Autochthonous Lactic Acid Bacteria Research Articles

Compensative role of autochthonous lactic acid bacteria in physical properties and taste profiles of dry sausage with partial substitution of NaCl by KCl

The defects of quality and taste caused by the partial substitution of NaCl by KCl limit the widespread use of KCl as salt substitute in fermented sausages. The objective of this study was to investigate the compensative role of four autochthonous lactic acid bacteria (LAB) strains, including Latilactobacillus curvatus, Latilactobacillus sakei, Weissella hellenica, and Lactiplantibacillus plantarum, in the physicochemical properties and taste profiles of the dry sausages with 40% substitution of NaCl by KCl. The results demonstrated that LAB inoculated reduced the pH, moisture content, and water activity and increased the organic acid contents and certain free amino acid (FAA) contents (P < 0.05). Higher contents of total acids was detected in the L. curvatus-inoculated and L. sakei-inoculated sausages (P < 0.05). Sweet FAAs contents in L. sakei-inoculated sausages increased by 36.90% and bitter FAAs contents decreased by 18.18%. According to the results of the electronic tongue, the sausage inoculated with L. sakei reduced the response value of bitterness by 5.7% (P < 0.05) and exhibited the most similar overall taste profile to traditional sausages. Overall, L. sakei could be used as a potential starter culture to enhance the physicochemical and compensate for the flavour deficiencies of NaCl-substituted dry sausages.

Autochthonous cultures to improve the quality of PGI Castellano cheese: Impact on proteolysis, microstructure and texture during ripening

The aim of this research was to find out the effect of different combinations of starter and non-starter cultures on the proteolysis of Castellano cheese during ripening. Four cheese batches were prepared, each containing autochthonous lactobacilli and or Leuconostoc, and were compared with each other and with a control batch, that used only a commercial starter. To achieve this, nitrogen fractions (pH 4.4-soluble nitrogen and 12 % trichloroacetic acid soluble nitrogen, polypeptide nitrogen and casein nitrogen), levels of free amino acids and biogenic amines were assessed. Texture and microstructure of cheeses were also evaluated. Significant differences in nitrogen fractions were observed between batches at different stages of ripening. The free amino acid content increased throughout the cheese ripening process, with a more significant increase occurring after the first 30 days. Cheeses containing non-starter lactic acid bacteria exhibited the highest values at the end of the ripening period. Among the main amino acids, GABA was particularly abundant, especially in three of the cheese batches at the end of ripening. The autochthonous lactic acid bacteria were previously selected as non-producers of biogenic amines and this resulted in the absence of these compounds in the cheeses. Analysis of the microstructure of the cheese reflected the impact of proteolysis. Additionally, the texture profile analysis demonstrated that the cheese's hardness intensified as the ripening period progressed. The inclusion of autochthonous non-starter lactic acid bacteria in Castellano cheese production accelerated the proteolysis process, increasing significantly the free amino acids levels and improving the sensory quality of the cheeses.

Cereus jamacaru DC. (mandacaru) fruit as a source of lactic acid bacteria with in vitro probiotic-related characteristics and its protective effects on Pediococcus pentosaceus during lyophilization and refrigeration storage

This study isolated and identified autochthonous lactic acid bacteria (LAB) from mandacaru fruit and evaluated their potential probiotic and technological aptitudes in vitro, as well as the protective effects of freeze-dried mandacaru fruit on the most promising LAB isolate during lyophilization and refrigeration storage. Initially, 212 colonies were isolated from mandacaru fruit, and 34 were preliminarily identified as LAB. Thirteen isolates identified by 16S-rRNA sequencing as Pediococcus pentosaceus were negative for DNase, gelatinase, hemolytic, and biogenic amine production. The selected isolates showed proteolytic activity, diacetyl and exopolysaccharide production, and good tolerance to different NaCl concentrations while having low cellular hydrophobicity and antagonistic activity against pathogens. The survival of isolates sharply decreased after 3 h of exposure to pH 2 and had a good tolerance to 1 % bile salt. A principal component analysis selected P. pentosaceus 57 as the most promising isolate based on the examined technological and probiotic-related physiological properties. This isolate was lyophilized with mandacaru fruit and stored under refrigeration for 90 days. P. pentosaceus 57 lyophilized with mandacaru fruit had high viable cell counts (9.69 ± 0.03 log CFU/mL) and >50 % of physiologically active cells at 90 days of refrigeration storage. The results indicate that mandacaru fruit is a source of P. pentosaceus with aptitudes to be explored as potential probiotic and technological characteristics of interest for the food industry, besides being a good candidate for use in lyophilization processes and refrigeration storage of LAB due to its cryoprotective effects.

