Molecular markers for the identification of resistance genes and marker-assisted selection in breeding wheat for leaf rust resistance

The resistance genes Lr9, Lr24, Lr25, Lr29, Lr35 and Lr37, which were not previously utilised in Hungary, have been incorporated into four Martonvásár winter wheat cultivars using marker-assisted selection with PCR-based markers. In the course of a backcross programme, the genes were transferred into Martonvásár wheat varieties and various BC generations were produced. Work aimed at pyramiding resistance genes is currently underway in Martonvásár, and plants containing the gene combinations Lr9 + Lr24, Lr9 + Lr25 and Lr9 + Lr29 are now available. From the BC₂F₄ generation of the ‘Mv Emma’*3/’R.L.6010’ combination (‘R.L.6010’ is the donor of the Lr9 gene) 287 lines were tested for leaf rust resistance in an artificially inoculated nursery. A co-dominant primer combination was designed to identify both resistant and susceptible offsprings. The results of resistance tests and molecular marker detection agreed in most cases. Designated leaf rust resistance genes were identified with molecular markers in wheat varieties and breeding lines. The Lr26 and Lr34 resistance genes occur frequently in the Martonvásár gene pool, and the presence of the Lr37 gene has also been detected in a number of Hungarian genotypes.     

Effect of fulvic and humic acids on performance, immune response and thyroid function in rats

Effect of humic acid (HA) and fulvic acid (FA) on production parameters, immune response and thyroid function of rats were investigated in two experiments. First experiment: control or 0.1%, 0.2%, 0.4%, 0.8% HA- or FA-supplemented diets. Second experiment: control and 0.4% HA- or FA-supplemented diets. The feeding period lasted 26 days in both trials. No significant changes were observed in production parameters. Ovalbumine antibody titre of rats on HA- or FA-supplemented diets showed dose-dependent (at 0.4% supplementation) and significant (p < 0.05) increase (350 and 418% respectively) over the control (100%). Dose-related increase of plasma TSH (r = 0.99), and decrease of the T(4)/T(3) ratio (r = -0.97) was observed in FA-supplemented rats. Second experiment: both FA and HA stimulated the immune response by the 14th day (mean values: control: 685.79; FA: 1131.37; HA: 1055.6099) and 26th day (control: 544.31; FA: 1969.83; HA: 1600.00). No significant differences were noted with lymphocyte stimulation test. Diameter of the 'B'-dependent lymphoid tissues in the ileum and spleen were significantly (p < 0.05) larger in both the FA- and HA-treated animals. Humic acid and FA supplementation resulted in strong humoral immune stimulation. Our data also indicate that FA content is responsible for the mild hypothyroid effect of humic substances.     

Manufacture and properties of bifidogenic saccharides derived from wood mannan

Pinus pinaster wood samples were subjected to double hydrothermal processing. The liquors coming from the second stage, containing soluble saccharides of polymeric or oligomeric nature from hemicelluloses (POHs), were subjected to membrane processing (operating in discontinuous diafiltration) for refining and fractionation. Refined POH fractions were characterized by matrix-assisted laser desorption/ionization time of flight mass spectrometry and chromatographic techniques. The most complex POH component was made up of 14 hexoses and contained 4 acetyl groups. The fermentability of purified POHs by human fecal inocula was assessed by measuring both carbon source consumption and formation of short-chain fatty acids. The bifidogenic ability of POHs was confirmed by fluorescence in situ hybridization. The stimulatory effects on the bifidobacterial population reached by POHs were of the same order as those obtained with commercial fructooligosaccharides.     

Human exposure to perfluorinated compounds in Catalonia, Spain: contribution of drinking water and fish and shellfish

In this study, the concentrations of 15 perfluorinated compounds (PFCs) were analyzed in 30 water samples collected in Catalonia (Spain) at three stages of the drinking water treatment process in several water purification plants. In addition, the concentrations of 13 PFCs were determined in samples of fish and shellfish collected from coastal areas of Catalonia. The intake of PFCs through both pathways, drinking water intake and fish and shellfish consumption, was also estimated. In water samples, the highest mean concentrations corresponded to perfluorooctane sulfonate (PFOS) and perfluorooctanoate (PFOA) (1.81 and 2.40 ng/L, respectively), whereas perfluorodecanosulfonate (PFDS) and perfluorotetradecanoic acid (PFTDA) were under their respective limits of detection in all analyzed samples. The results show that although the current treatment processes caused slight reductions in PFC concentrations, these processes did not mean significant changes in the amounts of PFCs already contained in the raw water. Among the analyzed PFCs in fish and shellfish, only seven compounds could be detected in at least one composite sample. PFOS showed the highest mean concentration (2.70 ng/g fw), being detected in all species with the exception of mussels. With regard to PFOA (mean, 0.074 ng/g fw), the highest concentrations were detected in prawn and hake (0.098 and 0.091 ng/g fw, respectively). The current exposure to PFCs through consumption of fish and shellfish indicates that it should not be of concern for the consumers. The amounts ingested are well below the recommended tolerable daily intakes, at least for those PFCs for which information is available.     

