Effect of the diets with pumpkin silage and synthetic β‐carotene on the carotenoid,immunoglobulin and bioactive protein content and fatty acid composition of colostrum

Forty Simmental cows were divided into four groups with 10 per group to investigate the effect of a transition diet supplemented with synthetic β‐carotene and natural carotenoid‐rich feed (pumpkin silage) on chemical composition of the colostrum. The control group (I) was fed grass and maize silage and supplemental concentrates, group II additionally received 400 mg/day/cow of synthetic β‐carotene, in group, III 40% DM of maize silage was replaced with pumpkin silage to increase β‐carotene intake by 400 mg in relation to group I, and in group IV 60% DM of maize silage was replaced with pumpkin silage without adjusting for β‐carotene. Colostrum was collected from the cows within 5 hr of calving to determine colostrum composition, the content of immunoglobulins and carotenoids, total antioxidant status (TAS), the content of some bioactive proteins and the composition of fatty acids. The study showed that the experimental diets had no effect on the gross composition of colostrum. The content of α‐carotene, β‐carotene and violaxanthin was significantly higher in group IV than in group I and that of lutein higher in group IV than in groups I and II. Total antioxidant status of colostrum in group IV was significantly higher than in group I. Groups III and IV were characterized by a higher concentration of immunoglobulin IgG in relation to group I. IgM level in groups II, III and IV was significantly higher than in group I and that in groups III and IV was also higher than in group II. The highest IgA concentration was observed in group IV and the lowest in group I. Lysozyme concentration was higher in group IV compared to groups I and II. The diets had no effect on the total content of SFA, MUFA and PUFA in colostrum.     

GLP1 and GIP are involved in the action of synbiotics in broiler chickens

Background: In order to discover new strategies to replace antibiotics in the post-antibiotic era in meat-type chicken production, two new synbiotics were tested:(Lactobacillus salivarius IBB3154 plus galactooligosaccharide(Syn1) and Lactobacillus plantarum IBB3036 plus raffinose family oligosaccharides(Syn2).Methods: The synbiotics were administered via syringe, using a special automatic system, into the egg air chamber of Cobb 500 broiler chicks on the 12 thday of egg incubation(2 mg of prebiotics + 105 cfu bacteria per egg).Hatched roosters(total 2,400) were reared on an experimental farm, kept in pens(75 animals per pen), with free access to feed and water. After 42 d animals were slaughtered. Blood serum, pancreas, duodenum and duodenum content were collected.Results: Syn2 increased trypsin activity by 2.5-fold in the pancreas and 1.5-fold in the duodenal content. In the duodenum content, Syn2 resulted in ca 30% elevation in lipase activity and 70% reduction in amylase activity.Syn1 and Syn2 strongly decreased expression of m RNA for GLP-1 and GIP in the duodenum and for GLP-1 receptors in the pancreas. Simultaneously, concentrations of the incretins significantly diminished in the blood serum(P 0.05). The decreased expression of incretins coincides with changed activity of digestive enzymes in the pancreas and in the duodenal content. The results indicate that incretins are involved in the action of Syn1 and Syn2 or that they may even be their target. No changes were observed in key hormones regulating metabolism(insulin, glucagon, corticosterone, thyroid hormones, and leptin) or in metabolic indices(glucose,NEFA, triglycerides, cholesterol). Additionally, synbiotics did not cause significant changes in the activities of alanine and aspartate aminotransferases in broiler chickens. Simultaneously, the activity of alkaline phosphatase and gamma glutamyl transferase diminished after Syn2 and Syn1, respectively.Conclusion: The selected synbiotics may be used as in ovo additives for broiler chickens, and Syn2 seems to improve their potential digestive proteolytic and lipolytic ability. Our results suggest that synbiotics can be directly or indirectly involved in incretin secretion and reception.     

