Thrombocytopenia in Cats: A Retrospective Study of 41 Cases

The prevalence of feline thrombocytopenia (<200,000 platelets/L) at North Carolina State University, College of Veterinary Medicine Teaching Hospital, from January 1985 to March 1990, was 1.2% (41/3300). Cats were divided into six categories based on clinical diagnoses: 29% (12/41) had infectious disease, 20% (8/41) had neoplasia, 7% (3/41) had cardiac disease, 2% (1/41) had primary immune-mediated disease, 22% (9/41) had multiple diseases, and 20% (8/41) had disorders of unknown etiology. The mean platelet count for all thrombocytopenic cats was 52,000/μL ± 46,000/μL (1 SD) with a range of 1000–190,000/μL. No significant differences were found between groups with respect to platelet count, packed cell volume, or white blood cell count, though anemia and leukopenia were common among the cats as a whole. Bleeding disorders (hemorrhage or thrombosis) were observed in 29% (12/41) of thrombocytopenic cats and were more likely to be associated with neoplasia, cardiac disease, and platelet counts less than or equal to 30,000/μL. Disseminated intravascular coagulopathy was diagnosed in 12% (5/41) of the cats. Infections and/or neoplasia affecting the bone marrow were the most common diseases associated with thrombocytopenia. Feline leukemia virus and myeloproliferative neoplasia accounted for approximately 44% (18/41) of the specific diagnoses in thrombocytopenic cats. (Journal of Veterinary Internal Medicine 1993; 7:261–265. Copyright © 1993 by the American College of Veterinary Internal Medicine.)     

Cypermethrin inhibits egg development in the ixodid tick, Amblyomma hebraeum

The pyrethroid insecticide, cypermethrin (CyM), stimulates vitellogenesis in Ornithodoros moubata (an argasid tick) by stimulating the release of the normal vitellogenesis-inducing factor (a neuropeptide) and subsequent release of the vitellogenic hormone [19]. Here we examine the effects of CyM on egg development in the ixodid tick, Amblyomma hebraeum. Ovary weight, oocyte size, and vitellin content of the ovary were measured after CyM treatment; in partially fed ticks, none of these parameters were affected significantly. However, CyM treatment caused an inhibition of ovary development, as well as reduction of both hemolymph 20-hydroxyecdysone (20E; the vitellogenic hormone in this species) and vitellogenin (Vg)-concentrations in engorged ticks. In addition, the degree of salivary gland degeneration (which is triggered by 20E) was slightly reduced in CyM-treated engorged ticks. These results indicate that CyM acts differently in Amblyomma compared to Ornithodoros. Instead of stimulating vitellogenesis, CyM inhibits egg development perhaps in part as a result of inhibiting release of 20E.     

Randomly primed,strand-switching,MinION-based sequencing for the detection and characterization of cultured RNA viruses

RNA viruses rapidly mutate, which can result in increased virulence, increased escape from vaccine protection, and false-negative detection results. Targeted detection methods have a limited ability to detect unknown viruses and often provide insufficient data to detect coinfections or identify antigenic variants. Random, deep sequencing is a method that can more fully detect and characterize RNA viruses and is often coupled with molecular techniques or culture methods for viral enrichment. We tested viral culture coupled with third-generation sequencing for the ability to detect and characterize RNA viruses. Cultures of bovine viral diarrhea virus, canine distemper virus (CDV), epizootic hemorrhagic disease virus, infectious bronchitis virus, 2 influenza A viruses, and porcine respiratory and reproductive syndrome virus were sequenced on the MinION platform using a random, reverse primer in a strand-switching reaction, coupled with PCR-based barcoding. Reads were taxonomically classified and used for reference-based sequence building using a stock personal computer. This method accurately detected and identified complete coding sequence genomes with a minimum of 20× coverage depth for all 7 viruses, including a sample containing 2 viruses. Each lineage-typing region had at least 26× coverage depth for all viruses. Furthermore, analyzing the CDV sample through a pipeline devoid of CDV reference sequences modeled the ability of this protocol to detect unknown viruses. Our results show the ability of this technique to detect and characterize dsRNA, negative- and positive-sense ssRNA, and nonsegmented and segmented RNA viruses.     

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.