Performance evaluation of canine-associated Bacteroidales assays in a multi-laboratory comparison study

The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contributions. However, data about the performance of available assays are scarce. The results of a multi-laboratory study testing two assays for the determination of dog-associated Bacteroidales (DogBact and BacCan-UCD) on 64 single and mixed fecal source samples created from pooled fecal samples collected in California are presented here. Standardization of qPCR data treatment lowered inter-laboratory variability of sensitivity and specificity results. Both assays exhibited 100% sensitivity. Normalization methods are presented that eliminated random and confirmed non-target responses. The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment.     

Multi-laboratory evaluations of the performance of Catellicoccus marimammalium PCR assays developed to target gull fecal sources

Here we report results from a multi-laboratory (n = 11) evaluation of four different PCR methods targeting the 16S rRNA gene of Catellicoccus marimammalium originally developed to detect gull fecal contamination in coastal environments. The methods included a conventional end-point PCR method, a SYBR^® Green qPCR method, and two TaqMan^® qPCR methods. Different techniques for data normalization and analysis were tested. Data analysis methods had a pronounced impact on assay sensitivity and specificity calculations. Across-laboratory standardization of metrics including the lower limit of quantification (LLOQ), target detected but not quantifiable (DNQ), and target not detected (ND) significantly improved results compared to results submitted by individual laboratories prior to definition standardization. The unit of measure used for data normalization also had a pronounced effect on measured assay performance. Data normalization to DNA mass improved quantitative method performance as compared to enterococcus normalization. The MST methods tested here were originally designed for gulls but were found in this study to also detect feces from other birds, particularly feces composited from pigeons. Sequencing efforts showed that some pigeon feces from California contained sequences similar to C. marimammalium found in gull feces. These data suggest that the prevalence, geographic scope, and ecology of C. marimammalium in host birds other than gulls require further investigation. This study represents an important first step in the multi-laboratory assessment of these methods and highlights the need to broaden and standardize additional evaluations, including environmentally relevant target concentrations in ambient waters from diverse geographic regions.     

Comparative Assay of Antioxidant Packages for Dimer of Estolide Esters

A series of 26 different antioxidants and commercial antioxidant packages designed for petroleum‐based materials, containing both natural and synthetic‐based materials, were evaluated with dimeric coconut‐oleic estolide 2‐ethylhexyl ester (2‐EH), a bio‐based material. The different antioxidants were categorized into different classes of phenolic, aminic, and blended/others materials. The oxidation onset temperatures (OT) using non‐isothermal pressurized differential scanning calorimetry (PDSC) were measured and recorded under previously reported standard conditions. The aminic series gave the best resistance to oxidation as defined by the PDSC method with OT of 246.6 and 244.7 °C for the best two performers, which was a 38 °C improvement over the uninhibited or unformulated dimer estolide material. The phenolic series, containing most of the naturally occurring antioxidants, was the least successful formulation package for the dimer estolide. The blended/other materials, which were specifically designed for petroleum‐based lubricants, did not have the best OT, since the estolides and other bio‐based materials interact differently than their petroleum counterparts. A number of potential antioxidants have been identified as useful additives for the estolides esters. The OT of the estolide and formulated materials correlated well with other bio‐based materials such as biodiesel.     

Misalignment-Induced Bending in Pin-Loaded Tensile-Creep Specimens

In this paper, we have examined the possibility that elastic bending induced by load misalignment can affect creep measurements on pin-loaded tension specimens of silicon nitride (Si₃N₄). We have shown that elastic bending at room temperature can be as great as 42% of the axial strain when clean pins were used but was reduced to ∼3% when lubricated pins were used. Creep tests at the same applied stress and temperature were conducted on two groups of Si₃N₄ test specimens: one group used clean pins, and the other group used lubricated pins. By measuring the shapes of the specimens before and after the creep tests, we have determined that the loading holes were slightly misaligned before the creep tests and that small amounts of bending were induced by the creep tests. Bending occurred both in the gauge section of the specimen and in the transition region between the gauge section and the heads of the specimen (the latter phenomenon is defined as hinge bending). Our study indicated that the secondary creep rate, the hinge bending, and the bending of the gauge section was not dependent on pin lubrication, because the results from the clean and lubricated groups were statistically indistinguishable. Hinge bending was dependent on initial misalignment of the loading holes, whereas bending of the gauge section was independent of such factors.     

Electrochemical characterization of electrolytes for lithium-ion batteries based on lithium difluoromono(oxalato)borate

The salt lithium difluoromono(oxalato)borate (LiDFOB) showed some promising results for lithium-ion-cells. It was synthesized via a new synthetic route that avoids chloride impurities. Here we report the properties of its solutions (solvent blend ethylene carbonate/diethyl carbonate (3:7, mass ratio), including its conductivity, cationic transference number, hydrolysis, Al-current collector corrosion-protection ability and its cycling performance with some electrode materials. Some Al-corrosion studies were also performed with the help of our recently developed computer controlled impedance scanning electrochemical quartz crystal microbalance (EQCM) that proofed to be a useful tool for battery material investigations.     

Oxidation-abrasion of TiC-based cermets in SiC medium

The aim of the current study was to investigate the effect of oxidation on abrasive wear behaviour of TiC based cermets at temperatures ranging from 20 to 900 °C. Three types of material performance maps were constructed: oxidation rate maps, wear rate maps and maps showing the effect of oxidation on abrasion. Discussion on the performance of different cermet grades is supported by the SEM images combined with EDS and XRD analysis. The results should facilitate the selection of TiC-based cermets providing optimum composition of cermets for high temperature applications.