Morphologic and immunoperoxidase study of neurologic lesions in naturally acquired rabies of raccoons.

Histopathologic (hematoxylin and eosin [HE]) and immunoperoxidase (streptavidin-biotin complex) methods were used for examination of formalin-fixed tissues of rabid raccoons from an enzootic area of Pennsylvania. Extensive morphologic lesions of rabies encephalitis were present in the cerebrum and the brain stem regions. Negri bodies were detected by both methods and were present in the brain (cerebral cortex, hippocampus, brain stem, cerebellum, and cervical spinal cord) and in the ganglia of the trigeminal nerves. The viral inclusions were also seen in ganglion cells in the tongue, parotid salivary glands, pancreas, intestines, and adrenal glands. These sites were not associated with any inflammatory cellular infiltrate. The immunoperoxidase method was superior to HE for the detection of Negri bodies. Because lesions of rabies encephalitis were consistently observed in the cerebrum, brain stem, and cervical spinal cord regions, these areas of the brain should be included when raccoons are examined by the fluorescent antibody test for rabies.     

Benefits and costs of using an orally absorbed vaccine to control rabies in raccoons.

In November 1989, the epizootic of rabies affecting raccoons in the mid-Atlantic states reached New Jersey. An economic evaluation was conducted in 2 counties first affected by the epizootic to estimate the costs of the epizootic and to assess the costs and benefits of orally administering a newly developed recombinant rabies vaccine to prevent further spread of the disease. Data on expenditures associated with prevention of rabies in human beings and domestic animals and laboratory testing of suspect animals were collected and analyzed for 1988 (before the epizootic) and 1990 (first full year of the epizootic). Benefit-cost ratios were calculated and used to evaluate the economic advisability of the vaccine at various vaccination program alternatives. Two indices of capital investment analysis, payback period and net present value, were used to evaluate the economic benefits of the rabies vaccine. Expenditures were estimated to be $1,952,014 in 1990 (primarily for pet animal vaccinations), compared with $768,488 in 1988. Benefit-cost ratios ranged from 2.21 for the most expensive vaccination program alternative to 6.80 for the least expensive alternative. The payback period varied from 0.69 to 2.11 years, and the net present value ranged from $2,105,453 to $4,877,452. The high costs of this epizootic necessitated the reallocation of scarce public health resources to various rabies prevention activities, particularly the vaccination of dogs. This study also demonstrated the usefulness of benefit-cost analysis in developing public health strategies. Although the mass application of this recombinant vaccine was found to be economically beneficial, other qualitative considerations must be used to supplement these findings.     

Use of infrared thermography to detect signs of rabies infection in raccoons (Procyon lotor).

Infrared thermography was evaluated as a technique to determine if raccoons (Procyon lotor) experimentally infected with rabies virus could be differentiated from noninfected raccoons. Following a 10-day adjustment period, raccoons (n = 6) were infected with a virulent rabies street strain raccoon variant by injection into the masseter muscle at a dose of 2 x 10(4) tissue-culture infectious dose (TCID50) in 0.2 ml (n = 4) or 10(5) TCID50 in 1 ml (n = 2). Five of the six raccoons developed prodromal signs of rabies 17 to 22 days postinoculation (PI) and distinctive clinical signs of furious rabies between 19 and 24 days PI. At the time of euthanasia, which occurred 2 days after the onset of clinical signs of rabies, these five raccoons tested positive for rabies virus in brain tissue. Infrared thermal images of each raccoon were recorded twice daily during the preinoculation and PI periods. No apparent differences were identified among thermal temperatures compared among days for the eye, average body surface, and body temperature recorded from subcutaneous implants throughout the experiment for any of the six raccoons. However, increases in infrared surface temperature of the noses and differences in the visual thermal images of the noses were detected when animals began showing clinical signs of rabies. Differences were detected among the mean infrared nose temperatures for the disease progression intervals (F3.12 = 70.03, P < 0.0001). The mean nose temperature in the clinical rabies stage (30.4 +/- 3.5 degrees C) was significantly elevated over the prodromal stage (F1,12 = 151.85, P < 0.0001). This experiment provides data indicating that infrared thermography can be used in an experimental setting to detect raccoons in the infectious stage and capable of exhibiting clinical signs of rabies.     

