A simplified procedure for terminal restriction fragment length polymorphism analysis of the soil bacterial community to study the effects of pesticides on the soil microflora using 4,6-dinitroorthocresol as a test case

We have studied the structural effects of application to the soil of a potentially detrimental herbicide, 4,6-dinitroorthocresol (DNOC) by analysing amplified ribosomal DNA restriction analysis (ARDRA) and terminal restriction fragment length polymorphism (T-RFLP) signatures of 16S rDNA fragments of culturable bacterial communities isolated from diluted soil suspensions. This approach has the potential to reveal changes induced by stressing the soil microflora with DNOC. This paper shows that, whereas only few changes of the ARDRA and T-RFLP profiles result from ageing of the soil, treatment of the soil with DNOC induces major modifications of these profiles. Therefore, for the practical purpose of pesticide registration, ARDRA and T-RFLP analysis performed on the dominant culturable fraction of the soil bacteria, implemented using conventional gel electrophoresis, offers the means of a routine, simple and meaningful test for detecting some of the changes affecting the structure of the soil microflora in response to pesticide application.     

A specialist-generalist classification of the arable flora and its response to changes in agricultural practices

Background  

Theory in ecology points out the potential link between the degree of specialisation of organisms and their responses to disturbances and suggests that this could be a key element for understanding the assembly of communities. We evaluated this question for the arable weed flora as this group has scarcely been the focus of ecological studies so far and because weeds are restricted to habitats characterised by very high degrees of disturbance. As such, weeds offer a case study to ask how specialization relates to abundance and distribution of species in relation to the varying disturbance regimes occurring in arable crops.     

GIS Coop: networks of silvicultural trials for supporting forest management under changing environment

Key message

The diversity of forest management systems and the contrasted competition level treatments applied make the experimental networks of the GIS Coop, a nationwide testing program in the field of emerging forestry topics within the framework of the ongoing global changes.

Context

To understand the dynamics of forest management systems and build adapted growth models for new forestry practices, long-term experiment networks remain more crucial than ever.

Aims

Two principles are at the basis of the experimental design of the networks of the Scientific Interest Group Cooperative for data on forest tree and stand growth (GIS Coop): contrasted and extreme silvicultural treatments in diverse pedoclimatic contexts.

Methods

Various forest management systems are under study: regular and even-aged stands of Douglas fir, sessile and pedunculate oaks, Maritime and Laricio pines, mixed stands of sessile oak, European silver fir, and Douglas fir combined with other species. Highly contrasted stand density regimes, from open growth to self-thinning, are formalized quantitatively.

Results

One hundred and eighty-five sites representing a total of 1206 plots have been set up in the last 20 years, where trees are measured regularly (every 3 to 10 years). The major outputs of these networks for research and management are the calibration/validation of growth and yield models and the drawing up of forest management guides.

Conclusion

The GIS Coop adapts its networks so that they can contribute to develop growth models that explicitly integrate pedoclimatic factors and thus also contribute to research on the sustainability of ecosystems under environmental and socio-economic changes.

     

Canine visceral leishmaniasis: comparison of in vitro leishmanicidal activity of marbofloxacin, meglumine antimoniate and sodium stibogluconate

The control of canine leishmaniasis largely depends on the success of treatment. Drugs currently available to treat this disease are toxic and partially effective. The curative effect of marbofloxacin, a third-generation fluoroquinolone developed for veterinarian individual treatment, was evaluated in vitro in the presence of Leishmania infantum promastigotes and dog-monocyte-derived macrophages; meglumine antimoniate and sodium stibogluconate were used as comparative treatments. We observed that the killing of Leishmania promastigotes and intracellular amastigotes by marbofloxacin was dose-dependent. We demonstrated that successful treatment of canine infected macrophages for 48 h was possible with 500 microg/ml of marbofloxacin. Leishmanicidal activity acted through a TNF-alpha and nitric oxide pathway and correlated with the generation of nitric oxide (NO(2)) production by monocytes derived macrophages from infected (23+/-5 microM) or healthy (21+/-6 microM) dogs, in comparison with NO(2) concentration in infected/non-treated macrophages (< 3 microM, P<0.01). This significant induced parasiticidal effect correlated with extensive elimination of amastigotes by macrophages derived from infected (11+/-5) and healthy dogs (6+/-2), when compared to infected/non-treated macrophages (530+/-105 and 472+/-86 amastigotes, respectively, P< 0.01). Marbofloxacin was shown to be non-toxic at 500 microg/ml in vitro and no cell apoptosis was observed. The molecule was able to induce a parasitic process after significant elimination of amastigotes in leishmania-infected dog macrophages. We propose that marbofloxacin, compared to standard chemotherapeutic agents (meglumine antimoniate and sodium stibogluconate), could be an effective and pragmatic oral route alternative to treat canine leishmaniasis.     

