Fast and reliable identification of the microorganisms causing mastitis is important for management of the disease and for targeting antimicrobial treatment. Methods based on PCR are being used increasingly in mastitis diagnostics. Comprehensive field comparisons of PCR and traditional milk bacteriology have not been available. The results of a PCR kit capable of detecting 11 important etiological agents of mastitis directly from milk in 4h were compared with those of conventional bacterial culture (48h). In total, 1,000 quarter milk samples were taken from cows with clinical or subclinical mastitis, or from clinically healthy quarters with low somatic cell count (SCC). Bacterial culture identified udder pathogens in 600/780 (77%) of the clinical samples, whereas PCR identified bacteria in 691/780 (89%) of the clinical samples. The PCR analysis detected major pathogens in a large number of clinical samples that were negative for the species in culture. These included 53 samples positive for Staphylococcus aureus by PCR, but negative by culture. A total of 137 samples from clinical mastitis, 5 samples from subclinical mastitis, and 1 sample from a healthy quarter were positive for 3 or more bacterial species in PCR, whereas culture identified 3 or more species in 60 samples from clinical mastitis. Culture identified a species not targeted by the PCR test in 44 samples from clinical mastitis and in 9 samples from subclinical mastitis. Low SCC samples provided a small number of positive results both in culture (4/93; 4.3%) and by PCR (7/93; 7.5%). In conclusion, the PCR kit provided several benefits over conventional culture, including speed, automated interpretation of results, and increased sensitivity. This kit holds much promise as a tool to complement traditional methods in identification of pathogens. In conventional mastitis bacteriology, a sample with 3 or more species is considered contaminated, and resampling of the cow is recommended. Further study is required to investigate how high sensitivity of PCR and its quantitative features can be applied to improve separation of relevant udder pathogens from likely contaminants in samples where multiple species are detected. Furthermore, increasing the number of species targeted by the PCR test would be advantageous.

Intramammary infection (IMI), also known as mastitis, is the most frequently occurring and economically the most important infectious disease in dairy cattle. This study provides a validation of the analytical specificity and sensitivity of a real-time PCR-based assay that identifies 11 major pathogen species or species groups responsible for IMI, and a gene coding for staphylococcal beta-lactamase production (penicillin resistance). Altogether, 643 culture isolates originating from clinical bovine mastitis, human, and companion animal samples were analyzed using the assay. The isolates represented 83 different species, groups, or families, and originated from 6 countries in Europe and North America. The analytical specificity and sensitivity of the assay was 100% in bacterial and beta-lactamase identification across all isolates originating from bovine mastitis (n = 454). When considering the entire culture collection (including also the isolates originating from human and companion animal samples), 4 Streptococcus pyogenes, 1 Streptococcus salivarius, and 1 Streptococcus sanguis strain of human origin were identified as Streptococcus uberis, and 3 Shigella spp. strains were identified as Escherichia coli, decreasing specificity to 99% in Strep. uberis and to 99.5% in E. coli. These false-positive results were confirmed by sequencing of the 16S rRNA gene. Specificity and sensitivity remained at 100% for all other bacterial targets across the entire culture collection. In conclusion, the real-time PCR assay shows excellent analytical accuracy and holds much promise for use in routine bovine IMI testing programs. This study provides the basis for evaluating the assay's diagnostic performance against the conventional bacterial culture method in clinical field trials using mastitis milk samples.

