Analysis of three-dimensional biofilms on different material surfaces.

Biofilms cause complications and high costs in both industry and medicine. Of particular interest are bacterial infections of prosthetic materials, which usually cannot be eliminated due to the high antibiotic resistance known for bacteria forming biofilms. The search for new materials and coatings with lower colonization potential and antibacterial activity is of great importance to reduce biofilm formation. However, there is no standardized procedure to examine the colonization characteristics of bacteria in the biofilm state in situ. Here, we describe an automated epifluorescence microscopy system for the semi-quantitative analysis of three-dimensional (3D) biofilms on various surfaces. To analyze adherent bacteria, three materials (glass, steel and titanium) were incubated with bacteria in a flow chamber system. After fluorescence staining of the bacteria, automated image capturing, quantification of the bacteria, measurement of the colonized area and determination of the 3D biofilm height were carried out by using novel software. Furthermore, the materials were examined for their surface topography using white light scanning interferometry. Titanium compared to glass showed a significantly higher number of adherent bacteria. We argue that this was due to the higher microroughness of titanium. The colonized area was in accordance with the number of adherent bacteria and was also significantly larger on titanium coupons compared to glass. Maximum 3D biofilm height on glass coupons was significantly lower compared to the ones on steel and titanium. This novel method enables the standardized, automated investigation of the colonization with bacteria on different materials. This approach can considerably support the characterization of new material surfaces and their innovative coatings by analyzing the amount of attached bacteria and thickness of biofilms in situ and eliminates the need of conventional cultivation.

Bacterial Adhesion on Femtosecond Laser-Modified Polyethylene.

In this study, femtosecond laser-induced sub-micrometer structures are generated to modify polyethylene (PE) surface topographies. These surfaces were subjected to bacterial colonization studies with Escherichia coli and Staphylococcus aureus as test strains. The results reveal that the nanostructures do not influence S. aureus coverage, while the adhesion of E. coli is reduced.

Flow Chamber System for the Statistical Evaluation of Bacterial Colonization on Materials.

Biofilm formation on materials leads to high costs in industrial processes, as well as in medical applications. This fact has stimulated interest in the development of new materials with improved surfaces to reduce bacterial colonization. Standardized tests relying on statistical evidence are indispensable to evaluate the quality and safety of these new materials. We describe here a flow chamber system for biofilm cultivation under controlled conditions with a total capacity for testing up to 32 samples in parallel. In order to quantify the surface colonization, bacterial cells were DAPI (4`,6-diamidino-2-phenylindole)-stained and examined with epifluorescence microscopy. More than 100 images of each sample were automatically taken and the surface coverage was estimated using the free open source software g'mic, followed by a precise statistical evaluation. Overview images of all gathered pictures were generated to dissect the colonization characteristics of the selected model organism Escherichia coli W3310 on different materials (glass and implant steel). With our approach, differences in bacterial colonization on different materials can be quantified in a statistically validated manner. This reliable test procedure will support the design of improved materials for medical, industrial, and environmental (subaquatic or subaerial) applications.

Integration of ecosystem services into the carbon footprint of milk of South German dairy farms.

Allocation of greenhouse gas emissions (GHG) in Life Cycle Assessments (LCA) is challenging especially when multi-functionality of dairy farms, which do not only produce milk but also meat is considered. Moreover, some farms fulfill a wide range of additional services for society such as management of renewable natural resources as well as preservation of biodiversity and cultural landscapes. Due to the increasing degradation of ecosystems many industrialized as well as developing countries designed payment systems for environmental services. This study examines different allocation methods of GHG for a comparatively large convenience sample of 113 dairy farms located in grassland-based areas of southern Germany. Results are carbon footprints of 1.99 kg CO2eq/kg of fat and protein corrected milk (FPCM) on average if "no allocation" for coupled products is performed. "Physical allocation" results in 1.53 kg CO2eq/kg FPCM and "conventional economic allocation" in 1.66 kg CO2eq/kg FPCM on average if emissions are apportioned between milk and meat. Economic allocation which includes ecosystem services for society based on the farm net income as a new aspect in this study results in a carbon footprint of 1.5 kg CO2eq/kg FPCM on average. System expansion that puts greater emphasis on coupled beef production accounts for a carbon footprint of 0.68 kg CO2eq/kg FPCM on average. Intense milk production systems with higher milk yields show better results based on "no allocation", "physical allocation" and "conventional economic allocation". By contrast, economic allocation, which takes into account ecosystem services favors extensive systems, especially in less favored areas. This shows that carbon footprints of dairy farms should not be examined one-dimensionally based on the amount of milk and meat that is produced on the farm. Rather, a broader perspective is necessary that takes into account the multi-functionality of dairy farms especially in countries where a wide range of ecosystem services is provided.

The effect of feed demand on greenhouse gas emissions and farm profitability for organic and conventional dairy farms.

The reduction of product-related greenhouse gas (GHG) emissions in milk production appears to be necessary. The reduction of emissions on an individual farm might be highly accepted by farm owners if it were accompanied by an increase in profitability. Using life cycle assessments to determine the product carbon footprints (PCF) and farm-level evaluations to record profitability, we explored opportunities for optimization based on analysis of 81 organic and conventional pasture-based dairy farms in southern Germany. The objective of the present study was to detect common determining factors for low PCF and high management incomes (MI) to achieve GHG reductions at the lowest possible operational cost. In our sample, organic farms, which performed economically better than conventional farms, produced PCF that were significantly higher than those produced by conventional farms [1.61 ± 0.29 vs. 1.45 ± 0.28 kg of CO2 equivalents (CO2eq) per kg of milk; means ± SD)]. A multiple linear regression analysis of the sample demonstrated that low feed demand per kilogram of milk, high grassland yield, and low forage area requirements per cow are the main factors that decrease PCF. These factors are also useful for improving a farm's profitability in principle. For organic farms, a reduction of feed demand of 100 g/kg of milk resulted in a PCF reduction of 105 g of CO2eq/kg of milk and an increase in MI of approximately 2.1 euro cents (c)/kg of milk. For conventional farms, a decrease of feed demand of 100 g/kg of milk corresponded to a reduction in PCF of 117 g of CO2eq/kg of milk and an increase in MI of approximately 3.1 c/kg of milk. Accordingly, farmers could achieve higher profits while reducing GHG emissions. Improved education and training of farmers and consultants regarding GHG mitigation and farm profitability appear to be the best methods of improving efficiency under traditional and organic farming practices.