Brazilian indigenous nonstarter lactic acid bacteria enhance the diversification of volatile compounds in short-aged cheese.

There is growing interest in using autochthonous lactic acid bacteria (LAB) that provide unique sensory characteristics to dairy products without affecting their safety and quality. This work studied the capacity of three Brazilian indigenous nonstarter LABs (NSLAB) to produce biogenic amines (BAs) and evaluated their effect on the volatile organic compounds (VOCs), microbial LAB communities, and physicochemical profile of short-aged cheese. Initially, the strain's potential for biosynthesis of BAs was assessed by PCR and in vitro assays. Then, a pilot-scale cheese was produced, including the NSLAB, and the microbial and VOC profiles were analyzed after 25 and 45 days of ripening. As a results, the strains did not present genes related to relevant BAs and did not produce them in vitro. During cheese ripening, the Lactococci counts were reduced, probably in the production of alcohols and acid compounds by the NSLAB. Each strain produces a unique VOC profile that changes over the ripening time without the main VOCs related to rancid or old cheese. Particularly, the use of the strain Lacticaseibacillus. paracasei ItalPN16 resulted in production of ester compounds with fruity notes. Thus, indigenous NSLAB could be a valuable tool for the enhancement and diversification of flavor in short-aged cheese.

New Genetic Determinants for qPCR Identification and the Enumeration of Selected Lactic Acid Bacteria in Raw-Milk Cheese.

Lactic acid bacteria (LAB) play an important role in the ripening of cheeses and contribute to the development of the desired profile of aroma and flavor compounds. Therefore, it is very important to monitor the dynamics of bacterial proliferation in order to obtain an accurate and reliable number of their cells at each stage of cheese ripening. This work aimed to identify and conduct a quantitative assessment of the selected species of autochthonous lactic acid bacteria from raw cow's milk cheese by the development of primers and probe pairs based on the uniqueness of the genetic determinants with which the target microorganisms can be identified. For that purpose, we applied real-time quantitative PCR (qPCR) protocols to quantify Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, and Lactococcus lactis subsp. cremoris cells in cheese directly after production and over three-month and six-month ripening periods. While L. lactis subsp. cremoris shows good acidification ability and the ability to produce antimicrobial compounds, L. delbrueckii subsp. bulgaricus has good proteolytic ability and produces exo-polysaccharides, and S. thermophilus takes part in the formation of the diacetyl flavor compound by metabolizing citrate to develop aroma, they all play an important role in the cheese ripening. The proposed qPCR protocols are very sensitive and reliable methods for a precise enumeration of L. delbrueckii subsp. bulgaricus, S. thermophilus, and L. lactis subsp. cremoris in cheese samples.

Technological traits and mitigation activity of autochthonous lactic acid bacteria from mediterranean fermented meat-products

The production of safe and standardized fermented sausages with typical characteristics linked to traditional origin is highly desirable. The use of autochthonous starter cultures that provide peculiar flavor, texture and color to the fermented products, while maintaining the meat-product safe can be a feasible strategy for producers. In this study, 45 strains of Latilactobacillus sakei and 1 Latilactobacillus curvatus isolated from natural Mediterranean fermented sausages, were screened as potential protective cultures for their use in the fermented sausage industry. Technological properties, inhibitory activity towards Escherichia coli and Listeria innocua and the presence of genes coding for bacteriocins, were investigated. All tested strains showed an antagonistic effect by growing, while inhibiting the growth of target harmful microorganisms, in a strain-specific manner. At least one bacteriocin encoding genes was present in 25 strains, mainly sakacin X and sakacin P. The technological performances of the strains highlighted a great variability in the behavior, confirming the phenotypic diversity already reported for LAB species highly adapted to meat environment. Results highlight the potentiality of these strains to be used as protective starters in fermented meat products to improve food quality and microbiological safety, as well as giving peculiar characteristics to the final product.

