Proteome and Peptidome Changes and Zn Concentration in Chicken after In Ovo Stimulation with a Multi-Strain Probiotic and Zn-Gly Chelate: Preliminary Research.

The aim of the study was to determine differences in the proteome and peptidome and zinc concentrations in the serum and tissues of chickens supplemented with a multi-strain probiotic and/or zinc glycine chelate in ovo. A total of 1400 fertilized broiler eggs (Ross × Ross 708) were divided into four groups: a control and experimental groups injected with a multi-strain probiotic, with zinc glycine chelate, and with the multi-strain probiotic and zinc glycine chelate. The proteome and peptidome were analyzed using SDS-PAGE and MALDI-TOF MS, and the zinc concentration was determined by flame atomic absorption spectrometry. We showed that in ovo supplementation with zinc glycine chelate increased the Zn concentration in the serum and yolk sac at 12 h post-hatch. The results of SDS-PAGE and western blot confirmed the presence of Cu/Zn SOD in the liver and in the small and large intestines at 12 h and at 7 days after hatching in all groups. Analysis of the MALDI-TOF MS spectra of chicken tissues showed in all experimental groups the expression of proteins and peptides that regulate immune response, metabolic processes, growth, development, and reproduction.

Pyrrolizidine Alkaloids-Pros and Cons for Pharmaceutical and Medical Applications.

Heterocyclic organic compounds named pyrrolizidine alkaloids (PAs) belong to a group of alkaloids and are synthesized by either plants or microorganisms. Therefore, they are naturally occurring secondary metabolites. They are found in species applied in the pharmaceutical and food industries, thus a thorough knowledge of their pharmacological properties and toxicology to humans is of great importance for their further safe employment. This review is original because it synthesizes knowledge of plant and microbial PAs, which is unusual in the scientific literature. We have focused on the Boraginaceae family, which is unique due to the exceptional richness and diversity of its PAs in plant species. We have also presented the microbial sources of PAs, both from fungi and bacteria. The structure and metabolism of PAs have been discussed. Our main aim was to summarize the effects of PAs on humans, including both negative, toxic ones, mainly concerning hepatotoxicity and carcinogenicity, as well as potentially positive ones for pharmacological and medical applications. We have collected the results of studies on the anticancer activity of PAs from plant and microbial sources (mainly Streptomyces strains) and on the antimicrobial activity of PAs on different strains of microorganisms (bacteria and fungi). Finally, we have suggested potential applications and future perspectives.

Effect of In Ovo Administration of a Multi-Strain Probiotic and Zinc Glycine Chelate on Antioxidant Capacity and Selected Immune Parameters in Newly Hatched Chicks.

The aim of this study was to determine the effect of in ovo co-supplementation of chicken embryos with a multi-strain probiotic containing effective microorganisms and zinc glycine chelate on total antioxidant capacity; concentrations of sulfhydryl groups, bityrosine bridges, formylkynurenines, hydroperoxides, proteins, corticosterone, pro- and anti-inflammatory cytokines and heat shock proteins; and the activity of catalase and superoxide dismutase in the serum, yolk sac and tissues of broiler chickens at 12 h and at 7 days after hatching. The results indicate high SOD activity in the small and large intestines of chicks at 12 h post-hatch in the groups receiving the multi-strain probiotic and in the small intestine and yolk sac of birds receiving the multi-strain probiotic and Zn-Gly chelate. High concentrations of TNF-α and IFN-γ in the yolk sac and serum after in ovo administration of Zn-Gly chelate were observed 12 h after hatching. The use of a probiotic and a probiotic with Zn-Gly chelate increased the total antioxidant capacity in the tissues of chickens. It can be concluded that in ovo administration of a multi-strain probiotic and Zn-Gly chelate can maintain the oxidant/antioxidant balance in chickens and increase the defense capacity against oxidative stress.

Effect of combined in ovo administration of zinc glycine chelate (Zn-Gly) and a multistrain probiotic on the modulation of cellular and humoral immune responses in broiler chickens.

