Profiling excretory/secretory proteins of Trichinella spiralis muscle larvae by two-dimensional gel electrophoresis and mass spectrometry

Infection of mammalian skeletal muscle with the intracellular parasite Trichinella spiralis results in profound alterations in the host cell and a realignment of host cell gene expression. The role of parasite excretory/secretory (E/S) products in mediating these effects is unknown, largely due to the difficulty in identifying and assigning function to individual proteins. In this study, we have used two-dimensional electrophoresis to analyse the profile of muscle larva excreted/secreted proteins and have coupled this to protein identification using MALDI-TOF mass spectrometry. Interpretation of the peptide mass fingerprint data has relied primarily on the interrogation of a custom-made Trichinella EST database and the NemaGene cluster database for T. spiralis. Our results suggest that this proteomic approach is a useful tool to study protein expression in Trichinella spp. and will contribute to the identification of excreted/secreted proteins.     

Systemic fungal infection in a dog: a unique case in Ireland

A three year old male entire Staffordshire bull terrier was referred to University College Dublin Veterinary Hospital, with a two week history of fever, inflammation of the right hock, lameness on the right hindlimb, peripheral lymphadenopathy and gastrointestinal signs (vomiting and diarrhoea). For the preceding three months the dog had been treated for atopic dermatitis with oral ciclosporin (5 mg/kg, PO, q 24 hours).Cytological analysis of the affected lymph nodes demonstrated fungal-like organisms predominantly contained within macrophages. Subsequent fungal culture and microscopic identification confirmed the presence of a Byssochlamys sp. This fungus is a saprophytic organism which has been associated with mycotoxin production. It has not previously been identified as a cause of systemic infection in animals or humans.Ciclosporin was discontinued, and a second generation triazole, voriconazole prescribed at a dose of 6 mg/kg for the first two doses, and continued at 3 mg/kg every 12 hours for six months. There was an excellent response. Follow-up examination five weeks after treatment was completed confirmed remission of the disease. The dog remains alive and well three years later.The present case represents an unusual fungal infection in a dog secondary to immunosuppressive therapy with ciclosporin. Such a possibility should be considered in animals presenting with signs consistent with systemic infection when receiving immunosuppressive medication.     

Enzyme-linked immunosorbent assay-based detection of free trenbolone in bovine bile

A rapid antibody-based detection system has been developed for the presence of free trenbolone in bovine samples. Polyclonal antibodies were produced that showed specificity toward epitopes located around the steroidal A-ring of the trenbolone molecule. These antibodies were shown to have little or no recognition for many closely related compounds. The antibodies were utilized as the specific biorecognition molecules in competitive and inhibitive enzyme-linked immunosorbent assay systems. While both assays were able to detect low nanogram concentrations of trenbolone in bovine bile, the competitive format was more sensitive (2.41 vs 17.15 ng/mL for TRAb2 and 3.31 vs 30.73 ng/mL for TRAb1). This format was also more accurate and the data produced by this assay fitted more closely to the four parameter equation used to calculate the standard curve. This was a common finding with both of the polyclonal antibodies, suggesting that this was a characteristic of the format used.     

Assessment of ruminal degradation, oral bioavailability, and toxic effects of anticoagulant rodenticides in sheep

OBJECTIVE: To assess the rate and extent of ruminal degradation of warfarin, chlorophacinone, and bromadiolone in vitro and determine the oral availability and clinical and hemostatic effects of each anticoagulant rodenticide in adult sheep. ANIMALS: 3 Texel sheep. PROCEDURE: Samples of ruminal fluid were incubated with each of the anticoagulants to assess the kinetics of ruminal degradation over 24 hours. To determine the plasma kinetics of the anticoagulants, each sheep received each of the anticoagulants IV or via a rumenimplanted cannula at 2-month intervals (3 rodenticide exposures/sheep). At intervals during a 240- to 360- hour period after treatment, prothrombin time (PT) was measured, plasma anticoagulant concentration was assessed, and clinical signs of rodenticide poisoning were monitored. In plasma and rumen extracts, anticoagulant concentrations were determined via high-performance liquid chromatography. RESULTS: In the rumen extracts, anticoagulants were slightly degraded (< 15%) over 24 hours. In vivo, oral availability of warfarin, chlorophacinone, and bromadiolone was estimated at 79%, 92%, and 88%, respectively. Although maximum PT was 80 seconds after chlorophacinone and bromadiolone treatments, no clinical signs of toxicosis were detected; PT returned to baseline values within 2 weeks. CONCLUSIONS AND CLINICAL RELEVANCE: In sheep, warfarin, chlorophacinone, and bromadiolone were not degraded in the rumen but their bioavailabilities were high after oral administration; the kinetics of these compounds in sheep and other mammals are quite similar. These data suggest that the lack of susceptibility of ruminants to these anticoagulant rodenticides cannot be explained by either ruminal degradation or the specific toxicokinetics of these anticoagulants.