The Influence of Enzymatic Hydrolysis on Bee Pollen Antioxidant and Antibacterial Activities.

Bee pollen is one of the most valuable apitherapeutic products with high nutritional value. To obtain a higher diversity of compounds, higher bioactivity, and improve the release of nutrients from bee pollen, additional processing of the raw material may be applied: fermentation using microorganisms or hydrolysis using selective enzymes. This research aimed to determine the impact of enzymatic hydrolysis on the antioxidant and antibacterial activities of bee pollen. Bee pollen samples from Sweden, Spain, Netherlands, Italy, Poland, Denmark, Slovakia, Malta, and Lithuania were hydrolyzed using pure enzymes, including lipase, cellulase, protease, and amyloglucosidase, as well as enzyme mixtures such as Viscozyme® L and Clara-diastase. Total phenolic content, total flavonoid content, and antioxidant activity were analyzed spectrophotometrically. Antibacterial activity against Staphylococcus aureus, Listeria monocytogenes, Staphylococcus enteritidis, and Salmonella typhimurium was evaluated using the agar well diffusion assay. Obtained results revealed a positive effect of enzymatic hydrolysis on biologically active compound content and activity: total phenolic content increased by 1.1 to 2.5 times, total flavonoid content by 1.1 to 3.0 times, radical scavenging activity by 1.1-3.5 times, and antibacterial activity by 1.1 to 3.3 times. K-means clustering analysis grouped samples into 5-9 clusters and was dependent on the measured characteristic used as an input-total phenolic compounds content, total flavonoid content, antioxidant activity, and antibacterial activity against four different bacteria. Chemometrics showed, that the enzyme used for the hydrolysis had a higher impact on clustering results than the geographical origin of the samples.

Ultrasonic needle hydrophone calibration in air by a parabolic off-axis mirror focused beam using three-transducer reciprocity.

An acoustic field distribution investigation in air requires a small receiving sensor. Needle hydrophones seem to be an attractive solution, and it has previously been demonstrated that needle hydrophones designed for use in water can be used in air. The metrology problem is that an absolute sensitivity calibration is needed, because needle hydrophones are not characterized in air, especially for frequencies below 1 MHz, which is of interest for air-coupled ultrasound. Conventional, three-transducer/microphone reciprocity calibration requires measurements to be done in the far field. However, when transducer diameter is large and the frequency is high, the required measurement distance becomes very large: 3 m for a 20 mm source, transmitting at 1 MHz. Large propagation distance leads to high attenuation and nonlinear effects in air propagation, and distortion and losses accumulate. Small needle hydrophones have low sensitivity, so that high excitation amplitudes would be required, which can lead to transducer heating and increase nonlinearity effects. A derivative of the three-transducer reciprocity calibration method is proposed, where a large aperture transducer is focused onto a hydrophone, using hybrid of plane wave and spherical wave reciprocity. Use of a focused source minimizes the frequency-dependent diffraction effects, and the spherical wave approximation is valid at the focal distance, and low level excitation signals can be used. Focusing is accomplished using a parabolic off-axis mirror. Calibration is in transmission, which reduces the complexity of the electrical measurements. The corresponding equations have been derived for this setup. Calibration of the transducer and needle hydrophone absolute sensitivity is obtained.

The Impact of Fermentation on Bee Pollen Polyphenolic Compounds Composition.

Bee-collected pollen is one of the most valuable natural products. However, the pollen cell walls limit the digestibility and release of nutrients to the human body. Solid-state lactic acid fermentation can be used to ease the release of bioactive compounds from the pollen cell. The aim of this research was to determine the impact of a solid-state lactic acid fermentation process on biologically active compound composition and antioxidant activity of bee-collected pollen from various European regions (Italy, Netherlands, Lithuania, Poland, Sweden, Denmark, Malta, Slovakia, and Spain). Spontaneous fermentation and fermentation using an L. rhamnosus culture were performed. The total content of phenolic compounds, total content of flavonoids, and radical (DPPH) scavenging activity were measured by spectrophotometric tests, while UPLC was employed for quantification of phenolic compounds. The determined fermentation positive effects included an increase of total phenolic content by 1.4-2.3 times, total flavonoid content by 1.1-1.6 times, and radical scavenging activity by 1.4-2.3 times. Naringenin (21.09-135.03 µg/g), quercetin (6.62-78.86 µg/g), luteolin (29.41-88.90 µg/g), and rutin (21.40-89.93 µg/g) were the most abundant flavonoids in all samples; however, their variation level was both geographical in origin and fermentation-type dependent. Fermentation increased the content of phenolic acids with high antioxidant potentials such as ellagic, ferulic and caffeic, while reduction of chlorogenic acid was determined.

Bioactive Compounds, Antioxidant, and Antibacterial Properties of Lonicera caerulea Berries: Evaluation of 11 Cultivars.

The aim of the study was to evaluate 11 cultivars of blue honeysuckle (Lonicera caerulea L.) for bioactive compounds, antioxidant capacity, and the antibacterial activity of berries. Total phenolic contents (TPCs) and total anthocyanin contents (TACs) were established by using ethanolic extracts. For contents of organic acids and saccharides, aqueous extracts were used, and vitamin C was determined by using oxalic acid solution. DPPH• radical scavenging capacity was evaluated by using ethanolic extracts; antibacterial activity was assessed by using both ethanolic and aqueous extracts. The TPC varied from 364.02 ± 0.41 mg/100 g in 'Vostorg' to 784.5 ± 0.3 mg/100 g in 'Obilnaja', and TAC ranged from 277.8 ± 1.1 mg/100 g in 'Celnocnaja' to 394.1 ± 8.4 mg/100 g in 'Nimfa'. Anthocyanins comprised 53.8% of total phenolic contents on average. Among organic acids, citric acid was predominant, averaging 769.41 ± 5.34 mg/100 g, with malic and quinic acids amounting to 289.90 ± 2.64 and 45.00 ± 0.37 mg/100 g on average, respectively. Contents of vitamin C were 34.26 ± 0.25 mg/100 g on average. Organic acids were most effective in the inhibition of both Gram-positive and Gram-negative bacteria tested. In conclusion, berries of L. caerulea are beneficial not only for fresh consumption, but also as a raw material or ingredients of foods with high health-promoting value.

