Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks

A reverse transcriptase - polymerase chain reaction based assay for Borrelia species detection in ticks was developed. The method was based on amplification of 552 nucleotide bases long sequence of 16S rRNA, targeted by Borrelia specific primers. In the present study, total RNA extracted from Ixodes ricinus ticks was used as template. The results showed higher sensitivity for Borrelia detection as compared to standard dark-field microscopy. Method specificity was confirmed by cloning and sequencing of obtained 552 base pairs long amplicons. Phylogenetic analysis of obtained sequences showed that they belong to B. lusitaniae and B. afzelii genospecies. RT-PCR based method presented in this paper could be very useful as a screening test for detecting pathogen presence, especially when in investigations is required extraction of total RNA from ticks

Investigation of Chlamydophila spp. in dairy cows with reproductive disorders

<p>Abstract</p> <p>Background</p> <p>Reports worldwide indicate high prevalence of <it>Chlamydophila </it>spp. infection in cattle. To assess the prevalence in Sweden, 525 cows in 70 dairy herds with reproductive disorders was investigated.</p> <p>Methods</p> <p>To detect antibodies two commercially available kits were used. Moreover, 107 specimens, including vaginal swabs, organ tissues and milk were analysed by Polymerase Chain Reaction (PCR).</p> <p>Results</p> <p>Two (0.4%) cows were seropositive in the Pourquier <it>Cp. abortus </it>ELISA. The seroprevalence with the Chekit ELISA was 28% with no difference between cases and controls. Five specimens were positive in real-time PCR and further analysed by nested PCR. <it>Cp. pecorum </it>was confirmed by partial <it>omp1 </it>DNA sequencing of the nested PCR product of vaginal swabs from control cows.</p> <p>Conclusion</p> <p>The results suggest that <it>Cp. abortus </it>infection is absent or rare in Swedish cows whereas <it>Cp. pecorum </it>is probably more spread. They also suggest that <it>Chlamydophila </it>spp. are not related to reproduction disorders in Swedish cattle.</p

OmpA and antigenic diversity of bovine Chlamydophila pecorum strains

Infections with the intracellular bacterium Chlamydophila (C.) pecorum are highly prevalent worldwide in cattle. These infections cause significant diseases such as polyarthritis, pneumonia, enteritis, genital infections and fertility disorders, and occasionally sporadic bovine encephalomyelitis. Subclinical respiratory infections of calves with C. pecorum have been associated with airway obstruction, pulmonary inflammation, and reduced weight gains. This investigation examined four chlamydial strains with biological properties of C. pecorum isolated from feces of clinically normal cattle, from calves with pneumonia, and from bulls with posthitis. The objective was to characterize the evolutionary relationships of these bovine chlamydial isolates to other chlamydiae by genetic analysis of the ompA gene, and by the immunological cross-reactivities in Western immunoblot analysis. PCR typing of the ompA gene identified these isolates as C. pecorum. The OmpA-deduced amino acid dissimilarities between these four strains spanned 10-20%. In phylogenetic analysis, the four isolates clustered with C. pecorum ruminant, porcine, and koala strains of different geographic origins rather than with each other. All four isolates showed different patterns of Western immunoblot reactivity with antiserum against bovine C. pecorum strain LW63, and, interestingly, no cross-reactivity of the OmpA proteins with the anti-LW613 OmpA antibodies. These data underscore the polyphyletic population structure of C. pecorum and suggest that the spectrum of C. pecorum OmpA proteins in a host species can occupy the entire evolutionary bandwidth within C. pecorum. The variant immunoblot reactivities support the notion of considerable genomic plasticity of C. pecorum

Antibiotikaresistenz von Staphylococcus aureus und Koagulase-negativen Staphylokokken isoliert aus Mastitismilchproben von Schafen und Ziege

Im Rahmen dieser Arbeit wurden aus Mastitismilchproben von Schafen und Ziegen von über 40 Betrieben der Schweiz Staphylococcus aureus und Koagulase-negative Staphylokokken (CNS) isoliert. Bei den CNS konnte mittels Restriktionsprofil-Analyse des groEL-Gens und 16S rDNA Sequenzierung gezeigt werden, dass verschiedene Spezies vorkamen, wenn auch einzelne dominierten. Die Resistenzprüfung der Mastitiserreger gegen ausgewählte Antibiotika erfolgte mittels des Mikrodilutionsverfahrens. Von den 67 S. aureus- und 208 CNS-Stämmen waren 31.3 % und 8.2 % resistent gegen Penicillin, 29.9 % und 1.0 % resistent gegen Ampicillin, 1.5 % und 10.6 % resistent gegen Erythromycin sowie 3.0 % und 7.7 % resistent gegen Tetracyclin. Zudem waren 10 CNS-Stämme (4.8 %) resistent gegen Oxacillin und ein CNS-Stamm gegen Sulfamethoxazol/Trimethoprim. Die vorliegenden Daten stellen eine erste Bestandesaufnahme zur Resistenzsituation bei Mastitiserregern von Schafen und Ziegen in der Schweiz dar. Zur Mastitisbekämpfung sind aber bei Kleinwiederkäuern auch flankierende und präventive Massnahmen von entscheidender Bedeutung. Abstract: Staphylococcus aureus and coagulase negative staphylococci (CNS) were isolated from ovine and caprine mastitis milk samples originating from more than 40 Swiss farms. CNS dominated as causal microorganisms of mastitis in small ruminants. By restriction fragment length polymorphism (RFLP) analysis of the groEL gene and sequencing of 16S rDNA, various CNS species were identified, albeit certain of them predominated. For susceptibility testing of mastitis pathogens to selected antibiotics, minimal inhibitory concentrations were determined. Of the 67 S. aureus and 208 CNS strains, 31.3 % and 8.2 % were resistant to penicillin, 29.9 % and 1.0 % to ampicillin, 1.5 % and 10.6 % to erythromycin, and 3.0 % and 7.7 % to tetracycline, respectively. Moreover, 10 CNS strains (4.8 %) were resistant to oxacillin and one CNS strain to sulfamethoxazole/trimethoprim. The results obtained describe for the first time the resistance situation of mastitis pathogens from sheep and goats in Switzerland. However, accompanying and preventing measures are also of importance in mastitis control of small ruminants

Accounting for financial instruments in the banking industry: conclusions from a simulation model

The paper analyses the effects of three sets of accounting rules for financial instruments - Old IAS before IAS 39 became effective, Current IAS or US GAAP, and the Full Fair Value (FFV) model proposed by the Joint Working Group (JWG) - on the financial statements of banks. We develop a simulation model that captures the essential characteristics of a modern universal bank with investment banking and commercial banking activities. We run simulations for different strategies (fully hedged, partially hedged) using historical data from periods with rising and falling interest rates. We show that under Old IAS a fully hedged bank can portray its zero economic earnings in its financial statements. As Old IAS offer much discretion, this bank may also present income that is either positive or negative. We further show that because of the restrictive hedge accounting rules, banks cannot adequately portray their best-practice risk management activities under Current IAS or US GAAP. We demonstrate that - contrary to assertions from the banking industry - mandatory FFV accounting adequately reflects the economics of banking activities. Our detailed analysis identifies, in addition, several critical issues of the accounting models that have not been covered in previous literature.