genes. This study monitors the changes in the antimicrobial-resistance gene pool in raw milk and cheese, from farm to consumer, utilizing next-generation sequencing. Five parallel sampling runs were conducted to assess the resistance gene pool, as well as phage or plasmid carriage and potential mobility. In terms of taxonomic composition, in raw milk the Firmicutes phylum made up 41%, while the Proteobacteria phylum accounted for 58%. In fresh cheese, this ratio shifted to 93% Firmicutes and 7% Proteobacteria. In matured cheese, the composition was 79% Firmicutes and 21% Proteobacteria. In total, 112 antimicrobial-resistance genes were identified. While a notable reduction in the resistance gene pool was observed in the freshly made raw cheese compared to the raw milk samples, a significant growth in the resistance gene pool occurred after one month of maturation, surpassing the initial gene frequency. Notably, the presence of extended-spectrum beta-lactamase (ESBL) genes, such as OXA-662 (100% coverage, 99.3% identity) and OXA-309 (97.1% coverage, 96.2% identity), raised concerns; these genes have a major public health relevance. In total, nineteen such genes belonging to nine gene families (ACT, CMY, EC, ORN, OXA, OXY, PLA, RAHN, TER) have been identified. The largest number of resistance genes were identified against fluoroquinolone drugs, which determined efflux pumps predominantly. Our findings underscore the importance of monitoring gene pool variations throughout the product pathway and the potential for horizontal gene transfer in raw products. We advocate the adoption of a new approach to food safety investigations, incorporating next-generation sequencing techniques.

Consumer knowledge, attitudes, and behaviour regarding food safety - A perspective from Laos.

Primary research data on food chain safety at consumer level is needed to help policymakers to decrease the risk of foodborne infections in the Lao People's Democratic Republic. Therefore, the objectives of this study are to 1) determine Lao people's food safety knowledge, attitudes, and intentions and identify the most influential factors that shape their food safety behaviour, and 2) to support the government in the development of more efficient risk communication programmes. A paper-based consumer survey was implemented to collect quantitative data on the selected topics about Laotian consumers, as well as on the equipment of households regarding the conditions and tools necessary for safe food preparation and storage. A model was developed based on the extended Theory of Planned Behaviour (TPB), and partial least squares structural equation modelling (PLS-SEM) was applied to analyse the factors that affect food safety behaviour. The elements of Knowledge and Perceived Behavioural Control positively influence behaviours, such as using separate chopping boards for raw and ready-to-eat foods. According to the model, only 10 % of the Intentions would be translated into Behaviours, which gap can be explained by the lack of a hygienic environment. Although the survey indicated that Laotian people intend to follow food safety guidelines, participants felt they lacked sufficient knowledge. This perception was validated by the knowledge test and behavioural part of the study. In the light of these findings, it is essential to raise awareness and promote food safety knowledge and interventions, such as enhancing food safety education in childhood. This would help to facilitate and encourage adopting safe practices to improve overall food safety at the consumer level, delivering positive impacts also to the food business sector.

Metal Load of Potentially Toxic Elements in Tuna (Thunnus albacares)-Food Safety Aspects.

The consumption of marine fishes has a positive effect on a consumer's health; however, it poses a potential risk due to their level of heavy metals in their body. Heavy metals can be naturally found in the environment, but their concentration can be increased with anthropogenic activities. Samples of tuna (Thunnus albacares) were collected at a fishery market. The potentially toxic elements (arsenic, cadmium, lead, and mercury) were determined with a validated method in the flesh of fish using inductively plasma optical emission spectrometry after microwave digestion. Generally, the average concentration of them was below the official limit values regulated by the European Union, except for lead. Based on the concentrations of arsenic (inorganic derivates: 0.05 ± 0.02 mg/kg) and cadmium (0.03 ± 0.01 mg/kg) in the tuna fish samples, and their calculated EDI values (As: 0.03-0.09 µg/kg/day; Cd: 0.05-0.07 µg/kg/day), the investigated food could be declared safe for human consumption. Generally, mercury content was below the official regulated limit, and the calculated EDI value was below the dietary reference value (0.3 µg/kg/day) in most of the samples (90%), exceeding it only in two samples (0.69 and 0.82 µg/kg/day); thus, they may not be harmful to the consumer. The concentration of lead above the official maximum limit (0.30 mg/kg) in 40% of tuna samples (0.30-1.59 mg/kg), as well as the exceeding of the dietary reference value for lead (adult: 0.16 µg/kg/day; children: 0.26 µg/kg/day) based on the calculated EDI values (0.28-1.49 µg/kg/day), draw attention to the importance of environmental pollution and the protection of consumers' health.