Effect of Geogenic Lead on Fungal and Collembolan Communities in Garden Topsoil

Geogenic lead (Pb) is considered to be less bioavailable than anthropogenic Pb and exerts less effect on the soil fauna. However, Pb contamination in vegetables has been reported in the case of geogenic anomalies, even at moderate concentrations (around 170 mg kg^–1). In this study, we investigated collembolan communities using both taxonomic- and trait-based approaches and observed fungal communities to assess the effects of a moderate geogenic Pb anomaly on collembolans and fungi in an urban vegetable garden soil. Results indicated that geogenic Pb indeed modified fungi communities and altered the functional structure of collembolan communities in garden soils. Although geogenic Pb presented low bioavailability, it affected soil fauna and vegetables similar to anthropogenic Pb.     

Climate data challenges in the 21st century

Climate data are dramatically increasing in volume and complexity, just as the users of these data in the scientific community and the public are rapidly increasing in number. A new paradigm of more open, user-friendly data access is needed to ensure that society can reduce vulnerability to climate variability and change, while at the same time exploiting opportunities that will occur.     

Automatisierte Unterscheidung von Unkrautarten nach Formparametern mit Hilfe der quantitativen Bildanalyse

Automatic distinction of weed species using form parameters by means of digital image processing
In former investigations the possibility to distinguish weed from soil background by computerized quantitative image analysis has been reported. In the present studies image analysis technique are applied to identify different weed species. From six typical winter weed species including various growth stages, 300 color slides were taken at the research station in Bonn on the 4th of February, 1988. From each slide a number of form parameters were measured automatically out of the digitized shape of each weed plant. Form parameters including area, perimeter, minimum and maximum diameter of the plants are taken from the digitized shape of each weed as well as figures representing the relative proportions of these parameters. For automatic discrimination up to 9 of the parameters were selected to compute five so-called canonical variables. The values of these variables are typical for each weed species and can be interpreted as their "finger prints". The average rate of correct identification was 82.3 %. The scientific and practical implications of the results are discussed.     

Biological responses to porcine respiratory and reproductive syndrome virus in pigs of two genetic populations

One hundred pigs from the NE Index Line (NEI) and 100 Hampshire-Duroc cross pigs (HD) were inoculated intranasally with porcine respiratory and reproductive syndrome virus (PRRSV 97-7895 strain) at 26 d of age to determine whether genetic variation in response to PRRSV exists. An uninfected littermate to each infected pig served as a control. Pigs were from 163 dams and 83 sires. Body weight and rectal temperature were recorded, and blood samples were drawn from each pig on d 0 before inoculation and on d 4, 7, and 14 after inoculation. Pigs were sacrificed on d 14. Lung and bronchial lymph nodes were collected, placed in optimal cutting temperature compound, and frozen at -80 degrees C. The presence of PRRSV in serum and in lung tissue and bronchial lymph nodes was determined by isolation in cell culture. The presence of antibodies in serum collected on d 14 was determined by a commercial ELISA test. Lung tissue was examined microscopically and scored for incidence and severity of lesions (score of 1 to 3; 1 = no or few lesions, and 3 = severe interstitial pneumonia). Data were analyzed with a mixed model that included random sire and dam effects. The interaction of line x treatment was significant (P < 0.001) for weight change and rectal temperature. Un-infected HD pigs gained 0.67 kg more from d 0 to 14 and averaged 0.32 degrees C higher rectal temperature than uninfected NEI pigs (P < 0.001), whereas infected NEI pigs gained 0.34 kg more and had -0.54 degrees C lower temperature than infected HD pigs (P < 0.001). Viremic titer (cell culture infectious dose 50%/mL) was greater (P < 0.05) in HD than NEI at d 4 (10(4.52) vs. 10(4.22)), 7 (10(4.47) vs. 10(3.99)), and 14 (10(3.49) vs. 10(3.23)). Viral titer loads in lung (P = 0.11) and bronchial lymph nodes tended (P = 0.07) to be greater in HD than NEI pigs. Antibody signal-to-positive (S/P) ELISA ratios in infected pigs ranged from 0.18 to 3.38, and 88% had levels > or = 0.40, which is the positive threshold for this ELISA. The S/P range in uninfected pigs was 0 to 1.11, and 99% had levels < or = 0.40. Mean S/P ratio for infected pigs was 0.23 units higher in HD than in NEI (P < 0.001). The HD pigs had a greater incidence of interstitial pneumonia and 0.65 higher mean lesion scores than NEI pigs (P < 0.001). In summary, responses of pigs of the two lines to infection with PRRSV differed, indicating that underlying genetic variation existed.