The aim of the study was to determine the effect of in ovo administration of zinc glycine chelate (Zn-Gly), and a multistrain probiotic on the hatchability and selected parameters of the cellular and humoral immune response of chickens. The study was conducted on 1,400 fertilized eggs from commercial broiler breeders (Ross x Ross 708). Material for the study consisted of peripheral blood and spleens of chicks taken 12 h and 7 d after hatching. The results showed that both combined and single in ovo administration of the multistrain probiotic and zinc glycine chelate significantly reduced hatchability of chicks. The flow cytometry study showed that the highest percentage of CD4+ T cells, CD4+CD25+, and high expression of KUL01 in the serum were obtained in the group supplemented with probiotic and Zn-Gly both 12 h and 7 d after hatching. In birds supplemented with probiotic and zinc chelate, a high percentage of TCRγδ+ cells was found in serum and spleen 12 h after hatching and in serum after 7 d. The percentage of Bu-1A+ lymphocytes in serum and spleen 12 h and 7 d after hatching was the highest in the group supplemented with probiotic and Zn-Gly. The highest expression of CD79A was observed in the group supplemented only with zinc chelate. There were no significant differences in the percentage of CD4+ cells in the spleens of birds in the groups receiving the multistrain probiotic at 12 h after hatching, and after 7 d, the percentage of CD4+ T cells was lower in the experimental groups than in the control group. The percentage of CD8+ cells in the serum of birds after hatching was lower in the group supplemented with multistrain probiotic and Zn-Gly than in the control group, but reached the highest value on d 7 after hatching. The obtained results confirm the strong effect of the combined administration of a multistrain probiotic and Zn-Gly chelate on lymphocyte proliferation and stimulation of cellular immune mechanisms in birds.

Chemical Composition and Antimicrobial Activity of New Honey Varietals.

Due to a widespread occurrence of multidrug-resistant pathogenic strains of bacteria, there is an urgent need to look for antimicrobial substances, and honey with its antimicrobial properties is a very promising substance. In this study, we examined for the first time antimicrobial properties of novel varietal honeys, i.e., plum, rapeseed, Lime, Phacelia, honeydew, sunflower, willow, and multifloral-P (Prunus spinosa L.), multifloral-AP (Acer negundo L., Prunus spinosa L.), multifloral-Sa (Salix sp.), multifloral-Br (Brassica napus L.). Their antimicrobial activity was tested against bacteria (such as Escherichia coli, Bacillus circulans, Staphylococcus aureus, Pseudomonas aeruginosa), yeasts (such as Saccharomyces cerevisiae and Candida albicans) and mold fungi (such as Aspergillus niger). In tested honeys, phenolic acids constituted one of the most important groups of compounds with antimicrobial properties. Our study found phenolic acids to occur in greatest amount in honeydew honey (808.05 µg GAE/g), with the highest antifungal activity aiming at A. niger. It was caffeic acid that was discovered in the greatest amount (in comparison with all phenolic acids tested). It was found in the highest amount in such honeys as phacelia-356.72 µg/g, multifloral (MSa) and multifloral (MBr)-318.9 µg/g. The highest bactericidal activity against S. aureus was found in multifloral honeys MSa and MBr. Additionally, the highest amount of syringic acid and cinnamic acid was identified in rapeseed honey. Multifloral honey (MAP) showed the highest bactericidal activity against E. coli, and multifloral honey (MSa) against S. aureus. Additionally, multifloral honey (MBr) was effective against E. coli and S. aureus. Compounds in honeys, such as lysozyme-like and phenolic acids, i.e., coumaric, caffeic, cinnamic and syringic acids, played key roles in the health-benefit properties of honeys tested in our study.

Differentiation and Propagation Potential of Arnica montana L. Achenes as a Consequence of the Morphological Diversity of Flowers and the Position of Flower Heads on the Plant.

