Recorded Marked Changes in the Haematological and Immune Responses of Two Non-Transgenic Rodents Inoculated Orally and Intraperitoneally with <i>Trypanosoma brucei brucei</i>

Objectives: The digestive track of mice and humans has always been an integral part of the pathogenesis of the Trypanosomes but is constantly overlooked. This realization opens up completely new strategies for the development of trypanosomes vaccines, allowing approaches that parenteral delivery forms would not permit. The target of the study was to compare the haematological changes and immunological responses of trypanosomiasis model systems (mice and rats) inoculated orally and intraperitoneally and to observe the afterward effect of a controlled drug [Isometamidium chloride (ISM)] in the restoration of these initial parameters. Methods: To achieve this, a total of 40 rodents (20 rats and 20 mice) were purchased, then grouped into two [sixteen younger (1 - 5 weeks) and older (7 - 15 weeks) groups each]. They were further sub-grouped into five each. Body weights, Parasitaemia and Packed Cell Volume (PCV) were taken before, after inoculation and after treatment with ISM at 4 mg/kg. Results: Based on presumptive clinical diagnosis, all rodents inoculated intraperitoneally showed clinical signs of fluctuations in weight, PCV and parasitaemia levels before, after inoculations and after treatment compared to those inoculated orally with a significant difference (P < 0.05) observed. Both young and older rodents also responded differently to the inoculants and to the different methods of inoculation. But more deaths were recorded among the mice when compared to the rats. Conclusion: Although these non-transgenic models would not have offered a completely new methods to vaccine development, their differences in response to various methods of inoculations is an indication of an exciting research processes and could offer desired results, particularly where transgenic rodents are employed.

Engineered biocontainable RNA virus vectors for non-transgenic genome editing across crop species and genotypes

CRISPR/Cas genome-editing tools provide unprecedented opportunities for basic plant biology research and crop breeding.However,the lack of robust delivery methods has limited the widespread adoption of these revolutionary technologies in plant science.Here,we report an efficient,non-transgenic CRISPR/Cas delivery platform based on the engineered tomato spotted wilt virus(TSWV),an RNA virus with a host range of over 1000 plant species.We eliminated viral elements essential for insect transmission to liberate genome space for accommodating large genetic cargoes without sacrificing the ability to infect plant hosts.The resulting non-insect-transmissible viral vectors enabled effective and stable in planta delivery of Cas12a and Cas9 nucleases as well as adenine and cytosine base editors.In systemically infected plant tissues,the deconstructed TSWV-derived vectors induced efficient somatic gene mutations and base conversions in multiple crop species with little genotype dependency.Plants with heritable,bi-allelic mutations could be readily regenerated by culturing the virus-infected tissues in vitro without antibiotic selection.Moreover,we showed that antiviral treatment with ribavirin during tissue culture cleared the viral vectors in 100%of regenerated plants and further augmented the recovery of heritable mutations.Because many plants are recalcitrant to stable transformation,the viral delivery system developed in this work provides a promising tool to overcome gene delivery bottlenecks for genome editing in various crop species and elite varieties.