The Role of Autochthonous Levilactobacillus brevis B1 Starter Culture in Improving the Technological and Nutritional Quality of Cow’s Milk Acid-Rennet Cheeses—Industrial Model Study

In the study, an attempt was made to develop an innovative technology for cheese manufacturing. It was hypothesized that selected autochthonous lactic acid bacteria as a starter culture are more suitable for the production of acid-rennet cheeses of good technological and sensory quality. The study aimed to assess the possibility of using the strain Levilactobacillus brevis B1 (L. brevis B1) as a starter culture to produce acid-rennet cheeses using raw cow’s milk. Two variants of cheese were manufactured. The control variant (R) was coagulated with microbial rennet and buttermilk, and the other variant (B1) was inoculated with rennet and L. brevis B1 starter culture. The effect of the addition of these autochthonous lactic acid bacteria on selected physicochemical characteristics, durability, the composition of fatty acids, cholesterol, Iipid Quality Indices, and microbiological and sensory quality of acid-rennet cheeses was determined during a 3-month period of storage. The dominant fatty acids observed in the tested cheeses were saturated fatty acids (SFA) (68.43–69.70%) and monounsaturated fatty acids (MUFA) (25.85–26.55%). Significantly higher polyunsaturated fatty acid (PUFA) content during storage was observed for B1 cheeses. The B1 cheeses were characterized by lower cholesterol content compared to cheese R and showed better indexes, including the Index of atherogenicity, Index of thrombogenicity, DFA, OFA, H/H, and HPI indexes, than the R cheese. No effect of the tested L. brevis B1 on sensory quality was observed in relation to the control cheeses during 3 months of storage. The results of the research indicate the possibility of using the L. brevis B1 strain for the production of high-quality, potentially probiotic acid-rennet cheeses.

Selecting autochthonous lactic acid bacteria for co-inoculation in Chinese bayberry wine production: Stress response, starter cultures application and volatilomic study

This study focused on isolating and characterising autochthonous lactic acid bacteria (LAB) from spontaneously fermented Chinese bayberry (CB) and their potential application in CB wine fermentation in co-inoculation with yeast starter cultures. Numerous LAB, including Lactiplantibacillus (Lp.) plantarum (9), Limosilactobacillus (Lb.) fermentum (6), Lactococcus (Lc.) lactis (3), Enterococcus (Ec.) hirae (1), Leuconostoc (Le.) mesenteroides (1), and Weissella (Ws.) cibaria (1), were isolated and identified. The isolated strains Lp. plantarum ZFM710 and ZFM715, together with Lb. fermentum ZFM720 and ZFM722, adapted well to unfavourable fermentation environment, including ethanol, osmolality, and acidity stresses, were selected for producing CB wine by co-inoculation with Saccharomyces cerevisiae. During fermentation, the presence of LAB promoted the development of S. cerevisiae, while the population dynamics of LAB in different groups at different stages showed strain-specific differences. Fermentation trials involving LAB yielded a lower ethanol concentration except for Lp. plantarum ZFM715. Compared to the pure S. cerevisiae fermented sample, the addition of LAB led to a clear modulation in organic acid composition. Lb. fermentum strains in co-fermentation led to significant decreases in each classified group of aroma compounds, while Lp. plantarum ZFM715 significantly increased the complexity and intensity of aroma compounds, as well as the intensities of fruity and floral notes. The study selects interesting strains for the design of starter cultures for use in CB wine production, underlining the interest in the selection of autochthonous LAB in fruit wines, with the aim of improving the adaptation of bacteria to specific environmental conditions and shaping the unique traits of the finished products.

