High Prevalence of Antibiotic Resistance in Traditionally Fermented Foods as a Critical Risk Factor for Host Gut Antibiotic Resistome.

This study aimed to assess the suitability of fermented food interventions to replenish damaged gut microbiota. Metagenomic assessment of published sequencing data found that fermented food interventions led to a significant increase in the gut antibiotic resistome in healthy human subjects. Antibiotic resistome and viable antibiotic-resistant (AR) bacteria were further highly prevalent in retail kimchi and artisan cheeses by metagenomic and culture analyses. Representative AR pathogens of importance in nosocomial infections, such as Klebsiella pneumoniae, Serratia marcescens, and vancomycin-resistant Enterococcus (VRE), as well as commensals and lactic acid bacteria, were characterized; some exhibited an extremely high minimum inhibitory concentration (MIC) against antibiotics of clinical significance. Exposing fermented food microbiota to representative antibiotics further led to a boost of the corresponding antibiotic and multidrug-resistance gene pools, as well as disturbed microbiota, including the rise of previously undetectable pathogens. These results revealed an underestimated public health risk associated with fermented food intervention at the current stage, particularly for susceptible populations with compromised gut integrity and immune functions seeking gut microbiota rescue. The findings call for productive intervention of foodborne AR via technology innovation and strategic movements to mitigate unnecessary, massive damages to the host gut microbiota due to orally administered or biliary excreted antibiotics.

Consumers' Segmentation Influences Acceptance and Preferences of Cheeses with Sanitary Inspection and Artisanal Seals.

Food labeling serves as a crucial medium for industries to communicate product qualities to consumers. Sanitary inspection and artisanal seals are significant markers for traditional cheeses, yet current information on this topic is limited. Therefore, this study aims to evaluate the impact of sanitary inspection and the ARTE seal on the acceptance of artisanal cheese. To achieve this objective, four hypothetical cheese labels featuring all combinations of sanitary inspection and ARTE seals were presented to 404 consumers. These consumers rated their acceptance of each label, a conjoint analysis was conducted, and the relative importance of each seal was calculated. Subsequently, consumers were segmented using hierarchical cluster analysis. Their socio-demographic profiles were statistically correlated to the clusters through a chi-squared method. The results revealed the existence of three distinct consumer groups: those who strongly prefer cheeses with a sanitary seal (assigning a relative importance of 80.2% to the seal), those who favor cheeses with an artisanal seal (assigning a relative importance of 52.5% to the seal), and those for whom the presence of either seal did not significantly affect acceptance. Consumers residing in metropolitan areas generally placed less value on both seals, whereas frequent purchasers of artisanal foods and residents of rural areas showed a preference for the artisanal seal. Other socio-demographic variables did not statistically correlate with cluster membership. In conclusion, consumer segmentation based on preferences for sanitary inspection and artisanal seals in food labeling is vital for developing effective marketing strategies and food safety education policies.

Relationship between Volatile Organic Compounds and Microorganisms Isolated from Raw Sheep Milk Cheeses Determined by Sanger Sequencing and GC-IMS.

Recently, the interest of consumers regarding artisan cheeses worldwide has increased. The ability of different autochthonous and characterized lactic acid bacteria (LAB) to produce aromas and the identification of the volatile organic compounds (VOCs) responsible for flavor in cheeses are important aspects to consider when selecting strains with optimal aromatic properties, resulting in the diversification of cheese products. The objective of this work is to determine the relationship between VOCs and microorganisms isolated (Lacticaseibacillus paracasei, Lactiplantibacillus plantarum, Leuconostoc mesenteroides and Lactococcus lactis subsp. hordniae) from raw sheep milk cheeses (matured and creamy natural) using accuracy and alternative methods. On combining Sanger sequencing for LAB identification with Gas Chromatography coupled to Ion Mobility Spectrometry (GC−IMS) to determinate VOCs, we describe cheeses and differentiate the potential role of each microorganism in their volatilome. The contribution of each LAB can be described according to their different VOC profile. Differences between LAB behavior in each cheese are shown, especially between LAB involved in creamy cheeses. Only L. lactis subsp. hordniae and L. mesenteroides show the same VOC profile in de Man Rogosa and Sharpe (MRS) cultures, but for different cheeses, and show two differences in VOC production in skim milk cultures. The occurrence of Lactococcus lactis subsp. hordniae from cheese is reported for first time.