Role of wild ruminants in the epidemiology of bluetongue virus serotypes 1, 4 and 8 in Spain

ABSTRACT: Although the importance of wild ruminants as potential reservoirs of bluetongue virus (BTV) has been suggested, the role played by these species in the epidemiology of BT in Europe is still unclear. We carried out a serologic and virologic survey to assess the role of wild ruminants in the transmission and maintenance of BTV in Andalusia (southern Spain) between 2006 and 2010.A total of 473 out of 1339 (35.3%) wild ruminants analyzed showed antibodies against BTV by both ELISA and serum neutralization test (SNT). The presence of neutralizing antibodies to BTV-1 and BTV-4 were detected in the four species analyzed (red deer, roe deer, fallow deer and mouflon), while seropositivity against BTV-8 was found in red deer, fallow deer and mouflon but not in roe deer. Statistically significant differences were found among species, ages and sampling regions. BTV RNA was detected in twenty-one out of 1013 wild ruminants (2.1%) tested. BTV-1 and BTV-4 RNA were confirmed in red deer and mouflon by specific rRT-PCR.BTV-1 and BTV-4 seropositive and RNA positive wild ruminants, including juveniles and sub-adults, were detected years after the last outbreak was reported in livestock. In addition, between the 2008/2009 and the 2010/2011 hunting seasons, the seroprevalence against BTV-1, BTV-4 and BTV-8 increased in the majority of provinces, and these serotypes were detected in many areas where BTV outbreaks were not reported in domestic ruminants. The results indicate that wild ruminants seem to be implicated in the dissemination and persistence of BTV in Spain.     

Echocardiographic and electrocardiographic examination of clinically healthy, conscious ferrets

Objectives : To assess the feasibility of performing electrocardiography (ECG) and echocardiography on conscious ferrets and to establish reference values for this species under these conditions. Methods : End‐systolic and end‐diastolic left ventricular (LV) diameters, LV and interventricular septal thicknesses were measured by M‐mode echocardiography, while left atrial, aortic and pulmonary trunk diameters were determined by two‐dimensional echocardiography in 46 ferrets. In some animals the maximum velocities of blood flow in the aorta and the pulmonary trunk were also measured by spectral Doppler. ECG examinations were performed by using limb leads both in right lateral recumbency and in the so‐called “hanging” position. In the course of the ECG examinations (n=43), the occurrences, directions, durations and amplitudes of the P, QRS and T waves and the duration of the PR‐ and QT intervals were recorded for all six leads. Results : ECG and echocardiography were successfully performed on non‐sedated ferrets in 90 and 96% of the cases, respectively. Males were significantly heavier and had larger cardiac dimensions compared to females. Clinical Significance : Performance of both echocardiography and ECG are feasible techniques in conscious ferrets, and reference values are now available for future comparisons.     

Claudin-7-positive synchronous spontaneous intrahepatic cholangiocarcinoma, adenocarcinoma and adenomas of the gallbladder in a Bearded dragon (Pogona vitticeps)

In this study, synchronous spontaneous, independent liver and gallbladder tumours were detected in a Bearded dragon (Pogona vitticeps). The multiple tumours consisted of intrahepatic cholangiocarcinoma as well as in situ adenocarcinoma and two adenomas of the gallbladder. The biliary epithelial cells and the cholangiocarcinoma showed membranous cross-immunoreactivity for claudin-7. The gallbladder epithelial cells, its adenoma and adenocarcinoma showed basolateral cross-reactivity for claudin-7. We think that the humanised anti-claudin-7 antibody is a good marker for the detection of different primary cholangiocellular and gallbladder tumours in Bearded dragons. The cholangiocytes, the cholangiocarcinoma, the endothelial cells of the liver and the epithelial cells and gallbladder tumours all showed claudin-5 cross-reactivity. The humanised anti-cytokeratin AE1-AE3 antibody showed cross-reactivity in the biliary epithelial cells, cholangiocarcinoma cells, epithelial cells and tumour cells of the gallbladder. It seems that this humanised antibody is a useful epithelial marker for the different neoplastic lesions of epithelial cells in reptiles. The humanised anti-α-smooth muscle actin (α-SMA) antibody showed intense cross-reactivity in the smooth muscle cells of the hepatic vessels and in the muscle layer of the gallbladder. The portal myofibroblasts, the endothelial cells of the sinusoids and the stromal cells of the cholangiocarcinoma and gallbladder tumours were positive for α-SMA. The antibovine anti-vimentin and humanised anti-Ki-67 antibodies did not show crossreactivity in the different samples from the Bearded dragon.     

