Metric and Spectral Insight into Bee-Pollen-to-Bee-Bread Transformation Process.

Due to numerous bioactive constituents, both bee pollen (BP) and bee bread (BB) represent valuable food supplements. The transformation of BP into BB is a complex biochemical in-hive process that enables the preservation of the pollen's nutritional value. The aim of this study was to determine the depth of the honeycomb cells in which bees store pollen and to provide a spectral insight into the chemical changes that occur during the BP-to-BB transformation process. This study was carried out on three experimental colonies of Apis mellifera carnica, from which fresh BP was collected using pollen traps, while BB samples were manually extracted from the cells two weeks after BP sampling. The samples were analyzed using infrared (FTIR-ATR) spectroscopy, and the depth of the cells was measured using a caliper. The results showed that the average depth of the cells was 11.0 mm, and that the bees stored BB up to an average of 7.85 mm, thus covering between ⅔ and ¾ (71.4%) of the cell. The FTIR-ATR analysis revealed unique spectral profiles of both BP and BB, indicating compositional changes primarily reflected in a higher water content and an altered composition of the carbohydrate fraction (and, to a lesser extent, the lipid fraction) in BB compared to BP.

Proteolytic Lactococcus lactis and Lipolytic Enterococcus durans of Dairy Origin as Meat Functional Starter Cultures.

RESEARCH BACKGROUND: As fermentation is an integral feature of both, dry sausage and cheese production, this has led to the evaluation of bacterial cultures Lactococcus lactis ssp. cremoris (LL8307) and Enterococcus durans (ED0207) originally isolated from artisanal Croatian hard type cheese to diversify the range of flavours of dry fermented sausages and to increase their microbiological safety. Both strains were chosen for their high or medium acidifying, proteolytic and/or lipolytic activity and bioprotective potential after step-by-step selection of wild isolates. Therefore, this study aims to evaluate the survival rate of selected starter cultures in wild boar meat sausages during the ripening period of 40 days at a local small-scale facility under artisanal conditions as well as their influence on sausage quality parameters. EXPERIMENTAL APPROACH: Safety, biotechnological and probiotic properties of twenty-three enterococcal and lactococcal isolates of dairy origin were studied. Based on the results, two best candidates were selected and added to the meat batter during the artisanal wild boar meat sausage preparation where their survival rate, effect on physicochemical, microbiological and sensorial properties and histamine content were evaluated. RESULTS AND CONCLUSIONS: As revealed by repetitive element-polymerase chain reaction (rep-PCR), native starter cultures survived up to 15 days of ripening and were either absent from (LL8307) or reduced by 80% (ED0207) in final products. The application of native starter cultures rapidly decreased pH (p<0.05), leading to the significantly lower load of E. coli, coliforms and Enterobacteriaceae in ready-to-eat sausages prepared by the addition of starter cultures (3.04-3.94 log CFU/g) than in the control (3.88-5.00 log CFU/g). Analysis of hedonic test data revealed that some of the sensory traits (odour, flavour, juiciness) of treatments with starter cultures were highly liked by the higher percentage of consumers. The results suggest that these starter cultures would represent a valuable tool to improve the homogeneity of artisanal manufacture and hygienic quality of fermented sausages and can be safely used for food application. NOVELTY AND CCIENTIFIC CONTRIBUTION: This is the first study to explore in depth the biotechnological potential of bacterial cultures isolated from artisanal Croatian cheese as functional starter cultures for high-quality game meat sausage production.

Chemical Profiling and Antimicrobial Properties of Honey Bee (Apis mellifera L.) Venom.

The incidence of antibiotic resistance in pathogenic bacteria has become an alarming clinical and social problem. Therefore, the demand for alternative antimicrobial compounds has increased. In this study, a chemical profile of honey bee (Apis mellifera L.) venom (HBV) has been determined by HPLC and FTIR-ATR spectroscopy, and tested for antibacterial activity, as well as efficiency with regard to conventional antibiotics. The investigated HBV was of high quality with melittin and total protein contents of 70.10 ± 7.01%, and 84.44 ± 3.12 g/100 g, respectively. The purity of HBV was confirmed by FTIR-ATR spectral profiling, which revealed a unique pattern of absorption bands that are characteristic of its major fractions. In addition, HBV showed a broad spectrum of activity against all three tested biomasses of potentially pathogenic Gram-positive and Gram-negative bacteria with MIC values ranging between 12.5 and 200 µg/mL, and MBC between 12.5 and 400 µg/mL. When compared to conventional antibiotics, HBV (400 µg) showed up to 27.8% efficiency of tetracycline (30 µg), 52.2% erythromycin (15 µg), 21.2% ciprofloxacin (5 µg), and 34.6% of ampicillin-sulbactam (20 µg). The overall results demonstrate the therapeutic potential of the analyzed HBV.

The survival rate and efficiency of non-encapsulated and encapsulated native starter cultures to improve the quality of artisanal game meat sausages.

