Viral respiratory disease of the horse.

The diagnosis of any viral respiratory disease relies on laboratory procedures to isolate the virus and demonstrate a significant rise in serum antibody titers. To isolate viruses from the upper respiratory tract, it is imperative that nasopharyngeal swabs are obtained from animals in the early acute stage of illness, i.e., during the pyrexic phase when the virus is replicating. Nasopharyngeal swabs must be placed in a virus transport medium and forwarded immediately to the laboratory at refrigerated temperature. Equine influenza, rhinopneumonitis, and equine viral arteritis are the three viral infections causing outbreaks of respiratory disease in North America. African horse sickness, although foreign to North America, could be introduced despite stringent horse importation regulations. Specific antiviral therapy is not available to treat viral respiratory disease in the horse. A variety of inactivated and modified live vaccines, however, are available to prevent clinical disease and the spread of infection caused by the common viral respiratory pathogens. A considerable amount of research is underway to enhance the potency and duration of immunity of the present vaccines against influenza and rhinopneumonitis. This research is directed at defining and characterizing the importance of specific glycoprotein antigens on the surface of the virus, which trigger the various host immune responses, and determining whether they are stimulatory or suppressive.     

Determination of plasma lysosomal enzyme activity and its relationship to severity of injury following blunt vehicular trauma in dogs

Objective: To determine plasma β‐d ‐glucuronidase (βG) activity in the first 4 hours following injury in dogs struck by a motor vehicle, and to evaluate whether the degree of enzyme activity is correlated with the severity of injury. Design: A prospective clinical study. Setting: Veterinary Medical Teaching Hospital. Animals: Thirteen client‐owned dogs that were presented to the Veterinary Hospital of the University of Pennsylvania between June and August 1999 for blunt vehicular trauma. Ten healthy student and staff‐owned dogs served as controls. Interventions: None. Measurements: Plasma was analyzed for βG enzyme activity at the time of presentation (n=13), and 1 and 4 hours (n=7) following presentation to the Emergency Service for blunt vehicular trauma. The results were compared with enzyme activity from healthy controls evaluated serially over 4 hours. Fluorometric analysis using 96‐well microtiter plates was used to perform the enzyme assays. The relationships between presentation (n=13) and 4 hours (n=7) of enzyme activity and 3 indices of metabolic and physical disturbance (serum pH, serum lactate and Animal Trauma Triage (ATT) score) at the time of presentation were also investigated. Main results: Of the 13 dogs, 7 fulfilled the inclusion criteria for comparison of enzyme activity of the trauma over time. A statistically significant difference in βG activity was found in the trauma group (mean 75.6±10.4 U) at 4 hours following presentation compared with controls (mean 48.0±6.4 U). This difference was suggested by 1 hour following presentation (trauma group, mean 70.4±10.9 U; control group, mean 49.8±5.5 U), although it did not reach statistical significance. Thirteen dogs fulfilled the inclusion criteria for comparison of only presentation enzyme activity with trauma severity score, serum lactate, and serum pH. No statistically significant relationship was found between the βd ‐glucuronidase activity and the presenting ATT score, serum lactate concentration, or serum pH at either presentation or 4 hours, although the power of these analyses was low. Conclusions: These results demonstrate that the activity of βG, a lysosomal enzyme, increases significantly in the systemic circulation in dogs 4 hours following blunt trauma. Additional research to include more severely injured dogs, a larger number of dogs, and to follow the course of injury for a longer period of time would be beneficial to further characterize βG activity following blunt trauma.     

C-reactive protein concentrations in canine acute pancreatitis

Objective: To determine if C‐reactive protein (CRP) concentration is elevated in spontaneously occurring canine acute pancreatitis (AP), and to measure changes in CRP during the course of hospitalization. Design: Prospective study. Setting: Tufts University School of Veterinary Medicine Foster Hospital for Small Animals. Animals: Sixteen client‐owned dogs with AP and 16 healthy controls. Interventions: Blood samples were obtained from the AP group on the day of diagnosis (Day 1), and on Days 3 and 5, unless the dog died or was discharged from the hospital. Blood was obtained from the control dogs once. Measurements and main results: Serum CRP was measured using a commercial immunoassay for each dog with AP and for healthy controls. Day 1 CRP concentrations were significantly higher in the AP group (56.1±12.7 μg/mL) compared with controls (2.8±1.3 μg/mL; P<0.001). For the 7 dogs that had samples collected on all 3 days, the mean CRP concentrations decreased significantly (P=0.043) over the 5 days of measurement. Of the 16 dogs with AP, 14 were discharged from the hospital and 2 were euthanized. Conclusions: Serum CRP concentrations were elevated in this group of 16 dogs with spontaneously occurring AP. In the 7 dogs that had measurements on all 3 days, the mean CRP concentration decreased from the day of diagnosis to the measurement made 5 days later.     