Effect of conjugated linoleic acids on the activity and mRNA expression of 5- and 15-lipoxygenases in human macrophages

Lipoxygenases are a family of non-heme enzyme dioxygenases. The role of lipoxygenases is synthesis of hydroperoxides of fatty acids, which perform signaling functions in the body. Studies on conjugated linoleic acids (CLAs) as fatty acids with a potential anti-atherosclerotic function have recently been initiated. The aim of the study was to test the effect of CLAs and linoleic acid on 5- and 15-lipoxygenase (5-LO, 15-LO-1) enzyme activity, their mRNA expression, and concentration in the cells. It was also desired to determine whether the CLAs are substrates for the enzymes. For the experiments monocytic cell line (THP-1) and monocytes obtained from human venous blood were used. Monocytes were differentiated to macrophages: THP-1 (CD14+) by PMA administration (100 nM for 24 h) and monocytes from blood (CD14+) by 7-day cultivation with the autologous serum (10%). After differentiation, macrophages were cultured with 30 microM CLAs or linoleic acid for 48 h. The 15- and 5-lipoxygenase products were measured by HPLC method. mRNA expression and protein content were analyzed by real-time PCR and Western blot analysis. The in vitro studies proved that both CLA isomers are not substrates for 15-LO-1; in ex vivo studies hydroxydecadienoic acid (HODE) concentration was significantly reduced (p = 0.019). The trans-10,cis-12 CLA isomer reduced HODE concentration by 28% (p = 0.046) and the cis-9,trans-11 CLA isomer by 35% (p = 0.028). In macrophages obtained from THP-1 fatty acids did not change significantly mRNA expression of the majority of the investigated genes. CLAs did not change the content of 5-LO and 15-LO-1 proteins in macrophages obtained from peripheral blood. Linoleic acid induced 15-LO-1 expression (2.6 times, p < 0.05). CLAs may perform the function of an inhibitor of lipoxygenase 15-LO-1 activity in macrophages.     

Differential Tolerance to Lead and Cadmium of Micropropagated <Emphasis Type="Italic">Gypsophila fastigiata</Emphasis> Ecotype

In vitro techniques may provide a suitable tool for effective propagation and conservation of plant species representing various ecological niches. The elaboration of such protocols is also prerequisite for selection of heavy-metal-tolerant plant material that could be afterwards used for restoration or remediation of polluted sites. In this study, culture protocol for Gypsophila fastigiata propagation was developed. The highest multiplication coefficient, which reached 6.5, and the best growth parameters were obtained on modified MS medium supplemented with 1.0 mg L^?1 2iP and 0.2 mg L^?1 IAA. The obtained cultures were treated with different concentrations of lead nitrate (0.1, 0.5, and 1.0 mM Pb(NO₃)₂) or cadmium chloride (0.5, 2.5, and 5.0 μM CdCl₂). The growth parameters, photosynthetic pigments, and phenolic compound content were examined in order to evaluate whether tested metal salts can have an adverse impact on studied culture. It was ascertained that Pb ions induced growth disturbances and contributed to shoot wither. On the contrary, the proliferative shoot cultures were established on media containing Cd ions and the multiplication coefficients and shoot length increased on all media enriched with CdCl₂. Chlorophylls and carotenoid contents were negatively affected by application of 5.0 μM of cadmium; nevertheless, in shoots treated with 2.5 μM CdCl₂, increased accumulation of photosynthetic pigments occurred and their amount was similar to untreated culture. Adaptation to Cd was associated with stimulation of phenolic compound synthesis. Hence, we have reported on unambiguous positive result of in vitro selection procedure to obtain vigorous shoot culture tolerant to cadmium.     

Does single-layer centrifugation with Bovicoll improve sperm quality of frozen-thawed semen in Fleckvieh bulls?

