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.     

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.     

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.     

Biochemical composition and quality of captive-spawned cobia Rachycentron canadum eggs

Interest in the commercial production of cobia Rachycentron canadum continues to rise as additional insight is gained into the hardy and fast growing nature of this species. However, research regarding the biochemical composition of captive-spawned eggs and egg and larval quality remains scarce. Such data is essential as a common bottleneck to production is a steady supply of fingerlings for grow-out. This study quantified the biochemical composition and quality of cobia eggs produced over 2 spawning seasons by broodstock on a traditional ‘trash fish’ diet which is commonly fed to tank spawning cobia. Throughout the study, batch fecundity, proportion of floating eggs and percent hatch averaged > 1 million eggs,  0.8 and 70%, respectively. Batch fecundity was significantly higher during the second spawning season as a result of the increased size of the females which weighed 18/22 kg and 22/26 kg at the beginning of each season. A positive correlation was found between the proportion of floating eggs and hatch rate for both spawning seasons. No correlations were found between egg composition (total lipid (30.0 ± 1.1% dry wt), protein (25.4 ± 2.2% dry wt), carbohydrate (2.4 ± 0.3% dry wt), vitamin E (10.2 ± 0.6 μg/g wet wt) or dry weight (119.1 ± 5.5 μg/egg)) and egg quality (proportion of floating eggs, hatch rate, larval growth and larval survival). Further, no differences in egg composition were noted between seasons or over the course of each season. The fatty acid composition of cobia eggs varied between seasons possibly due to changes in the quality of frozen feed (fish, shrimp, squid) given to the broodstock. The only correlation between the fatty acid profile and egg quality was a decrease in the proportion of floating eggs as the total amount of n-3 highly unsaturated fatty acids increased. No relationship between egg quality and amino acid content was noted with the most prominent amino acids being glutamate, leucine, alanine, proline, lysine and aspartate nor were any differences detected between spawning seasons.     

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.     

Frequent mutation of BAP1 in metastasizing uveal melanomas

Metastasis is a defining feature of malignant tumors and is the most common cause of cancer-related death, yet the genetics of metastasis are poorly understood. We used exome capture coupled with massively parallel sequencing to search for metastasis-related mutations in highly metastatic uveal melanomas of the eye. Inactivating somatic mutations were identified in the gene encoding BRCA1-associated protein 1 (BAP1) on chromosome 3p21.1 in 26 of 31 (84%) metastasizing tumors, including 15 mutations causing premature protein termination and 5 affecting its ubiquitin carboxyl-terminal hydrolase domain. One tumor harbored a frameshift mutation that was germline in origin, thus representing a susceptibility allele. These findings implicate loss of BAP1 in uveal melanoma metastasis and suggest that the BAP1 pathway may be a valuable therapeutic target.     

Treatment of anovulatory anoestrous postpartum dairy cows with a gonadotropin-releasing hormone (GnRH), prostaglandin F2 alpha, GnRH regimen or with progesterone and oestradiol benzoate

AIM: To compare 2 treatments for anovulatory anoestrus (AA) in postpartum dairy cows. The treatments were combinations of gonadotropin-releasing hormone (GnRH) and prostaglandin F2 (PG) or progesterone (P4) and oestradiol benzoate (ODB). METHODS: Forty AA cows from each of 5 herds were blocked by age (2 or 2 years old) and randomly assigned to 1 of 2 treatments. The first group (GPG) were treated with 250 mug of a GnRH analogue, gonadorelin, followed 7 days later by 15 mg of the PG analogue, luprostiol. Two days later the cows were injected with 250 mug of gonadorelin. Cows were artificially inseminated 16-24 h after the second GnRH injection. The second group (P4+ODB) were treated with an intravaginal P4 releasing device for 6 days, followed 24 h after device removal by injection of 1 mg of ODB. Cows were pregnancy tested 35-40 days after the initial insemination and twice again at 6-8 week intervals thereafter. RESULTS: There was no significant difference between P4+ODB and GPG groups in the percentage of cows submitted for insemination in the first 7 days (94.0% vs 100% for P4+ODB vs GPG, respectively; p>0.3), in conception rate to first insemination within the first 7 days (43.6% vs 35.0% for P4+ODB vs GPG, respectively; p>0.2), in the percentage of cows conceiving in the first 28 days of the breeding period (68.0% vs 58.3%, P4+ODB vs GPG, respectively; p>0.1), or in median interval from the end of treatment to conception (20 vs 21 days; p>0.1). CONCLUSIONS: No differences in the reproductive performance of AA cows treated with either P4+ODB or GPG were detected. However, given the small number of animals enrolled, further data are required before the GPG protocol can be recommended for treatment of AA cows.