Oral immunization against rabies: afterthoughts and foresight

The article contains personal views on some issues that are frequently addressed in discussions about rabies control, and on some related topics that are often overlooked. The first field applications of oral wildlife rabies immunization in the Swiss Rhone Valley were preceded by many years of international cooperative studies on efficacy and safety. They were significant "faits accomplis" that facilitated similar endeavors in other countries. Some aspects of the residual pathogenicity of oral rabies vaccines are discussed. The field efficacy of oral wildlife immunization is the outcome of complex interactions between vaccine and bait attributes, bait distribution procedures, and habitat properties. Significant difficulties hinder the interpretation of field observations on efficacy. Though oral wildlife immunization is not an animal welfare act and not a conservationist achievement, it is an attempt at zoonosis control intended to protect human health and prevent economic losses.     

Oral vaccination of foxes against rabies in Turkey between 2008 and 2010

Following a sustained spill-over event from dogs to foxes, fox rabies spread rapidly in the Aegean region, Turkey. In order to control the outbreak a program of oral vaccination of foxes against rabies was introduced. In the selected vaccination area three annual campaigns between 2008 and 2010 were undertaken during the winter months whereby the vaccine baits were distributed exclusively by plane using a density of 18 baits per km2. Subsequently, fox rabies cases were reported only from locations bordering the non-vaccinated areas. Hence, it was shown that fox rabies control by means of oral rabies vaccination is feasible in Turkey. However, for the progress towards the elimination of fox-mediated rabies in Turkey to be maintained, it is necessary that political and financial support is secured to extend oral vaccination where infected foxes remain.     

Gene immunization of mice with plasmid DNA expressing rabies virus glycoprotein

Gene immunization can be an effective vaccine strategy eliciting both humoral and cell-mediated immune responses. We constructed plasmid vectors expressing the full-length Vnukovo-32 rabies virus glycoprotein G under the control of CMV IE promoter and enhancer, adenovirus tripartite leader sequences and poly A signal of SV40. The gene vaccines were evaluated for the ability to elicit neutralizing antibodies and to protect BALB/c mice against lethal rabies virus challenge. First, mice were injected intramuscularly (i.m.) into the left hind leg and by the intradermoplantar (i.d.p.) route with equal amounts of plasmid DNA (0.25-0.1 mg). Two weeks later, immunization was boosted with an additional dose of the DNA. The immunized mice were challenged by intracerebral (i.c.) inoculation of CVS-27 (10-50 LD50) rabies virus. All mice produced anti-rabies virus neutralizing antibodies with a titre of > or = 1:45 after immunization with 0.1-0.4 mg of DNA. In challenge experiments, 83 to 91.6% protection was observed. These results confirm that a DNA vaccine could be a simple and effective solution for preventing the spread of rabies.     

The development of strategies for the field application of oral immunization of foxes against rabies

Preconditions for successful field application of the idea to orally immunize foxes against rabies were a) the development of an efficacious and safe vaccine and an attractive bait and b) a suitable strategy to reach a high proportion of the vector population with vaccine-ladden baits. A geographical and epidemiological concept on how to implement the vaccination strategy and how to achieve a sufficiently high level of surveillance were further cornerstones contributing to the success of the method. Finally, political decisions and a legal basis for the application of the method of oral immunization were required to eradicate fox rabies in Switzerland.     

Efficacy of a baiting system for vaccinating foxes against rabies with vaccinia-rabies recombinant virus

The efficacy of a vaccinia-rabies recombinant virus (10(8) TCID50) contained in a machine-made baiting system has been tested in 22 captive young foxes which were divided into three experimental groups of six and a control group of four foxes. Each fox in groups 1, 2 and 3 were fed one, two and three vaccine-baits, respectively, on successive days. The four unvaccinated foxes were housed separately. As shown by the incorporation of a tetracycline biomarker into their bones, all the baited foxes ingested at least one bait. Thirty days after baiting seroconversion to rabies was observed in 15 (83 per cent) of the foxes and seroconversion to vaccinia in 14 (78 per cent). Sixteen of the 18 (89 per cent) baited foxes resisted a rabies challenge 30 days after baiting. One cub was protected against rabies despite the absence of detectable anti-rabies antibody. The results demonstrate that the bait-sachet system permits a good release of the virus suspension into the mouth.     

Oral rabies vaccination of foxes with one or two delayed distributions of SAG2 baits during the spring

During the spring of 1997, various protocols of rabies vaccine bait (SAG2) distribution for foxes were compared: in the first test zone, a first distribution was organised at the end of April, followed by a second distribution two weeks later; in the second test zone, there was a first distribution at the same period as for the previous zone, followed by a second distribution four weeks later, at the end of May. In two control zones, a classical single bait distribution was organised during the same periods as for the second distribution in the respective test zones. No statistical differences were observed for adult foxes or fox cubs sampled in the test and control zones neither for baits uptake nor for seroconversion rate. However, seroconversion rates observed in fox cubs population were significantly higher (P < 0.01) in areas vaccinated at the end of May (43 and 56%) compared with those vaccinated at mid-May (24 and 20%). The vaccinal efficacy of baits was also significantly (P < 0.05) increased for the fox cubs in the areas vaccinated at the end of May (46 and 57%) compared with those vaccinated at mid-May (24 and 25%). This increase in immunological response by fox cubs when vaccinating in late spring must be related to their development. In the early spring, fox cubs are generally too young to have access to baits or to be vaccinated when eating them. For most of these fox cubs, a second distribution will not constitute a booster. Therefore, in order to increase the efficient access of fox cubs to vaccine baits, Spring distribution of baits should preferably be organised during May or June rather than in April.     

