Estimation of reproducibility and repeatability in microbiological ring trials--robust versus conservative methods

During the last years there was a lot of progress to be seen in the development of standardized methods for microbiological ring trials. The statistical analyzing strategies, in particular the calculation of estimations for the parameters repeatability and reproducibility, will be considered in this paper. Apart from the conservative method of the variance analysis robust methods are increasingly discussed. We will compare and discuss these methods using data of recently realized ring trials. If we can assume a normal distribution of our data, then all estimations are theoretically precise and efficient. But up to now, we know very little about the character of the robust estimations, if the normal distribution cannot be assumed. In addition to this, we have to mention once more, that the use of robust estimators is unreasonable without taking a critical look on the data themselves. Thus, we will show the possibilities of graphical presentation of all data to identify laboratories with critical results.     

Functional risk assessment of handling of sheep housing and pasture areas at a scrapie-infested farm in Mecklenburg-West Pomerania

In the wake of a scrapie monitoring programme 2002 in Mecklenburg-Westpomerania a total of 10 positive cases were identified after culling at a suffolk-farm with about 1000 animals. Scientific based recommendations are difficult to judge as longitudinal studies and subsequent quantitative risk factors regarding the epidemiology of scrapie are either not available or conflicting. Given this background a risk assessment was developed that is based on the probability of validated risk factors considering special husbandry and circumstances at the very farm.     

Molecular markers for detection of pathogenic Escherichia coli strains belonging to serogroups O 138 and O 139

Escherichia coli strains belonging to O-serogroup 138 and 139 are important as disease agents in pigs causing post-weaning diarrhea and edema disease. Several types of shiga toxin-producing O 138 and O 139 strains were isolated from diarrheic humans and from cattle and food of bovine origin. Serotyping is the current method for detection of O 138 and O 139 strains but its applicability can be limited due to the presence of capsules and capsular-like bacterial surface antigens and in the case of rough LPS. To overcome these difficulties for diagnosis, we have developed a specific PCR method suitable for detection of different types of O 138 and O 139 strains. The O-antigen gene clusters of E. coli O 138 and O 139 type strains were sequenced, and the genes were identified on the basis of homology. By screening against 186 E. coli and Shigella type strains, two genes specific to each of E. coli O 138 and O 139 were identified, respectively, and were tested on 15 clinical and environmental isolates of those two serogroups in a double-blind test. The sensitivity of the PCR assays was determined, and the detection limits were 2 pg per mul of chromosomal DNA and 2 CFU per 10 g of water or pork samples. PCR-based detection of O-antigen specific genes of E. coli O 138 and O 139 was shown to be accurate, highly sensitive and rapid, and is suggested as a new diagnostic tool for investigations of infections and outbreaks with these strains in animals and humans and for control of food.     

Monitoring megabenthic communities in abyssal manganese nodule sites of the East Pacific Ocean in association with commercial deep-sea mining

Marine mining of manganese nodules will affect the deep sea benthos in a harmful and longterm way. The near complete removal of manganese nodules will result in a change of the megabenthic community from one that is rich in both diversity and density to a soft bottom community with greatly reduced diversity. Pre-pilot mining tests and pilot mining operations, which have to demonstrate the feasibility of deep-sea mining, should be accompanied by environmental impact studies before commercial mining begins. The photographic assessment of megabenthic communities represents one cost-effective possibility of monitoring large-scale impacts on the deep sea floor. A method based on the analysis of video and photomaterial was used to obtain basic data on community structure and density of the abyssal megabenthos of undisturbed manganese nodule sites in the North (Clarion-Clipperton Fracture Zone) and South (Peru Basin) Pacific Ocean. A comparison of the results with data from the literature demonstrates that the different methods used result in different density values. Total density varied between less than 300 to more than 1600 individuals/10000 m². There is a clear need for standardization of methods. A system which combines video and photographic capabilities is useful. It will give the best results when the distance of the camera to the sea floor is between 2 and 3 m, and the area covered by one photograph is within 3 to 5 m².     

Multi-site validation of a soil organic matter model for arable fields based on generally available input data

The paper describes a simplified carbon balance model, derived from the CANDY model, which works in annual time steps requiring only clay content, soil type of the German classification system “Reichsbodenschätzung”, average air temperature and rainfall as site characteristics, a value for organic carbon content as the initial value as well as crop yield and organic matter amendments as management data.The Candy Carbon Balance (CCB) model has been validated using a dataset from 40 long-term experiments situated in Central Europe including 391 treatments with a total number of 4794 C_org observations. Statistical measures to prove model validity were mean error (ME = − 0.001) and root mean square error (RMSE = 0.119). In addition a number of tests were performed to make sure that the model has no systematic error for different types of site conditions and management activities. After this successful validation the CCB model is considered applicable for advisory service for arable fields on a wide range of site conditions. Due to the poor representation of clay soils in this study some more model tests on these soils would be recommendable.     