Arnica montana L. is a very important medicinal plant and simultaneously a European endemic endangered plant species. The morphological features and details of seed development and achene variability are poorly recognized. The aim of this study was to determine the impact of the achene position in the infructescence and the location of the inflorescence on the plant on the (i) morphological characteristics and germination ability of achenes, and (ii) recruitment of seedlings and their biometric features. Infructescences containing fully ripe achenes were randomly collected from A. montana individuals for the measurements and the germination experiment. Scanning electron microscopy, fluorescence microscopy, and light microscopy were used for characterization of flowers and achenes. The morphological traits of achenes and reproductive characteristics of A. montana were determined by the position of the achenes in the infructescence and the location of the inflorescence on the plant. The surface of arnica achenes is equipped with non-glandular and glandular trichomes, which is very rarely presented in species of the family Asteraceae. It is possible that the fluid-containing glandular trichomes are a source of essential oils. The peripherally located achenes were longer, thinner, and lighter. They were characterized by lower embryo weight, lower embryo/achene weight ratio, and lower germination capacity in comparison to the centrally located ones. The results presented in this article fill the gap in the knowledge of the morphology of achenes and the biology of the species, and provide information that can help in breeding programs, active protection, and field cultivation.

Allantoin overaccumulation enhances production of metabolites under excess of metals but is not tightly regulated by nitric oxide.

The aln-3 mutant overaccumulating allantoin and respective wild type (WT) strain of Arabidopsis thaliana were exposed to cadmium (Cd) or mercury (Hg) with or without nitric oxide (NO) donor (sodium nitroprusside, SNP) to study cross-talk, metabolic and oxidative changes between these nitrogen sources (organic vs. inorganic). The aln-3 accumulated over 10-fold more allantoin than WT with the effect of Cd and Hg differing in leaf and root tissue: aln-3 contained more ascorbic acid and phytochelatins when treated with Cd or Hg and more Cd in both organs. SNP depleted leaf Cd and root Hg accumulation in aln3 but had a positive impact on the amount of metabolites typically in WT plants, indicating potentially negative relation between allantoin and NO. In agreement, aln-3 roots showed lower NO signals in control or metal treatments, but higher ROS signal, and SNP had more pronounced impact in WT roots. Flavonol glycosides were more abundant in aln-3 and were affected more by metals than by SNP. Malate was the most affected Krebs acid with strong reaction to SNP and Hg treatment. Data indicate that allantoin overaccumulation influences the accumulation of specific metabolites but nitric oxide has a greater impact on the metabolite profile in WT.

The Effect of Feed Supplementation with EM Bokashi® Multimicrobial Probiotic Preparation on Selected Parameters of Sow Colostrum and Milk as Indicators of the Specific and Nonspecific Immune Response.

The aim of the study was to determine the effect of EM Bokashi® on selected parameters of the specific and nonspecific immune response of sows by in colostrum and milk samples. The percentage of cells with expression of CD19+, CD5+CD19+, CD21+, SWC3a (macrophage/monocyte), and CD11b+ molecules on the monocytes and granulocytes as well as the concentrations of lysozyme and acute phase proteins - serum amyloid-A (SAA) and haptoglobin (Hp) were evaluated. The study was carried out on a commercial pig farm, including 150 sows (Polish Large White × Polish Landrace) at the age of 2-4 years. Sixty female sows were divided into two groups: I - control and II - experimental. For the experimental group, a probiotic in the form of the preparation EM Bokashi® in the amount of 10 kg/tonne of feed was added to the basal feed from mating to weaning. The material for the study consisted of colostrum and milk. The samples were collected from all sows at 0, 24, 48, 72, 96, 120, 144, and 168 h after parturition. The study showed that exposure of the pregnant sow to the probiotic microbes contained in EM Bokashi® significantly affects the immunological quality of the colostrum and milk and caused an increase in the percentage of the subpopulations of B cells with CD19+, CD21+, and CD5+CD19+ expression in the colostrum and milk, which demonstrates an increase in the protective potential of colostrum and indicates stimulation of humoral immune mechanisms that protect the sow and the piglets against infections.

High Temperature Alters Secondary Metabolites and Photosynthetic Efficiency in Heracleum sosnowskyi.

