The occurrence of rabies in pre-Columbian Central America: an historical search.

Rabies is considered one of the oldest infectious diseases known to humans. However, the first written reports on rabies cases in the Americas did not appear until the first decade of the 18th century from Mexico. In an attempt to clarify if the disease was already present in pre-Columbian times, we searched for evidence in the Maya and Aztec cultures. Other sources of information were early manuscripts written by the conquistadors and early explorers. We did not identify any unequivocal direct evidence that the disease rabies was known in pre-Columbian Central America but sufficient circumstantial evidence is available suggesting that (bat) rabies was already present in these early times.

Evaluation of an animal rabies vaccine by use of two types of potency tests.

OBJECTIVE: To evaluate the potency of an inactivated animal rabies vaccine for domestic animals by use of 2 types of potency tests after challenge exposure with a laboratory standard virus or 1 of 5 viruses obtained from various wildlife species. ANIMALS: 384 mice vaccinated twice intraperitoneally; 384 mice vaccinated once IM. PROCEDURE: Mice vaccinated with an inactivated, adjuvanted rabies vaccine for domestic animals were challenge exposed with the common fixed challenge virus or 1 of 5 rabies viruses obtained from wild animal species (street viruses) that most commonly transmit the virus in the United States and Canada. Potency tests included 2 types of antigen extinction tests: the National Institutes of Health (NIH) test and the Centers for Disease Control test. RESULTS: Results of both tests indicated that protection was highest against raccoon and bat viruses. Marked differences were detected in the relative potency ratios for the NIH versus the Centers for Disease Control tests, though the relative potencies themselves (against the street viruses) did not differ markedly. CONCLUSIONS: The markedly reduced potency against the street viruses indicated by the NIH test results was suggestive of an inherent bias associated with double intraperitoneal vaccination and intracerebral challenge exposure, whereas the single IM vaccination and IM challenge exposure reduced that bias.

Acceptance of candidate baits by domestic dogs for delivery of oral rabies vaccines.

Protocols for evaluating oral rabies vaccine baits for domestic dogs were field tested in central Mexico, after which dog-food manufacturers and suppliers to the pet-food industry were advised as to potential ingredients for use in prototype dog baits. Bait-preference trials in which confined dogs were used were then undertaken, followed by field tests of free-ranging farmer-owned dogs in three towns in the Nile River Delta region of Egypt. Both confined and free-ranging dogs showed strong preferences for certain baits or bait coatings (poultry, beef tallow, cheese, egg and a proprietary product). Fish-meal polymer baits, widely used for wildlife species, were less preferred. In Egypt, a commercial dog-food-meal bait coated with beef tallow and dry cheese, was consumed at a rate approaching that of a chicken-head bait. The percentage baits that were actually eaten after they had been offered to dogs, ranged from 71-96% for household dogs tested in Mexico, 65-91% for confined dogs (beagles and mixed breeds) tested in the United States, and 32-88% for farmer-owned dogs tested in Egypt.

Evaluation of the safety of two attenuated oral rabies vaccines, SAG1 and SAG2, in six Arctic mammals.

The safety of two attenuated oral rabies vaccines was evaluated in mink and in five species of rodents which occur in the Arctic. A 0.03 ml sample of liquid vaccine was installed directly into the mouth of voles and lemmings and 0.1 ml into the mouth of Arctic ground squirrels and mink. Animals were euthanized at 36 and 46 days postexposure; brain tissue was analyzed by FAT and serum by RFFIT. No rabies deaths occurred in 47 animals tested. Four animals representing three rodent species seroconverted, the highest titer being 0.5 IU ml-1. The absence of rabies virus in brain tissue indicates the safety of these vaccines in these species. The replacement of arginine with glutamic acid at position 333 reduces the pathogenicity of these vaccines, thereby presumably preventing the deleterious effect of viral entry into CNS neurons.

Immunogenicity of rabies vaccines used during an urban epizootic of rabies in Mexico.

