Fatty acid pattern, oxidation product development, and antioxidant loss in muscle tissue of rainbow trout and Dicentrarchus labrax during growth

The levels of hydrophilic, lipophilic, and enzymatic antioxidants, the oxidative damage to lipids and proteins, and the fatty acid patterns of triglyceride and phospholipid fractions were assayed in fresh muscle tissue of rainbow trouts (Oncorhynchus mykiss) and sea basses (Dicentrarchus labrax) during aging, to investigate the correlation between oxidative stress and aging processes in fish. The present studies suggests that lipid peroxidation and accumulation of oxidized proteins during in vivo aging are most likely to be linked with an age-dependent decline of lipophilic antioxidants (CoQH(2), CoQ, and vitamin E) and vitamin C contents in muscle tissue, whereas fish aging is not linked to a decline in antioxidant enzymes and reduced glutathione levels. Lipophilic antioxidant and vitamin C levels represent a reliable marker of oxidative stress during aging, and their determination might be useful for the assessment of fish age.     

Mortality,progeny production and preference of Sitophilus zeamais adults to wheat from integrated and alternative production systems

The objectives of this study were to assess the effects of the production systems of wheat from different production systems on the mortality, progeny production and preference of Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae). The factors tested were production system (integrated [INT], organic [ORG], biodynamic [BD] and control), which differed in plant protection and fertiliser procedures during plant growth and development; exposure interval (7, 14 and 21 d); relative humidity (r.h.) (55% and 75%) and temperature (20°C, 25°C and 30°C). Mortality after 7 d increased with the temperature increase and decreased with the increase in r.h. in most of the tested combinations. The mortality of weevils was higher in ORG compared to INT-produced wheat after 7 d. Progeny production was recorded 56 d after removal of parental adults and was higher at 75% r.h. in comparison to 55% r.h. At 55% r.h. and 20°C, progeny was 60.8% higher when S. zeamais were exposed to ORG in comparison to INT-produced wheat. Wheat from different production systems influenced mortality rates which were higher in alternative compared to INT production systems under optimal conditions for wheat storage (low temperature and r.h.). The reverse was recorded for temperature and r.h. increase. Progeny was not affected by wheat from various production systems. Significantly more S. zeamais adults were found in traps containing wheat from BD and control in comparison to INT. An understanding of the agricultural processes, biotic and abiotic factors which alter the post-harvest response of storage pests could be useful for the development of efficient post-harvest strategies for ORG and BD farms and the processing industry.     

Decrease of heat shock protein levels and cell populations by wine phenolic extracts

The effect of red and white wine total extracts and phenolic fractions on heat shock protein (Hsp) levels in tumor cells and on tumor and endothelial cell populations in vitro has been investigated. Total extracts of red wines decreased Hsp70 and Hsp27 levels and the numbers of tumor and endothelial cells. Several red and white wine fractions significantly decreased Hsp27 levels, and some of them had also an effect on Hsp70 levels. A red wine fraction rich in polymeric flavanols and a white wine one rich in phenolic acids, flavonols, and tyrosol strongly lowered Hsp27 levels. Some red and white wine fractions strongly reduced tumor cell numbers, whereas most of them decreased endothelial cell numbers to variable extents. The present results indicate that wine phenolics decrease Hsp levels in tumor cells and tumor and endothelial cell populations. These properties may be important in the potent anticarcinogenic action of wine phenolics.     

Amino acid variation in the 10 kDa Oryza prolamin seed storage protein

The deduced amino acid variability for the 10 kDa prolamin was determined for 16 Oryza species, both cultivated (rice) and wild. Prolamin, a seed storage protein and site of nitrogen and sulfur accumulation, is sequestered in the subaleurone layer of the starchy endosperm for use during seedling germination. The 10 kDa prolamin amino acid distribution for the cultivated species (O. sativa and O. glaberrima) was determined and compared to those of wild and, hitherto unknown, noncultivated Oryza species. Four wild species (O. granulata, O. australiensis, O. brachyantha, and O. meyeriana) exhibited the greatest residue heterogeneity in both the signal and mature peptide regions. A breakdown of the essential amino acid variance among three Central/South American and one African endemic wild species is also presented and compared with those of rice.     

Persistence of benomyl and thiophanate compounds in soil and various plants following soil application

Control ofSclerotinia sclerotiorum in muskmelons was obtained by a single soil drench with benomyl. The material was found in the apical parts of the plant and in the soil until the end of the growing season. Benomyl [methyl l-(butylcarbamoyl)-2-benzi-midazolecarbamate] and thiophanate methyl-NF 44 [1.2- bis (3 methoxycarbonyl-2-thioureido) benzene], applied to seedbeds, were taken up by and then persisted in tomato, pepper and eggplant for approximately 8 weeks after application. Thiophanate-NF 35 [1.2-bis (ethoxycarbonyl-2-thioureido) benzene] could be detected only in the bottom leaves of pepper and eggplant, for up to 4 weeks after application.     