This study addresses the application of native, multiple strain starter cultures for standardization of game meat sausages production. The designed starter cultures consisting of two indigenous Lactobacillus sakei and one Leuconostoc mesenteroides strains. These strains were used in both, the encapsulated and non-encapsulated form, in the game meat dough, individually or in combination, with eight treatments in total. Microbiological and physicochemical characteristics of the sausages were monitored throughout the manufacturing process, while sensory properties, biogenic amine content, and volatile compounds were evaluated in the final products. As revealed by rep-PCR, native starter cultures, encapsulated or non-encapsulated, had survived the whole sausage production process; however, to varying degrees. The application of indigenous decarboxylase negative Lb. sakei strains significantly (P < 0.05) reduced tyramine content, rapidly decreased pH and promoted the number reduction of Enterobacteriaceae and elimination of E. coli, L. monocytogenes and coliforms in ready-to-eat products. A total of 84 volatile compounds were identified by SPME-GC-MS in the eight treatment batches of game meat sausages, with only minor differences between the treatments. No significant differences in sensory traits (P > 0.05) between tested treatments were found, although treatment with the Lb. sakei strains received the highest scores for the sensory traits including cross-section, odour, hardness, aroma, and overall acceptability. Combination of multi-strain Lb. sakei starter cultures resulted in growth prevention of undesirable microbiota, reduction of tyramine content and increased the acceptability parameters of full-ripened sausages, which make them good candidates for industrial as well as artisanal application.

Microbial Key Players Involved in P Turnover Differ in Artificial Soil Mixtures Depending on Clay Mineral Composition.

Nutrient turnover in soils is strongly driven by soil properties, including clay mineral composition. One main nutrient is phosphorus (P), which is known to be easily immobilized in soil. Therefore, the specific surface characteristics of clay minerals might substantially influence P availability in soil and thus the microbial strategies for accessing P pools. We used a metagenomic approach to analyze the microbial potential to access P after 842 days of incubation in artificial soils with a clay mineral composition of either non-expandable illite (IL) or expandable montmorillonite (MT), which differ in their surface characteristics like soil surface area and surface charge. Our data indicate that microorganisms of the two soils developed different strategies to overcome P depletion, resulting in similar total P concentrations. Genes predicted to encode inorganic pyrophosphatase (ppa), exopolyphosphatase (ppx), and the pstSCAB transport system were higher in MT, suggesting effective P uptake and the use of internal poly-P stores. Genes predicted to encode enzymes involved in organic P turnover like alkaline phosphatases (phoA, phoD) and glycerophosphoryl diester phosphodiesterase were detected in both soils in comparable numbers. In addition, Po concentrations did not differ significantly. Most identified genes were assigned to microbial lineages generally abundant in agricultural fields, but some were assigned to lineages known to include oligotrophic specialists, such as Bacillaceae and Microchaetaceae.

Encapsulation of two fermentation agents, Lactobacillus sakei and calcium ions in microspheres.

Alginate microspheres loaded with two fermentation active agents, calcium cations and strain LS0296 identified as Lactobacillus sakei, have been prepared and characterized. The role of calcium cation is twofold, it acts as gelling cation and as fermentation active agent. Encapsulation and the presence of calcium ions in the same compartment do not inhibit the activity of LS0296. Molecular interactions in microspheres are complex, including mainly hydrogen bonds and electrostatic interactions. In vitro calcium cations and strain LS0296 release profiles were fitted to the Korsmeyer-Peppas empirical model. The calcium cation release process is driven at first by Fickian diffusion through microspheres and then by anomalous transport kinetics. The in vitro LS0296 release process is driven by Fickian diffusion through microspheres showing a much slower releasing rate than calcium cations. The release of LS0296 strain is followed by a decrease in the pH value. Results obtained give us a new insight into complex interactions between bacterial cultures and microsphere constituents. Prepared formulations of calcium alginate microspheres loaded with LS0296 could be used as a new promising tool and a model for different starter cultures encapsulation and use in the production of fermented foods.

Characterization of Enterococcal Community Isolated from an Artisan Istrian Raw Milk Cheese: Biotechnological and Safety Aspects.

In this study, prevalence, biotechnological and safety profiles of 588 Enterococcus isolates isolated from raw milk and Istrian cheese during different stages of ripening were analyzed. Despite the low and variable presence of enterococci in milk ((3.65±2.93) log CFU/mL), highly comparable enterococcal populations were established after 30 days of cheese ripening ((7.96±0.80) log CFU/g), confirming Enterococcus spp. as a major part of the core microbiota of Istrian cheese. The dominant species were E. faecium (53.8%) and E. faecalis (42.4%), while minor groups, consisting of E. durans (2.84%) and E. casseliflavus (0.95%), also occurred. A pronounced intraspecies variability was noticed based on molecular fingerprinting, with 35 strains (genotypes) detected. Most of the genotypes were farm-specific with one third being shared between the farms. This genotype variability reflected particular differences of Istrian cheese production, mainly variable salt concentration, ripening temperature and air humidity as well as microclimatic or vegetation conditions. There was considerable variation between the strains of the same species regarding wide range of biotechnologically important traits as well as their ability to survive in simulated gastrointestinal conditions. A considerable number of strains were resistant to critically important antibiotics such as tetracycline (43.56%), erythromycin (35.79%) and vancomycin (23.48%). Polymerase chain reaction-based detection did not identify any of the common genetic determinants for vancomycin and erythromycin resistance; for tetracycline tetM gene was detected. The presence of virulence genes including agg, efaAfs, gelE, cylM, cylB, cylA, esp, efaAfm, cob and cpd was frequently recorded, especially among E. faecalis strains.