From 1 July 1987 to 31 December 1988, 30% of 247 rabid dogs in Hermosillo, Mexico had a positive history of rabies vaccination. Serosurveys suggested that inactivated suckling mouse brain vaccine (INACT-SMBV) and inactivated tissue culture vaccine (INACT-TC) used before and during the epizootic were poor immunogens. Prospective studies showed that only about one-third of dogs vaccinated with INACT-SMBV were seropositive 5 weeks after vaccination. Lack of vaccine potency was the most likely cause of poor immunogenicity. Rabies vaccines should be evaluated periodically by measuring antibody responses in animals. In some circumstances, minimum seroconversion rates and antibody titres in vaccinated animals may be better measures of immunogenicity than relative potency.

Sickness and recovery of dogs challenged with a street rabies virus after vaccination with a vaccinia virus recombinant expressing rabies virus N protein.

Dogs were vaccinated intradermally with vaccinia virus recombinants expressing the rabies virus glycoprotein (G protein) or nucleoprotein (N protein) or a combination of both proteins. The dogs vaccinated with either the G or G plus N proteins developed virus-neutralizing antibody titers, whereas those vaccinated with only the N protein did not. All dogs were then challenged with a lethal dose of a street rabies virus, which killed all control dogs. Dogs vaccinated with the G or G plus N proteins were protected. Five (71%) of seven dogs vaccinated with the N protein sickened, with incubation periods 3 to 7 days shorter than that of the control dogs; however, three (60%) of the five rabid dogs recovered without supportive treatment. Thus, five (71%) of seven vaccinated with the rabies N protein were protected against a street rabies challenge. Our data indicate that rabies virus N protein may be involved in reducing the incubation period in dogs primed with rabies virus N protein and then challenged with a street rabies virus and, of more importance, in subsequent sickness and recovery.

Oral rabies vaccination of arctic foxes (Alopex lagopus) with an attenuated vaccine.

Arctic foxes were immunized with the SAG1 oral rabies vaccine. The effectiveness was determined by the serological response and by the survival to a challenge dose of rabies virus from an Alaskan fox. Vaccine virus was isolated from saliva 1 h after the liquid vaccine was placed directly into the mouth but not subsequently (tested up to 1 week postvaccination). Two weeks after vaccination, protective antibody levels were present in all foxes and all vaccinated foxes survived challenge at 9 weeks postvaccination. At 26 weeks postvaccination (17 weeks postchallenge) all but one fox had detectable antibody levels. Neural tissue harvested from surviving foxes was negative for rabies virus by direct immunofluorescent testing. One of the foxes vaccinated with SAG1 seroconverted and survived challenge even though the titre of the vaccine used was almost 4 logs less than that used to vaccinate the other foxes. These results suggest that the avirulent SAG1 oral rabies vaccine is very effective in protecting arctic foxes.

Oral vaccination of skunks with raccoon poxvirus recombinants expressing the rabies glycoprotein or the nucleoprotein.

Twenty nine skunks (Mephitis mephitis) were vaccinated orally with raccoon poxvirus (RCN) recombinants: 10 with a recombinant expressing the rabies virus glycoprotein (RCNRG), 10 with RCNRG mixed with a recombinant expressing the rabies virus nucleoprotein (RCNRN) and nine with RCN alone. Rabies virus neutralizing antibodies were detected in six of the 20 skunks; five skunks (three given RCNRG, two given a mixture of recombinants) survived a rabies challenge that was lethal for nine skunks vaccinated with RCN alone.

The protective role of humoral neutralizing antibody in the NIH potency test for rabies vaccines.

Intraperitoneal vaccination of mice with rabies vaccine results in both dosage-dependent rabies virus neutralizing antibody titres and protection from lethal intracerebral (i.c.) challenge with fixed strain CVS rabies virus. Pre-exposure adoptive intravenous transfer of naive or immune cells did not significantly protect naive Balb/c mice from lethal i.c. CVS challenge, but immune serum and anti-rabies glycoprotein monoclonal antibodies (individually and in combination) did confer significant protection when administered before or up to 24 h after lethal i.c. rabies virus challenge.