Cytauxzoon felis infection in cats in the mid-Atlantic states: 34 cases (1998-2004)

OBJECTIVE: To describe the demographic and clinical characteristics of feline cytauxzoonosis in the mid-Atlantic states and compare the Cytauxzoon felis 18S rRNA gene sequences from affected cats with sequences reported from affected cats in other regions. DESIGN: Retrospective case series. ANIMALS: 34 cats with C. felis infection. PROCEDURE: Medical records of cats in which C. felis infection was diagnosed from May 1998 through June 2004 were reviewed; data collected included signalment, month of diagnosis, geographic location, clinicopathologic abnormalities, medical treatments, outcome, and necropsy findings when applicable. Cytauxzoon felis DNA was amplified, cloned, and sequenced from 4 of these cats and compared with previously reported C. felis DNA sequences. RESULTS: Of 34 C. felis-infected cats, 28 resided in North Carolina, 3 resided in South Carolina, and 3 resided in Virginia; in 32 cats, a diagnosis of C. felis infection was made in April through September. Pancytopenia and icterus were the most common clinicopathologic abnormalities. Thirty-two cats either died or were euthanatized, and 2 cats survived. At 5 veterinary hospitals, multiple cases were identified, and 4 multicat households had > 1 cat infected with C. felis. The 18S rRNA gene sequences characterized in organisms obtained from 4 cats were nearly identical to C. felis DNA sequences reported from other US regions. CONCLUSIONS AND CLINICAL RELEVANCE: Data indicate that veterinarians in the mid-Atlantic region of the United States should consider C. felis infection in cats that become ill with fever, icterus, and pancytopenia or bicytopenia, especially in the spring and summer months.     

Porcupine quills in raccoons as an indicator of rabies, distemper, or both diseases: disease management implications

A relationship was detected between the presence of embedded porcupine quills and the diagnosis of rabies in raccoons in eastern Canada during 1999-2004. No relationship was found between the presence of quills in raccoons and the diagnosis of canine distemper. Raccoons with embedded quills should be submitted for rabies testing.     

Control of terrestrial animal rabies in Anne Arundel County, Maryland, after oral vaccination of raccoons (1998-2007)

Objective-To evaluate the effectiveness of an oral rabies vaccination (ORV) project conducted from 1998 through 2007 in Anne Arundel County, Md, for the control of rabies in terrestrial animals. Design-Retrospective analysis of surveillance data (1997 through 2007). Animals-Free-ranging raccoons (Procyon lotor) and other terrestrial mammals. Procedures-Vaccinia-rabies glycoprotein recombinant virus oral rabies vaccine-bait units were distributed annually by aircraft and ground teams targeting free-ranging raccoons. Approximately 2 to 4 weeks following the vaccine-bait placement, raccoons were live trapped, sedated, processed, and then released. Serologic samples were tested for the presence of rabies virus-neutralizing antibodies (RVNAs). Bait acceptance was estimated by analysis of tetracycline biomarking of sampled teeth. Rabies incidence was determined by the passive identification of rabid terrestrial animals. Results-The incidence of rabies in terrestrial animals decreased 92% between 1997 (the year prior to the start of the ORV project) and 2007. The mean RVNA prevalence across all years was 33% among trapped raccoons in areas baited with a fish meal polymer bait type, whereas the mean bait acceptance was 30%. Adult raccoons had a seropositivity rate twice that of juvenile raccoons, whereas the bait acceptance rate between adults and juveniles did not differ significantly. For areas baited with a coated sachet bait, adults and juveniles had the same seroprevalence. Juveniles had better seroprevalence when the annual campaign started in September and October, compared with August. Conclusions and Clinical Relevance-The ORV project contributed to a significant decrease in annual incidence of terrestrial animal rabies in Anne Arundel County, Md, during the 10-year project period. For fish meal polymer baits, juvenile raccoons accessed bait at the same rate as adult raccoons but had a significantly lower prevalence of RVNAs. For coated sachet baits, seroprevalence was the same in both age groups. The time of year the bait distribution occurred and the bait type used may be partial explanations for the difference in RVNA seroprevalence between adults and juvenile raccoons.     