Biocontrol of L. monocytogenes with Selected Autochthonous Lactic Acid Bacteria in Raw Milk Soft-Ripened Cheese under Different Water Activity Conditions.

The effect of selected autochthonous Lactic Acid Bacteria (LAB) against Listeria monocytogenes was evaluated in two elaborations of soft-ripened cheese performed under high and low relative humidity (RH) elaborations, to achieve aw ranging from 0.97 to 0.94 in ripened cheeses. Two selected autochthonous strains of Lacticaseibacillus casei 31 and 116 were used. In each elaboration, 8 batches were physicochemically and microbiologically evaluated throughout the ripening process. The aw and pH decreased during ripening to final values ranging from 0.944 to 0.972 aw and 5.0 to 5.3 pH, respectively. LAB was the only microbial group that increased throughout the ripening in high and low RH elaborations. In batches that were uninoculated with LAB strains, L. monocytogenes was either maintained at the initial inoculation level or showed a slight reduction by the end of the ripening process. However, in LAB-inoculated batches in the two elaborations, steady decreases of L. monocytogenes were observed throughout maturation. L. casei 31 alone or in combination with strain 116 provoked reductions of 2 to 4 log CFU/g in L. monocytogenes over 60 days of ripening, which could be enough as a strategy for biocontrol to deal with the usual contamination by L. monocytogenes during cheese processing.

Antilisterial activity of raw sheep milk from two native Epirus breeds: Culture-dependent identification, bacteriocin gene detection and primary safety evaluation of the antagonistic LAB biota

Raw milk from native small ruminant breeds in Epirus, Greece, is a valuable natural source of autochthonous lactic acid bacteria (LAB) strains with superior biotechnological properties. In this study, two bulk milks (RM1, RM2) from two local sheep yards, intended for traditional Kefalotyri cheese production, were preselected for bacteriocin-like antilisterial activity by in vitro tests. Their antagonistic LAB biota was quantified followed by polyphasic (16S rRNA gene sequencing; IGS for Enterococcus; a multiplex-PCR for Leuconostoc) identification of 42 LAB (RM1/18; RM2/24) isolates further evaluated for bacteriocin encoding genes and primary safety traits. Representative isolates of the numerically dominant mesophilic LAB were Leuconostoc mesenteroides (10) in both RMs, Streptococcus parauberis (7) in RM2, and Lactococcus lactis (1) in RM1; the subdominant thermophilic LAB isolates were Enterococcus durans (8), E. faecium (6), E. faecalis (3), E. hirae (1), E. hermanniensis (1), Streptococcus lutetiensis (2), S. equinus (1) and S. gallolyticus (1). Based on their rpoB, araA, dsr and sorA profiles, six Ln. mesenteroides strains (8 isolates) were atypical lying between the subspecies mesenteroides and dextranicum, whereas two strains profiled with Ln. mesenteroides subsp. jonggajibkimchii that is first-time reported in Greek dairy food. Two RM1 E. faecium strain biotypes (3 isolates) showed strong, enterocin-mediated antilisterial activity due to entA/entB/entP possession. One E. durans from RM1 possessed entA and entP, while additional nine RM2 isolates of the E. faecium/durans group processed entA or entP singly. All showed direct (cell-associated) antilisterial activity only, as also both S. lutetiensis strains from RM2 did strongly. Desirably, no LAB isolate was β-hemolyrtic, or cytolysin-positive, or possessed vanA, vanB for vancomycin resistance, or agg, espA, hyl, and IS16 virulence genes. However, all three E. faecalis from RM2 possessed gelE and/or ace virulence genes. In conclusion, all Ln. mesenteroides strains, the two safe, enterocin A-B-P-producing E. faecium strains, and the two antilisterial S. lutetiensis strains should be validated further as potential costarter or adjunct cultures in Kefalotyri cheese. The prevalence of α-hemolytic pyogenic streptococci in raw milk, mainly S. parauberis in RM2, requires consideration in respect to subclinical mastitis in sheep and the farm hygiene overall.