Comparison of Physicochemical Properties of Bee Pollen with Other Bee Products.

The aim of this study was to compare the physichochemical composition of various bee products, namely, bee pollen, beebread, propolis, honey, and royal jelly. The samples (37 out of 53) were collected in Lithuania, several samples from other Europe countries (Italy, Denmark, Sweden, Slovakia, Poland, Spain, Republic of Malta, The Netherlands, Latvia, Ukraine) were used for comparison. Various quantities, such as pH, electrical conductivity, oxidation-reduction potential, NaCl content, refraction index, Brix value, total phenolic compound content, total flavonoid content and antiradical activity were measured. Together with the mentioned, the content of micro- and macroelements (As, Ba, Ca, Cd, Co, Cr, Cu, Fe, K, Mg, Mn, Na, Ni, P, Pb, Se, Sr, V and Zn), ultraviolet-visible spectroscopy absorption spectra were analysed. To our knowledge, the literature data about comprehensive comparison of various characteristics of bee products are scarce. Also, to the best of our knowledge, this is the first study revealing mineral content in Lithuanian bee pollen, beebread and royal jelly. The study exposed that bee pollen not only showed the highest values of pH, electrical conductivity and content of soluble solids, but also distinguished from the other samples by the highest flavonoid content (up to 48.3 mg/10 g), the absence of Cr, the presence of Co (0.011-0.100 mg/kg) and Sr (0.73-5.37 mg/kg) and the highest content of Ca (997-2455 mg/kg) and Mg (644-1004 mg/kg). Hierarchical clustering analysis was applied to group the tested samples according to the physicochemical analysis results and mineral content. The clustering analysis revealed that bee pollen formed separate group with the highest distance from the other samples in both cases.

Evaluation of Fresh Cheese Quality Prepared with Newly Isolated Nisin Z-Producing Lactococcus lactis Bacteria.

The main task of the present study was to evaluate an impact of three nisin Z-producing Lactococcus lactis bacteria newly isolated from raw goat milk for some fresh cow cheese characteristics during the storage. Microbiological evaluation for Listeria monocytogenes, Staphylococcus aureus, and viable lactic acid bacteria counts and determination of pH, titratable acidity, and lactic acid concentration of produced cheese were performed after 0, 24, 48, 72, and 96 h. Sensory analysis for the evaluation of acidity, flavor intensity, color intensity, bitterness, and crumbliness of prepared cheese was performed. The changes of volatile compounds in fresh cheese were evaluated using headspace solid phase microextraction (SPME) coupled with gas chromatography-mass spectrometry. Chemometric methods were applied for the data analysis. Study showed that tested bacteria are suitable for the manufacturing of fresh cheese and possible application for fresh cheese biopreservation, as pathogenic bacteria did not grow during 4 days (96 h); chemometric analysis revealed that L. lactis strain LL56 was the most similar to commercially available L. lactis ATCC11454.

Evaluation of phytochemical composition of fresh and dried raw material of introduced Chamerion angustifolium L. using chromatographic, spectrophotometric and chemometric techniques.

Due to the wide spectrum of biological activities, Chamerion angustifolium L. as medicinal plant is used for the production of food supplements. However, it should be kept in mind that quality (biological activity) of the herb depends on its geographic origin, the way of raw material preparation or extraction and chemotype. The purpose of this study was to evaluate and compare the compositions of volatile, non-volatile compounds and antioxidant activities of C. angustifolium grown in Kaunas Botanical Garden after the introduction from different locations in Lithuania. The compositions of fresh and air-dried samples were compared. The profile of volatile compounds was analyzed using headspace solid phase microextraction coupled with GC/MS. trans-2-Hexenal (16.0-55.9% of all volatiles) and trans-anethole (2.6-46.2%) were determined only in the dried samples, while cis-3-hexenol (17.5-68.6%) only in fresh samples. Caryophyllenes (α- and ß-) were found in all analyzed samples, contributing together from 2.4% to 52.3% of all volatiles according to the origin and preparation (fresh or dried) of a sample. Total amount of phenolic compounds, total content of flavonoids and radical scavenging activity (using 2,2-diphenyl-1-picrylhydrazyl (DPPH)) were determined using spectrophotometric assays. The variation of total phenolic compounds content was dependent on the sample origin, moreover, drying reduced amount of phenolics 1.5-3.5 times. The DPPH free radical scavenging activity was in the range of 238.6-557.1mg/g (expressed in rutin equivalents) in the fresh samples and drastically reduced to 119.9-124.8 mg/g after drying. The qualitative analysis of phenolic compounds in the aqueous methanolic extracts of C. angustifolium was performed by means of HPLC with UV detection. Oenothein B and rutin were predominant in the samples; also caffeic and chlorogenic acids, and quercetin were determined. Chemometric methods, namely principal component analysis, hierarchical cluster analysis and K-means clustering analysis, were applied for evaluation of the results. Chemometric analysis showed existence of different chemotypes of C. angustifolium L. and their relation to the geographic origin.