Antifungal effects of iron sulfate on grapevine fungal pathogens

The present study aimed to determine the most efficient experimental conditions of iron sulfate use leading to optimal inhibition in the development of fungal pathogens. Assays have been focused on fungal species inducing severe grapevine diseases. FeSO₄ directly inhibited the in vitro mycelial growth of Botrytis cinerea, Eutypa lata, Phaeomoniella chlamydospora, Phaeoacremonium aleophilum, Diplodia seriata, and Neofusicoccum parvum with variable efficiency in the range of 0.5–10 mM. The development was always completely inhibited at 20 mM. This inhibitory effect was greatly increased at acidic pH values. The anionic moiety of the molecule was of importance since bromide, chloride and sulfate were highly active, whereas acetate and oxalate showed a small effect. Electron microscope observations on E. lata and B. cinerea showed that a treatment with FeSO₄ induced dramatic changes in the hyphal organization leading to cell death. No toxicity was observed on grapevine leaves following repeated FeSO₄ sprays in the antifungal concentration range. Therefore, FeSO₄ may be proposed to effectively replace the long-term pollutant use of CuSO₄ as an antifungal agent, with the additional advantage of iron being an important plant micronutrient.     

Sensitivity in the estimation of parameters fitted by simple linear regression models in the ratio of blueberry buds to fruits in Chile using percentage counting

Blueberry exporting is an important activity in Chile, with fresh blueberries commanding the highest prices and being among the most exported products to the European and North American markets. To maintain quality in the centres of consumption, farmers must continuously improve the logistics of harvesting and shipping the blueberries. Thus every year they must calculate the production of the orchard well in advance in order to hire staff and ensure the logistic cold chain. For this calculation they use a count of flower buds and a simple linear model of which the slope parameter represents the number of fruits per bud. However, due to the cost of the counting procedure, some producers count only a fraction of each plant (25%, 50% or 100%), and furthermore they do not know what effect the variety and productive age of the plants may have on the estimation. The objective of this work is to measure the impact of the cultivated variety, the age of the plant in productive years, and the percentage of fruits counted in estimating the parameter fruits per bud. The study involved monitoring 310 plants of different varieties and ages distributed in northern, central and southern Chile (over an area of approximately 700 km × 200 km). The parameter was estimated by fitting simple linear regression models (SLRM) as a function of the number of fruits and flower buds. To evaluate the impact on the parameter, the SLRM was fitted considering the variables observed in all the plants, by percentage counted, by variety and by variety-age of the plant. The major findings indicate significant differences in the estimation of the parameter, suggesting that in order to estimate fruits per bud the whole plant must be counted and its age and variety taken into account.     

<Emphasis Type="Italic">In silico</Emphasis> genomic subtraction guides development of highly accurate,DNA-based diagnostics for <Emphasis Type="Italic">Ralstonia solanacearum</Emphasis> race 3 biovar 2 and blood disease bacterium

New rapid diagnostic methods are urgently needed to discriminate the quarantine pathogen Ralstonia solanacearum (Rs) race 3 biovar 2 (R3B2) from other populations of Rs that lack the adaptation to cause bacterial wilt disease in temperate regions. We used an in silico bioinformatic approach to identify several genome sequences potentially specific to R3B2 strains. Primer sets were designed to PCR-amplify sequences in these regions, and four sets were ultimately shown to be >99% accurate for detection of R3B2 strains. On the basis of these results, several primers were designed to enable development of a loop-mediated isothermal amplification assay that was rapid, technologically simple, and essentially 100% accurate for identification of R3B2 when applied to a comprehensive collection of geographically diverse Rs strains. We fortuitously found that a sequence in one of the “R3B2-specific” regions has ~90% identity to a sequence present in strains of the blood disease bacterium (BDB), a member of the Rs species complex that infects banana. Alignments of these sequences allowed design of a second PCR primer set that proved 100% accurate for identification of BDB strains when tested on the 22 BDB strains available to us. These results demonstrate the power of in silico genomic subtraction for rapid identification of population-specific DNA sequences and for the development of simple, reliable detection methods for Rs subpopulations.     