Graduate Student Literature Review: Enterotoxigenic potential and antimicrobial resistance of staphylococci from Brazilian artisanal raw milk cheeses.

More than 30 types of artisanal cheeses are known in Brazil; however, microorganisms, such as Staphylococcus spp., can contaminate raw milk cheeses through different sources, from milking to processing. Staphylococcal food poisoning results from the consumption of food in which coagulase-positive staphylococci, mostly Staphylococcus aureus, have developed and produced enterotoxins. In addition, an emerging public health concern is the increasing antimicrobial resistance of some Staphylococcus strains. Furthermore, the ability of Staphylococcus spp. in sharing antibiotic resistance-related genes with other bacteria increases this problem. In light of these observations, this review aims to discuss the presence of, enterotoxins of, and antibiotic-resistant of Staphylococcus spp. in Brazilian artisanal cheese produced with raw milk.

Caracterización de bacterias ácido lácticas con actividad antimicrobiana aisladas del queso crema de Chiapas, México / Characterization of lactic acid bacteria with antimicrobial activity isolated from cream cheese from Chiapas, Mexico

Resumen El queso crema de Chiapas es un producto artesanal fabricado en diferentes regiones de México. La capacidad de las bacterias lácticas, de producir grandes cantidades de ácido láctico y acético, tiene como efecto la disminución del pH, considerado uno de los factores primarios en la inhibición de microorganismos indeseables, como patógenos y bacterias coliformes, en alimentos lácteos. El objetivo de este trabajo fue aislar bacterias ácido lácticas y evaluar su efecto antagónico contra bacterias patógenas in vitro y contra patógenos presentes en la leche cuando se adicionan durante la elaboración del queso crema. Se aislaron cepas de bacterias lácticas del queso crema de Chiapas, México. Los aislados fueron identificados mediante pruebas bioquímicas API 50CH y secuencias del gen ribosomal 16S. Las bacterias de interés se usaron en pruebas de inhibición del crecimiento con cepas patógenas. Se cuantificó la producción de ácido acético y láctico por cromatografía de gases acoplado a masas en los quesos, al inicio y a los 10 d posteriores a su elabo ración. Se aislaron 203 cepas bacterianas, de las cuales 82 tuvieron la capacidad de inhibir el crecimiento de 7 cepas bacterianas patógenas. Las cepas fueron identificadas por la secuencia parcial del gen ribosomal 16S como pertenecientes al género Lactobacillus; la adición de estas cepas en la elaboración del queso crema redujo o eliminó coliformes, Staphylococcus aureus, mohos y levaduras, mientras que en el queso elaborado con leche pasteurizada se detectaron 290 UFC/mL de mohos y levaduras. Estos resultados sugieren que las cepas de Lactobacillus aisladas del queso crema artesanal de Chiapas tienen potencial para su uso como cultivo iniciador con actividad de bioconservación en este y productos similares.

Abstract Chiapas cream cheese is an artisanal product manufactured in different regions of Mexico. The ability of lactic bacteria to produce large amounts of lactic and acetic acid has the effect of decreasing the pH, considered one of the primary factors in the inhibition of undesirable microorganisms such as pathogens and coliform bacteria, in dairy foods. The objective of this work was to isolate lactic acid bacteria and evaluate their antagonistic effect against pathogenic bacteria in vitro, as well as their effect against pathogens that are present in milk when they are added in the preparation of cream cheese. Strains of lactic bacteria were isolated from cream cheese from Chiapas, Mexico. Isolates were identified by API 50CH biochemical tests and 16S ribosomal gene sequences. The bacteria under study were used in pathogen growth inhibition tests. The production of acetic and lactic acid was quantified by gas chromatography coupled to masses in the cheeses at the beginning and 10 d after its elaboration. 203 bacterial strains were isolated, 82 of which had the ability to inhibit the growth of 7 pathogenic bacterial strains. The strains were identified by the partial sequence of the 16S ribosomal gene as belonging to the genus Lactobacillus; The addition of these strains in the elaboration of cream cheese reduces or eliminates coliforms, Staphylococcus aureus, molds and yeasts, while in the cheese made with pasteurized milk 290 CFU/mL of molds and yeasts were detected. These results suggest that Lactobacillus strains isolated from Chiapas artisanal cream cheese has the potential to be used as a starter culture with bioconservation activity in this and similar products.

