Cocoa Mucilage as a Novel Ingredient in Innovative Kombucha Fermentation.

Cocoa tree plantations aim to harvest grains found in the cob to produce cocoa and chocolate. There has been a growing interest in valorizing the secondary components of the cocoa fruit, such as the peel, placenta, and mucilage/pulp, as valuable sources of nutrients for healthy food preparation. In other words, by-products derived from these raw materials are an exploitable source of nutrients in the preparation of healthy food. In the present study, two varieties of cocoa, National Cocoa Fino de Aroma (NCFA) and Colección Castro Naranjal 51 (CCN-51), were evaluated and harvested during both dry and rainy seasons. This evaluation was based on the profiling of the cob, peel, grain, placenta, and mucilage in different stages of ripeness (underripe, ripe, and overripe). Also, from the ripe raw material, a fermented beverage prototype was developed, such as kombucha, with different concentrations of mucilage (40, 60, 80, and 100 g/L). Physicochemical analyses, such as acidity, °Brix, pH, moisture, ash, protein, fat, fiber, vitamins, sugars, and polyphenols of the raw mucilage material and acidity, °Brix, and pH values of the fermented kombucha, were carried out. The best performances were obtained with the CCN-51 variety in the rainy season. Among the fermented drink panelists, the CN40 treatment (Nacional Mucilage + 40 g/L of sugar) received the highest acceptability and was considered the best. Given its efficiency, nutritional content, and potential applications, this product presents a promising strategy to address Sustainable Development Goals related to zero hunger, health and well-being, and climate action.

Prevalence and coexistence of type 2 inflammatory diseases.

BACKGROUND: Type 2 inflammation has been described as a pathophysiological basis common to some diseases, such as atopic dermatitis (AD), chronic rhinosinusitis with nasal polyps, and asthma (CRSwNP). OBJECTIVE: The present study used population-based prevalence in Catalonia to analyse the coexistence of type 2 inflammatory diseases in patients primarily diagnosed with the above mentioned conditions. RESULTS: We found a high degree of coexistence of type 2 inflammatory diseases among these patients, with the prevalence being higher in the severe forms, except for AD. For the severe forms of primary diseases, the proportion of patients with coexisting type 2 inflammatory diseases (severe or non-severe) was 16.2% for AD, 19.8% for asthma, and a striking 62.4% for CRSwNP. This patient population has the highest proportion of coexisting type 2 inflammatory diseases, both severe (48.9%) and non-severe (13.5%). CONCLUSION: Our findings have significant implications for the management of patients with AD, asthma, and CRSwNP.

Meta-analysis of antimicrobial activity of Allium, Ocimum, and Thymus spp. confirms their promising application for increasing food safety.

Biopreservation strategies such as the use of Mediterranean plant extracts to ensure food safety are promising to deal with the emergence of antimicrobial resistances and the overreliance on food chemical additives. In the last few decades, antimicrobial susceptibility testing (AST) for evaluating the in vitro antibacterial potential of plant extracts against the most relevant foodborne pathogens has been widely reported in the literature. The current meta-analysis aimed to summarise and analyse the extensive evidence available in the literature regarding the in vitro antimicrobial capability of Allium, Ocimum and Thymus spp. extracts against foodborne pathogens. A systematic review was carried out to gather data on AST results of these extracts against Listeria monocytogenes, Staphylococcus aureus, Salmonella spp., Escherichia coli and Bacillus cereus, including inhibition diameters (ID) and minimum inhibitory concentrations (MIC). A total of 742 records were gathered from a raw collection of 2,065 articles. Weighted mixed-effect linear models were adjusted to data to obtain pooled ID, pooled MIC and the relationship between both model estimations and observations. The pooled results revealed B. cereus as the most susceptible bacteria to Allium sativum (pooled ID = 20.64 ± 0.61 mm) by diffusion methods and S. aureus (pooled MIC = 0.146 mg/mL) by dilution methods. Diffusion methods did not yield conclusive results for Ocimum spp. extracts; however, the lowest pooled MIC was obtained for S. aureus (0.263 mg/mL). Among the foodborne pathogens evaluated, B. cereus showed the highest sensitivity to Thymus spp. extracts by both diffusion and dilution methods (pooled ID = 28.90 ± 2.34 mm and MIC = 0.075 mg/mL). The methodology used for plant extraction was found to not significantly affect MIC values (p > 0.05). Overall, the antimicrobial effectiveness of the studied extracts against Gram-positive and Gram-negative bacteria was demonstrated. Finally, the robustness of the meta-regression model was confirmed, also revealing an inversely proportional correlation between the ID and MIC measurements (p < 0.0001). These results provide a robust scientific basis on the factors affecting the in vitro antimicrobial efficacy of extracts from Mediterranean plants. They also provide valuable information for stakeholders involved in their industrial application in food, including producers, regulatory agencies and consumers which demand green-labelled foods.

