A longitudinal study on transmission of Staphylococcus aureus genotype B in Swiss communal dairy herds.

Staphylococcus aureus is a common mastitis causing pathogen of dairy cattle. Several S. aureus genotypes exist, of which genotype B (GTB) is highly prevalent in Swiss dairy herds. Dairy farming in mountainous regions of Switzerland is characterised by the movement of dairy cattle to communal pasture-based operations at higher altitudes. Cows from different herds of origin share pastures and milking equipment for a period of 2 to 3 months during summer. The aim of this longitudinal observational study was to quantify transmission of S. aureus GTB in communal dairy operations. Cows (n=551) belonging to 7 communal operations were sampled at the beginning and end of the communal period. Transmission parameter ß was estimated using a Susceptible-Infectious-Susceptible (SIS) model. The basic reproduction ratio R0 was subsequently derived using previously published information about the duration of infection. Mean transmission parameter ß was estimated to be 0.0232 (95% CI: 0.0197-0.0274). R0 was 2.6 (95% CI: 2.2-3.0), indicating that S. aureus GTB is capable of causing major outbreaks in Swiss communal dairy operations. This study emphasized the contagious behaviour of S. aureus GTB. Mastitis management in communal dairy operations should be optimized to reduce S. aureus GTB transmission between cows and back to their herds of origin.

Primary bovine synoviocyte cultures: a useful tool for in vitro drug testing?

The aim of the study was to evaluate bovine synoviocyte culture as an in vitro model to test new intra-articular drugs. The inflammatory reaction pattern of synoviocytes as compared to fibroblasts was studied over nine passages. Expression of pro-inflammatory cytokines was assessed after stimulation with lipopolysaccharide. Immunohistochemical markers were used to identify synoviocyte populations. Primary synoviocytes expressed markedly higher amounts of interleukin-1ß mRNA and tumour necrosis factor-α mRNA than fibroblasts after stimulation. This difference was lost over two passages. CD68-positive macrophage-like synoviocytes diminished over three passages, which may explain the reduced pro-inflammatory cytokine response. Primary bovine synoviocytes appear to be an appropriate and optimised model for testing novel drugs for cattle, because their response may more closely reflect in vivo tissue responses compared to cultured cell lines.

Slow release antibiotics for treatment of septic arthritis in large animals.

The search for an effective treatment for septic arthritis is ongoing. Current therapies are expensive since they require repeated joint lavage and long term antibiotic treatment. Local application of antimicrobial drugs is advantageous because high concentrations can be attained at the infection site, although repeated injections increase the risk of superinfection of the joint. Thus, slow release formulations, which have the advantage of local treatment yet single application of the drug, are appealing. Antibiotics used in slow release formulations are selected for tissue compatibility, an appropriate antibacterial spectrum, and stability both during the mixing procedure and within the carrier during the release period. Ideally the carriers should be bioresorbable. Promising reports on the clinical use of poly(methyl methacrylate) (PMMA) mixed with several different antibiotics, and of collagen sponges impregnated with gentamicin, should encourage the search for formulations optimally adapted to veterinary medical requirements.

In vitro cell compatibility and antibacterial activity of microencapsulated doxycycline designed for improved localized therapy of septic arthritis.

OBJECTIVES: For the treatment of septic arthritis in large animals, the local application of antibiotics as a slow release system may be an appropriate means to reach high local bioactivity and low systemic side effects and drug residues. In this study, doxycycline microspheres were developed and tested in vitro for their drug-release properties, suitability for intra-articular application and antimicrobial activity. METHODS: The development of a slow release system was achieved by microencapsulation of the drug into poly(lactide-co-glycolide) microspheres by a novel ultrasonic atomization method. Drug elution was evaluated from microspheres dispersed in elution medium at pre-defined time points by HPLC. Joint-tissue compatibility was tested on cultured bovine synoviocytes by evaluating the expression of pro-inflammatory cytokine mRNA and the production of nitric oxide (NO). Finally, the antimicrobial activity of the released antibiotic was assessed with gram-negative and gram-positive bacteria exposed to release medium sampled at days 1, 7 and 12 after microsphere suspension. RESULTS: An adequate size of the microspheres, sufficient stabilization of doxycycline in aqueous environment and drug release (25 mg microspheres in 4 mL medium) above MIC for bacteria usually isolated in bovine and equine joints were obtained over 15 days. Although the cytokine mRNA expression reflected the excellent tissue compatibility, the results with NO yielded contradictory results. Antimicrobial tests of the release medium proved to match perfectly the activity of non-encapsulated, free doxycycline as reported in the literature. CONCLUSIONS: The newly developed doxycycline delivery system achieved the target specifications and is ready for in vivo testing.

Intra-articular administration of doxycycline in calves.

Objective-To evaluate local tissue compatibility of doxycycline hyclate (DOX) in antebrachiocarpal joints of calves. Animals-10 healthy calves between 80 and 110 kg. Procedures-Calves were assigned to 2 treatment groups. Calves in groups DOX(low) and DOX(high) were administered 5 and 10 mg of DOX, respectively, locally in 1 antebrachiocarpal joint. The contralateral joint served as a control joint and was injected with 0.9% NaCl solution. General and local clinical findings were scored. Several variables were assessed in blood and synovial fluid for 9 days. Calves were euthanatized and pathologic changes and drug residues evaluated. Results-Throughout the study, none of the calves had clinical changes or abnormal hematologic values. Significant differences between treatment and control joints were evident only for matrix metalloproteinases at 0.5 hours after injection, with less activity for the DOX-treated joints in both treatment groups. Values for all synovial fluid variables, except nitric oxide, increased significantly during the first 12 to 72 hours after arthrocentesis in control and DOX-treated joints. Histologic examination revealed minimal infiltration of inflammatory cells independent of the treatment. No drug residues were detected 9 days after arthrocentesis in any tissues obtained from the liver, kidneys, fat, and skeletal muscles. Conclusions and Clinical Relevance-DOX had excellent intra-articular compatibility in healthy calves. Arthrocentesis induced a mild transient increase of inflammatory mediators in the synovial fluid. Significant decreases in matrix metalloproteinase activity in DOX-treated joints may indicate a potential chondroprotective effect of DOX.