Molecular typing of Staphylococcus aureus based on PCR-RFLP of coa gene and RAPD analysis

The aim of this study was molecular identification of S. aureus strains isolated from mastitic milk samples and establishing the genetic relationship between strains isolated from cows belonging to the same herd. In all 43 isolated strains the gap gene (930 bp) was amplified, which enabled their affiliation to the Staphylococcus genus to be established. PCR-RFLP with AluI endonuclease of the gap gene as well as nuc (450 bp) and coa (1130 bp) gene amplification allowed precise S. aureus species identification. One hundred percent of the genetic relationship between strains was established via RAPD-PCR and coa-typing.     

Dose-dependent in vivo effect of Rhodiola and Echinacea on the mitogen-induced lymphocyte proliferation in mice

Echinacea purpurea (EP) and Echinacea angustifolia (EA) are ones of the most important world's herbs with immunotropic activity. They were traditional medicinal plants used by North American Indians for the treatment of various illnesses. Now they are cultivated in many countries and are used mainly to treat respiratory tract infections. Rhodiola rosea (RR) and Rhodiola quadrifida (RQ) are medicinal plants originated from Asia and used traditionally as adaptogens, antidepressants, and anti-inflammatory remedies. We previously reported, that extracts of underground parts of RR and RQ exhibited immunotropic activity. We have demonstrated in pigs that in vitro RR or RQ supplementation of blood lymphocyte cultures stimulated T cell proliferative response to Con A in lower, and inhibited it in higher Rhodiola extract concentrations. The aim of this work was to evaluate the in vivo effect of these herbal remedies on the in vitro proliferative response of mouse splenic lymphocytes to another T-cell mitogen- Phaseolus vulgaris haemagglutinin (PHA). We have found significant stimulation of proliferative response, in comparison to the controls, in mice fed lower doses of tested remedies, and inhibition, no effect or lower stimulation, in mice fed higher doses of these drugs.     

Slaughter value, meat quality, creatine kinase activity and cortisol levels in the blood serum of growing-finishing pigs slaughtered immediately after transport and after a rest period

The experimental materials comprised 44 hybrid [female (Polish Large White x Polish Landrace) x male Duroc] growing-finishing pigs. The animals were randomly divided into two groups: 24 pigs were slaughtered immediately after transport and 20 pigs were slaughtered after a 24-hour rest period in the lairage. The meat content of pork carcasses, carcass dressing percentage, the proximate chemical composition, physicochemical and sensory properties of meat and shear force values were determined. Serum creatine kinase activity and cortisol levels were determined in blood samples collected before transport and during carcass bleeding. Pigs slaughtered immediately after transport, compared with those slaughtered after a 24-hour rest period, were characterized by a higher meat content of the carcass and a higher carcass dressing percentage. Pre-slaughter handling had no effect on pork quality. The incidence of normal-quality meat, partially PSE (pale, soft, exudative) meat and PSE meat was similar in both groups. Chemical analysis showed that the content of dry matter, total protein, fat and minerals in meat was comparable in both groups. As regards the functional properties of the pork, samples from the carcasses of pigs that had rested before slaughter had a higher contribution of the red color component. Meat from pigs slaughtered immediately after transport had more desirable sensory properties. Pre-slaughter resting had a significant effect on those analyzed physiological parameters which were found to be good indicators of pre-slaughter stress. Serum creatine kinase activity and cortisol levels were higher in blood samples collected after transport (during carcass bleeding) than in samples collected before transport, pointing to a strong stress response of animals to pre-slaughter treatment. The decrease in serum cortisol levels in blood samples collected during bleeding from the carcasses of pigs slaughtered after a 24-hour rest period, compared with samples collected from animals slaughtered immediately after transport, suggests that rest before slaughter alleviated stress induced by pre-slaughter handling operations.     