Characterization of Lactic Acid Bacteria and Yeast from Grains as Starter Cultures for Gluten-Free Sourdough.

With the increasing number of people affected by gluten consumption-related diseases, adhering to a gluten-free (GF) diet is the most effective preventive measure. Herein, we aimed to isolate and characterize the functional properties of autochthonous lactic acid bacteria (LAB) and yeast from various GF sourdoughs to determine their suitability in starter cultures for sourdough preparation. Three LAB, Weissella confusa BAQ2, Lactobacillus brevis AQ2, Leuconostoc citreum YC2, and Saccharomyces cerevisiae BW1, were identified. The isolated LAB exhibited greater TTA, faster acidification rates, and higher acid tolerance than commercial LAB. W. confusa BAQ2 exhibited the highest EPS production, W. confusa BAQ2 and L. brevis AQ2 showed high maltose utilization, and S. cerevisiae BW1 exhibited the highest CO2 production rate. Accordingly, all four microbial strains were mixed for the starter culture. The sourdough prepared with starter cultures exhibited differences in gas production depending on fermentation time, which influenced the volume of GF bread dough. GF bread prepared with fermented sourdough exhibited a 16% higher specific volume and enhanced crumb firmness and elasticity than that prepared using non-fermented sourdough. Thus, autochthonous LAB strains isolated from various GF sourdoughs can be used together to improve the quality of sourdough bread, demonstrating their potential for use in starter cultures for GF sourdough production.

Identification, Viability, and Membrane Potential during the Cryopreservation of Autochthonous Lactic-Acid Bacteria Isolated from Artisanal Adobera Cheese from Los Altos de Jalisco

Lactic acid bacteria (LAB) comprise a group of microorganisms responsible for developing the sensory and chemical characteristics of several foods and fermented products, particularly cheese. For this reason, after isolation and identification of LAB, validated protocols and procedures for their long-term preservation without compromising its integrity and technological properties, as well as methodologies aiming to assess their viability and integrity are paramount. This study aimed to isolate and identify autochthonous LAB from artisanal Adobera cheese and determine the effect of LAB cryopreservation with thioglycolate broth and glycerol on their viability, membrane integrity, and kinetics. Sixteen LAB were isolated and genetically identified from artisanal cheese samples; eleven of those strains were selected (genus Lactobacillus, Leuconostoc, Streptococcus, and Lactococcus) and included in the cryo-preservation assay. The initial average concentration of the bacterial suspensions was 6.89 log10 CFU mL−1; increasing to 8.9 log10 CFU mL−1 21 days later and slightly reduced at day 42 post-preservation (losses below one logarithm). About 77% of the cells maintained their membrane potential 180 days after their preservation and showed normal Kinetic parameters, maintaining normal adaptation times (Lag phase) and Log phases (9 h average), before reaching the stationary phase. The proposed protocol constitutes a viable alternative to the long-term preservation of different LAB genera because it keeps their viability and integrity. Using flow cytometry allowed the enumeration of viable LAB and provide evidence of the integrity of their membrane.

Combination approach of paired starter culture and lactic acid on inhibiting autochthonous lactic acid bacteria for extending kimchi shelf life

Excessive kimchi fermentation by lactic acid bacteria (LAB) during distribution and storage may negatively influence the organoleptic properties of kimchi. Thus, this study aimed to extend kimchi shelf life by inhibiting the growth of autochthonous lactic acid bacteria by applying paired starter culture, consisting of nisin-producing Lactococcus lactis WiKim0098 and nisin-resistant Leuconostoc citreum WiKim0096, and lactic acid. To maximize nisin production in the L. lactis WiKim0098 culture medium, the food grade medium composition and culture conditions were optimized using response surface methodology. Leu. citreum WiKim0096 exhibited resistance to nisin owing to high positive surface charge and was successfully used as a paired starter. In the kimchi application experiments, paired starter culture addition delayed the over-acidification (more than 1% as acidity) of kimchi for approximately 5–6 days, and when combined with lactic acid, delayed the ripening for more than 8 days compared to control kimchi fermented at 10 °C. The results of bacterial community and metabolite analysis via principal component analysis showed that paired starter culture and lactic acid inhibited the LAB growth at the early stage of kimchi fermentation. Our results suggest that combining paired starter culture with lactic acid can effectively extend the kimchi shelf life. Specifically, it is expected to be effectively applicable to kimchi products intended for export, which demand extended shelf life.

