Relatedness of cytotoxins from geographically diverse isolates of Moraxella bovis

To determine whether amino acid sequence variation exists in the Moraxella bovis (M. bovis) cytotoxin (MbxA) from geographically diverse M. bovis isolated in the United States, mbxA was amplified and sequenced. The MbxA deduced amino acid sequence from M. bovis originally isolated in California, Washington, North Carolina, and Georgia, as well as reference strains of M. bovis isolated at the National Animal Disease Laboratory, Ames, IA, USA, all encoded a nearly identical 927 amino acid protein. MbxA from two of the four California isolates (SFS 9a and SFS 100a) differed from all other isolates at two sites at which the polar amino acids glutamine (position 666) and asparagine (position 823) were replaced by ionized amino acids glutamic acid and aspartic acid, respectively. Rabbit antiserum to the expressed carboxy terminus (amino acids 590-927) of MbxA from M. bovis (Tifton I) neutralized the hemolytic activity of SFS 9a and SFS 100a. The M. bovis cytotoxin appears to be conserved amongst geographically diverse isolates of M. bovis from the USA. Antiserum against the carboxy terminus of MbxA common to the majority of isolates neutralized the hemolytic activity of two strains with a divergent MbxA deduced amino acid sequence. Vaccines against IBK that incorporate MbxA as antigen may offer protection against geographically diverse strains of M. bovis.     

Molecular cloning of a Mycoplasma meleagridis-specific antigenic domain endowed with a serodiagnostic potential

A recombinant phage library harbouring Mycoplasma meleagridis (MM) genomic DNA fragments was generated in the bacteriophage lambda gt11 expression vector. The library was screened for expression of MM specific antigens with a polyclonal antiserum that had been preadsorbed with antigens of the most common unrelated avian mycoplasma species. A 49-amino acid antigenic domain unique to MM was isolated, expressed in Escherichia coli, and its serodiagnostic potential was demonstrated. An antiserum raised against this MM-specific antigenic domain recognized a cluster of seven membrane-associated MM proteins with molecular masses ranging from 34 to 75 kDa. Overall, this study resulted in the identification of a potent serodiagnostic tool and revealed the complex antigenic nature of MM.     

Multiplex real-time PCR for the detection and differentiation of equid herpesvirus 1 (EHV-1) and equid herpesvirus 4 (EHV-4)

A multiplex real-time PCR was designed to detect and differentiate equid herpesvirus 1 (EHV-1) and equid herpesvirus 4 (EHV-4). The PCR targets the glycoprotein B gene of EHV-1 and EHV-4. Primers and probes were specific to each equine herpesvirus type and can be used in monoplex or multiplex PCRs, allowing the differentiation of these two closely related members of the Alphaherpesvirinae. The two probes were minor-groove binding probes (MGB) labelled with 6-carboxy-fluorescein (FAM) and VIC for detection of EHV-1 and EHV-4, respectively. Ten EHV-1 isolates, six EHV-1 positive clinical samples, one EHV-1 reference strain (EHV-1.438/77), three EHV-4 positive clinical samples, two EHV-4 isolates and one EHV-4 reference strain (EHV-4 405/76) were included in this study. EHV-1 isolates, clinical samples and the reference strain reacted in the EHV-1 real-time PCR but not in the EHV-4 real-time PCR and similarly EHV-4 clinical samples, isolates and the reference strain were positive in the EHV-4 real-time PCR but not in the EHV-1 real-time PCR. Other herpesviruses, such as EHV-2, EHV-3 and EHV-5 were all negative when tested using the multiplex real-time PCR. When bacterial pathogens and opportunistic pathogens were tested in the multiplex real-time PCR they did not react with either system. The multiplex PCR was shown to be sensitive and specific and is a useful tool for detection and differentiation of EHV-1 and EHV-4 in a single reaction. A comprehensive equine herpesvirus disease investigation procedure used in our laboratory is also outlined. This procedure describes the combination of alphaherpesvirus multiplex real-time PCR along with existing gel-based PCRs described by other authors.     

Assessment of mild hindlimb lameness during over ground locomotion using linear discriminant analysis of inertial sensor data

REASONS FOR PERFORMING STUDY: Hindlimb lameness is common and can be difficult to diagnose or quantify in evaluating response to nerve blocks. An objective measure of lameness can also be used to evaluate the effectiveness of the treatment's contribution to evidence-based medicine. The inertial sensor system can be used to capture 6 degree of freedom movement during over ground locomotion and here was used to quantify tuber coxae movement in nonlame and lame horses. HYPOTHESIS: Tuber coxae movement is useful for discriminating between nonlame and lame horses. OBJECTIVES: To measure left and right tuber coxae movement in lame and nonlame horses during over ground locomotion and to implement a linear discriminant analysis to discriminate between lame and nonlame horses. METHODS: Two inertial sensors were attached to the skin over left and right tuber coxae of 21 horses (9 mildly and 12 not lame). Horses were trotted on a hard surface. A total of 1021 strides were collected. For each stride 34 features were extracted from the dorsoventral and craniocaudal movement and used in 2 different classification scenarios (lame vs. nonlame or left lame, right lame and nonlame) using linear discriminant analysis. RESULTS: Six degree of freedom inertial sensors were successfully used to collect kinematic data continuously from left and right tuber coxae in horses during over ground locomotion. These data were used for an automated classification of lameness. In the first scenario, a sensitivity of 89% was achieved with a specificity of 75%. In the second scenario, all horses could be correctly assigned to the correct class in a simple 3 class reclassification test. POTENTIAL RELEVANCE: A mobile system that reliably detects and quantifies hindlimb lameness in horses during unconstrained locomotion could be a valuable tool to perform an evidence-based assessment of lameness in horses in a clinical setting, e.g. before and after nerve blocks or before and after surgery.     