Growth factor and cytokine interactions in myogenesis. Part II. Expression of IGF binding proteins and protein kinases essential for myogenesis in mouse C2C12 myogenic cells exposed to TNF-alpha and IFN-gamma

The aim of the study was to examine potential interactions among IGF-I and proinflammatory cytokines, TNF-alpha and IFN-gamma, in the regulation of local IGF-I bioavailability and cellular proteins mediating myogenic signals. We investigated levels of IGFBP-4, -5, -6, protein kinase Czeta (PKC zeta), p38 and extracellular signal-regulated kinase (ERK1/2) in differentiating mouse C2C12 myoblasts. IGF-I significantly stimulated expression of IGFBP-5. TNF-alpha and IFN-gamma attenuated the expression of IGFBP-4 and -6 under basal conditions and in the presence of IGF-I, and inhibited IGF-I-induced IGFBP-5 expression during 5-day myogenesis. TNF-alpha and IFN-gamma markedly attenuated p38 expression in the presence of IGF-I on the 5th day of myogenesis. When combined with IGF-I the cytokines exerted opposite effects on the PKC zeta level, i.e. TNF-alpha caused an increase, whereas IFN-gamma reduced the cellular content of this kinase. Exposition of C2C12 myoblasts to IGF-I or cytokines led to the stimulation of ERK1/2 phosphorylation; however, both TNF-alpha and IFN-gamma exerted an inhibitory effect on the activation of ERK1/2 in myoblasts cultured in the presence of IGF-I. We concluded as follows: i) TNF-alpha and IFN-gamma present in the extracellular environment of differentiating C2C12 myoblasts can alter the local bioavailability of IGF-I by inhibiting the expression of IGFBP-4, -5, and -6, ii) the decrease in p38 expression and ERK1/2 phosphorylation in C2C12 myoblasts exposed to cytokines can lead to disturbances in IGF-I-regulated myogenesis.     

The effects of prenatal exposure to methylxanthines

This review discusses epidemiology and laboratory studies on the effects of prenatal methylxanthine administration on some systems developing organisms. They are mainly absorbed from coffee, tea and cocoa products such as cola beverages and chocolate bars. Prenatal methylxanthine exposure can induce several unfavourables changes in the developing organism, which are persistent even in later phases of life. Based on results obtained from animal studies, the effect on embryogenesis is not only poorly understood but also controversial. It is therefore important to study interspecies differences as results may differ depending on animals used and administration methods.     

The effect of diets supplemented with fish broth and fish oil on the health of weaners

The aim of this study was to determine the effect of fish-based feed materials, as a source of readily available protein contained in fish broth and essential polyunsaturated fatty acids (PUFAs) found in fish oil, on the health of piglets and rearing results. The experiment was conducted on a commercial pig fattening farm. The study involved a total of 80 weaners with an approximate body weight of 15 kg. The experiment was carried out over a period of 40 days. Feed samples were subjected to laboratory analyses. Blood samples were collected from experimental group animals to determine serum biochemical and immunological parameters. The body weight gains of weaners, mortality rates and average feed intake per animal were calculated for the entire experimental period. The addition of fish broth and fish oil significantly improved the n3:n6 fatty acid ratio in diets. The presence of EPA and DHA in the experimental diet could have had a positive health effect on piglets, comparable with that exerted by therapeutic doses of zinc often administered to pigs of this age group. During the experiment, feed conversion ratio (FCR) gain was considerably reduced in the experimental group, with similar daily gains in the control and experimental group.     

New natural host plants of ‘Candidatus Phytoplasma pini’ in Poland and the Czech Republic

The presence of phytoplasmas in seven coniferous plant species (Abies procera, Pinus banksiana, P. mugo, P. nigra, P. sylvestris, P. tabuliformis and Tsuga canadensis) was demonstrated using nested PCR with the primer pairs P1/P7 followed by R16F2n/R16R2. The phytoplasmas were detected in pine trees with witches’ broom symptoms growing in natural forest ecosystems and also in plants propagated from witches’ brooms. Identification of phytoplasmas was done using restriction fragment length polymorphism analysis (RFLP) of the 16S rDNA gene fragment with AluI, MseI and RsaI endonucleases. All samples showed RFLP patterns similar to the theoretical pattern of ‘Candidatus Phytoplasma pini’, based on the sequence of the reference isolate Pin127S. Nested PCR‐amplified products, obtained with primers R16F2n/R16R2, were sequenced. Comparison of the 16S rDNAs obtained revealed high (99·8–100%) nucleotide sequence identity between the phytoplasma isolates. The isolates were also closely related to four other phytoplasma isolates found in pine trees previously. Based on the results of RFLP and sequence analyses, the phytoplasma isolates tested were classified as members of the ‘Candidatus Phytoplasma pini’, group 16SrXXI.     

