The ecological conditions during the ripening of dry-cured ham favour the development of moulds on its surface, being frequently the presence of Penicillium nordicum, a producer of ochratoxin A (OTA). Biocontrol using moulds and yeasts usually found in dry-cured ham is a promising strategy to minimize this hazard. The aim of this work is to evaluate the effect of previously selected Debaryomyces hansenii and Penicillium chrysogenum strains on growth, OTA production, and relative expression of genes involved in the OTA biosynthesis by P. nordicum. P. nordicum was inoculated against the protective cultures individually and combined on dry-cured ham for 21 days at 20 °C. None of the treatments reduced the growth of P. nordicum, but all of them decreased OTA concentration. The lower production of OTA could be related to significant repression of the relative expression of otapksPN and otanpsPN genes of P. nordicum. The efficacy of the combined protective cultures was tested in 24 dry-cured hams in industrial ripening (an 8 month-long production). OTA was detected in nine of the 12 dry-cured hams in the batch inoculated only with P. nordicum. However, in the batch inoculated with both P. nordicum and the combined protective culture, a considerable reduction of OTA contamination was observed. In conclusion, although the efficacy of individual use P. chrysogenum is great, the combination with D. hansenii enhances its antifungal activity and could be proposed as a mixed protective culture to control the hazard of the presence of OTA in dry-cured ham.

Toxigenic moulds can develop on the surface of dry-cured meat products during ripening due to their ecological conditions, which constitutes a risk for consumers. A promising strategy to control this hazard is the use of antifungal microorganisms usually found in these foods. However, to date, the effectiveness of gram-positive catalase-positive cocci (GCC+) has not been explored. The aim of this work was to select GCC+ isolates with antifungal activity to study its effectiveness in a dry-cured ham model system at the environmental conditions reached during the ripening. Forty-five strains of GCC+ were evaluated and the isolate Staphylococcus xylosus Sx8 was selected to assess its efficacy at two different concentrations (10⁶ and 10⁴ cfu/mL) against Penicillium nordicum, Aspergillus flavus, Aspergillus parasiticus, and Penicillium griseofulvum at 15, 20, and 25 °C. The results showed that the inoculation of 10⁶ cfu/mL of S. xylosus completely inhibited the growth of most fungi. In addition, in the presence of this strain at 10⁴ cfu/mL, a significant reduction in fungal growth and mycotoxins production was observed at the three temperatures studied. In conclusion, S. xylosus Sx8 possesses great potential as a biological agent to control toxigenic moulds in dry-cured meat products.

Ripened foods of animal origin comprise meat products and dairy products, being transformed by the wild microbiota which populates the raw materials, generating highly appreciated products over the world. Together with this beneficial microbiota, both pathogenic and toxigenic microorganisms such as Listeria monocytogenes, Salmonella enterica, Staphylococcus aureus, Clostridium botulinum, Escherichia coli, Candida spp., Penicillium spp. and Aspergillus spp., can contaminate these products and pose a risk for the consumers. Thus, effective strategies to hamper these hazards are required. Additionally, consumer demand for clean label products is increasing. Therefore, the manufacturing sector is seeking new efficient, natural, low-environmental impact and easy to apply strategies to counteract these microorganisms. This review gathers different approaches to maximize food safety and discusses the possibility of their being applied or the necessity of new evidence, mainly for validation in the manufacturing product and its sensory impact, before being implemented as preventative measures in the Hazard Analysis and Critical Control Point programs.

The role of indigenous fungi in the appropriate development of sensory properties and the safety of dry-fermented sausages has been widely established. Nonetheless, their applications as probiotic agents have not been elucidated in such products yet, despite their promising functional features. Thus, it should be interesting to evaluate the probiotic potential of native Debaryomyces hansenii isolates from dry-fermented sausages and their application in the meat industry, because it is the most frequently isolated yeast species from these foodstuffs and its probiotic effects for animals as well as its possible probiotic activity for human beings have been demonstrated. Within the functional ability of foodborne yeasts, anti-inflammatory, antioxidant, antimicrobial, antigenotoxic, and immunomodulatory properties have been reported. Similarly, the use of dry-fermented sausages as vehicles for probiotic moulds remains a challenge because the survival and development of moulds in the gastrointestinal tract are still unknown. Nevertheless, some moulds have been isolated from faeces possibly from their spores as a form of resistance. Additionally, their beneficial effects on animals and humans, such as the decrease in lipid content and the anti-inflammatory activity, have been reported, although they seem to be more related to their postbiotic capacity due to the generated bioactive compounds with profunctional attributes than to their role as probiotics. Therefore, further studies providing knowledge useful for generating dry-fermented sausages with improved functionality are fully necessary.

The ripening process of dry-cured meat products is characterised by the development of fungi on the product’s surface. This population plays a beneficial role, but, uncontrolled moulds represent a health risk, since some of them may produce mycotoxins, such as ochratoxin A (OTA). The aim of the present work is to assess the potential of near-infrared spectroscopy (NIRS) for the detection of OTA-producing mould species on dry-cured ham-based agar. The collected spectra were used to develop Support Vector Machines–Discriminant Analysis (SVM-DA) models by a hierarchical approach. Firstly, an SVM-DA model was tested to discriminate OTA and non-OTA producers; then, two models were tested to discriminate species among the OTA producers and the non-OTA producers. OTA and non-OTA-producing moulds were discriminated with 85% sensitivity and 86% specificity in the prediction. Furthermore, the SVM-DA model could differentiate non-OTA-producing species with a 95% sensitivity and specificity. Promising results were obtained for the prediction of the four OTA-producing species tested, with a 69% and 90% sensitivity and specificity, respectively. The preliminary approach demonstrated the high potential of NIR spectroscopy, coupled with Chemometrics, to be used as a real-time automated routine monitorization of dry-cured ham surfaces.

Competition for resources is a common microbial interaction in the gut microbiome. Inulin is a well-studied prebiotic dietary fiber that profoundly shapes gut microbiome composition. Several community members and some probiotics, such as Lacticaseibacillus paracasei, deploy multiple molecular strategies to access fructans. In this work, we screened bacterial interactions during inulin utilization in representative gut microbes. Unidirectional and bidirectional assays were used to evaluate the effects of microbial interactions and global proteomic changes on inulin utilization. Unidirectional assays showed the total or partial consumption of inulin by many gut microbes. Partial consumption was associated with cross-feeding of fructose or short oligosaccharides. However, bidirectional assays showed strong competition from L. paracasei M38 against other gut microbes, reducing the growth and quantity of proteins found in the latter. L. paracasei dominated and outcompeted other inulin utilizers, such as Ligilactobacillus ruminis PT16, Bifidobacterium longum PT4, and Bacteroides fragilis HM714. The importance of strain-specific characteristics of L. paracasei, such as its high fitness for inulin consumption, allows it to be favored for bacterial competence. Proteomic studies indicated an increase in inulin-degrading enzymes in co-cultures, such as β-fructosidase, 6-phosphofructokinase, the PTS D-fructose system, and ABC transporters. These results reveal that intestinal metabolic interactions are strain-dependent and might result in cross-feeding or competition depending on total or partial consumption of inulin. Partial degradation of inulin by certain bacteria favors coexistence. However, when L. paracasei M38 totally degrades the fiber, this does not happen. The synergy of this prebiotic with L. paracasei M38 could determine the predominance in the host as a potential probiotic.