Insects as food - Changes in consumers' acceptance of entomophagy in Hungary between 2016 and 2021.

Public interest in entomophagy (consumption of insects) has developed significantly over the past several years. Possible nutritional benefits are perceived by consumers according to several recent studies, as well as sustainability and food security. However, most European communities, including the Hungarian, do not embrace entomophagy, despite the widespread practice elsewhere globally. This study aims to evaluate the changes in the perception of entomophagy among the Hungarian population between 2016 and 2021, together with the factors differentiating between acceptive and dismissive consumers. The results of the two representative quantitative surveys indicate that more than 70% of Hungarian consumers are not willing to try entomophagy, which had not changed significantly in the observed period, despite the high media coverage of this topic in recent years. Some groups open to insect consumption can still be identified. According to the socioeconomic segmentation of the data collected in 2021, consumers who accept insect-based foods can be found in high numbers among men between 18 and 39 years old (49.3%). Positive attitudes are less likely to be observed among females; however, 27.6% of highly educated women between 18 and 59 years demonstrated a certain level of interest. Those consumers willing to consume insects are driven mainly by curiosity, and also value high protein content and sustainability, and perceive insect-based food as nutritious. Consumers who prefer local and national food tend to refuse to eat insects in a higher ratio.

Carry-Over of Aflatoxin B1 from Feed to Cow Milk-A Review.

The conversion of aflatoxin B1 in feed consumed by cows into aflatoxin M1 in their milk poses a challenge to food safety, with milk being a popular staple food and because of the harmful effects of these compounds. This study aimed at reviewing scientific information about the extent of carry-over of AFB1 from feed to milk. A range of studies reported about correlations of carry-over with different factors, particularly with milk yield and AFB1 intake. The extent of carry-over considerably varies, being 1-2% on average, which may be as high as 6% in the case of increased milk production. Specific factors influencing transfer rates, including milk yield, somatic cell counts, aflatoxin B1 intake, source of contamination, seasonal effects, particle size of feed, and the effects of certain interventions, namely vaccination and the use of adsorbents, are identified as the most important and are discussed in this review. The different mathematical formulas describing carry-over and instances of their application are reviewed as well. These carry-over equations may lead to largely different results, and no single carry-over equation can be suggested as the best one. While the exact quantification of carry-over is difficult as the process depends on many factors, including individual variabilities between animals, the intake of aflatoxin B1 and milk yield seem to be the most important factors influencing the excreted amount of aflatoxin M1 and the rate of carry-over.

Probabilistic modeling and risk characterization of the chronic aflatoxin M1 exposure of Hungarian consumers.