Fermentative optimization and characterization of exopolysaccharides from Enterococcus faecium F58 isolated from traditional fresh goat cheese.

This study focused on optimizing the fermentation-based production of Exopolysaccharides (EPS) from Enterococcus faecium F58 initially isolated from traditional Moroccan Jben, a fresh goat cheese. Using the central composite design, yeast extract, MnSO4, and time affect EPS concentration. The highest experimental and predicted EPS production yields were 2.46 g/L ± 0.38 and 2.86 g/L, respectively. Optimal concentrations of yeast extract (4.46 g/L) and MnSO4 (0.011 g/L) were identified after 26 h at 30 °C. Characterization of EPS was conducted using SEM with EDX, XRD, and FTIR analyses. These tests revealed a specific morphology and an amorphous structure. Additionally, thermogravimetric analysis indicated adequate EPS stability up to 200 °C with anti-adhesion properties against different pathogens. This study offers valuable insights into the optimized production of EPS from Enterococcus faecium F58, which exhibits significant structural and functional properties for various applications in the food and biotechnology industries. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-023-01424-9.

Optimal experimental design (OED) for the growth rate of microbial populations. Are they really more "optimal" than uniform designs?

Secondary growth models from predictive microbiology can describe how the growth rate of microbial populations varies with environmental conditions. Because these models are built based on time and resource consuming experiments, model-based Optimal Experimental Design (OED) can be of interest to reduce the experimental load. In this study, we identify optimal experimental designs for two common models (full Ratkowsky and Cardinal Parameters Model (CPM)) for a different number of experiments (10-30). Calculations are also done fixing one or more model parameters, observing that this decision strongly affects the layout of the OED. Using in silico experiments, we conclude that OEDs are more informative than conventional (equidistant) designs with the same number of experiments. However, OEDs cluster the experiments near the growth limits (Xmin and Xmax) resulting in impractical designs with aggregated experimental runs ~10 times longer than conventional designs. To mitigate this, we propose a novel optimality criterion (i.e., the objective function) that accounts for the aggregated time. The novel criterion provides a reduction in parameter uncertainty with respect to the conventional design, without an increase in the experimental load. These results underline that an OED is only based on information theory (Fisher information), so the results can be impractical when actual experimental limitations are considered. The study also emphasizes that most OED schemes identify where to measure, but do not give an indication on how many experiments should be made. In this sense, numerical simulations can estimate the parameter uncertainty that would be obtained for a particular experimental design (OED or not). These results and methodologies (available in Open Code) can guide the design of future experiments for the development of secondary growth models.

Comparison of Long-term Response and Remission to Omalizumab and Anti-IL-5/IL-5R Using Different Criteria in a Real-life Cohort of Severe Asthma Patients.