Control of rabies: epidemiology of rabies in Asia and development of new-generation vaccines for rabies

Rabies is an enzootic viral disease widespread throughout the world. Although it is a vaccine-preventable disease, the annual number of human deaths caused by rabies is estimated to be 32,000 in Asia. Phylogenetic analysis based on sequence data of the partial N gene of rabies viruses in Asia has shown that the viruses are divided into five genogroups, distributed in Middle East, South Asia, South East Asia, Malay, and Arctic regions. The genetic relationships among these rabies viruses agree basically with the results of previous studies. Meanwhile, new types of vaccines are being developed by applying gene manipulation techniques to rabies virus in order to overcome the disadvantages of current vaccines. This article reviews the molecular epidemiology of rabies in Asia and progress made in the development of new-generation rabies vaccines with the goal of elimination or control of rabies in Asia.     

Survey of the dog population of Zimbabwe and its level of rabies vaccination

A random sample survey using personal interviews was conducted in Zimbabwe in 1986 to determine the size and structure of the national dog population and its level of rabies vaccination. There was an average of 0.91 dogs per household in Zimbabwe giving an extrapolated total population of 1,308,577 dogs. There was a dog to people ratio of 1:6.5 and an average of 3.4 dogs per square kilometre. The ratio of adult male to female dogs was 0.56:0.44, with 20 per cent of the dog population being less than three months old. In the period 1950 to 1986 there was a 4.7 per cent per annum growth rate of the dog population. In one of the provinces, Manicaland, dogs were found to have an average age of 2.3 years and a life expectancy at birth of 4.6 years. An estimated 40 per cent of the dog population three months old and above had been vaccinated against rabies in 1985/1986. With the rabies incidence in Zimbabwe still unacceptably high this level of vaccination is clearly inadequate and measures designed to increase it are discussed.     

Hematotropic parasites of translocated raccoons in the southeast.

Ninety-four raccoons (Procyon lotor) from 6 southeastern states were examined for hematotropic parasites, to evaluate the carrier potential of these animals as they relate to translocation and release for hunting purposes. Four species of hematotropic parasites were identified, viz, Babesia procyonis, Haemobartonella procyoni, Hepatozoon procyonis, and Trypanosoma cruzi. Lesions in infected raccoons were associated only with H procyonis. Babesia procyonis was enzootic in populations of raccoons used as sources for exportation and in raccoons resident in typical release areas. Haemobartonella procyoni, H procyonis, and T cruzi were prevalent only in locales of raccoon origin. Due to the potential pathogenicity of H procyonis and T cruzi in raccoons or other hosts, the practice of translocating raccoons from enzootic to nonenzootic areas was considered biologically hazardous.     

Serological survey for antibodies to Leptospira in dogs and raccoons in Washington State