An inactivated vaccine for the control of bluetongue virus serotype 16 infection in sheep in Italy

Because no suitable products are at the moment available to safely control the spread of BTV-16 in Europe, an inactivated vaccine was produced from the reference field isolate of bluetongue virus serotype 16. One group of six sheep was vaccinated subcutaneously with the inactivated vaccine twice, on days 0 and 28, whereas a second group of eight sheep was inoculated with saline solution and used as mock-vaccinated control animals. Seventy-eight days after the first vaccination, all sheep were inoculated subcutaneously with a suspension containing 10(6.3) TCID(50) of a virulent reference BTV-16 isolate. Apart from a transient inflammatory reaction at the injection site, no adverse effects were reported following vaccination. All vaccinated animals developed high titres (7.3-9.3log(2)(ED50%/50 microl)) of virus-specific neutralising antibodies and were resistant to challenge with BTV-16. Conversely, following challenge, control animals developed hyperthermia and long lasting high-titre viraemia.     

Variation in E(rns) viral glycoprotein associated with failure of immunohistochemistry and commercial antigen capture ELISA to detect a field strain of bovine viral diarrhea virus

Bovine viral diarrhea virus (BVDV) affects cattle populations causing clinical signs that range from subclinical immunosuppression to severe reproductive and respiratory problems. Detection and removal of persistently infected (PI) calves is the single most important factor for control and eradication of BVDV. Current testing strategies to detect PI calves rely heavily on immunohistochemistry (IHC) and a commercially available antigen capture ELISA (ACE) assay. These viral assays depend on 1 or 2 monoclonal antibodies which target the E(rns) glycoprotein of BVDV. The sensitivity and specificity of these two tests have been reported previously. The purpose of this research was to characterize a strain of BVDV (AU501) that was undetectable using IHC and ACE based on a single monoclonal antibody, but was consistently detected in samples from a Holstein steer using virus isolation and PCR testing. Sequencing of this AU501 viral isolate revealed a unique mutation in the portion of the genome coding for the E(rns) glycoprotein. This unique field strain of BVDV demonstrates the risk of relying on a single monoclonal antibody for detection of BVDV. Multiple testing strategies, including polyclonal or pooled monoclonal antibodies that detect more than one viral glycoprotein may be necessary to detect all PI calves and facilitate eradication of BVDV.     

Oral and intravenous administration of nimesulide in the horse: rational dosage regimen from pharmacokinetic and pharmacodynamic data

REASONS FOR PERFORMING STUDY: The selective COX-2-inhibitor nimesulide is used extra-label in equine veterinary practice as an anti-inflammatory agent. However, there are no data on which to base the rational use of the drug in this species. OBJECTIVES: To determine the effective COX selectivity of nimesulide in the horse, and suggest a suitable dosing schedule. METHODS: The pharmacokinetics of nimesulide in the horse after oral administration (1 mg/kg bwt), and oral and i.v. administration (1.5 mg/kg bwt) were investigated, effects of feeding status on bioavailability determined, and plasma protein binding of the drug and its principal metabolites measured. Compartmental and noncompartmental pharmacokinetic analyses were performed. The plasma concentration-time profile was used, together with in vitro literature data on nimesulide inhibition of COX isoforms, to determine the effective COX selectivity of nimesulide in the horse, and suggest a suitable dosing schedule. RESULTS AND CONCLUSIONS: The findings suggest that 1.5 mg/kg bwt may produce adequate clinical effects, and the dosing interval should be 12-24 h depending on condition severity. However, at that dose, the concentration in the animal exceeds the in vitro IC50 for both isoforms, so that COX-1/COX-2 selectivity is lost and side-effects due to COX-1 inhibition are a possibility. Nimesulide should therefore be used with caution in equine clinical practice.     

Factors associated with practice decisions of Nebraska veterinarians regarding type of practice and community size

Nebraska veterinary practitioners were surveyed to collect data about background characteristics and other factors related to veterinarians' decision to include or not include food animals in their practices and to practice in rural versus urban communities. Background characteristics that were significantly (p < or = 0.05) associated with choosing food-animal practice included growing up on a working farm or ranch; having parents who owned livestock; growing up in a town with a population of less than 10,000; majoring in animal science at university; being male; and having a primary interest, at the time of entering veterinary college, in food animal-exclusive or mixed-animal veterinary practice. The primary factor for choosing the community in which to practice was rural/urban lifestyle for rural veterinarians, while this factor was second for urban veterinarians. For all groups of veterinarians, the primary consideration in selecting their current practice was the species orientation of the practice. The primary reason for not choosing food-animal practice was better working conditions and lifestyle in companion-animal practice, followed by greater interest elsewhere.     

Helping students learn veterinary physiology through the use of concept maps

Concept maps have become a ubiquitous tool in education in the last 20 years and are used extensively to teach scientific concepts. A concept map is a tool learners can use to organize information about a topic in terms of key concepts and the relationships among them. As a graphical representation, the map provides an overall picture of a particular concept or topic. This article describes how we have used concept maps to teach physiology to veterinary medical and veterinary technician students enrolled in our on-campus and distance-learning programs.     

Efficacy of topical eprinomectin in the treatment of Chorioptes sp. infestation in alpacas and llamas

Chorioptes sp. mite infestation is increasingly recognized as a cause of skin disease in New World camelids and there is a need for an effective treatment protocol to eliminate herd infestation. In this field trial, eprinomectin applied topically at the rate of 0.5 mg kg(-1) weekly for 10 weeks was found to be ineffective in a herd of 12 llamas and 16 alpacas.