American artisan cheese quality and spoilage: A survey of cheesemakers' concerns and needs.

Production of artisan cheeses, including surface-ripened cheeses, has increased in the United States over the past 2 decades. Although many of these cheesemakers report unique quality and spoilage problems during production, a systematic assessment of the quality concerns facing this sector of specialty cheese production has not been conducted. Here we report the effects of microbial spoilage and quality issues on US artisan cheese production. In a survey of 61 cheesemakers, the most common issues reported were undesirable surface molds (71%) and incorrect or unexpected colors or pigments on rinds (54%). When asked, 18% of participants indicated that they were extremely concerned about quality and spoilage problems, and they indicated that their quality standards are frequently not met, either annually (39%) or monthly (33%). Although most of the respondents (62%) said that just 0 to 5% of their cheese was lost or rendered less valuable due to quality issues annually, a small number (7% combined) reported large losses of 20 to 30% or >30% of their product lost or rendered less valuable. Almost all respondents (95%) agreed that improved quality would reduce waste, increase profits, and improve production. The survey respondents indicated in open response questions that they want access to more online resources related to quality issues and digital forums to discuss issues with experts and peers when problems arise. These findings represent the first attempt to document and estimate the effect of quality and spoilage on the American artisan cheese industry. Future work should investigate what technologies, interventions, or information could reduce losses from these problems.

Correlation between carcass condemnations in slaughterhouses and official notifications of cattle testing positive for tuberculosis.

The objective of this study was to analyze the correlation between condemnations in slaughterhouses situated in Minas Gerais State and notifications to the Brazilian Official Veterinary Service of cattle that were positive for intradermal tuberculin tests. Data were obtained from three slaughterhouses under different surveillance conditions and from the Brazilian Agriculture and Livestock Health Agency for 2011 to 2017. During this period, there was an increase in the number of condemnations of females aged over 36 months, despite the number of females testing with intradermal tests and being reported as positive decreasing. Therefore, there is a discrepancy between the analyzed variables. Since there is a belief that slaughter condemnations can be used as tools for epidemiological surveys in beef and dairy farms, it is advisable for there to be a greater integration of the Brazilian Health Inspection Services in slaughterhouses and Brazilian Agriculture and Livestock defense department. This will ensure safe animal products.

Using PacBio sequencing to investigate the bacterial microbiota of traditional Buryatian cottage cheese and comparison with Italian and Kazakhstan artisanal cheeses.

Traditional fermented dairy foods including cottage cheese have been major components of the Buryatia diet for centuries. Buryatian cheeses have maintained not only their unique taste and flavor but also their rich natural lactic acid bacteria (LAB) content. However, relatively few studies have described their microbial communities or explored their potential to serve as LAB resources. In this study, the bacterial microbiota community of 7 traditional artisan cheeses produced by local Buryatian families was investigated using single-molecule, real-time sequencing. In addition, we compared the bacterial microbiota of the Buryatian cheese samples with data sets of cheeses from Kazakhstan and Italy. Furthermore, we isolated and preserved several LAB samples from Buryatian cheese. A total of 62 LAB strains (belonging to 6 genera and 14 species or subspecies) were isolated from 7 samples of Buryatian cheese. Full-length 16S rRNA sequencing of the microbiota revealed 145 species of 82 bacterial genera, belonging to 7 phyla. The most dominant species was Lactococcus lactis (43.89%). Data sets of cheeses from Italy and Kazakhstan were retrieved from public databases. Principal component analysis and multivariate ANOVA showed marked differences in the structure of the microbiota communities in the cheese data sets from the 3 regions. Linear discriminant analyses of the effect size identified 48 discriminant bacterial clades among the 3 groups, which might have contributed to the observed structural differences. Our results indicate that the bacterial communities of traditional artisan cheeses vary depending on geographic origin. In addition, we isolated novel and valuable LAB resources for the improvement of cottage cheese production.