Lithium phosphate, Li 3 PO 4 , has been considered a potential electrolyte material for lithium ion batteries and CO 2 sensors in particular if the films can be made dense and of high quality already at low thickness. In this work, Li 3 PO 4 thin films were deposited by atomic layer deposition (ALD) between 225 and 350 • C using trimethyl phosphate and either of the two lithium sources, namely lithium hexamethyldisilazide or lithium tert-butoxide. The deposited films showed slightly crystalline Li 3 PO 4 structure in X-ray diffraction and the elastic recoil detection analysis confirmed this stoichiometry with some carbon and hydrogen impurities. The crystallinity and thermal stability of the films at elevated temperatures in N 2 were also examined. The long term stability of the deposited Li 3 PO 4 films under ambient air may be an issue for the applicability of these processes. Lithium phosphate, Li 3 PO 4 , can be applied as an electrolyte in solid state lithium ion batteries 1-3 and CO 2 gas sensors. 4-6 Also optical humidity sensors based on Li 3 PO 4 have been presented recently. 7 Thin Li 3 PO 4 film has also been used as an interfacial layer between LiCoO 2 electrode and solid polymer electrolyte in solid state lithium ion battery. 8 Crystallization of Li 3 PO 4 greatly decreases the ionic conductivity: amorphous Li 2.7 PO 3.9 has an ionic conductivity of 6.6 × 10 −8 S/cm while the polycrystalline γ-Li 3 PO 4 has been extrapolated to be highly resistive with as low conductivity as 4.2 × 10 −18 S/cm at 25 • C. 9 Even if the ionic conductivity is only moderate, amorphous Li 3 PO 4 is still a potential electrolyte material provided it can be made dense already at low thickness.In lithium ion battery applications the 3D structuring of the materials would be a way to effectively increase the energy storage capacity by increasing the specific surface area. 10, 11 Atomic layer deposition (ALD) 12-15 may prove to be useful for depositing thin films on demanding 3D structures as films grown by ALD inherently possess good conformality and uniformity because of the alternating, saturative precursor doses and self-limiting surface reactions. The resulting accurate thickness controllability and repeatability of the films is a major benefit of the layer by layer approach used in ALD. Some of the electrolyte material candidates for 3D batteries include for example Li 3 PO 4 and nitrogen mixed lithium phosphate known as LiPON. 10,11 ALD processes for phosphate films are very few in number, although aluminum phosphate thin films have been deposited already in the 90's. 16,17 Besides aluminum phosphate, only ALD of calcium phosphate has been published. 18 Also lithium containing ALD processes were absent until recent reports on ALD of lithium hydroxide, lithium carbonate, lithium aluminate, lithium silicate, lithium lanthanate and lithium lanthanum titanates. [19][20][21][22][23] In addition, ALD of LiFePO 4 and Ca:LaPO 4 materials have been recently presented. 24,25 Here we report ALD growth and characterization of lithium...

The associations between quantitative bacteriological results from a real-time PCR test and concentrations of acute-phase proteins (APP) and N-acetyl-β-d-glucosaminidase (NAGase) activity in milk in naturally occurring clinical mastitis were investigated. Milk APP concentrations and NAGase activity in clinical mastitis caused by different udder pathogens were studied. The associations between the severity of the clinical signs and concentrations of APP and NAGase activity were estimated. Milk samples from 281 cases of clinical mastitis were collected from 3 Estonian dairy farms and analyzed by PCR to identify pathogens. Twenty-seven samples out of 281 (9.6%) were PCR negative. Milk samples containing 4 or more bacterial species (n=28) were considered possibly contaminated and excluded from all further analyses. In total, 443 bacterial identifications were made from the remaining 226 milk samples. A single bacterial species was detected in 68 samples (30.1%), 2 species were detected in 99 samples (43.8%), and 3 species were detected in 59 (26.1%) samples. To determine the inflammatory response in the udder, the concentrations of milk amyloid A (MAA) and haptoglobin (Hp) and NAGase activity in the milk were analyzed. A significant positive association was found between the severity of the clinical signs and inflammatory markers in the milk. Milk amyloid A and Hp concentrations and NAGase activity were significantly higher in samples with large quantities of bacterial DNA from Escherichia coli or Streptococcus dysgalactiae compared with milk samples not containing those species. Large quantities of bacterial DNA from Trueperella pyogenes or Streptococcus uberis in the milk were associated with elevated concentrations of Hp and high NAGase activity, but not with increased MAA concentrations. Milk samples containing Corynebacterium bovis and coagulase-negative staphylococci had significantly lower concentrations of MAA and Hp and lower NAGase activity compared with samples where these species were not detected. It can be concluded that concentrations of APP and NAGase activity in the milk were associated with the quantity of bacterial DNA in the milk samples.

A needleless electrospinning setup named 'Needleless Twisted Wire Electrospinning' was developed. The polymer solution is electrospun from the surface of a twisted wire set to a high voltage and collected on a cylindrical collector around the wire. Multiple Taylor cones are simultaneously self-formed on the downward flowing solution. The system is robust and simple with no moving parts aside from the syringe pump used to transport the solution to the top of the wire. The structure and process parameters of the setup and the results on the preparation of polyvinyl pyrrolidone (PVP), hydroxyapatite (HA) and bioglass fibers with the setup are presented. PVP fiber sheets with areas of 40 × 120 cm(2) and masses up to 1.15 g were prepared. High production rates of 5.23 g h(-1) and 1.40 g h(-1) were achieved for PVP and HA respectively. The major limiting factor of the setup is drying of the polymer solution on the wire during the electrospinning process which will eventually force to interrupt the process for cleaning of the wire. Possible solutions to this problem and other ways to develop the setup are discussed. The presented system provides a simple way to increase the production rate and area of fiber sheet as compared with the conventional needle electrospinning.