Porcine nephropathy induced by long-term ingestion of ochratoxin A.

Nine pigs were fed crystalline ochratoxin A in their feed at a concentration of about 1 mg/kg. Three pigs and their controls were killed after 3 months and 6 pigs and controls were killed after 2 years. A decrease of the ratio TmPAH/CIn, increased urinary glucose excretion and decreased ability to concentrate urine, occurred within a few weeks and aggravated slightly during the 2-year period. Changes in renal structure, characterized by degeneration and atrophy of proximal tubules, interstitial fibrosis and hyalinization of glomeruli, were progressive during time of exposure, but terminal renal failure was not reached. The kidney, liver, muscular and adipose tissue contained 3 to 27 microgram ochratoxin A/kg after 3 months of exposure. No further accumulation of ochratoxin A residue was found after 2 years of exposure.     

Rabies in a 10-week-old puppy

A 10-week-old, male, border collie-cross puppy was examined for an acute onset of unilateral vestibular signs. Neurologic deterioration was rapid over the next 12 hours and the puppy was euthanized. Rabies was diagnosed by histopathologic and immunohistochemical examination.     

The transferrin receptor of Actinobacillus pleuropneumoniae: quantitation of expression and structural characterization using a peptide-specific monoclonal antibody

When Actinobacillus pleuropneumoniae (A. pp) is grown under iron-restricted conditions in vitro, transferrin binding proteins (Tbps) are induced. The functional transferrin receptor of A. pp is composed of two outer membrane proteins (Tbp1 and Tbp2) and shows an exquisite specificity for porcine transferrin. This complex was studied using a monoclonal antibody (Mab 1.48) raised against a synthetic peptide corresponding to a hydrophilic domain of Tbp2 common to several A. pp serotypes. The antibody reacted specifically with a 60-70kDa Tbp2-antigen found in all serotypes of A. pp obtained from iron-restricted culture. It was found that Tbp2 was not expressed in iron replete medium by any serotype except serotypes 5a, 5b and 6 where a weak expression was seen. There was a weak expression of related antigens in Actinobacillus indolicus and Actinobacillus suis under iron-depleted conditions while no similar antigens were detected with the Mab in iron-starved Actinobacillus lignieresii, Actinobacillus porcinus, Actinobacillus minor, Haemophilus influenzae, and Haemophilus parasuis.Using an enzyme-linked immunosorbent assay (ELISA) based on the Mab 1.48, Tbp2 could be detected in both recombinant E. coli expressing Tbp2 and in wild type A. pp grown under iron restricted conditions. The subcellular location of Tbp2 in A. pp was studied by immunoelectron microscopy using the Mab 1.48. Interestingly, all antibody binding was found inside the A. pp cells, while Tbp2 expressed in recombinant E. coli was found both in the cytosol and on the outer membrane. These results indicate that the Mab 1.48-reactive epitope of Tbp2 is surface exposed when it is expressed without Tbp1 in E. coli while the inaccessibility of this epitope of Tbp2 in A. pp could be due to shading by the association between Tbp2 and Tbp1.     

Dendroecological assessment of the complex causes of decline and recovery of the growth of silver fir (Abies alba Mill.) in Southern Germany

Causes of decline and silver fir (Abies alba Mill.) mortality in Central Europe have been controversially discussed for a long time. This contribution details a comprehensive dendroecological investigation of two basic and alternative hypotheses that explain the decline by either climatic influences or by SO₂ pollution as the primary factors. Growth reduction between the 1960s and 1980s as well as the recovery of growth during recent decades have be seen as a response to complex interactions of multiple factors. Herein, we undertake a multidisciplinary evaluation of a broad database, representing the long-term growth of 1010 firs from 51 sites, long-term climate records as well as data assessing the long-term pollution situation in Southern Germany. The results provide clear evidence that SO₂-immissions play a key role in decline and that tree growth corresponds with the regional and temporal pattern of SO₂ pollution. Calculations with a new complex growth model indicate that tree-ring widths of fir at many sites were more influenced by SO₂ than by any other growth-determining factor during the second half of the last century. Strongest coherence between measured and modelled ring widths are found if – in addition to temperature, available soil water content, heavy frost events and long-term growth trends – the site-specific trend of SO₂ pollution is included. Both the immediate response to SO₂ pollution and the quick recovery of fir since the beginning of the 1980s after the decrease of SO₂ emissions indicate a direct impact of SO₂ in gaseous form and not via the soil in terms of soil acidification. Tree-ring widths document that radial growth response occurs at air concentrations above 10 μg SO₂ m⁻³ (annual average), which can be thought of as a critical threshold for silver fir health. Since the beginning of the 1980s, increasing growth trends have been observed which predominantly result from decreased SO₂ pollution in most parts of Central Europe, although climate change, fertilization by nitrogen deposition and a reduced tree-to-tree competition are also relevant factors. The drastic reduction of SO₂ emission during the recent decades significantly improved the environmental conditions for fir. Consequently, an increased cultivation of silver fir on suitable sites is recommended.     