BACKGROUND: Evaluation of biologic therapy response is vital to monitor its effectiveness. Authors have proposed various response criteria including good responder, super-responder, non-responder, and clinical remission. OBJECTIVES: To ascertain the prevalence of response and clinical remission after long-term treatment (>6 months) of anti-IgE and anti-IL-5/IL-5Rα biologics, compare these results with existing criteria, and identify predictors for non-responders and clinical remission. METHODS: A multicenter, real-life study involving severe asthma patients in Spain. Various outcomes were assessed to gauge response and clinical remission against established criteria. RESULTS: The study included 429 patients, 209 (48.7%) omalizumab, 112 (26.1%) mepolizumab, 19 (4.4%) reslizumab and 89 (20.7%) benralizumab, with a mean treatment duration of 55.3±38.8 months. In the final year of treatment, 218 (50.8%) were super-responders, 173 (40.3%) responders, 38 (8.9%) non-responders, and clinical remission in 116 (27%), without differences among biologics. The short-term non-responders (<6 months) were 25/545 (4.6%). Substantial variations in response and clinical remission were observed when applying different published criteria. Predictors of non-response included higher BMI (OR:1.14; 95% CI:1.06-1.23; p<0.001), admissions at ICU (2.69; 1.30-5.56; p=0.01), high count of SAE (1.21; 1.03-1.42; p=0.02) before biologic treatment. High FEV1% (0.96; 0.95-0.98; p<0.001), a high ACT score (0.93; 0.88-0.99; p=0.01) before biologic treatment or NSAID-ERD (0.52; 0.29-0.91; p=0.02) showed strong associations with achieving clinical remission. CONCLUSION: A substantial proportion of severe asthma patients treated long-term with omalizumab or anti-IL5/IL-5Rα achieved a good response. Differences in response criteria highlight the need for harmonization in defining response and clinical remission in biologic therapy to enable meaningful cross-study comparisons.

Upregulation of Platelet-Activating Factor Receptor Expression and Lyso-Platelet-Activating Factor Isoforms in Human Nasal Polyp Tissues.

BACKGROUND: The Platelet-Activating Factor (PAF)/receptor (PAFR) system is involved in asthma and allergic rhinitis; however, its role in chronic rhinosinusitis (CRS) is still unclear. This study aimed to assess the expression of PAFR and the concentration of Lyso-PAF isoforms in the nasal polyps (NP) of patients suffering from CRS with/without comorbidities such as asthma and NSAID-exacerbated respiratory disease (N-ERD) compared to healthy nasal mucosa (NM) from control subjects. METHODS: NM (n = 8) and NP tissues were obtained from patients undergoing surgery for septal deviation/turbinate hypertrophy or endoscopic sinus surgery, respectively. Three phenotypes were studied: CRSwNP with no asthma (n = 6), CRSwNP with non-steroidal anti-inflammatory drug (NSAID)-tolerant asthma (n = 6), and CRSwNP with NSAID-exacerbated respiratory disease (n = 6). PAFR protein and mRNA were assessed via immunochemistry, immunofluorescence, Western blot, and real-time quantitative PCR. Lyso-PAF isoforms (C16, C18, and C18:1) were quantified via mass spectrometry. RESULTS: PAFR protein was expressed in the NM and NP, concretely in epithelial cells and submucosal glands. Compared to NM, PAFR mRNA expression was higher in all NP phenotypes (p < 0.05) while all Lyso-PAF isoform concentrations were higher in the NP from asthmatic patients (p < 0.05). Lyso-PAF C16 and C18 concentrations were higher in the NP from asthmatic patients than in the NP from patients without asthma. CONCLUSIONS: The PAF/PAFR system could play a pathophysiological role in CRSwNP pathogenesis.

The provenance of the stones in the Menga dolmen reveals one of the greatest engineering feats of the Neolithic.

The technical and intellectual capabilities of past societies are reflected in the monuments they were able to build. Tracking the provenance of the stones utilised to build prehistoric megalithic monuments, through geological studies, is of utmost interest for interpreting ancient architectures as well as to contribute to their protection. According to the scarce information available, most stones used in European prehistoric megaliths originate from locations near the construction sites, which would have made transport easier. The Menga dolmen (Antequera, Malaga, Spain), listed in UNESCO World Heritage since July 2016, was designed and built with stones weighting up to nearly 150 tons, thus becoming the most colossal stone monument built in its time in Europe (c. 3800-3600 BC). Our study (based on high-resolution geological mapping as well as petrographic and stratigraphic analyses) reveals key geological and archaeological evidence to establish the precise provenance of the massive stones used in the construction of this monument. These stones are mostly calcarenites, a poorly cemented detrital sedimentary rock comparable to those known as 'soft stones' in modern civil engineering. They were quarried from a rocky outcrop located at a distance of approximately 1 km. In this study, it can be inferred the use of soft stone in Menga reveals the human application of new wood and stone technologies enabling the construction of a monument of unprecedented magnitude and complexity.