Effect of farm characteristics and practices on hygienic quality of ovine raw milk used for artisan cheese production in central Italy.

A survey on ovine dairy farms directly transforming own-produced milk, in the Italian Marche region, was carried out to assess flock and milking practices that may influence milk hygienic-sanitary conditions. A census survey established that 24 dairy farms were located in this region. Bulk milk samples were collected throughout the milking period in each dairy farm in 2013. Analyzed variables were: (i) chemical parameters such as fat, protein and lactose content, dry matter and pH; and (ii) total bacterial (TBC) and somatic cell counts (SCC). Chemical parameter values were in agreement with published data while, geometric mean (GM) log10 SCC was 5.91 and TBC GM was 57 978 colony forming units/mL, in compliance with Eropean Union criteria. A positive correlation was found between SCC and TBC when GMs of all farm data were considered (Spearman's rho = 0.7925; P = 0.0001). Statistical analysis did not show significant correlation between SCC or TBC GM and dairy farm principal characteristics. Although SCC levels detected in the present study should suggest the need to implement mastitis control programs, Marche's dairy sheep flocks revealed a good hygienic condition level. This is an important aspect in implementing safety for end users of the final product.

A decision-making tool to determine economic feasibility and break-even prices for artisan cheese operations.

Artisan cheese makers lack access to valid economic data to help them evaluate business opportunities and make important business decisions such as determining cheese pricing structure. The objective of this study was to utilize an economic model to evaluate the net present value (NPV), internal rate of return, and payback period for artisan cheese production at different annual production volumes. The model was also used to determine the minimum retail price necessary to ensure positive NPV for 5 different cheese types produced at 4 different production volumes. Milk type, cheese yield, and aging time all affected variable costs. However, aged cheeses required additional investment for aging space (which needs to be larger for longer aging times), as did lower yield cheeses (by requiring larger-volume equipment for pasteurization and milk handling). As the volume of milk required increased, switching from vat pasteurization to high-temperature, short-time pasteurization was necessary for low-yield cheeses before being required for high-yield cheeses, which causes an additional increase in investment costs. Because of these differences, high-moisture, fresh cow milk cheeses can be sold for about half the price of hard, aged goat milk cheeses at the largest production volume or for about two-thirds the price at the lowest production volume examined. For example, for the given model assumptions, at an annual production of 13,608kg of cheese (30,000 lb), a fresh cow milk mozzarella should be sold at a minimum retail price of $27.29/kg ($12.38/lb), whereas a goat milk Gouda needs a minimum retail price of $49.54/kg ($22.47/lb). Artisan cheese makers should carefully evaluate annual production volumes. Although larger production volumes decrease average fixed cost and improve production efficiency, production can reach volumes where it becomes necessary to sell through distributors. Because distributors might pay as little as 35% of retail price, the retail price needs to be higher to compensate. An artisan cheese company that has not achieved the recognition needed to achieve a premium price may not find distribution through distributors profitable.

Start-up and operating costs for artisan cheese companies.

Lack of valid economic data for artisan cheese making is a serious impediment to developing a realistic business plan and obtaining financing. The objective of this study was to determine approximate start-up and operating costs for an artisan cheese company. In addition, values are provided for the required size of processing and aging facilities associated with specific production volumes. Following in-depth interviews with existing artisan cheese makers, an economic model was developed to predict costs based on input variables such as production volume, production frequency, cheese types, milk types and cost, labor expenses, and financing. Estimated values for start-up cost for processing and aging facility ranged from $267,248 to $623,874 for annual production volumes of 3,402 kg (7,500 lb) and 27,216 kg (60,000 lb), respectively. First-year production costs ranged from $65,245 to $620,094 for the above-mentioned production volumes. It is likely that high start-up and operating costs remain a significant entry barrier for artisan cheese entrepreneurs.