华北农业高产粮区地下水硝酸盐污染环境价值损失评估技术研究——以山东省桓台县为例

应用人力资本法、防护支出法和支付意愿调查法评估了华北高产粮区桓台县地下水农业面源硝酸盐污染的环境经济损失,并分析了不同评估技术对农业面源硝酸盐污染地下水环境价值损失评估的适用性,其适用性优先序为防护支出法的深井工程法>脱硝工程法≥人力资本法>支付意愿调查法。2 0 0 2年该县由农业地下水硝酸盐污染造成的年环境价值损失约为880万元(86 0 .8～1170 .1万元/浕) ,占当年农业总产值的1.1%～1.5 %。     

Modified method for the analysis of free fatty acids in fish

The objective of this study was the development of a method for the quantification of free fatty acids (FFA) using less aggressive reactants against the handler and the environment than those used in the classic method of Lowry and Tinsley. The modified procedure is a variation of the Lowry and Tinsley method employing cyclohexane in place of benzene. The use of benzene is prohibited in certain work processes and laboratories, and the competent authority in each country is actively promoting research into harmless or less harmful products that could replace benzene. A comparison with the traditional AOCS titration method for oil analysis was performed. FFA content in mackerel frozen at -10 degrees C was measured according to the three methods over a 12 month period. The results showed similar values, and good correlations were obtained.     

Abundant PrP(CWD) in tonsil from mule deer with preclinical chronic wasting disease.

A monoclonal antibody dot-blot assay was used to evaluate detergent lysates of tonsil tissue from mule deer to detect PrP(CWD), the marker for the cervid transmissible spongiform encephalopathy chronic wasting disease (CWD). Samples of formalin-fixed brain and tonsil tissues from mule deer were examined for PrP(CWD) using immunohistochemistry (IHC) with Mab F99/97.6.1, the gold standard for diagnosis of preclinical CWD. The contralateral tonsil from each of the 143 deer was prepared for confirmatory IHC and as a 10% (wt/vol) detergent lysate without purification or enrichment steps for monoclonal antibody dot-blot assay. PrP(CWD) was detected by dot-blot assay in 49 of 50 samples considered positive by IHC. Forty-eight of the positive samples were evaluated with a quantitative dot-blot assay calibrated with recombinant PrP. Tonsillar PrP(CWD) concentrations ranged from 34 to 1,188 ng per 0.5 mg starting wet weight of tissue. The abundant PrP(CWD) in mule deer tonsil will facilitate development and validation of high-throughput screening tests for CWD in large populations of free-ranging deer.     

The relationship between carcass characteristics,plasma hormones and metabolites in young fattening bulls

Six Belgian Blue bulls (double-muscled type) and six Friesian bulls were offered a fattening diet for 34 weeks. Plasma samples were obtained once a week and also every 20 min over a 24 h period, 7 weeks before slaughter.No differences were observed between the breeds in plasma glucose, urea and free amino nitrogen concentrations, while creatinine was significantly higher in the Belgian Blue bulls. Tri-iodothyronin, tetra-iodothyronin, insulin-like growth factor 1, insulin and testosterone concentrations were higher in the Holstein group. In contrast, the Belgian Blue bulls appeared to produce more growth hormone. The slaughter weight, carcass weight, dressing percentage and proportion of lean meat were significantly higher in the Belgian Blue group. The characteristics of muscle mass (carcass weight, dressing percentage and proportion of lean meat) were positively correlated with creatinine and with the total peak area or peak amplitude of growth hormone. The insulin concentration was positively correlated with the proportion of adipose tissue in the carcass and negatively correlated with the proportion of muscle. There were no correlations between the carcass characteristics and insulin-like growth factor 1 or testosterone. No further information was provided when the ratios of the hormones were correlated with carcass characteristics.     