Metabolism of amitrole in apple: III. Model systems

Excised shoots from apple trees and cell suspension cultures were used as model systems to study the metabolism of [3,5-¹⁴C]amitrole in Malus domestica Borkh. Significant differences in the metabolism of the compound applied were observed with excised shoots, cultured cells and whole apple trees. The major metabolite in excised shoots was aminotriazolylalanine which occurred both in the free form and as conjugates. The major metabolite from whole plants. triazolylalanine, was detected in shoots in minor amounts only. In cell suspension cultures, the type of metabolism strongly depended on the concentration of amitrole when initially applied. At 10 ^?3 m or lower, mainly aminotriazolylalanine was formed. Depending on the concentration of the active ingredient, this metabolite predominantly occurred in free form or as glycosides. At concentrations above 5 × 10^?4 M a new metabolite, 3,5-dihydroxytriazole, was detected which was the only metabolite found at 5 × 10^?3M. Significant amounts of nonmetabolized amitrole remained in the medium.     

Studies on the effect of different cyclohexane-1,3-diones on de-novo fatty acid biosynthesis in poaceae

Cyclohexane-1,3-diones such as the herbicides cycloxydim, sethoxydim, alloxydim and clethodim are known to be specific inhibitors of the plastid-located acetyl-CoA carboxylase (ACCase) in Poaceae, a key enzyme of de-novo fatty acid biosynthesis in higher plants. Using several new cyclohexane-1,3-dione derivatives and known herbicides, the relationships between chemical structure and enzyme inhibition have been studied. The basic cyclohexane-1,3-dione structure was modified at three different positions. These compounds were tested for inhibition of the de novo fatty-acid biosynthesis in test systems of etioplasts isolated from Avena sativa L. and Hordeum vulgare L. seedlings and also for inhibition of the isolated barley ACCase. The I₅₀ values of these cyclohexane-1,3-diones were determined. The influence of the modification of alkyl chains (length and type of substituent) on the degree of ACCase-inhibition is discussed. Several new compounds were found that were about two orders more active than the known herbicides cycloxydim or sethoxydim in the etioplast and ACCase test systems but not necessarily on the level of whole plants.     

Pymetrozine,a Fast-Acting and Selective Inhibitor of Aphid Feeding. In-situ Studies with Electronic Monitoring of Feeding Behaviour

Pymetrozine, a pyridine azomethine compound, represents a novel insecticide with a selective activity against homopteran insects. It acts in a unique way: aphids are not knocked down on contact but seem to die of starvation. This implies an effect of pymetrozine on feeding behaviour. The aim of the present work was to elucidate how and at which step pymetrozine interferes with the complex mechanisms underlying phloem feeding. The effect of pymetrozine, applied in various ways, on different phases of stylet penetration and feeding activity of individual aphids was studied using the Electrical Penetration Graph technique (EPG). Initial choice experiments indicated that pymetrozine does not have a deterrent or antifeedant action. Topical application (150 ng pymetrozine mg^-1 fresh weight) inhibited stylet insertion into the plant. When injected, less than 30 ng mg^-1 was sufficient to produce the same effect. When pymetrozine was systemically applied via plant spraying or root uptake, aphids started feeding normally. After some time, however, they withdrew their stylets from the phloem and walked around with unaffected locomotion. At low doses aphids eventually recovered and resumed feeding. High doses, however, irreversibly disrupted feeding and prevented stylet reinsertion. Aphid motility was not affected up to an estimated haemolymph concentration of 1 mM pymetrozine. Aphids which eventually stopped feeding on pymetrozine-treated plants showed EPGs with distorted salivation/ingestion patterns. It is concluded that pymetrozine does not have a general toxic effect on aphids but selectively interferes with the nervous regulation of feeding behaviour which consequently results in death due to starvation after a few days. © 1997 SCI.     

Effect of grapefruit juice, naringin, naringenin, and bergamottin on the intestinal carrier-mediated transport of talinolol in rats

The effect of two varieties of grapefruit juice (white and ruby red) and its selected components (naringin, naringenin, and bergamottin) was investigated on the activity of the P-glycoprotein (P-gp) in male Sprague-Dawley rats. Talinolol, a nonmetabolized P-gp substrate, was used as a marker compound. The white grapefruit juice (GFJ) had a minor effect on talinolol pharmacokinetics, but the ruby red GFJ reduced the C max and the AUC (0-infinity) by 60% and 50% of the control, respectively. However, among the GFJ constituents tested, bergamottin (0.22 mg/kg) was the most potent component augmenting the C max and the AUC (0-infinity) of talinolol by 2.4- and 1.8-fold, respectively, if compared to the control group. The flavonoids naringenin (0.7 mg/kg) and naringin (2.4 and 9.4 mg/kg) had a similar effect increasing the talinolol C max and AUC (0-infinity) by 1.5- to 1.8-fold, respectively. In conclusion, the effect of GFJ on P-gp activity seems to depend on the variety, the concentration of compounds in the juice, and the composition of different ingredients.