Distribution of the rabies virus in the central nervous system of naturally infected foxes]

Quantitative distribution of rabies virus and the character of immunofluorescence in various parts of the central nervous system [CNS] and salivary glands were investigated in 21 naturally infected foxes. A mean death period was recorded in mice infected with the virus obtained from various parts of the CNS and salivary glands of the investigated foxes. The highest average titer of the virus was found out in the salivary glands - 4.46 log MLD50 per 0.03 ml i. c. The average titer was 2.30 log in the cornu ammonis, 1.99 log in the lobus olifactorius, 1.80 log in the spinal cord of the sacro-lumbar region, 1.73 log in the cortex, 1.71 log in the medulla oblongata, and 1.64 log in the cerebellum. The highest intensity of specific fluorscence was recorded in the thalamus, lobus piriformis, and cornu ammonis. In these regions, numerous round fluorescent inclusion bodies similar to Babes-Negri bodies occurred. Babes-Negri bodies in the cornu ammonis of foxes were proved in 81% of cases. Mice infected with the virus obtained from the salivary glands showed the shortest mean death period - 12.2 days and from the cornu ammonis it was 14.6 days. In virus infection from the other parts of the CNS the mean death period ranged from 16.0 to 17.4 days.     

Stability of attenuated live virus rabies vaccine in baits targeted to wild foxes under operational conditions

The viability of an attenuated live virus rabies vaccine in a bait targeted to red foxes was examined under various operational conditions in a series of experiments in Ontario. The virus was relatively stable over a 28-day period in the field, losing a mean 0.5, s = 0.2 log10 of virus titer. The micro-environment into which the bait was placed (open cultivated field, grassy meadow, wooded grove, sun or shade) did not make an appreciable difference in the viability of the virus. There was no significant difference (P < or = 0.05) between mean ambient temperatures and the temperature of fluids in blister packs of baits placed in sun or shade. Sixty-three percent of foxes fed baits exposed to sun and shade conditions for 21 days (titer 10(6.2) tissue culture infective doses per 1 mL) developed rabies virus-neutralizing antibodies. Storage of vaccine baits at -30 degrees C prior to bait distribution was important in maintaining virus viability.     

Optimising spring oral vaccination campaigns of foxes against rabies

To optimise spring vaccination campaigns the following set of data has been analysed; (i) population dynamics of the red fox, (ii) onset and progress of the reproductive season, and (iii) maternal immunity and the immune response of fox cubs to oral vaccination. The field data originated from foxes caught in Bavaria, Germany. The results of our analysis clearly demonstrate that certain periods during spring are less suitable for bait distribution. If the objective of a vaccination campaign is to reach only the adult foxes, it is suggested to conduct the campaign during the first half of March. If also young foxes are to be vaccinated, baits should not be distributed before the end of May in previously baited areas, because a large segment of the young fox population can not be vaccinated effectively before this date as a result of maternally transferred immunity. In areas vaccinated for the first time, baits can be distributed earlier, since 5 weeks old cubs are already immunocompetent.     

Attempted immunization of cats against feline infectious peritonitis, using avirulent live virus or sublethal amounts of virulent virus

Kittens vaccinated with an avirulent biotype of the Black strain of feline infectious peritonitis virus (FIPV; given oronasally) developed both indirect fluorescent and virus-neutralizing antibodies, but were not protected against oronasal challenge exposure with virulent virus. In fact, kittens vaccinated with avirulent virus were more readily infected than were nonvaccinated cats. A proportion of kittens could be immunized to FIPV by giving sublethal amounts of virulent virus. This technique, however, was too inconsistent and hazardous to have clinical relevance. The results of these studies indicated that humoral immunity was not protective in FIPV infection. There was no correlation between fluorescent and virus-neutralizing antibodies and either disease or immunity. Immune serum from FIPV-resistant cats failed to passively protect susceptible animals against virulent virus given intraperitoneally or oronasally, and as expected, actually sensitized them to infection. It was concluded that cell-mediated immunity was probably responsible for protection.     

Elimination of terrestrial rabies in Germany using oral vaccination of foxes

Oral rabies vaccination (ORV) has become the method of choice in fox rabies control in Europe. During the past three decades fox-mediated rabies virtually disappeared from Western and Central Europe. Following Switzerland, Germany was the second European country to launch ORV field trials on its territory in 1983. This paper provides a historical overview on the emergence of fox rabies in Germany; describing the basic principles and milestones of the German rabies eradication programme and presenting results of two decades of efforts to control the disease in foxes. Also, setbacks as well as country-specific differences and particularities on Germany's long way to rabies elimination in comparison to other European countries are addressed. Since the first field trials in Germany the number of rabies cases steadily decreased from 10 484 in 1983 to three cases recorded in 2006. On February 3rd 2006 the last case of terrestrial rabies in Germany was detected in a fox near the town of Mainz, Rhineland-Palatinate. In 2008, ORV ceased after 25 years and Germany was officially declared as free from terrestrial rabies. The German rabies eradication programme did cost approximately 100 million euro of which 37 million euro were covered by the EU. For the future, efforts should focus on maintaining a rabies free status by implementing measures to prevent reintroduction of terrestrial rabies from endemic countries.