Development of a Multiplex Polymerase Chain Reaction for the Differentiation of Bovine Herpesvirus‐1 and ‐5

Bovine herpesvirus‐1 (BHV‐1) and bovine herpesvirus‐5 (BHV‐5) are closely related viruses which exhibit some important differences at the genetic and immunogenic levels which may explain the differences in their pathogenicity and epidemiological characteristics. A multiplex polymerase chain reaction (M‐PCR) was developed to detect and differentiate between BHV‐1 and BHV‐5. In this M‐PCR two pairs of primers (TK1, TK2 and GD1, GD2) were used in the same reaction mix to amplify a thymidine kinase genomic region (183 bp) of BHV‐1 and one genomic region of the glycoprotein D (564 bp) of BHV‐5. The specificity of the M‐PCR was demonstrated when using both primers pairs simultaneously with BHV‐1 and BHV‐5 templates. The two expected bands were amplified without the apparition of non‐specific products. However, when other herpesvirus strains were used, there was no amplification. To evaluate the sensitivity of the assay, dilutions of purified viral DNA were made for M‐PCR amplification. The detection limit was 7 pg for BHV‐1 and 22 pg for BHV‐5. It was also determined by comparing the M‐PCR with viral isolation. M‐PCR was able to detect one log₁₀ more than viral isolation for BHV‐1 and for BHV‐5 was two logarithms lower. The applicability of M‐PCR was demonstrated on different specimens. Twenty isolates from field samples (11 BHV‐1 and nine BHV‐5) were positive by M‐PCR, and the results were completely coincident with previous characterization using the immunoperoxidase assay. M‐PCR could detect viral DNA in organ samples from natural infections, such as semen and brain. In addition, M‐PCR detected more positive samples than observation of the citophatic effect in cell culture of nasal swabs from experimentally infected animals in two different assays. Owing to the difference in size of the M‐PCR products which allows easy identification in an electrophoretic run, it is not necessary to use extra blotting and hybridization steps or a second round of amplification to differentiate clearly between BHV‐1 and BHV‐5.     

Effect of mixed inoculations with Fusarium oxysporum f. sp. lycopersici and F. oxysporum f. sp. dianthi on the phenols content of tomato plants

Forms ofFusarium oxysporum specific on hosts other than tomato induce in this plant greater initial increases of the phenols content than the pathogenic f. sp.lycopersici. Mixed inoculations of f. sp.lycopersici and f. sp.dianthi are on the contrary no more effective in inducing the phenol accumulation 24 h after the infection than f. sp.lycopersici alone. This observation suggests that the pathogen can suppress the phenolic response that is typical of the incompatible combinations.Samenvatting Vormen vanFusarium oxysporum welke pathogeen zijn voor andere planten dan de tomaat induceren in deze plant aanvankelijk een grotere toename van het fenolgehalte dan de pathogene f. sp.lycopersici. Inoculaties met een gemengd inoculum van de f. sp.lycopersici en f. sp.dianthi hebben daarentegen geen groter effect op de toename van het fenolgehalte 24 uur na infectie dan de inoculaties met f. sp.lycopersici alleen. Verondersteld wordt dat het pathogeen de toename van het fenolgehalte, dat typerend is voor de incompatibele combinatie, kan onderdrukken.     

Prevalence of Campylobacter spp. in poultry meat and raw milk using PCR, conventional cultural methods and Fourier transform infrared spectroscopy

The identification of thermotolerant campylobacters in official food control in the state of Baden-Wuerttemberg has been traditionally performed using the cultural procedure as described in the ISO-Norm 10272:1995. Analysis thus took 5-6 days to complete. Additionally diagnostic problems caused by the accompanying flora as well as the resistance to nalidixic acid occured. Within the scope of this study these problems could be solved by introducing a filtration step for the reduction of the accompanying flora and by performing the indoxyl acetate-hydrolysis-test in addition to the antibiotic-resistance-test. Besides various PCR protocols for the identification of thermotolerant campylobacters from food were established as an alternative to the cultural procedure, providing reliable results within two days. Furthermore, infrared spectroscopy was tested for the identification of Campylobacter isolates. Using this technique and with the help of a suitable data base, bacterial pure cultures can be differentiated within 2 hours. Among others 356 samples of raw poultry meat were tested with the newly established procedures as well as with the classical cultural method, showing that 32% of the samples were Campylobacter spp. positive. 37% of these isolates were resistant against nalidixic acid. This indicates that the development of resistances in Campylobacter spp. in Germany follows the same trend described for other European countries.     

New polyphenol derivative in Ipomoea batatas tubers and its antioxidant activity

Four different polyphenolic compounds were isolated by chromatographic methods from methanolic and hydromethanolic extracts of Ipomoea batatas tuber flour. On the basis of UV, mass, and NMR analysis procedures, the structure of the isolated compounds were determined as 4,5-di-O-caffeoyldaucic acid (1), 4-O-caffeoylquinic acid (2), 3,5-di-O-caffeoylquinic acid (3), and 1,3-di-O-caffeoylquinic acid (4). To the best of our knowledge, this is the first report of isolation and characterization of compound 1. Then, we evaluated the antioxidant activity of daucic acid derivative by using DPPH and FRAP methods together with authentic antioxidant standards, l-ascorbic acid, tert-butyl-4-hydroxy toluene (BHT), and gallic acid. The activity of compound 1 in both methods was higher than that of all standards used at the same molar concentration.