Application of multi-functional lactic acid bacteria strains in a pilot scale feta cheese production.

Feta cheese is the most recognized Greek Protected Designation of Origin (PDO) product in the world. The addition of selected autochthonous lactic acid bacteria (LAB) strains to cheese milk as adjunct cultures is gaining more attention, since they can impact the nutritional, technological and sensory properties of cheeses, as well as improve the safety of the product. The aim of this study was to produce Feta cheese with enhanced quality and safety, and distinctive organoleptic characteristics by applying autochthonous lactic acid bacteria (LAB) with multi-functional properties as adjunct cultures. Feta cheeses were produced with the commercial lactococcal starter culture and the addition of 9 LAB strains (Lactococcus lactis SMX2 and SMX16, Levilactobacillus brevis SRX20, Lacticaseibacillus paracasei SRX10, Lactiplantibacillus plantarum FRX20 and FB1, Leuconostoc mesenteroides FMX3, FMX11, and FRX4, isolated from artisanal Greek cheeses) in different combinations to produce 13 cheese trials (12 Feta trials with the adjunct LAB isolates and the control trial). In addition, Feta cheese manufactured with FMX3 and SMX2 and control Feta cheese were artificially inoculated (4 log CFU/g) with Listeria monocytogenes (a cocktail of 4 acid or non-acid adapted strains). Cheese samples were monitored by microbiological and physicochemical analyses during ripening, and microbiological, physicochemical, molecular and sensory analyses during storage at 4°C. The results showed that after manufacture, the LAB population was ca. 9.0 log CFU/g at all samples, whereas during storage, their population declined to 6.5-7.0 log CFU/g. In the Listeria inoculated samples, Listeria was absent after 60 days (end of ripening) and after 90 days in the adjunct culture, and in the control trials, respectively. Moreover, the addition of selected strains, especially Lcb. paracasei SRX10, led to cheeses with desirable and distinctive organoleptic characteristics. Furthermore, randomly amplified polymorphic PCR (RAPD-PCR) molecular analysis confirmed that the multi-functional LAB strains were viable by the end of storage. Overall, the results of this study are promising for the use of autochthonous strains in various combinations with the commercial starter culture to satisfy industry requirements and consumer demands for traditional and high added value fermented products.

Growth Performance and Fecal Microbiota of Dairy Calves Supplemented with Autochthonous Lactic Acid Bacteria as Probiotics in Mexican Western Family Dairy Farming.

Probiotic supplementation in dairy cattle has achieved several beneficial effects (improved growth rate, immune response, and adequate ruminal microbiota). This study assessed the effects on the growth parameters and gut microbiota of newborn dairy calves supplemented with two Lactobacillus-based probiotics, individually (6BZ or 6BY) or their combination (6BZ + 6BY), administrated with the same concentration (1 × 109 CFU/kg weight) at three times, between days 5 and 19 after birth. The control group consisted of probiotic-unsupplemented calves. Growth parameters were recorded weekly until eight weeks and at the calves' ages of three, four, and five months. Fecal microbiota was described by high-throughput sequencing and bioinformatics. Although no significant effects were observed regarding daily weight and height gain among probiotic-supplemented and non-supplemented calves, correlation analysis showed that growth rate was maintained until month 5 through probiotic supplementation, mainly when the two-strain probiotics were supplied. Modulation effects on microbiota were observed in probiotic-supplemented calves, improving the Bacteroidota: Firmicutes and the Proteobacteria ratios. Functional prediction by PICRUSt also showed an increment in several pathways when the two-strain probiotic was supplemented. Therefore, using the three-administration scheme, the two-strain probiotic improved the growth rate and gut microbiota profile in newborn dairy calves. However, positive effects could be reached by applying more administrations of the probiotic during the first 20 days of a calf's life.

