Effects of cyclophosphamide myelosuppression in adult dogs with latent canine herpesvirus-1 infection

Latent canine herpesvirus-1 (CHV-1) infection is common in domestic dogs, but triggers for viral reactivation and recrudescent CHV-1 disease are poorly understood. Cyclophosphamide is a potent immunosuppressive and myelosuppressive agent used for the therapy of a variety of neoplastic and immune-mediated canine disorders. Cyclophosphamide (200mg/m(2)) was administered to mature dogs latently infected with CHV-1 to determine its potential to induce recurrent CHV-1 disease and viral shedding. Non-infected dogs and dogs recovered from experimental primary ocular CHV-1 infection with experimentally confirmed latent CHV-1 infection were divided into groups and administered cyclophosphamide or placebo. Dogs were monitored for myelosuppression and viral reactivation for 28days using clinical and virological outcome measures. Clinical ophthalmic and in vivo ocular confocal microscopic examinations were performed at intervals. Samples were collected for CHV-1 polymerase chain reaction (PCR), CHV-1 virus neutralizing (VN) antibody, and hemogram assays. Myelosuppression (i.e., decreased total leukocyte, segmented neutrophil, and erythrocyte counts) was detected on study day 7 in dogs administered cyclophosphamide, but not dogs administered placebo. There were no abnormalities suggestive of recurrent CHV-1 ocular disease during clinical ophthalmic or in vivo confocal microscopic examination in any dogs during the study. Ocular CHV-1 shedding was not detected by PCR and CHV-1 VN titers remained stable in all dogs. Following study conclusion, the presence of reactivatable latency was reconfirmed in the infected dogs by administering systemic prednisolone. Myelosuppression elicited by a single dose of cyclophosphamide does not result in detectable recurrent ocular CHV-1 infection in adult dogs with experimentally induced latent CHV-1 infection.     

Recent developments in genome editing and applications in plant breeding

Increasing genetic variation beyond natural variation is an important aim in plant breeding. In the past 70 years, random mutagenesis by irradiation or by chemicals has created numerous mutants which have been frequently used in breeding. However, their application is hampered by the mutational load due to many background mutations. In the past 10 years, new techniques for site‐directed mutagenesis have been introduced to plant breeding which are commonly referred to as “genome editing.” Among these, the CRISPR/Cas9 system turned out to be the most efficient and easy to apply. DNA is cleaved by a nuclease precisely at a target site where a mutation is likely to be beneficial. The DNA is healed by the cellular repair system either by error‐prone non‐homologous end joining or by homologous recombination, by which small DNA fragments can be inserted at the target site. In this review, we describe the application of targeted mutagenesis to crop plants and the modification of agronomically important traits, which could have direct impacts on plant breeding.