Efficacy of Ontario Rabies Vaccine Baits (ONRAB) against rabies infection in raccoons.

In the US, rabies lyssavirus (RABV) only circulates in wildlife species and the most significant reservoir from a public and animal health perspective is the raccoon (Procyon lotor). Management of wildlife rabies relies principally on oral rabies vaccination (ORV) strategies using vaccine-laden bait delivery to free-ranging target hosts, in order to reduce the susceptible population to prevent the spread of and eliminate RABV circulation. Our objective was to evaluate efficacy of the Ontario Rabies Vaccine Bait (ONRAB) against a lethal RABV challenge in captive raccoons. Sham or live vaccine baits were offered to 50 raccoons and efficacy was evaluated in 46, split into two trials of 17 and 29 raccoons. Raccoons were challenged with a lethal dose of RABV 180 days post-vaccination and observed for 90 days post-infection. Raccoon bait interactions were assigned increasing integer scores for approach, oral manipulation, puncture, and consumption behaviors. Higher bait interaction scores were observed in the fall compared to the spring trial, indicating that more raccoons consumed baits in the fall. Although animal age did not explain variation in bait interaction scores, the geometric mean rabies virus antibody titers among juvenile vaccinates were higher than adults at all pre-challenge time points. The prevented fraction associated with ONRAB delivery was 0.73 (8/11, 95% CI 0.39-0.94) in the spring trial and 0.91 (21/23, 95% CI 0.72-0.99) in the fall trial. All sham-vaccinated raccoons (12/12) succumbed to rabies infection, in contrast to 15% (5/34) mortality among vaccinated raccoons. Our results indicate a high efficacy of ONRAB bait vaccination in protecting adult and juvenile raccoons against RABV infection for a minimum of six months. These data complement experimental field trials that have also demonstrated the potential of ONRAB for the control and prevention of RABV circulation in free-ranging raccoon populations in the US.

FLAVOR PREFERENCE AND EFFICACY OF VARIABLE DOSE ONTARIO RABIES VACCINE BAIT (ONRAB) DELIVERY IN STRIPED SKUNKS ( MEPHITIS MEPHITIS).

In North America, terrestrial wildlife rabies control is achieved by oral rabies vaccination programs that principally target mesocarnivores. Success at rabies control in striped skunks ( Mephitis mephitis) has been more limited and may require additional enhancements to existing bait products or novel bait designs and attractants. We evaluated preference among captive striped skunks for six different flavors of placebo Ontario Rabies Vaccine Bait (ONRAB®) "Ultralite" Baits (Artemis Technologies, Guelph, Ontario, Canada). Different doses and delivery methods of ONRAB vaccine were tested for efficacy in a subsequent experiment with the same skunks. Cheese-, egg-, and chicken-flavored baits were preferred over plain-flavored baits, but a strong preference for a singular flavor was not observed. Vaccine efficacy of 80-100% was observed among skunks challenged at 335 d postvaccination across a log range of doses tested by a direct instillation into the oral cavity route, respectively (109.3-1010.2 median tissue culture infective doses), in contrast to more-limited efficacy by bait delivery. Our results extended the duration of ONRAB vaccine efficacy in skunks and suggested that there may be limited flexibility to alter vaccine titer and volume in novel bait designs for skunks.

Process optimization and scale-up for production of rabies vaccine live adenovirus vector (AdRG1.3).

Rabies virus is an important causative agent of disease resulting in an acute infection of the nervous system and death. Although curable if treated in a timely manner, rabies remains a serious public health issue in developing countries, and the indigenous threat of rabies continues in developed countries because of wildlife reservoirs. Control of rabies in wildlife is still an important challenge for governmental authorities. There are a number of rabies vaccines commercially available for control of wildlife rabies infection. However, the vaccines currently distributed to wildlife do not effectively immunize all at-risk species, particularly skunks. A replication competent recombinant adenovirus expressing rabies glycoprotein (AdRG1.3) has shown the most promising results in laboratory trials. The adenovirus vectored vaccine is manufactured using HEK 293 cells. This study describes the successful scale-up of AdRG1.3 adenovirus production from 1 to 500 L and the manufacturing of large quantities of bulk material required for field trials to demonstrate efficacy of this new candidate vaccine. The production process was streamlined by eliminating a medium replacement step prior to infection and the culture titer was increased by over 2 fold through optimization of cell culture medium. These improvements produced a more robust and cost-effective process that facilitates industrialization and commercialization. Over 17,000 L of AdRG1.3 adenovirus cultures were manufactured to support extensive field trials. AdRG1.3 adenovirus is formulated and packaged into baits by Artemis Technologies Inc. using proprietary technology. Field trials of AdRG1.3 rabies vaccine baits have been conducted in several Canadian provinces including Ontario, Quebec and New Brunswick. The results from field trials over the period 2006-2009 demonstrated superiority of the new vaccine over other licensed vaccines in immunizing wild animals that were previously difficult to vaccinate.