Effect of Lactobacillus spp. strains on the microbiological, biochemical, and organoleptic properties of Moroccan goat’s cheese during ripening

The aim of the present work was to investigate the effect of adding autochthonous lactic acid bacteria strains (LAB) to Moroccan goat’s milk cheese, focusing on its microbiological and biochemical evolution during the ripening process, as well as on the product’s sensorial properties. We prepared three types of cheese: (i) a control cheese (CNT) made by adding a starter culture composed of a strain of Lactococcus lactis subsp. lactis var. diacetylactis and a strain of Lactobacillus lactis subsp. lactis; (ii) a cheese (“LP”) made with the same lactococcal starter culture, and adding a strain of Lactobacillus plantarum as an adjunct culture, and (iii) a cheese (“LPC”) made with the same lactococcal starter culture, and adding Lactobacillus paracasei as an adjunct culture. The counts of most bacterial groups decreased as the cheeses matured, except for lactic acid bacteria, which increased during ripening. Throughout the ripening period, LP cheese showed the highest values for total nitrogen, soluble nitrogen, and soluble nitrogen in 12% TCA. On the 60th day of ripening, the highest contents of diacetyl and acetoin were recorded in LPC cheese. A significant difference in FFA was observed among the LP, LPC, and CNT cheeses, thus indicating a change in lipolysis extension (C4:0-C18:3 FFA) that might be influenced by the starter employed. These suggested that both adjunct strains could produce high-quality Moroccan goat cheese. However, the addition of Lactobacillus paracasei culture had the particular advantage of improving aroma intensity and overall cheese quality. Moreover, the lipolysis process increased continually until the end of the ripening period, thereby confirming that the ripening phase is the main factor that affects these cheeses’ sensory properties. The cheeses made with adjunct cultures had the typical taste of Moroccan goat cheese, presented an acceptable flavour, and fulfilled the usual sensory requirements.

Selection of food cultures with protective properties for cooked ham.

Sliced cooked ham stored in modified atmosphere packaging (MAP) can be spoiled by lactic acid bacteria (LAB) which are dominating under psychrotrophic conditions. Depending on the strains, the colonization can result in a premature spoilage characterized by off-flavors, gas and slime production, discoloration, and acidification. The purpose of this study was the isolation, identification and characterization of potential food culture with protective properties, able to prevent or delay spoilage in cooked-ham. The first step was to identify by means of microbiological analysis, the microbial consortia both in unspoiled and in spoiled lots of sliced cooked ham by the use of media for the detection lactic acid bacteria and total viable count. Counts ranged from values lower than 1 Log CFU/g to 9 Log CFU/g in spoiled and unflawed samples. The interaction between consortia was then studied in order to screen for strains able to inhibit spoilage consortia. Strains showing antimicrobial activity were identified and characterized by molecular methods and tested for their physiological features. Among a total of 140 strains isolated, nine were selected for their ability to inhibit a large number of spoilage consortia, to grow and ferment at 4°C and to produce bacteriocins. The effectiveness of the fermentation made by food culture was evaluated, through challenge tests in situ, analysing the microbial profiles of artificially inoculated cooked-ham slices during storage by high throughput 16S rRNA gene sequencing. The native population in situ resulted competitive against the inoculated strains and only one strain was able to significantly reduce the native populations reaching about 46.7% of the relative abundance. The results obtained in this study provide information about the selection of autochthonous LAB on the base of their action against spoilage consortia, in order to select protective potential cultures able to improve the microbial quality of sliced cooked ham.