The aim of the present study was to evaluate the effect of sperm selection by single-layer centrifugation (SLC) performed before freezing on sperm quality after thawing of Fleckvieh bull semen. Ejaculates from 22 bulls were collected by artificial vagina and divided into two aliquots. One aliquot (control sample) was diluted with Steridyl^® and frozen over nitrogen vapour in a Digitcool freezer (IMV Technologies). Sperm from the second aliquot (SLC sample) was selected using the SLC technique with Bovicoll colloid and then frozen over nitrogen vapour in a Digitcool freezer. After thawing, both samples (control and SLC) were evaluated by computer-aided sperm analysis (CASA; SCA 6.4 System; Microptic S.L) for sperm motility parameters. Integrity of the plasma membrane (viability), high mitochondrial membrane potential (HMMP) and acrosome integrity were assessed using a Guava® easyCyte flow cytometer (IMV Technologies). Morphological examination of spermatozoa was performed by Differential Interference Contrast microscopy (Leica DMi8). Morphological examination of live, immobilized spermatozoa was analysed under high magnification (≥6,600×). After thawing, the mean sperm viability of the control sample was 51.57%, compared to 40.37% for the SLC sample (p < .01). HMMP was higher (p < .01) in the control sample (40.37% versus 28.96%), and the mean of live spermatozoa with damaged acrosome was significantly higher (p < .03) in the SLC sample (1.63% versus 1.95%). The mean percentage of motile spermatozoa was 80.17% in the control sample, compared to 75.14% in the SLC sample (p < .0195), and rapid subpopulation reduced from 20.08% to 8.99% (p < .0001) after SLC. Percentage of hyperactivated sperm decreased from 12.23% to 4.28% (p < .0001) after SLC. Given the overall results, the sperm quality of thawed Fleckvieh bull semen was not improved when sperm were selected by SLC before freezing.     

Synchronous fluorescence spectroscopy of edible vegetable oils. Quantification of tocopherols

The study demonstrates the application of front face and right angle synchronous fluorescence spectroscopy for the characterization of edible oils. The method enables monitoring of tocopherols, pheophytins, and other fluorescent components in edible oils. Principal component analysis of synchronous fluorescence spectra revealed sample clustering according to the type of oil. Partial least-squares regression was utilized to develop calibration models between fluorescence spectra and total tocopherol content determined by HPLC. The regression models showed a good ability to predict tocopherol content. The best fitting results were obtained for 1% v/v diluted oils and for bulk samples using the entire spectrum, yielding the regression coefficient, r, of 0.991, and root mean square error of cross-validation of approximately 8%. The results presented confirm the capabilities of the fluorescence techniques as a tool for the analysis of edible oils.     

Different flight behaviour of the endangered scarce large blue butterfly Phengaris teleius (Lepidoptera: Lycaenidae) within and outside its habitat patches

Understanding individual movements in heterogeneous environments is central to predicting how landscape changes affect animal populations. An important but poorly understood phenomenon is behavioural response to habitat boundaries and the way animals cross inhospitable matrix surrounding habitat patches. Here, we analyze movement decisions, flight behaviour, and activity of the endangered scarce large blue Phengaris (Maculinea) teleius, focusing on the differences among the patterns observed in patch interior, at patch boundaries and within matrix. The probability of crossing an external patch boundary, regardless of the land use in the adjacent area, was considerably lower than crossing a ‘control line’ within patch interior. Movement distances, flight durations and net squared displacement were largest in matrix, while similarly smaller at patch boundaries and in patch interior. The distribution of angles between successive movements was clearly clustered around 0° (indicating flight in a straight line) in matrix and at patch boundaries, but not in patch interior. There were no differences in time spent on foraging, resting and ovipositing between patch interior and boundaries, but the first two activities rarely, and oviposition never, happened in matrix. Our results suggest that although P. teleius adults do not avoid using the resources located in the boundaries of habitat patches, they often return to the interior of the patches when crossing their boundaries. However, having entered the matrix the butterflies perform relatively long and straight flights. The estimated probability of emigration and net squared distance implies that the dispersal between local populations is common in this species in the studied area.     

