The secreted lipase FGL1 is sufficient to restore the initial infection step to the apathogenic Fusarium graminearum MAP kinase disruption mutant Δgpmk1

Mitogen activated protein (MAP) kinases are key components in signalling networks. In Fusarium graminearum, one of the most devastating fungal plant pathogens, two MAP kinases are known to be involved in pathogenicity, Mgv1 and the Gibberella pathogenicity MAP kinase Gpmk1. ??gpmk1 mutants with a disrupted GPMK1 gene are unable to infect wheat spikelets. They exhibit altered secretion of several extracellular enzymes. Among those, the lipase FGL1 is known to be a major virulence factor of F. graminearum. FGL1 gene expression was decreased in ??gpmk1 strains during wheat head infection. To uncouple FGL1 expression from Gpmk1 activity, we generated ??gpmk1 strains that constitutively express FGL1 under control of the Cochliobolus heterostrophus glyceraldehyde 3-phosphate dehydrogenase (GPD) promoter. The level of extracellular lipolytic activity of these strains in culture was comparable to the wild-type. These mutants showed fully restored conidiation and partial complementation of defects in development that were reported from the ??gpmk1 mutant. They also partially complemented the apathogenic disease phenotype of the ??gpmk1 mutants causing lesions in directly inoculated wheat spikelets. But in contrast to wild-type, their growth was restricted to directly inoculated spikelets. A barrier-like formation was observed at the rachis node. Based on these results, we could show that the lipase FGL1 is necessary but not sufficient to restore complete pathogenicity to the apathogenic ??gpmk1 mutant. Hence, we hypothesize that the MAP kinase Gpmk1 is involved in the regulation of additional factors required for complete virulence of F. graminearum.     

Relationship between oxidant stress and milk productivity in dairy cows

An imbalance between formation and detoxification of oxygen radicals leads to oxidant stress that may increase in more intense oxidative metabolism caused by a high intake of metabolizable energy to provide metabolic intermediates for the milk synthesis and secretion. This hypothesis was tested using dairy cows and the concentration of hydroperoxides in lipids (LHP) extracted from circulative lipoprotein particles of low and very low density (LDL and VLDL/chylomicrons) as oxidant stress indicator. The particles were prepared by ultracentrifugation of serum obtained by coccygeal bleeding (13 cows, 1. parity, n=8 and 2. parity, n=5, lactation stage, 53 +/- 1.4 days post partum) and purified by precipitation. Concentrations of LHP-LDL/mg Lipoprotein correlated significantly with daily milk yield (r = 0.73, P = 0.004) or daily milk energy output (r = 0.77, P = 0.003) in contrast to LHP of VLDL/chylomicron particles. Thus, some evidence was obtained for an almost linear, positive relationship between milk productivity and oxidant stress occurring in LDL.     

Prevalence and association of porcine circovirus type 2 (PCV2), porcine parvovirus (PPV) and porcine reproductive and respiratory syndrome virus (PRRSV) in aborted fetuses, mummified fetuses, stillborn and nonviable neonatal piglets

Porcine circovirus type 2 (PCV2) seems to cause reproductive failure in sows not only in experimental studies. A retrospective study was made with a total of 252 aborted fetuses, mummified fetuses, stillborn and nonviable neonatal piglets to determine the presence of PCV2, porcine parvovirus (PPV) and porcine respiratory and reproductive syndrome virus (PRRSV) by PCR. PCV2 was found in all stages of gestation in 27.1 percent of samples examined. A statistically significant association could be shown between the detection of PCV2 and PRRSV. However, no significant association was seen between the detection of PCV2 and PPV and between PPV and PRRSV.     

Two cases of FeLV-associated dermatoses

Two cases of feline leukaemia virus (FeLV)-associated dermatosis are described. The first cat was affected by an ulcerative dermatitis identified as a giant-cell dermatosis. The second case was a cutaneous lymphoma. In both cases, FeLV antigens and FeLV genome were demonstrated in the affected skin immunologically and with polymerase chain reaction, respectively. The first case suggests that, like other retroviruses, at least some strains of FeLV can induce syncytium formation. As FeLV antigens and genome were demonstrated in a serologically negative cat, the second case suggests that focal skin FeLV replication may occur. FeLV-associated dermatoses are rare skin conditions that may be under-diagnosed.     

果树病毒RNA的二氧化硅提取法

针对果树组织含有较高的多酚和多糖物质,研究建立了适合果树病毒日常进行大量RT-PCR检测时,病毒RNA的快速、经济、高效的提取方法。该方法是基于在适宜条件下,二氧化硅对RNA的吸附和洗脱作用,避免了常规方法使用的酚、氯仿等有机溶剂。在2 h内同时可以进行数十个样品的提取,由于提取的是总RNA,1次提取,1次反转录得到的cDNA样品可以用于各种待检病毒的PCR检测。得到的总RNA和cDNA在-20℃条件下可以长期保存备用,大大提高了核酸的提取效率。     

P-32 Giant cell dermatosis in a FeLV-infected cat: detection of the virus in infected skin by immunohistology and PCR

The aim of this study was to report 20 cases of Lynxacarus radovskyi infestation in 20 cats . Lynxacarus radovskyi was diagnosed by physical examination and microscopic examination of plucked hair where countless adult mites and immature stages were identified. Microscopic examination was repeated every 7 days. The animals were observed daily for the first 7 days, and again on day 15 and day 30 for mites, pruritus and clinical signs. Clinical signs included salt-and-pepper appearance to a dull and opaque hair coat, alopecia of the dorsal and lateral areas of the hind limbs, and evidence of self-mutilation. One case had papules and crusts of the hind limbs. The pruritus was mild in animals with low infestation, but very intense in those with a heavy infestation. Most mites occurred in the lateral area of the hind limbs and sacral area. Twenty of 100 animals had confirmed L. radovskyi infestation. Therefore, we suggest that the transmission occurs through fomites. Treatment consisted of 0.5 mL application of Fipronil^® (Frontline TopSpot) per cat, with resolution in 100% of cases by day 15.
Funding: Merial.     