Autochthonous Cultures to Improve Safety and Standardize Quality of Traditional Dry Fermented Meats

Traditional dry fermented meat products are obtained artisanally in many countries, where they represent a gastronomic heritage well distinguished from industrial counterparts. This food category is most often obtained from red meat, a food commodity that is under attack because of evidence of increased risk of cancer and degenerative diseases with high consumption. However, traditional fermented meat products are intended for moderate consumption and gastronomic experience, and, as such, their production must be continued, which would also help safeguard the culture and economy of the geographical areas of origin. In this review, the main risks attributed to these products are considered, and how these risks are reduced by the application of autochthonous microbial cultures is highlighted by reviewing studies reporting the effects of autochthonous lactic acid bacteria (LAB), coagulase negative staphylococci (CNS), Debaryomyces hansenii and Penicillium nalgiovense on microbiological and chemical safety and on sensory attributes. The role of dry fermented sausages as a source of microorganisms that can be beneficial to the host is also considered. From the results of the studies reviewed here it appears that the development of autochthonous cultures for these foods can ensure safety and stabilize sensory characteristics and has the capacity to be extended to a larger variety of traditional products.

Implementation of Novel Autochthonous Microencapsulated Strains of Lactiplantibacillus plantarum, Lactococcus lactis, and Lamb’s Rennet in the Production of Traditional “Paški Sir” Cheese

This study aimed to implement a microencapsulated form of selected autochthonous lactic-acid bacteria (LAB) isolated from the cheese-production chain and natural rennet obtained from suckling lambs in the traditional production of hard sheep cheese, “Paški sir”, from the island of Pag, Croatia. Two different formulations of microparticles were prepared: (i) microparticles containing the strain of both Lactiplantibacillus plantarum and Lactococcus lactis (S2) and (ii) microparticles containing both strains and natural rennet (S3). These formulations were used in the production of Paški sir cheese simultaneously with standard production using non-encapsulated commercial starter cultures and commercial rennet (S1). The number of Lc. lactis isolates decreased at day 30 and were not isolated during the remaining ripening process, whereas the number of L. plantarum remained stable throughout the ripening process. The level of LAB and the release of the rennet from microsphere formulations at the end allowed for the production of cheese with the same characteristics as the commercial product, indicating no negative interactions of natural rennet, bacterial culture, and chemical components of microparticles. To our knowledge, this is the first report of a microencapsulated L. plantarum (isolated from the abomasum of lambs) coupled with natural lamb’s rennet used in the production of hard sheep cheese. This pilot study showed the great potential for maintaining authenticity in cheese production by combining traditional and sustainable innovative technologies.

In Vitro Probiotic Properties and In Vivo Anti-Ageing Effects of Lactoplantibacillus plantarum PFA2018AU Strain Isolated from Carrots on Caenorhabditis elegans.

Lactic acid bacteria (LAB) share and provide several beneficial effects on human health, such as the release of bioactive metabolites, pathogen competition, and immune stimulation. The two major reservoirs of probiotic microorganisms are the human gastro-intestinal tract and fermented dairy products. However, other sources, such as plant-based foods, represent important alternatives thanks to their large distribution and nutritive value. Here, the probiotic potential of autochthonous Lactiplantibacillus plantarum PFA2018AU, isolated from carrots harvested in Fucino highland, Abruzzo (Italy), was investigated through in vitro and in vivo approaches. The strain was sent to the biobank of Istituto Zooprofilattico Sperimentale della Lombardia ed Emilia Romagna in Italy for the purpose of patent procedures under the Budapest Treaty. The isolate showed high survival capability under in vitro simulated gastro-intestinal conditions, antibiotic susceptibility, hydrophobicity, aggregation, and the ability to inhibit the in vitro growth of Salmonella enterica serovar Typhimurium, Listeria monocytogenes, Pseudomonas aeruginosa, and Staphylococcus aureus pathogens. Caenorhabditis elegans was used as the in vivo model in order to analyse prolongevity and anti-ageing effects. L. plantarum PFA2018AU significantly colonised the gut of the worms, extended their lifespan, and stimulated their innate immunity. Overall, these results showed that autochthonous LAB from vegetables, such as carrots, have functional features that can be considered novel probiotic candidates.