Due to global warming, invasive species have spread across the world. We therefore studied the impact of short-term (1 day or 2 days) and longer (7 days) heat stress on photosynthesis and secondary metabolites in Heracleum sosnowskyi, one of the important invasive species in the European Union. H. sosnowskyi leaves exposed to short-term heat stress (35 °C/1 d) showed a decrease in chlorophyll and maximum potential quantum efficiency of photosystem II (Fv/Fm) compared to control, 35 °C/2 d, or 30 °C/7 d treatments. In turn, the high level of lipid peroxidation and increased H2O2 accumulation indicated that the 30 °C/7 d stress induced oxidative damage. The contents of xanthotoxin and bergapten were elevated in the 2 d and 7 d treatments, while isopimpinellin was detected only in the heat-stressed plants. Additionally, the levels of free proline and anthocyanins significantly increased in response to high temperature, with a substantially higher increase in the 7 d (30 °C) treatment. The results indicate that the accumulation of proline, anthocyanins, and furanocoumarins, but not of phenolic acids or flavonols, contributes to protection of H. sosnowskyi plants against heat stress. Further studies could focus on the suppression of these metabolites to suppress the spread of this invasive species.

Physiological Diversity of Spitsbergen Soil Microbial Communities Suggests Their Potential as Plant Growth-Promoting Bacteria.

The objective of the study was to assess the physiological diversity and metabolic activity of the soil bacterial communities inhabiting Spitsbergen soils in search of bacterial abilities facilitating plant growth promotion. In the soil, the total number of culturable microorganisms, the number of their individual physiological groups (including Siderophore Synthesizing; SSB and Phosphate Solubilizing Bacteria; PSB), the dehydrogenase (DH) activity, and the ability to utilize sources of C, N, P (EcoPlate) were analysed. In bacterial isolates, siderophores production, ACC (1-aminocyclopropane-1-carboxylate) deaminase (ACCD) activity, IAA (indole-3-acetic acid) synthesis were examined. The isolates were applied to the seeds of Phaseolus coccineus regarding their germination and root length. The results showed differences between copio- and oligotrophic bacteria. A usually high number of SSB was accompanied by the raised number of PSB. A bigger number of SSB was connected with low values of Fe in the soil. High DH activity was assisted by greater number of copio- and oligotrophic bacteria, raised average well color development value, and N and C contents in the soil. Germination index was more alike relative seed germination than relative root growth. IAA concentration and ACCD activity were conversely related. Synthesis of siderophores was matched with ACCD activity and its high level was combined with elevated germination index. In spite of different localization of soil samples, some isolates proved similar traits of activity. Distinct affiliation of isolates and their various localizations were displayed. Among all isolates tested, some possessed one main trait of activity, but most of them had two or more significant features for potential plant growth stimulation. These isolates could be an important source of useful bacteria.

Separation and determination of coumarins including furanocoumarins using micellar electrokinetic capillary chromatography.

The conditions of micellar electrokinetic capillary chromatography for separation and simultaneous measurement of coumarins (coumarin, scoparone, isoscopoletin, esculin, esculetin, umbelliferone) including furanocoumarins (xanthotoxin, byakangelicin, isopimpinellin, bergapten, phellopterin, xanthotoxol) have been elaborated. The influence of different parameters, such as the pH of the buffer, sodium cholate (SC) or methanol concentration in the buffer, on the migration time, peak resolution, peak asymmetry, and number of theoretical plates was investigated. The optimum separation of the compounds was achieved using 50-µm i.d. capillaries with a total length of 64.5 cm (56 cm effective length) and a buffer system at pH 9.00 consisting of 50 mM sodium tetraborate, 45 mM SC, and 20% of methanol (v/v). The developed method ensured good repeatability of corrected peak areas and migration times (the relative standard deviations were in the range of 2.8-6.1% and 0.8-4.0%, respectively). The average limit of detection for all studied compounds was below 1.3 µg mL. Moreover, good linearity of the relationship between the peak corrected area and the concentration of the compounds was observed (correlation coefficient >0.99). The method was successfully applied in the quantitative analysis of two different types of samples, i.e. Heracleum sphondylium herb and Aesculus hippocastanum cortex.