Variability in Physicochemical Parameters and Its Impact on Microbiological Quality and Occurrence of Foodborne Pathogens in Artisanal Italian Organic Salami.

Artisanal salami is produced in small-scale production plants, where the lack of full automation might result in higher variability in food intrinsic properties. The aim of the present study was to evaluate the inter- and intra-batch variability in physicochemical parameters and its impact on microbial quality and occurrence of foodborne pathogens on 480 samples collected from six batches of an artisanal Italian production of organic salami. Relatively high total bacterial counts (TBC) were found on the surface of the table in the stuffing room (4.29 ± 0.40 log cfu/cm2). High loads of Enterobacteriaceae in the meat mixture of batch 2 and TBC in batch 5 were associated with a higher occurrence of bacterial pathogens. During ripening, water activity (aw) and pH failed to reach values lower than 0.86 and 5.3, respectively. Six Staphylococcus aureus and four Listeria monocytogenes isolates were collected from the salami meat mixture during ripening and the processing environment. A total of 126 isolates of Enterobacteriaceae were characterized at a species level, with Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, and Citrobacter freundii isolated from the final products. Results suggest the relevance of first steps of production in terms of the hygiene of raw materials and handling during stuffing procedures, especially when the physicochemical parameters of the final products do not reach values that represent hurdles for foodborne pathogens.

LP309 a new strain of Lactiplantibacillus pentosus that improves the lactic fermentation of Spanish-style table olives.

Different varieties of table olives have suitable morphological characteristics that allow them to be processed as Spanish-style green table olives. However, the Campiñesa cultivar presents difficulties when submitted to a lactic fermentation, in spite of being inoculated with dedicated starter cultures such as OleicaStarter. The strategy followed in this study to facilitate the start of lactic fermentation was to reinforce the OleicaStarter culture with the use of the Lactoplantibacillus pentosus Lp309 a strain that enhanced the survival of lactic acid bacteria (LAB) at the beginning of fermentation, reaching final pH values (4.08 ± 0.01), free acidity (1.00 ± 0.01 g/100 mL of brine), LAB population (6.17 ± 0.09 log CFU/mL), nutrient depletion (0.80 ± 0.09 g/kg of pulp), and lactic acid production (11.85 ± 0.72 g/L). These values allowed stabilization of the final product, thus complying with the quality and food safety standards established by the Codex Alimentarius for table olives.

Modeling the combined resistance to microwave treatments and salt conditions of Escherichia coli and Staphylococcus aureus.

In the present study, the efficiency of the combined effect of microwave irradiation treatments together with salt concentration was assessed against Escherichia coli and Staphylococcus aureus. Microbial survival has been modeled through a one-step Weibull equation considering the non-isothermal profiles during the heating treatments. Three sodium chloride concentrations 0.5%, 3.5%, and 8.5% (w/v) treated under three microwave power levels (450, 600, and 800 W) were studied. Predictive models were validated using the determination coefficient (R2), root mean squared error and the acceptable prediction zone with external data obtained from ultra high temperature milk. The results obtained suggested that increasing microwave power levels and decreasing salt concentrations led to a higher microbial inactivation, being the δ values (time for achieving a first decimal reduction) for E coli of 19.57 s at 800 W and 0.5% NaCl. In contrast, experimental data of S aureus showed a higher variability since it presented more resistance to the microwave treatments. The results obtained and generated models can be used as decision-making tools to set specific guidelines on microwave treatments for assuring food safety.

The earliest basketry in southern Europe: Hunter-gatherer and farmer plant-based technology in Cueva de los Murciélagos (Albuñol).

Plant material culture can offer unique insights into the ways of life of prehistoric societies; however, its perishable nature has prevented a thorough understanding of its diverse and complex uses. Sites with exceptional preservation of organic materials provide a unique opportunity for further research. The burial site of Cueva de los Murciélagos in southern Iberia, uncovered during 19th-century mining activities, contained the best-preserved hunter-gatherer basketry in southern Europe, together with other unique organic artifacts associated with the first farming communities, such as sandals and a wooden hammer. We present 14 14C dates for the perishable artifacts (N = 76), situating the assemblage between the Early and Middle Holocene (c. 7500 to 4200 cal BCE). Our integrated analysis includes raw material determination and technological and chrono-cultural contextualization of this unique and important set of materials.