Aflatoxin contamination can appear in various points of the food chain. If animals are fed with contaminated feed, AFB1 is transformed-among others-to aflatoxin M1 (AFM1) metabolite. AFM1 is less toxic than AFB1, but it is still genotoxic and carcinogenic and it is present in raw and processed milk and all kinds of milk products. In this article, the chronic exposure estimation and risk characterization of Hungarian consumers are presented, based on the AFM1 contamination of milk and dairy products, and calculated with a probabilistic method, the two-dimensional Monte-Carlo model. The calculations were performed using the R plugin (mc2d package) integrated into the KNIME (Konstanz Information Miner) software. The simulations were performed using data from the 2018-2020 food consumption survey. The AFM1 analytical data were derived from the Hungarian monitoring survey and 1,985 milk samples were analyzed within the framework of the joint project of the University of Debrecen and the National Food Chain Safety Office of Hungary (NÉBIH). Limited AFM1 concentrations were available for processed dairy products; therefore, a database of AFM1 processing factors for sour milk products and various cheeses was produced based on the latest literature data, and consumer exposure was calculated with the milk equivalent of the consumed quantities of these products. For risk characterization, the calculation of hazard index (HI), Margin of Exposure, and the hepatocellular carcinoma incidence were used. The results indicate that the group of toddlers that consume a large amount of milk and milk products are exposed to a certain level of health risk. The mean estimated daily intake of toddlers is in the range of 0.008-0.221 ng kg-1 bw day-1; the 97.5th percentile exposure of toddlers is between 0.013 ng kg-1 bw day-1 and 0.379 ng kg-1 bw day-1, resulting in a HI above 1. According to our study, the exposure of older age groups does not pose an emergent health risk. Nevertheless, the presence of carcinogenic compounds should be kept to a minimum in the whole population.

A Systematic Review of the Efficacy of Interventions to Control Aflatoxins in the Dairy Production Chain-Feed Production and Animal Feeding Interventions.

The study presents a systematic review of published scientific articles investigating the effects of interventions aiming at aflatoxin reduction at the feed production and animal feeding phases of the milk value chain in order to identify the recent scientific trends and summarize the main findings available in the literature. The review strategy was designed based on the guidance of the systematic review and knowledge synthesis methodology that is applicable in the field of food safety. The Web of Science and EBSCOhost online databases were searched with predefined algorithms. After title and abstract relevance screening and relevance confirmation with full-text screening, 67 studies remained for data extraction, which were included in the review. The most important identified groups of interventions based on their mode of action and place in the technological process are as follows: low-moisture production using preservatives, acidity regulators, adsorbents and various microbiological additives. The results of the listed publications are summarized and compared for all the identified intervention groups. The paper aimed to help feed producers, farmers and relevant stakeholders to get an overview of the most suitable aflatoxin mitigation options, which is extremely important in the near future as climate change will likely be accompanied by elevated mycotoxin levels.

Protective Effects of Grape Seed Oligomeric Proanthocyanidins in IPEC-J2-Escherichia coli/Salmonella Typhimurium Co-Culture.

Intestinal epithelium provides the largest barrier protecting mammalian species from harmful external factors; however, it can be severely compromised by the presence of bacteria in the gastrointestinal (GI) tract. Antibiotics have been widely used for the prevention and treatment of GI bacterial infections, leading to antimicrobial resistance in human and veterinary medicine alike. In order to decrease antibiotic usage, natural substances, such as flavonoids, are investigated to be used as antibiotic alternatives. Proanthocyanidins (PAs) are potential candidates for this purpose owing to their various beneficial effects in humans and animals. In this study, protective effects of grape seed oligomeric proanthocyanidins (GSOPs) were tested in IPEC-J2 porcine intestinal epithelial cells infected with Escherichia coli and Salmonella enterica ser. Typhimurium of swine origin. GSOPs were able to alleviate oxidative stress, inflammation and barrier integrity disruption inflicted by bacteria in the co-culture. Furthermore, GSOPs could decrease the adhesion of both bacteria to IPEC-J2 cells. Based on these observations, GSOPs seem to be promising candidates for the prevention and treatment of gastrointestinal bacterial infections.

Distribution of cortactin in cerebellar Purkinje cell spines.

Dendritic spines are the primary sites of excitatory transmission in the mammalian brain. Spines of cerebellar Purkinje Cells (PCs) are plastic, but they differ from forebrain spines in a number of important respects, and the mechanisms of spine plasticity differ between forebrain and cerebellum. Our previous studies indicate that in hippocampal spines cortactin-a protein that stabilizes actin branch points-resides in the spine core, avoiding the spine shell. To see whether the distribution of cortactin differs in PC spines, we examined its subcellular organization using quantitative preembedding immunoelectron microscopy. We found that cortactin was enriched in the spine shell, associated with the non-synaptic membrane, and was also situated within the postsynaptic density (PSD). This previously unrecognized distribution of cortactin within PC spines may underlie structural and functional differences in excitatory spine synapses between forebrain, and cerebellum.