BTH-mediated antioxidant system responses in apple leaf tissues

BTH (S-methylbenzo-1,2,3-thiadiazole-7-carbothiate), an active compound of the commercial preparation Bion, has been studied as an elicitor of resistance to fire blight (Erwinia amylovora) in apple. However, the biochemical mechanisms of its action are not fully elucidated. Our study indicated that BTH at the best time of its protection activity (2–14 days after application) induced changes in prooxidant–antioxidant balance in the leaves of apple trees, but in different ways in the enzymatic and nonenzymatic antioxidants. Glutathione as low molecular antioxidant as well as superoxide anion radical and lipid peroxides as oxidants exhibited changes at the early phase of BTH action. Glutathione-dependent enzymes were strongly affected by the elicitor used. On the 2nd day glutathione transferase (GST) and glutathione peroxidase (GSH-Px) activities increased by about 70% and 30% above the control, respectively. GST activity normalized about the 14th day but GSH-Px at the same time showed 27% of the control value. Among enzymes utilising hydrogen peroxide only catalase showed increase (37%) at the early phase of experiment. Compared with the control, BTH-treated plants did not show changes in ascorbate peroxidase and phenylalanine ammonia-lyase activities. Tocopherol (TOC) level diminished starting from the 7th day after BTH treatment and on the 14th day it was only 28% of the control. It is proposed that extinguishing of BTH-mediated signal resulted from TOC and glutathione action. The diminished ascorbate level at all examined times may play a crucial role in BTH-mediated cell growth regulation. The direct influence of BTH on lipid metabolism should be also taken into consideration.     

Effect of semen extenders and storage time on quality of Muscovy duck (Cairina moschata) drake semen during the entire reproductive season

Avian semen dilution with appropriate extender allows to prolong the fertilizing ability of sperm stored in vitro. In the present study, the impact of extenders and time of storage on morphology of Muscovy duck (Cairina moschata) drake semen were examined. Semen was collected twice a week, using male stimulation by a female method, from 12 adults (29 weeks old) drakes kept individually in cages, under controlled environmental conditions. Freshly collected, pooled ejaculates were divided into three part: neat undiluted sample, and diluted 1:1 with Schramm (SCH) or Watanabe (W) extender and stored at 4°C. Morphological examination of all samples was conducted after dilution and then, after 3 and 6 hr of storage. The storage of undiluted semen caused decrease (p ≤ .01) in live morphologically normal sperm, from 79.73% in the freshly collected ejaculates to 55.75% and to 12.12% after 3 and 6 hr of storage, respectively (average calculated for the entire reproductive season). In the semen diluted with Schramm's extender the adequate values attained 86.84, 79.65 and 61.66%, and using Watanabe extender 84.77, 83.58 and 75.25%, respectively. The period of semen storage and the type of extender caused significant (p ≤ 0,05; p ≤ 0,01) changes in sperm morphology. The longer period of storage contributed to the decrease in number of morphologically normal sperm, whereas their content in Watanabe extender after 3 and 6 hr of storage was higher (p ≤ .01) than in semen diluted in Schramm extender.     

The effect of temperature on the phenotypic features and the maceration ability of <Emphasis Type="Italic">Dickeya solani</Emphasis> strains isolated in Finland,Israel and Poland

Pectinolytic bacteria from the genus Dickeya (former Erwinia chrysanthemi), belonging to Dickeya dianthicola and Dickeya solani species, are causative agents of blackleg and soft rot diseases in Europe. Recently, D. solani have been isolated most frequently from potato plants with the symptoms of blackleg and soft rot. D. solani strains were shown to cause more severe disease symptoms on potato plants than D. dianthicola especially at the higher temperature. They are also able to develop blackleg disease from lower inoculum levels. In the presented study we not only compared phenotypic features of fifteen D. solani strains isolated in countries having different climatic conditions, Poland, Finland and Israel, but also we examined three D. dianthicola strains. The comparison was performed to determine the influence of the strain origin and the temperature of incubation on the ability of the strains to macerate potato tissue and on their major virulence factors such as: pectinolytic, cellulolytic and proteolytic activities, siderophore production and motility. Polish D. solani strains showed higher activities of cell wall degrading enzymes than the Finnish and Israeli strains at all the tested temperatures: 18, 27, 37 °C. This observation is correlated with the higher ability of Polish D. solani strains to cause soft rot. In addition, D. solani strains exhibited higher activity of the above mentioned enzymes and caused more severe potato tuber maceration in laboratory tests than the tested D. dianthicola strains. The collected results indicate that although D. solani strains from different climatic conditions have identical Pulse Field Gel Electrophoresis (PFGE) profiles in addition to the same fingerprint profiles obtained by the repetitive sequence-based polymerase chain reaction (REP, ERIC and BOX repetitive sequences), they differ in the examined phenotypic features, especially in the activities of pectinolytic, cellulolytic and proteolytic enzymes and their capacity to macerate potato tuber tissue.