"A little information excites us." Consumer sensory experience of Vermont artisan cheese as active practice.

This research is concerned with explaining consumer preference for Vermont artisan cheese and the relationship between that preference and sensory experience. Artisan cheesemaking is increasingly an important part of Vermont's dairy sector, and this tracks a growing trend of artisan agricultural practice in the United States. In popular discourse and academic research into products like artisan cheese, consumers explain their preferences in terms of intrinsic sensory and extrinsic - supposedly nonsensory - food qualities. In laboratory sensory studies, however, the relationship between preference, intrinsic, and extrinsic qualities changes or disappears. In contrast, this study explains this relationship by adopting a social theory of sensory perception as a practice in everyday life. This theory is applied to a series of focus group interviews with Vermont artisan cheese consumers about their everyday perceptions. Based on the data, a conceptual framework for the sensory perception of Vermont artisan cheese is suggested: consumers combine information about producer practice, social context, and the materiality of the product through an active, learned practice of sensory perception. Particular qualities that drive consumer sensory experience and preference are identified from the interview data. Many of these qualities are difficult to categorize as entirely intrinsic or extrinsic, highlighting the need for developing new approaches of sensory evaluation in order to fully capture everyday consumer sensory perception. Thus, this research demonstrates that social theory provides new and valuable insights into consumer sensory preference for Vermont artisan cheese.

Recuento de Staphylococcus aureus y detección de enterotoxinas estafilocócicas en queso blanco venezolano artesanal tipo “telita” expendido en mercados de la ciudad de Caracas / Counts of Staphylococcus aureus and detection of staphylococcal enterotoxins in Venezuelan white artisan “telita” type cheese sold at Caracas city markets

La intoxicación alimentaria estafilocócica es producida por el consumo de alimentos contaminados con enterotoxinas estafilocócicas producidas por cepas de Staphylococcus aureus, constituyéndose en una de las principales enfermedades alimentarias a nivel mundial. En este estudió se investigó la presencia de S. aureus y la producción de enterotoxinas estafilocócicas en muestras de queso blanco artesanal tipo “telita” expendidas en diferentes mercados de la ciudad de Caracas. Se analizaron 80 muestras y el recuento de S. aureus se efectuó según Norma Venezolana COVENIN 1292:89. La presencia de enterotoxinas tipo A, B, C y D de S. aureus se determinó con la prueba de aglutinación de látex en fase reversa (SET RPLA Oxoid). Se encontró carga microbiana de 10³-10(4) UFC/g de S. aureus en el 40,0% de las muestras. Se detectaron enterotoxinas estafilocócicas en 34,2% de las muestras, resultando la enterotoxina tipo A la más frecuente (80,7%). El queso blanco artesanal tipo “telita” expendido en mercados de la ciudad de Caracas revela deficientes medidas sanitarias en su expendio y la presencia de enterotoxinas representa un potencial riesgo para la salud de los consumidores.

Staphylococcal food intoxication is produced by eating food contaminated with staphylococcal enterotoxins produced by Staphylococcus aureus, constituting one of the main diseases produced by food at a worldwide level. In this study we investigated S. aureus presence in white artisan “telita” type” cheese sold at various markets in Caracas city. Eighty samples were analyzed and the bacterial counts were done according to COVENIN Venezuelan guidelines 1292:89. Presence of S. aureus enterotoxins type A, B, C, and D was determined by the reverse phase latex agglutination test (SET RPLA Oxoid). We found S. aureus bacterial loads between 10³ - 10(4) CFU/g in 40.0% of the samples. We detected staphylococcal enterotoxins in 34.2% of the samples, enterotoxin type A being the most frequent (80.7%). The white artisan “telita” type cheese being sold at markets in Caracas city reveals deficient sanitary measures for its sale, and the presence of enterotoxins represents a potential health risk for consumers.