Characterization and therapeutic application of canine adipose mesenchymal stem cells to treat elbow osteoarthritis.

Visceral adipose tissue (AT) obtained from surgical waste during routine ovariectomies was used as a source for isolating canine mesenchymal stem cells (MSCs). As determined by cytofluorimetry, passage 2 cells expressed MSC markers CD44 and CD90 and were negative for lineage-specific markers CD34 and CD45. The cells differentiated toward osteogenic, adipogenic, and chondrogenic directions. With therapeutic aims, 30 dogs (39 joints) suffering from elbow dysplasia (ED) and osteoarthritis (OA) were intra-articularly transplanted with allogeneic MSCs suspended in 0.5% hyaluronic acid (HA). A highly significant improvement was achieved without any medication as demonstrated by the degree of lameness during the follow-up period of 1 y. Control arthroscopy of 1 transplanted dog indicated that the cartilage had regenerated. Histological analysis of the cartilage biopsy confirmed that the regenerated cartilage was of hyaline type. These results demonstrate that transplantation of allogeneic adipose tissue-derived mesenchymal stem cells (AT-MSCs) is a novel, noninvasive, and highly effective therapeutic tool in treating canine elbow dysplasia.

Du tissu adipeux viscéral (TA) obtenu de résidus chirurgicaux lors d'ovariectomies de routine a été utilisé comme source pour isoler des cellules souches mésenchymateuses canines (CSMs). Tel que déterminé par cytofluorométrie, les cellules du 2e passage exprimaient les marqueurs de CSM CD44 et CD90, et étaient négatives pour les marqueurs spécifiques de lignée CD34 et CD45. Les cellules se sont différenciées dans des directions ostéogéniques, adipogéniques, et chondrogéniques. À des fins thérapeutiques, 30 chiens (39 articulations) souffrant de dysplasie du coude (DC) et d'ostéoarthrite (OA) ont reçu une transplantation intra articulaire de CSMs allogéniques en suspension dans 0,5 % d'acide hyaluronique (AH). Une amélioration hautement significative a été obtenue sans aucune médication tel que démontré par le degré de boiterie durant la période de suivi de 1 an. Une arthroscopie de contrôle de un des chiens ayant reçu une transplantation montrait que le cartilage s'était régénéré. L'analyse histologique de la biopsie du cartilage a confirmé que le cartilage régénéré était de type hyalin. Ces résultats démontrent que la transplantation de cellules souches mésenchymateuses dérivées de tissu adipeux allogène est un outil thérapeutique novateur, non-invasif, et très efficace pour traiter la dysplasie du coude chez le chien.(Traduit par Docteur Serge Messier).

Depletion of gentamicin and its major components from various tissues of turkeys.

OBJECTIVE: To determine tissue depletion profiles for gentamicin and its 3 major components (C1, C1a, and C2) in turkeys. ANIMALS: Twenty 10-week-old male turkeys. PROCEDURE: birds were maintained as untreated controls. The remaining birds were treated with gentamicin sulfate at a dosage of 2 mg/kg, IM, once daily for 5 days. Treated birds were euthanatized 45, 60, 75, and 90 days (4 birds at each sample time) after the last dose of gentamicin was administered, and samples of muscle, liver, kidney, and skin and fat were collected. Control birds were euthanatized on day 45. Concentrations of the 3 major components of gentamicin were measured by means of reversed-phase high-performance liquid chromatography. RESULTS: Total gentamicin concentration (ie, sum of the concentrations of the 3 major components) was < 100 microg/kg for all muscle and skin and fat samples by day 45 and all liver samples by day 75. At all sample times, concentration of the gentamicin C1 component was higher than concentrations of the C1a and C2 components in all tissues. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that tissue depletion profiles of the 3 major components of gentamicin differ from each other. Withdrawal time, therefore, may depend on the ratio of the components in the pharmaceutical preparation used.