Evaluation of a commercial intravaginal thermometer to predict calving in a Hungarian Holstein-Friesian dairy farm

In this study, the utility of a commercial intravaginal thermometer was evaluated as an automated method for the prediction of calving in a total of 257 healthy pregnant Holstein–Friesian female cattle. The accuracy and the sensitivity of predicting calving within 48 hr before calving were also evaluated. The intravaginal temperature changes from 72 hr before and up to calving were significantly (p ≤ .001) affected by parity, season (summer vs. autumn), the time of day (8 a.m. or 8 p.m.) and the 6-hr time intervals (38.19°C: first interval 0 to 6 hr before calving vs. 38.78°C: twelfth interval 66 to 72 hr before calving), while the gender (p = .943), and the weight of the calf (p = .610), twinning (p = .300), gestation length (p = .186), foetal presentation (p = .123), dystocia (p = .197) and retention of foetal membranes (p = .253) did not affect it significantly. The sensitivity of the SMS of expecting calving within 48 hr and the positive predictive value were 62.4% and 75%, respectively, while the sensitivity and the positive predictive value for the SMS of expulsion reached 100%. It can be concluded that the investigated thermometer is not able to predict calving within 48 hr accurately; however, imminent calving can be accurately alerted.     

Antigenotoxicity and Antioxidant Activity of Acerola Fruit (Malpighia glabra L.) at Two Stages of Ripeness

Genotoxic and antigenotoxic effects of acerola fruit at two stages of ripeness were investigated using mice blood cells. The results show that no ripeness stage of acerola extracts presented any genotoxic potential to damage DNA (Comet assay) or cytotoxicity (MTT assay). When antigenotoxic activity was analyzed, unripe fruit presented higher DNA protection than ripe fruit (red color) extract. The antioxidant capacity of substances also showed that unripe samples inhibit the free radical DPPH more significantly than the ripe ones. The results about determination of compounds made using HPLC showed that unripe acerola presents higher levels of vitamin C as compared to ripe acerola. Thus, vitamin C and the complex mixture of nutrients of Malpighia glabra L., and especially its ripeness stages, influenced the interaction of the fruit extract with the DNA. Acerola is usually consumed when ripe (red fruit), although it is the green fruit (unripe) that has higher potential as beneficial to DNA, protecting it against oxidative stress.     

Occurrence, Distribution and Epidemiology of Grapevine Yellows in Spain

The main viticultural production areas in Spain were surveyed in 1994, 1995 and 1996 for the occurrence and incidence of Grapevine Yellows diseases associated to phytoplasmas. Samples from 300 plants showing symptoms of phytoplasma infection were collected from grapevine fields in the Spanish regions of Aragón, Catalonia and Navarra and analysed by PCR with specific primers for a non-ribosomal DNA of stolbur/Bois Noir (BN) and of Flavescence dorée (FD) phytoplasma. Nested PCR with universal primers P1/P7 and fU5/rU3 was also used. In the survey conducted in 1994 and 1995 only BN/stolbur phytoplasma was detected. The incidence of symptomatic plants was low in five plots of Catalonia from 3% to 18% in 1994 and 1995, respectively, and high in two plots of Navarra, from 60% to 80%. In the survey conducted in 1996 the incidence of symptomatic plants in Catalonia increased (6–80%) due to the presence of FD in five plots in the Northeastern Catalonia. An epidemiological study was carried out in two BN-affected plots of two regions from 1994 to 1997, with the evaluation of potential vectors and of host plants. The stolbur phytoplasma was found in individuals from different insect species belonging to the families Cicadellidae and Delphacidae. Some wild plants naturally infected with stolbur phytoplasma around the infected grapevines were: Convolvulus arvensis, Lavandula officinalis, Polygonum convolvulus, Solanum nigrum, and Thymus officinalis. The incidence of the disease in one BN-infected grapevine plot increased from 3.4% in 1994 to 18.40% in 1997.