In vitro antimicrobial activity of extracts and essential oils of Cinnamomum, Salvia, and Mentha spp. against foodborne pathogens: A meta-analysis study.

Essential oils (EOs) are a class of natural products that exhibit potent antimicrobial properties against a broad spectrum of bacteria. Inhibition diameters (IDs) and minimum inhibitory concentrations (MICs) are the typical measures of antimicrobial activity for extracts and EOs obtained from Cinnamomum, Salvia, and Mentha species. This study used a meta-analytical regression analysis to investigate the correlation between ID and MIC measurements and the variability in antimicrobial susceptibility tests. By utilizing pooled ID models, this study revealed significant differences in foodborne pathogens' susceptibility to extracts, which were dependent on both the plant species and the methodology employed (p < .05). Cassia showed the highest efficacy against Salmonella spp., exhibiting a pooled ID of 26.24 mm, while cinnamon demonstrated the highest efficacy against Bacillus cereus, with a pooled ID of 23.35 mm. Mint extract showed the greatest efficacy against Escherichia coli and Staphylococcus aureus. Interestingly, cinnamon extract demonstrated the lowest effect against Shiga toxin-producing E. coli, with a pooled ID of only 8.07 mm, whereas its EOs were the most effective against this bacterial strain. The study found that plant species influenced the MIC, while the methodology did not affect MIC measurements (p > .05). An inverse correlation between ID and MIC measurements was identified (p < .0001). These findings suggest that extracts and EOs obtained from Cinnamomum, Salvia, and Mentha spp. have the potential to inhibit bacterial growth. The study highlights the importance of considering various factors that may influence ID and MIC measurements when assessing the effectiveness of antimicrobial agents.

Assessing Listeria monocytogenes growth kinetics in rice pudding at different storage temperatures.

Rice pudding is a popular artisanal dairy dessert highly consumed in the main rice-producing countries, including Egypt. This study aimed to evaluate and model the growth of Listeria monocytogenes in rice pudding dessert stored at different temperatures (4-25 °C) over its shelf-life. Lab-scale rice pudding samples were prepared following a traditional Egyptian recipe and inoculated with a three-strain cocktail of L. monocytogenes (ca. 3 × 102 cfu/g). Inoculated rice pudding samples (pH = 6.67 ± 0.06 and aw = 0.99 ± 0.002) were stored at different isothermal conditions (4, 8, 12, 18, and 25 °C) and microbiologically analysed for up to 30 days for pathogen quantification by plate count methodology. Global regression analysis was used to fit the Baranyi model coupled with the Ratkowsky model to growth data, relating L. monocytogenes concentrations (N, log cfu/g) with storage temperature (°C) and times (d). Model validation was performed using published independent data. L. monocytogenes growth potential increased by increasing storage temperature. The estimated Ratkowsky model parameters were b = 0.0819 ± 0.0017 log cfu/d·°C and Tmin = -3.28 ± 0.20 °C. The indices RMSE = 0.39 and R2adj = 0.97 indicated a good agreement between the experimental data and the model predictions. The estimated maximum growth rate (µmax) values ranged between 0.355 and 5.363 log cfu/d from 4 to 25 °C. The model was successfully validated using published L. monocytogenes Scott A and California strains growth data in rice pudding samples stored at 5, 12 and 22 °C, as evidenced by the assessed statistical indices. The predictive model developed and validated in this study will aid in decision-making regarding the microbiological safety of rice pudding dessert with respect to L. monocytogenes growth, considering a wide range of storage temperatures.

Prevalence of atopic dermatitis in the adolescent population of Catalonia (Spain).