Prevalence of feline herpesvirus 1, Chlamydophila felis, and Mycoplasma spp DNA in conjunctival cells collected from cats with and without conjunctivitis

OBJECTIVE: To use PCR assays to determine the prevalence of feline herpesvirus 1 (FHV-1), Chlamydophila felis, and Mycoplasma spp DNA in conjunctival cells collected from cats with and without conjunctivitis; to compare results of conventional and real-time fluorogenic PCR assays for amplification of FHV-1 DNA; and to determine whether copy numbers of FHV-1 DNA are correlated with conjunctivitis. ANIMALS: 55 cats with active conjunctivitis, 39 healthy cats that never had conjunctivitis, and 32 cats with a history of conjunctivitis that had been resolved for at least 3 months. PROCEDURES: Samples were obtained by rolling cotton-tipped applicators on the ventral conjunctiva of awake cats treated topically with proparacaine. The DNA was extracted from the swab specimens and assessed in PCR assays to detect DNA of FHV-1 (fluorogenic PCR assay and conventional PCR assay), Mycoplasma spp (conventional PCR assay), and C felis (conventional PCR assay). RESULTS: Overall prevalence rates of FHV-1, C felis, and Mycoplasma spp as assessed by the conventional PCR assays were 6.7%, 3.2%, and 9.6%, respectively. Percentage concordance between conventional PCR and fluorogenic PCR assays for FHV-1 was 92.5%. There were no significant differences among the 3 groups of cats for the mean copy number of FHV-1 divided by the copy number of glyceraldehyde-3-phosphate dehydrogenase. CONCLUSIONS AND CLINICAL RELEVANCE: Mycoplasma spp were the most prevalent organism detected and was associated with conjunctivitis. This study could not confirm that there are increased copy numbers of FHV-1 DNA in cats with conjunctivitis, compared with the copy numbers for cats without conjunctivitis.     

Assessment of EST- and genomic microsatellite markers for variety discrimination and genetic diversity studies in wheat

It is likely that in the near future sequence information from sequencing programmes and EST libraries will generate an abundance of genic microsatellite markers. This study is focused on the assessment of their likely impact and performance vis-à-vis their genomic counterparts. Microsatellites from two sources were used to assess the genetic diversity in 56 old and new varieties of bread wheat on the UK Recommended List. A set of 12 microsatellite markers generated from genomic libraries and 20 expressed sequence tag (EST)-derived microsatellites were used in the study, and the performance of both marker sets assessed. The EST-derived or genic microsatellites delivered fingerprints of superior quality, amplifying clear products with few stutter bands. Diversity levels as revealed bygenic microsatellites are similar to the few published results. The PIC values for the genic markers were generally lower than those calculated for the genomic microsatellites, though advantages of both marker classes for variety identification applications are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Chemical comparison of goldenseal (Hydrastis canadensis L.) root powder from three commercial suppliers

The characterization of herbal materials is a significant challenge to analytical chemists. Goldenseal (Hydrastis canadensis L.), which has been chosen for toxicity evaluation by NIEHS, is among the top 15 herbal supplements currently on the market and contains a complex mixture of indigenous components ranging from carbohydrates and amino acids to isoquinoline alkaloids. One key component of herbal supplement production is botanical authentication, which is also recommended prior to initiation of efficacy or toxicological studies. To evaluate material available to consumers, goldenseal root powder was obtained from three commercial suppliers and a strategy was developed for characterization and comparison that included Soxhlet extraction, HPLC, GC-MS, and LC-MS analyses. HPLC was used to determine the weight percentages of the goldenseal alkaloids berberine, hydrastine, and canadine in the various extract residues. Palmatine, an isoquinoline alkaloid native to Coptis spp. and other common goldenseal adulterants, was also quantitated using HPLC. GC-MS was used to identify non-alkaloid constituents in goldenseal root powder, whereas LC-MS was used to identify alkaloid components. After review of the characterization data, it was determined that alkaloid content was the best biomarker for goldenseal. A 20-min ambient extraction method for the determination of alkaloid content was also developed and used to analyze the commercial material. All three lots of purchased material contained goldenseal alkaloids hydrastinine, berberastine, tetrahydroberberastine, canadaline, berberine, hydrastine, and canadine. Material from a single supplier also contained palmatine, coptisine, and jatrorrhizine, thus indicating that the material was not pure goldenseal. Comparative data for three commercial sources of goldenseal root powder are presented.     

Buyer beware! Disease testing newly arrived cattle to dairy farms in Ontario

The objective of this cross-sectional study was to evaluate the presence of infectious disease in newly arrived cattle on dairy farms in Ontario. Cattle that were more than 2 years old and arrived at dairy farms within the previous year were tested. A total 321 cattle from 56 dairy farms were sampled and had blood submitted to a diagnostic laboratory. Of all sampled cattle, 0.0%, 39.6%, 2.2%, and 1.3% tested positive for Anaplasma, bovine leukemia virus, Mycobacterium avium subsp. paratuberculosis, and Salmonella Dublin, respectively. Based on these results, it is imperative that dairy producers are vigilant to ensure they do not purchase animals with these important and untreatable infectious diseases.