Evolutionary history and phylogeography of Scots pine (<Emphasis Type="Italic">Pinus sylvestris</Emphasis> L.) in Europe based on molecular markers

In this review we summarized recent historical records and molecular studies on evolutionary history and phylogeography of Scots pine with focus on the European highly fragmented distribution area of the species. Fossilized pollen, plant micro- and macrofossil records provided evidences on the large-scale species’ range shifts and demographic changes during the Quaternary. Populations of Scots pine were documented both in the glacial (incl. full glaciation) and interglacial periods. Recolonization of Europe after the glaciation originated from the (Sub) Mediterranean areas like the Balkan Peninsula but also from around the Eastern Alps and the surroundings of the Danube plain. Fennoscandia and northern European Baltic regions were most probably colonized from two main directions, from Western Europe and from the Russian Plain. Modern history of Scots pine was hardly affected by anthropogenic activities that started to strengthen in the Bronze and Iron Age. Along with the fossil records, molecular genetic tools were used to infer the origin and putative history including migration, differentiation and demography of the species. In this paper we compiled the major publications (30) of molecular genetic studies of the past 20 years derived from distinctly inherited organelle genomes (mitochondrial, chloroplast, nuclear) revealed by different marker systems (mtDNA-cox1, -nad1, -nad3, -nad7, ISSR, cpSSR, nSSR, B-SAP, SNP). It is important to consider that different phylogeographic patterns can be drawn by the analysis of different DNA marker types. Accordingly the use of more than one marker simultaneously outlines the most sophisticated phylogeographical pattern on the genetic lineages and can reveal high differentiation of the European distribution. Combined marker systems and markers derived from coding sequences have also been used to detect species’ phylogeographic patterns, but these were rarely applied to Scots pine. Although new molecular techniques can provide higher resolution data for populations, the reviewed results can shape the direction of further studies.     

Determination of the chromium content of Hungarian winter wheat.

Bread and different winter wheat (Triticum aestivum) products are the main sources of chromium in the human diet in Hungary. Graphite furnace atomic absorption spectrometry (GF-AAS) is able to give reliable results for Cr. In this study the chromium content of some winter wheat species was determined after wet digestion. Contamination was the most critical part of chromium measurement; thus, appropriate sample preparation and the cleaning of kernels before digestion were required. The chromium content was 2-3 times higher than expected when contamination occurred. Samples came from a field plot experiment and, therefore, the effect of fertilization (150 kg/ha N, 112.5 kg/ha P(2)O(5), and 132.5 kg/ha K(2)O) on chromium content of grains was studied. The average measured values of different species were 114+/-3 and 134+/-4 microg/kg for the control and fertilized plots, respectively.     

Effect of different concentrations of Bacillus subtilis on growth performance, carcase quality, gut microflora and immune response of broiler chickens

1. An experiment was conducted to study the effects of different Bacillus subtilis concentrations on productivity, carcase quality, immune response and the gut microflora of broiler chickens. 2. There were 5 treatment groups: control, with no added Bacillus subtilis supplementation; and 4 treatment groups receiving feed supplemented with different concentrations of B. subtilis. 3. Weight gain was significantly higher and the feed conversion rate was significantly better in all groups receiving feed supplemented with B. subtilis, regardless of its concentration. The groups given B. subtilis supplementation produced relatively bigger breasts, and smaller carcases and thighs, compared with the control group. 4. The appearance of increased diffuse lymphohistiocytic infiltration and solitary lymphoid follicles in the mucosa, and a stronger response to NDV vaccination, indicate increased immunological responses in chickens fed with the B. subtilis supplemented diet. 5. The higher inclusion rate of B. subtilis did not increase Lactobacillus concentrations in the ileum or in the caecum, but decreased the E. coli population significantly.     

Comparison of feed preference and digestion of three different commercial diets for cats and ferrets

Diet preference and digestibility experiments were conducted using a total of 10 cats and 10 ferrets. The composition of the three different kinds of dry cat feed was as follows (each data are given in dry matter, DM): (i) normal diet (N): 95.3% DM, 33.7% crude protein (CP), 20.4% ether extract (EE), 37.6% nitrogen-free extract (NFE); (ii) 'light diet' (L): 94.2% DM, 31.6% CP, 10.7% EE, 52.2% NFE; (iii) 'veterinary diet' (D): 94.57% DM, 38.7% CP, 9.6% EE, 47.2% NFE. During the period of the preference test, the average daily dry matter intake (calculated with the mean of the three diets: 94.7% DM) was 98.0, 15.0 and 16.7 g DM in cats and 25.0, 7.3 and 8.1 g DM in ferrets. The preference rates of the three different diets, expressed in percentage of their total consumption, were as follows: 60.4% N (54.4 g DM), 12.4% L (12.1 g DM) and 27.2% D (26.6 g DM) in cats, and 46.2% N (11.6 g DM), 29.9% L (7.5 g DM) and 23.9% D (6.0 g DM) in ferrets. This indicates that cats and ferrets have a clear preference for diets of higher fat content. In all three diets, the digestibility of CP was significantly (p < 0.05) lower (70.1 +/- 5.4 vs. 75.9 +/- 5.8) while that of EE was significantly (p < 0.05) higher (95.6 +/- 1.5 vs. 89.4 +/- 5.3) in ferrets than in cats. The average digestible/metabolizable energy (DE/ME) ratio of feeds turned to be 95.6% for cats and 90.6% for the ferrets. From the data one can conclude that the ferret cannot be used as a model animal for cats either for preference or digestibility studies.     