One-year study of the 2-1-1 intramuscular postexposure rabies vaccine regimen in 100 severely exposed Thai patients using rabies immune globulin and Vero cell rabies vaccine.

The 2-1-1 rabies postexposure treatment schedule is an abbreviated regimen in which a tissue culture rabies vaccine is administered intramuscularly at two sites on day 0, and at one site on days 7 and 21. Compared to the standard five-dose intramuscular regimen, the 2-1-1 schedule reduces the number of clinic visits from five to three and the amount of vaccine used by 20%. One hundred Thai patients, who were severely exposed to rabies, were treated with rabies immune globulin and the 2-1-1 regimen using purified Vero cell rabies vaccine. They were followed for 1 year. Rabies antibody titres were measured in 10% of this group. All patients survived and adverse reactions were mild. A satisfactory antibody response (a titre greater than 0.5 IU ml-1) occurred in all ten patients studied at day 14, but persisted for 90 days in 80% and for 360 days in only 50%. The authors therefore do not recommend use of the 2-1-1 schedule in severely exposed patients who also need to receive rabies immune globulin.

Formulation and evaluation of baits for oral rabies vaccination of raccoons (Procyon lotor).

Captive raccoons were offered a variety of vaccine containers and bait components in a series of three-choice tests. Paraffin wax ampules were the most readily accepted vaccine container. Preferred bait components included corn and shellfish oils, deep fried corn meal batter, and egg, apple and buttermilk flavorings. These results, together with factors including ease of bait formulation, cost, and suitability for field use, were used to develop an experimental delivery system for an oral rabies vaccine. The developed system was composed of a polyurethane sleeve (1.5 x 5.5 cm) dipped in a commercial food batter mix together with corn meal, milk and egg. The sleeve was deep fried in corn oil and a 2.0 ml ampule containing a recombinant rabies vaccine was then inserted into the sleeve bait. These baits were presented to 10 captive raccoons. Nine of the 10 animals developed high levels of rabies virus neutralizing antibodies. Field tests are needed to determine if the delivery system developed also is effective for wild raccoons.

Postexposure prophylaxis for rabies with antiserum and intradermal vaccination.

The Thai Red Cross intradermal postexposure rabies treatment schedule was prospectively assessed in 100 Thai patients severely bitten by proven rabid animals. It consists of 0.1 ml of purified Vero cell rabies vaccine containing more than 2.5 IU of rabies antigen per 0.5 ml of reconstituted vaccine given intradermally at two sites on days 0, 3, and 7, followed by one 0.1 ml injection on days 30 and 90. The commercial vaccine used had an antigen content of 3.17 IU per 0.5 ml ampoule. Purified equine or human rabies immuno-globulin was also given on day 0 to patients with severe exposures. As much of the immunoglobulin as possible was infiltrated around the wounds. All patients were followed for 1 year post exposure. There were no deaths; the efficacy of the regimen was 100%. Antibody titre determination in a randomly selected subgroup showed seroconversion in all 10 patients.

Oral vaccination of dogs fed canine adenovirus in baits.

Six groups of 5 dogs each were fed dilutions of canine adenovirus-2, either as raw liquid or after insertion into cornmeal baits. By the fourth week after vaccination, 29 of the 30 dogs developed high titers of serum-neutralizing antibodies to the virus.

Acceptance of simulated oral rabies vaccine baits by urban raccoons.

In summer 1986, a study was conducted to evaluate raccoon (Procyon lotor) acceptance of oral baits that could be used for rabies vaccination. One thousand wax-coated sponge bait cubes were filled with 5 mg of a seromarker (iophenoxic acid), placed in polyethylene bags, and hand-distributed in an 80 ha area within an urban National Park in Washington, D. C. (USA). After 3 wk, target and nontarget animals were trapped and blood samples collected to evaluate bait uptake. Thirty-three of 52 (63%) raccoons had elevated blood iodine levels indicating they had eaten at least one bait, 13 (25%) were negative, and six (12%) had marginal values. These results indicate that sponge baits hand-placed at a density of 12.4/ha can reach a significant proportion of an urban raccoon population. Implications for oral rabies vaccination of raccoons are discussed.