Phosphorus use Efficiency of Safflower (Carthamus tinctorius L.) and Sunflower (Helianthus annuus L.)

This study comparatively evaluates phosphorus (P) use efficiency of safflower and sunflower in view of identifying low input oil crops. Sunflower accumulated more P than safflower at all equivalent P supplies. Sunflower was more efficient at their best P supplies to utilize absorbed P than safflower in terms of efficiency ratio and utilization index. Yield response curves interpreting the functional relationship between nutrient supply and yield using Michaelis-Menten equation indicate that sunflower is less sensitive to inadequate P supply and thus more P-efficient than safflower with respect to yield, as the term ‘Km’ is lower for sunflower. The minimal P amount per pot required to trigger yield formation (term ‘Cmin’) was lower in sunflower than safflower. Harvest Index in sunflower out-yielded that of safflower at all P supplies. Critical deficiency contents of sunflower were lower than that for safflower. It can be concluded that safflower cannot be considered as low-input crop.     

Bound amino acids in humic acids from arable cropping systems

We investigated the varying concentrations of bound amino acids in humic acids (HA) extracted from soils under both crop rotation and continuous cropping of rye. The experiment was created in 1957. Since then, winter rye had been grown continuously and also the sequence of the 7 yr rotation had been started: potato, spring barley, alfalfa, alfalfa, oil seed rape, winter rye, and winter rye. Soils were fertilized with NPK and manure. Continuous cropping of rye increased total acidity of soils and the contents of carboxylic and phenolic groups in HA. The total amounts of the bound amino acids in HA from soils under crop rotation were higher than from continuous cropping of rye. Fertilization with NPK increased the contents of bound amino acids more than manure. Neutral amino acids dominated in all samples of HA, and basic amino acids had the lowest concentrations. In both types of cultivation, glutamic acids, glycine, alanine, valine, and lysine dominated. The proline contents in HA from continuous rye cropping were higher than in HA from soils under crop rotation. The concentrations of β‐alanine and lysine were higher in HA from crop rotation indicating a higher microbial biomass since these compounds are typical constituents of bacteria cell walls.     

Changes in the chemical composition of water percolating through the soil profile in a moderately polluted catchment

Throughfall (TF), stemflow (SF), soil solution below the organic layer (SS_org) and at 50 cm depth (SS₅₀), and output with stream water (SW) were measured and analyzed for four years in a moderately polluted forest catchment in southern Poland. The input of water with stemflow was ca. 6% of input with TF. However, due to higher concentrations of most ions in SF, the input of most elements with SF was from 8% to 9%. Sulphate (SO₄ ^2–), chloride (Cl^–) and magnesium (Mg²⁺) were the only ions steadily increasing in concentrations in water percolating through the soil profile. Nitrogen reached the forest floor mainly as ammonium (NH₄ ⁺). In the soil organic layer the NH₄ ⁺ concentration decreased, while concentrations of nitrate (NO₃ ^–) and hydrogen (H⁺) increased, probably due to nitrification. For NO₃ ^–, sodium (Na⁺) and calcium (Ca²⁺), the highest concentrations were found in SS_org and SW. This indicates both efficient cycling in the biotic pool of the ecosystem and intensive weathering processes in the mineral soil below the plant rooting zone. The latter was especially pronounced for Mg and Ca. Concentrations of zinc (Zn), lead (Pb) and cadmium (Cd) were the highest in SS_org and SS₅₀. As this was accompanied by a low pH and constant input of H⁺, NH₄ ⁺ and heavy metal ions to the catchment area, it may pose a serious threat to forest health.