BACKGROUND: Studies on the prevalence of atopic dermatitis (AD) for the adolescent cohort in general-based large populations are scarce worldwide. We performed a retrospective population--based observational cohort study of 76,665 adolescent patients diagnosed with AD in Catalonia (Spain). We studied the prevalence of AD by age, gender, disease severity, comorbidities, serum total immunoglobulin E (tIgE) and appropriate medical treatment (AMT) for the Catalan population. METHODS: Adolescent individuals (12-17 years) diagnosed with AD by medical records at different health care levels (primary, hospital, emergency) from the Catalan Health System (CHS) were included. Statistical analyses evaluated sociodemographic characteristics, prevalence, comorbidities, serum tIgE and AMT. RESULTS: The overall diagnosed AD prevalence in the adolescent Catalan population (76,665) was 16.9%, being higher for the non-severe (16.7%) than for the severe (0.2%) populations. Topical corticosteroids were the most prescribed drug (49.5%), and the use of all prescribed treatments was higher in severe AD patients, especially systemic corticosteroids (49.7%) and immunosuppressants (45.4%). AD patients had, on average, a serum tIgE of 163.6 KU/L, which was higher for severe than non-severe disease (155.5 KU/L vs 101.9 KU/L, respectively). Allergic rhinitis (15.0%) and asthma (13.5%) were among the most frequent comorbid respiratory and allergy diseases. CONCLUSIONS: This is the first Spanish study reporting the overall diagnosed prevalence for a large-scale adolescent cohort (12-17 years old) from Catalonia. It provides new and robust evidence of AD's prevalence and related characteristics in this region.

Effect of Obesity on the Expression of Genes Associated with Severe Asthma-A Pilot Study.

Asthma is a complex condition resulting from the interaction of genes and environment. Obesity is a risk factor to develop asthma and contributes to poor response to asthma therapy and severity. The aim of the study was to evaluate the effect of obesity on the expression levels of genes previously associated with severe asthma. Three groups of subjects were studied: non-obese asthmatics (NOA), obese asthma patients (OA), and non-asthmatic obese subjects (O). Previously reported overexpressed (IL-10, MSR1, PHLDA1, SERPINB2, and CD86) and underexpressed genes (CHI3L1, CPA3, IL-8, and PI3) in severe asthma were analyzed by RT-qPCR in peripheral blood mononuclear cells (PBMCs). In the overexpressed genes, obesity significantly decreased the expression of MSR1 and PHLDA1 and had no effects on CD86, IL-10, and SERPINB2. In underexpressed genes, obesity did not affect PI3, CHI3L1, and IL-8 and significantly reduced CPA3 expression. The results of this study show that obesity should be included among the known factors that can contribute toward modifying the expression of genes associated with asthma and, in particular, severe asthma.

Tracking microbial quality, safety and environmental contamination sources in artisanal goat cheesemaking factories.

A harmonised microbiological survey was performed in two artisanal factories of raw goat milk cheeses (A and B) located in the Andalusian region (Spain). A total of 165 different control points or samples (raw materials, final products, food-contact surfaces [FCS], and air) were examined as microbial and pathogen sources of contamination of artisanal goat raw milk cheeses. For raw milk samples analysed from both producers, the concentrations of aerobic-mesophilic bacteria (AMB), total coliforms, coagulase-positive Staphylococcus spp. (CPS), lactic-acid bacteria (LAB) and moulds and yeasts ranged between 3.48 and 8.59, 2.45-5.48, 3.42-4.81, 4.99-8.59 and 3.35-6.85 log cfu/mL respectively. For the same microbial groups, the analysis of raw milk cheeses revealed concentrations ranging from 7.82 to 8.88, 2.00-6.82, 2.00-5.28, 8.11-9.57 and 2.00-5.76 log cfu/g, respectively. Although the raw material analysed from producer A presented higher microbial loads and between-batch variability, it was B the producer with the most loaded final products. Regarding the microbial air quality, the fermentation area, storage room, milk reception and packaging room were the most AMB loaded places, while the ripening chamber was the area with higher fungal loads in bioareosol from both producers. Conveyor belts, cutting machine, storage boxes and brine tank were highlighted as the most contaminated FCS evaluated. Staphylococcus aureus was the only pathogen detected within the set of 51 isolates from samples as revealed by MALDI-TOF and molecular PCR, with a prevalence of 12.5% for samples from the producer B. The public health risk attributed to the consumption of artisanal goat cheese should not be neglected, and may consider the whole cheesemaking processing chain, from microbiological quality of raw milk to the ready-to-eat final product, especially concerning the presence of S. aureus.