Estimating the mass of mango fruit (Mangifera indica, cv. Chok Anan) from its geometric dimensions by optical measurement

Uniform fruit mass and shapes of mangoes for export are required in order to ease handling and transportation, and to help satisfy consumer preferences. However, with its irregular shape, one which cannot be approximated by using standard geometrical figures, the mango is not as easy to model as round or oval-shaped fruits.In this study, more than 1000 mango ‘Chok Anan’ fruit were measured with respect to their length, maximum width and maximum thickness, to within an accuracy of 1.0  mm, and their weight was determined to within an accuracy of 0.1  g. Pictures were taken of 30 fruit, through the use of digital cameras, manually rectified and analyzed by use of “Measure 2.1” a computer code.An equation was proposed in order to calculate the mass of the mangoes based on three geometric dimensions, which then showed a high level of explanatory power of R² = 0.97. The root mean standard error of 12.22 was determined, and the mean relative deviation found was 0.05. Based on measurements taken from the digital photos, the equation was then used to determine the estimated mass based on three dimensions. Estimated and measured values were plotted against each other. A high correlation (R² = 0.96) was found between measured and calculated mass.It was shown that the mass can be estimated based on data obtained from two orthogonally taken photographs.     

Differential responses of germinating Venturia inaequalis conidia to kresoxim-methyl

A new class of agricultural fungicides derived from strobilurins act as respiration inhibitors by binding to mitochondrial cytochrome b. The effects of the strobilurin, kresoxim-methyl, on conidia germination, mycelial growth and the protection of apple leaves from scab development were investigated for two isolates of Venturia inaequalis randomly selected from a culture collection. Inhibition of mycelial growth required relatively high doses of kresoxim-methyl (ED₅₀=1 μg ml^-1) for both isolates. In comparison, germination of conidia was highly sensitive for one of the isolates (ED₅₀=0·005 μg ml^-1), while the level of inhibition achieved for the second isolate was 60-fold less (ED₅₀=0·3 μg ml^-1). As deduced from identical sequences of cytochrome b cDNAs prepared from both isolates, the different responses of germinating conidia to kresoxim-methyl were not caused by differences in the sequence of cytochrome b as the target site for strobilurin action. Strong synergistic effects of salicylhydroxamic acid on kresoxim-methyl inhibitory potency suggested that the reduced kresoxim-methyl sensitivity observed for germinating conidia was caused by interference of the alternative respiratory pathway with inhibitor action. However, this mechanism of target site circumvention in germinating conidia had no adverse effects on the protection of apple leaves from scab infection by kresoxim-methyl. © 1998 Society of Chemical Industry     

Impact of alternative respiration and target-site mutations on responses of germinating conidia of Magnaporthe grisea to Qo-inhibiting fungicides

Qo-inhibiting fungicides act as respiration inhibitors by binding to the Qo center of cytochrome b. Sensitivities of fungi to Qo inhibitors can be influenced by the induction of alternative respiration or by mutational changes of the cytochrome b target site. Previous studies on both mechanisms in Magnaporthe grisea (Hebert) Barr were focused on the mycelial stage of the pathogen. The present study describes the expression and impact of both resistance mechanisms during the stage of conidia germination. In the absence of a host, alternative respiration provided a >500-fold rescue from azoxystrobin during the germination of conidia derived from four wild-type isolates of M. grisea. This rescue potential during conidia gemination was substantially more pronounced than for mycelial growth. However, the pronounced effectiveness of alternative respiration during conidia germination was not apparent when barley leaves were protected with azoxystrobin prior to inoculation with conidia. In a comparison of a wild-type strain and an alternative respiration-deficient mutant, azoxystrobin efficacies in suppressing symptom development differed by a factor of two, with full disease control achieved for both genotypes at 1 microg ml(-1) azoxystrobin. In contrast, conidia derived from two QoI-resistant target site mutants were highly resistant to azoxystrobin and trifloxystrobin and fully capable of infecting leaf surfaces protected with 10 microg ml(-1) of azoxystrobin. Both target-site mutants had emerged spontaneously in the presence of high azoxystrobin doses when residual mycelial growth was supported by alternative respiration. The effective silencing of alternative respiration in protective applications of Qo-inhibiting fungicides might constitute a strategy of slowing the emergence of highly resistant target site mutants.     