A high number of reported canine leptospirosis cases occurred in Washington State from 2004 to 2006. This prompted a serosurvey of healthy dogs from around the state to determine the distribution of exposure risk and to provide insight into serovar epidemiology in the region. In addition, a convenience sample of sera from injured raccoons was also tested, and clinical serological data from the Washington Animal Disease Diagnostic Laboratory were examined. The proportion of dogs with an antibody titre (>or=1:100) to any serovar was 27/158 (17.1%, 95% CI 11.6-23.9), and that proportion among raccoons was 22/115 (19.1%, 95% CI 12.4-27.5) suggesting that the potential for exposure in Washington state is not uncommon. The most frequently detected serovars in healthy dogs were Autumnalis, Icterohemorrhagiae and Canicola, in clinical canine samples Autumnalis, Bratislava and Pomona were more frequent and in sick or injured raccoons Autumnalis, and Pomona were most frequently detected. Clinical canine serology demonstrated a late summer-fall seasonality that was consistent with other reports. An outbreak of canine leptospirosis occurred during 2004-2006 and was located primarily in western Washington counties, as were three reported human cases in 2005. Canine leptospirosis surveillance is an important tool for detecting human risk of exposure and may provide insights into which serovars are currently of clinical importance.     

Prevalence of Toxoplasma gondii infection in raccoons.

Serum samples from 427 raccoons (93 from Pennsylvania, 45 from New Jersey, 72 from South Carolina, 68 from Virginia, 30 from Iowa, and 119 from Ohio) were evaluated for Toxoplasma gondii antibodies in dilutions of 1:25, 1:50, and 1:500. The distribution of T gondii antibody titers was less than 1:25 for 212 raccoons (49.6%), 1:25 for 34 raccoons (7.9%), 1:50 for 117 raccoons (27.4%), and greater than or equal to 1:500 for 64 raccoons (14.9%). Tissue cysts were seen in the liver, and tachyzoites were in the brain of a raccoon with abnormal neurologic signs and concurrent infection with canine distemper virus. Organisms in the liver were stained with anti-T gondii serum, and the raccoon had a T gondii titer of 1:160 in the agglutination test.     

Economic implications of bovine leukemia virus infection in mid-Atlantic dairy herds

OBJECTIVE: To determine the baseline costs of bovine leukemia virus (BLV) infection, including costs of clinical disease and subclinical infection, in a dairy herd representative of the mid-Atlantic region and compare these costs with the cost of a test-and-manage BLV control program. DESIGN: Stochastic spreadsheet model. SAMPLE POPULATION: A commercial Holstein dairy herd with 100 milking cows. PROCEDURES: A spreadsheet model was developed. The overall cost of infection included the cost of clinical disease (ie, lymphosarcoma [LS]) and the effects of subclinical infection on milk production and premature culling. Model input values and distributions were designed to reflect economic conditions in the mid-Atlantic region. Relative costs of infection and control were calculated for infection prevalences of 20, 50, and 80%. RESULTS: Estimated mean cost to the producer per case of LS was 412 dollars; for a herd with a 50% prevalence of BLV infection, annual incidence of LS was 0.66. Mean annual cost of subclinical infection at a 50% prevalence of infection was 6,406 dollars. Mean annual cost of a test-and-manage control program was 1,765 dollars. The cost of clinical disease and subclinical infection varied substantially with the prevalence of infection, whereas the cost of control varied with herd size. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that a basic BLV control program may be economically beneficial in herds in which the prevalence of BLV infection is > or = 12.5%. Farm-specific considerations may factor prominently when weighing the costs and benefits of an individual BLV control program.     

A quantitative risk assessment model to evaluate effective border control measures for rabies prevention

Border control is the primary method to prevent rabies emergence. This study developed a quantitative risk model incorporating stochastic processes to evaluate whether border control measures could efficiently prevent rabies introduction through importation of cats and dogs using Taiwan as an example. Both legal importation and illegal smuggling were investigated. The impacts of reduced quarantine and/or waiting period on the risk of rabies introduction were also evaluated. The results showed that Taiwan’s current animal importation policy could effectively prevent rabies introduction through legal importation of cats and dogs. The median risk of a rabid animal to penetrate current border control measures and enter Taiwan was 5.33 × 10⁻⁸ (95th percentile: 3.20 × 10⁻⁷). However, illegal smuggling may pose Taiwan to the great risk of rabies emergence. Reduction of quarantine and/or waiting period would affect the risk differently, depending on the applied assumptions, such as increased vaccination coverage, enforced custom checking, and/or change in number of legal importations. Although the changes in the estimated risk under the assumed alternatives were not substantial except for completely abolishing quarantine, the consequences of rabies introduction may yet be considered to be significant in a rabies-free area. Therefore, a comprehensive benefit-cost analysis needs to be conducted before recommending these alternative measures.     