Relation of lactose levels in milk and indicators of mammary gland health in the first third of lactation]

A total of 662 bucket milk samples from cows of two breed groups were examined: red and white breed = Bohemian Pied cattle with different genetic proportions of Ayrshire and Red Holstein improvement breeds; black and white breed = Black and White Lowland breed and different degree of absorptive crossing with a genetic proportion of the Holstein breed. Samples of daily milk yields were taken in the first three months of lactation once a month within a year. A possibility of using lactose content as an auxiliary indicator for detection of the mammary gland secretion disorders in the initial lactation stage was evaluated. The average values of the different indicators and their variability are summarized in Tab. I showing also the significance in a statistical model of included effects. Lactose content (L) was 4.88 +/- 0.20%, chloride content (Cl-) 113.7 +/- 22.4 mg/100 ml, somatic cell (SC) count 474 +/- 805 thousand/ml, SC count log corresponds to the geometrical mean of 234 thousand/ml, titratable acidity (SH) 7.34 +/- 0.83 x 2.5 mmol/l, chloride-lactose ratio (ClL) 2.27 +/- 0.51, conductivity (gamma) 442.4 +/- 34.5 mS/m and mastitis test (MT-NK) 0.72 +/- 1.18. The efficiency of the used statistical model was highest for Cl- content (Tab. I, R2 = 0.41), and it was lowest for SC counts (R2 = 0.07), while it increased to the twofold value (R2 = 0.15) after logarithmic transformation of SC counts. The breed group exerted a significant effect on Cl-, SC, log SC, SH, ClL, gamma and MT-NK (Tab. I). The breed group of red and white cows (Tab. II) had higher component contents and better indicators of the udder health state (Cl-, SC, log SC, ClL, gamma and MT-NK). The month of lactation influenced significantly SC, log SC, SH and gamma (Tab. I). A decrease in SC counts with the accruing month of lactation was observed (Tab. II), the trend of gamma and SH was opposite. The effect of lactation number was found to be significant for L, SC, log SC, SH, ClL, gamma and MT-NK (Tab. I). A tendency of a gradual decrease with the lactation number was observed in these indicators: L, SH, proteins and solids-non-fat (Fig. 1), while Cl-, gamma and ClL showed an opposite tendency. The year season influenced significantly L, log SC, SH, ClL, gamma and MT-NK (Tab. I, Fig. 2).(ABSTRACT TRUNCATED AT 400 WORDS)     

Porcine epidemic abortion and respiratory syndrome (mystery swine disease). Isolation in Spain of the causative agent and experimental reproduction of the disease.

In March of 1991, a disease that affected pregnant sows and caused a high mortality in unweaned piglets was detected in Spain. Based on the clinical signs observed, mystery swine disease, which had been described recently in Germany, Holland and Belgium, was suspected. From the samples obtained from the affected farm, a filtrable agent (0.22 micron) was isolated on cell culture. It produced cytopathic effects, its replication was intracytoplasmic, it was sensitive to chloroform, and cross-reacted with a Lelystad reference serum. When inoculated into pregnant sows, the agent produced inappetence for 2-4 days, without hyperthermia. One of the sows aborted at 100 days of gestation; the two others had delayed parturitions (days 115 and 116). There was a mixture of healthy piglets, mummified fetuses, stillbirths and weak piglets. Microscopic examination of the lungs of healthy piglets killed at 8 and 12 days of life revealed the presence of interstitial pneumonia. The sera from the three sows at 39 days after infection cross-reacted with the Lelystad virus (titres > or = 1/640), whereas pre-inoculation sera did not recognize it (titres < or = 1/10). This is the first report from Spain of the isolation of an agent (antigenically related to the Lelystad virus), capable of reproducing the disease previously designated as mystery swine disease.     

Proteome analysis of the sarcoplasmic fraction of pig semimembranosus muscle: implications on meat color development

Two-dimensional electrophoresis was used to investigate sarcoplasmic protein expression in pig Semimembranosus muscles sampled 20 min after slaughter. Two groups (light and dark) of 12 animals were selected from 1000 pigs, based on meat L values measured 36 h postmortem. Twenty-two proteins or fragments (p < 0.05) were differentially expressed. Muscles leading to darker meat had a more oxidative metabolism, indicated by more abundant mitochondrial enzymes of the respiratory chain, hemoglobin, and chaperone or regulator proteins (HSP27, alphaB-crystallin, and glucose-regulated protein 58 kDa). Conversely, enzymes of glycolysis were overexpressed in the lighter group. Such samples were also characterized by higher levels of glutathione S-transferase omega, which can activate the RyR calcium channels, and higher levels of cyclophilin D. This protein pattern is likely to have severe implications on postmortem metabolism, namely, acceleration of ATP depletion and pH fall and subsequent enhanced protein denaturation, well-known to induce discoloration.