Escherichia coli, but not Staphylococcus aureus triggers an early increased expression of factors contributing to the innate immune defense in the udder of the cow

The outcome of an udder infection is influenced by the pathogen species. We established a strictly defined infection model to better analyze the unknown molecular causes for these pathogen-specific effects, using Escherichia coli and Staphylococcus aureus strains previously asseverated from field cases of mastitis. Inoculation of quarters with 500 CFU of E. coli (n = 4) was performed 6 h, 12 h, and 24 h before culling. All animals showed signs of acute clinical mastitis 12 h after challenge: increased somatic cell count (SCC), decreased milk yield, leukopenia, fever, and udder swelling. Animals inoculated with 10 000 CFU of S. aureus for 24 h (n = 4) showed no or only modest clinical signs of mastitis. However, S. aureus caused clinical signs in animals, inoculated for 72 h-84 h. Real-time PCR proved that E. coli inoculation strongly and significantly upregulated the expression of beta-defensins, TLR2 and TLR4 in the pathogen inoculated udder quarters as well as in mammary lymph nodes. TLR3 and TLR6 were not significantly regulated by the infections. Immuno-histochemistry identified mammary epithelial cells as sites for the upregulated TLR2 and beta-defensin expression. S. aureus, in contrast, did not significantly regulate the expression of any of these genes during the first 24 h after pathogen inoculation. Only 84 h after inoculation, the expression of beta-defensins, but not of TLRs was significantly (> 20 fold) upregulated in five out of six pathogen inoculated quarters. Using the established mastitis model, the data clearly demonstrate a pathogen-dependent difference in the time kinetics of induced pathogen receptors and defense molecules.     

Behavior of heavy metals from sewage sludge in a Chernozem of the dry belt in Saxony‐Anhalt/Germany

In a small‐plot trial different doses of sewage sludge (equivalent 82‐330 tons of dry matter per hectare) were incorporated in 0—25 cm depth (1982—1985). The aim of the investigations was to study the fate of the heavy metals Zn, Cd, Cu, Ni, Pb, and Cr, to determine their concentration in different soil fractions using a sequential extraction method and to ascertain their uptake by Zea mays L. plants. Eleven years after the last application the metals supplied with the sludge had moved as far as 50 cm in depth. The concentrations of Zn, Cd, Cu, Ni, and Cr in the saturation extract of the sampled soil layers were closely correlated with the concentrations of dissolved organic carbon (DOC). This result suggests that the heavy metal displacement was partly connected with the DOC movement in the soil. Considerable amounts of Zn and Cd coming from sewage sludge were found in the mobile fractions of the soil. Cu, Ni, and Pb were located especially in organic particles, and Cr was obviously bound by Fe‐oxides. Nine years after the last application the binding species of heavy metals were still different compared with those in the untreated soil. The whole withdrawal of heavy metals by plants yielded <1 % of the applied amounts. In the case of Zn the uptake from the sludge amended soil decreased during the experimental period. No similar tendency was observed for the other elements. In any case their annual variations of uptake exceeded the effect of sludge application.     

Correlates of fruit and vegetable intake among Norwegian schoolchildren: parental and self-reports

OBJECTIVES: To identify correlates of 6th and 7th graders' (age 10-12 years) fruit and vegetable intake, to investigate parent-child correlations of fruit and vegetable intake, and to compare parents' and children's reports of children's accessibility, skills and preferences with respect to fruit and vegetables. DESIGN: The results presented are based on the baseline survey of the 'Fruits and Vegetables Make the Marks Project', where 38 schools participated. SETTING: Fruit and vegetable intake was measured by food frequency questions. Theoretical factors, based on Social Cognitive Theory, potentially correlated to intake were measured, including behavioural skills, accessibility, modelling, intention, preferences, self-efficacy and awareness of 5-a-day recommendations. SUBJECTS: In total, 1950 (participation rate 85%) 6th and 7th graders and 1647 of their parents participated. RESULTS: Overall, 34% of the variance in the pupils' reported fruit and vegetable intake was explained by the measured factors. The strongest correlates to fruit and vegetable intake were preferences and accessibility. The correlation between the children's and their parents' fruit and vegetable intake was 0.23. The parents perceived their children's accessibility to be better than what was reported by the children (P<0.01), while the children reported their skills to be better than what was perceived by their parents (P<0.01). CONCLUSION: The results from this study clearly point to a need for nutrition interventions aimed at parents. An important next step will be to investigate whether the identified correlates predict future fruit and vegetable intake, and whether they mediate any changes in intake in an intervention study.