Risk factors for human rabies in China

In China, we have witnessed an increasing incidence of rabies in recent years and the number of deaths ranked first among the 39 notifiable infectious diseases. We conducted a case-control study to identify risk factors for human rabies infection and disease to recommend prevention and treatment among people exposed to rabies. Exposure site, pre-exposure prophylaxis and post-exposure prophylaxis were significantly associated with rabies infection. Exposure site at upper limb and trunk or at lower limb were at lower risk as compared with head-exposed patients. The OR was 0.09(95% CI: 0.009-0.93) and 0.01(95% CI: 0.001-0.115) respectively. Pre-exposure prophylaxis (OR = 0.05, 95% CI: 0.03-0.11) and post-exposure prophylaxis (OR = 0.02, 95% CI: 0.01-0.40) were both protective factors as compared with no prophylaxis. For patients who had post-exposure prophylaxis, dose for the first injection and immunity procedure were significantly associated with rabies infection.     

Postexposure prophylaxis for prevention of rabies in dogs

OBJECTIVE: To evaluate postexposure prophylaxis (PEP) in dogs experimentally infected with rabies. ANIMALS: 29 Beagles. PROCEDURE: Dogs were sedated and inoculated in the right masseter muscle with a salivary gland homogenate from a naturally infected rabid dog (day 0). Six hours later, 5 dogs were treated by administration of 2 murine anti-rabies glycoprotein monoclonal antibodies (mAb) and commercial vaccine; 5 received mAb alone; 5 received purified, heat-treated, equine rabies immune globulin (PHT-ERIG) and vaccine; 5 received PHT-ERIG alone; 4 received vaccine alone; and 5 control dogs were not treated. The mAb or PHT-ERIG was administered at the site of rabies virus inoculation. Additional vaccine doses for groups mAb plus vaccine, PHT-ERIG plus vaccine, and vaccine alone were administered IM in the right hind limb on days 3, 7, 14, and 35. RESULTS: All control dogs and dogs that received only vaccine developed rabies. In the PHT-ERIG and vaccine group, 2 of 5 dogs were protected, whereas none were protected with PHT-ERIG alone. Use of mAb alone resulted in protection in 4 of 5 dogs. Administration of mAb in combination with vaccine provided protection in all 5 dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Current national guidelines recommend euthanasia or a 6-month quarantine for unvaccinated animals exposed to rabies. Findings from this study document that vaccine alone following severe exposure was unable to provide protection from rabies. However, vaccine combined with mAb resulted in protection in all treated dogs, revealing the potential use of mAb in PEP against rabies in naive dogs.     

Naturally occurring spirochetes in the colonic mucosa of raccoons (Procyon lotor)

The large intestines of 21 raccoons (Procyon lotor; 11 wild caught, 10 laboratory confined) were examined for the presence of intestinal spirochetes. Light microscopy of sections stained with hematoxylin and eosin and Warthin-Starry stain showed the presence of spiral shaped organisms deep within the lumina of intestinal glands at the ileocolonic junction of 16 raccoons (76% prevalence). All laboratory-confined, group-housed raccoons harbored the organisms, but only 6/11 (55% prevalence) live-trapped raccoons were positive for these spirochetes. The organisms were free in the glandular lumina, and there were no microscopic lesions. Two types of spirochetes were identified in the colonic glands: a slender spirochete 10-13 microm in length, 0.3 microm in diameter, and possessing long, thin tapered ends and a larger, regularly waved spiral organism (0.5 microm in diameter). The slender spirochete did not resemble any of the known spirochete genera and failed to grow on medium used to propagate oral treponemes and members of the genus Brachyspira.