Use of GnRH in livestock production--current position and new research data

The hypothalamic gonadotropin-releasing hormone (GnRH) is seen as the key hormone of neuroendocrine regulation of reproduction. The ability of GnRH and its analogues to stimulate the release of the gonadotropins FSH and LH is world-wide utilized for various veterinary purposes, including treatment of certain hormone-dependent disturbances and stimulation of ovulation in controlled breeding programmes. A large difference is striking, however, when comparing the efficiencies reported. This may underline the importance of accurate treatment and reflect the manifold influences by animals and their environment on reproductive performance. During the last years, novel analytical methods have been established enabling a significant progress in reproductive research. The discovery and characterization of natural GnRH variants and their receptors in several vertebrate species may become more important.The reason is, that these GnRHs affect the release of the gonadotropins FSH and LH, but they may transmit, moreover, seasonal and nutritive signals to reproductive organs. It might be expected that the further exploration of these functions may serve as basis for the development of new and effective biotechnical methods in farm animal treatment.     

Ermittlung kinetischer Parameter der sauren Phosphatasen intakter Zuckerrübenwurzeln bei variierter Phosphaternährung

Determination of kinetic parameters of acid phosphatases in intact sugar beet roots of variable phosphorus nutrition Organically bound phosphorus has to be hydrolysed before its P can be taken up by plants. Both microbes and plant roots possess phosphatases, which could be of importance especially in soils with low concentrations of inorganic phosphorus in the soil solution. This could be the reason why nutrient uptake models underestimate the P-uptake by plants when P-mobilization by the phosphatases of roots is not taken into consideration. Therefore the activity of acid phosphatases (P_ase) was determined to answer the following questions: 1) To which extent does the root bound acid phosphatase (P_ase) follow Michaelis-Menten kinetics? 2) By which of the four linear transformations of the Michaelis-Menten equation (Lineweaver/Burk, Hanes, Eadie/Hofstee, Eisenthal/Cornish-Bowden) can plausible values of V_max and K_m be determined? 3) Which effect has the P nutrition of the plant on these kinetic parameters? Sugar beet plants were grown in full nutrient solution containing 1 and 100 μM P respectively. The P_ase activity of the intact roots was measured at pH 5 using p-nitrophenylphosphate (25—15000 μM p-NPP). V_max values were calculated per m root length. Acid phosphatase activity principally followed Michaelis-Menten kinetics. Transformations and calculations of V_max and K_m after Eadie/Hofstee and Eisenthal/Cornish-Bowden suggested the existence of at least two enzyme systems (P_{ase 1}, P_{ase 2}). The following kinetic parameters were found: P_{ase 1}: P deficient plants: V_max: 43—45 nmol m^—1 min^—1, K_m: 31—37 μM NPP; P sufficient plants: V_max: 7 nmol m^—1 min^—1, K_m: 47—53 μM NPP. P_{ase 2}: P deficient plants: V_max: 230—293 nmol m^—1 min^—1, K_m: 1579—3845 μM NPP; P sufficient plants: V_max: 123—171 nmol m^—1 min^—1, K_m: 3027—7000 μM NPP. Thus plants with sufficient P nutrition have a lower affinity to P_org and a lower hydrolysis of P_org. For P nutrition of crops P_{ase 1} might be the most important enzyme.     

Phosphordüngewirkung eisenhaltiger Klärschlämme

<?show $38#Bo;>Phosphorus fertilization effect of sewage sludges treated with iron compounds The result of waste water treatment with iron salts are sewage sludges with higher P but also Fe contents. The effect of such sludges on P availability in soils is not clear and was, therefore, compared with CaHPO₄ as a P mineral fertilizer. In experiments (Mitscherlich pots, 6 kg soil) two sandy soils (pH 4.3 and 4.9), two luvisols ( pH 6.3 and 7.2), a loess (13 % CaCO₃), and quartz sand all differing in their P status were treated with two sewage sludges which differed in their molar Fe:P ratios (sludge Gö: 1:0.3, sludge Sh: 1:1.2). For sludge Gö the P elimination had been carried out with FeSO₄ and the sludge was stabilized with FeCl₃ + Ca(OH)₂ (filter press). For sludge Sh P was eliminated by FeCl₃ in the aerobic basin. The first crop was maize (total shoot dry matter), the second red clover. In case of sludge Sh both crops took up in comparison to CaHPO₄ the same amount of P from all substrates with the exception of quartz sand. The effect of sludge Gö was quite different: from the neutral luvisol soils maize was able to take up only 64—82 % P and red clover 77—82 % P only as compared to CaHPO₄. On the sandy soils maize and red clover grew after sludge Gö as good or better than after CaHPO₄ application and P uptake was quite similar (83—106 %). This result was concomitant to an increase of pH values (from 4.3 to 4.9, 4.9 to 5.6, respectively). The negative efficiency of P uptake in heavy soils after application of sludge Gö resulted from an increase of P sorption and decrease of orthophosphate concentration in soil solution. This sludge contains a high proportion of iron hydroxides/oxides with free sorption sites for P. In future P elimination from waste water should be done without iron salts. At least during the process of conditioning no Fe salts should be used.<?show $6#>     

Die Genauigkeit von Messungsmethoden und Messungsergebnissen bei Holzmassenermittlungen

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