Seasonal fertility differences in synchronised dairy cows: ultrasonic, metabolic and endocrine findings

The aim of this study was to identify relationships among seasonal differences of fertility, metabolic parameters and appearance of irregular luteal forms in high-yielding dairy cows. Holstein-Friesian cows were put on the Provsynch regimen in winter (n = 10) and in summer (n = 10). Blood sampling (starting 35 days post partum) and rectal ultrasound examinations (starting post insemination) were carried out once a week in each examination period. Metabolic [plasma nonesterified fatty acid (NEFA) and beta-hydroxybutyrate (BHB) levels, ferric reducing ability of plasma (FRAP) and serum beta-carotene] and endocrine parameters [plasma thyroxine (T4), triiodothyronine (T3), insulin-like growth factor (IGF-I) and insulin levels] were measured. In summer, two cows were excluded from the study because of metritis and none of the remaining animals became pregnant, but 6 of the 8 cows had irregular luteal forms (ILF) on their ovaries. In winter, one cow was excluded because of metritis and 6 of the 9 cows became pregnant, while 2 of the 3 open cows had irregular luteal forms. In summer the mean plasma NEFA and BHB concentrations were significantly higher, while serum carotene and plasma IGF-I concentrations were significantly lower than in winter. The high plasma NEFA concentration found in summer seemed to be in association with the lower body condition score (BCS) caused by depressed appetite. In conclusion, statistical analysis supports the hypothesis that increased plasma NEFA and BHB and decreased plasma IGF-I concentrations may result in reduced fertility in summer. These changes may be associated with the more frequent appearance of ILFs and probably have a negative effect on ovarian function and/or oocyte quality.     

Emergent properties of reduced-genome Escherichia coli

With the use of synthetic biology, we reduced the Escherichia coli K-12 genome by making planned, precise deletions. The multiple-deletion series (MDS) strains, with genome reductions up to 15%, were designed by identifying nonessential genes and sequences for elimination, including recombinogenic or mobile DNA and cryptic virulence genes, while preserving good growth profiles and protein production. Genome reduction also led to unanticipated beneficial properties: high electroporation efficiency and accurate propagation of recombinant genes and plasmids that were unstable in other strains. Eradication of stress-induced transposition evidently stabilized the MDS genomes and provided some of the new properties.     

Periparturient endocrine and metabolic changes in healthy cows and in cows affected by mastitis

Transition from pregnancy to lactation in dairy cows involves considerable metabolic adaptation. Additional stress is incurred during infections such as periparturient mastitis. Multiparous Holstein-Friesian cows kept under normal production conditions (n = 15) were used to evaluate changes in circulating metabolite and hormone concentrations from 5 days before to 5 days after calving. Insulin-like growth factor binding protein (IGFBP) profiles were also monitored. Marked time-related changes were observed for plasma thyroid hormone, IGF, cortisol, insulin, beta-hydroxybutyrate (BHB) and non-esterified fatty acid (NEFA) concentrations but not for plasma leptin. A decrease in IGF-II concentration and maximal intensity of the putative IGFBP-1 band occurred at parturition. When compared with the five healthy cows,low IGF-II levels were prolonged to day 2 post-partum in five cows with Escherichia coli-associated mastitis. However, marked decreases in IGFBP-2 band intensity were evident only in two of the four cases examined. Individual total ligand (IGF-I + IGF-II) concentration and IGFBP pattern prepartum were largely regained 5 days post-partum in all cows. Hormone and metabolite concentrations in the two cows with Staphylococcus aureus-associated mastitis were very similar to those in the five healthy cows. Plasma thyroxine (T4) was lower 2 days prepartum in the cows, which later developed Gram-negative mastitis. Multiregression analysis showed that variance in T4 concentration was significantly and independently associated with triiodothyronine (T3) and IGF-I positively and with cortisol negatively (R2 = 0.648). This study confirms the close inter-relationship between the thyroid hormone and IGF axes in cattle and indicates possible effects of Gram-negative mastitis infection on IGF-II metabolism.     

Occurrence of atypical myxomatosis in Central Europe: clinical and virological examinations

An outbreak of the atypical form of myxomatosis struck a rabbit farm in Hungary. The animals had previously been vaccinated with a vaccine containing Shope rabbit fibroma virus strain. The disease appeared in winter when the presence of mosquitoes and fleas is not common. The virus was isolated from an eyelid specimen of a naturally infected rabbit. The surviving animals were observed for four weeks, blood samples were collected and, after euthanasia, organ specimens were also examined by morphological methods including pathology and electron microscopy. Serum samples were examined by virus neutralisation for antibodies. Genetic analysis of the isolated virus was carried out by polymerase chain reaction (PCR) and direct sequencing. The primers were designed on the basis of the major envelope gene (Env) of the Lausanne reference strain in the GenBank. The viral proteins were examined by SDS-PAGE. The isolated virus (ref. no.: BP04/2001) was able to infect the susceptible animals directly, by contact. The disease was characterised by respiratory symptoms of the upper tracheal tract, conjunctivitis and high mortality by the 11th-14th day. Aerogenic infection with strain BP04/2001 resulted in 100% morbidity among the susceptible animals. Sequencing of the amplified 400-bp-long DNA revealed 97% homology with the Env gene of the Lausanne strain, which proves that strain BP04/2001 is a variant of the Lausanne strain having been enzootic throughout Europe. The live vaccine strain used in Hungary against myxomatosis, which is also a Lausanne-derived strain, protected the animals. According to the protein analysis a protein of 200 kDa in size is not expressed in strain BP04/2001. This is the first report on atypical myxomatosis in Central Europe. The virus spreads by airborne transmission and may cause severe losses in the rabbit population.