Serologic survey of select infectious diseases in coyotes and raccoons in Nebraska

To obtain data about select zoonotic and other infectious diseases in free-ranging predators in five ecoregions in Nebraska, sera were collected from 67 coyotes (Canis latrans) and 63 raccoons (Procyon lotor) from November 2002 through January 2003. For coyotes, antibodies were detected against canine distemper virus (CDV, 61%), Francisella tularensis (32%), Rickettsia rickettsi (13%), and flaviviruses (48%). None of the coyote sera had antibodies to Borrelia burgdorferi, Brucella canis, or six serovars of Leptospira interrogans. Because serologic cross-reactivity exists among flaviviruses, 14 sera from flavivirus-positive coyotes were also tested for St. Louis encephalitis virus (SLE) antibodies and two (14%) were positive, suggesting that up to 48% of coyotes tested had antibodies against West Nile virus (WNV). For raccoons, antibodies were detected against CDV (33%), F. tularensis (38%), and three serovars of L. interrogans (11%).     

Serologic survey for selected disease agents in wolves (Canis lupus) from Alaska and the Yukon Territory, 1984-2000

Wolves (Canis lupus) were captured in several geographic areas of Alaska (USA) and the Yukon Territory (Canada) during 1984-2000. Blood was collected from 1,122 animals. Sera were tested for antibodies against infectious canine hepatitis virus (ICH), canine distemper virus (CDV), canine parvovirus (CPV), Francisella tularensis, and serovars of Leptospira interrogans. Antibody prevalence for ICH was >84% for all areas. Area-specific prevalences of antibodies ranged from 12% to 70% for CPV, from 0% to 41% for CDV, and from 4% to 21% for F. tularensis. There was no evidence of CDV exposure at the two southernmost locations in Alaska. Prevalence of antibodies for ICH increased slightly during the 16-yr course of the survey. There was essentially no evidence of exposure to L. interrogans. Prevalences of antibodies for both CPV and CDV were age-specific, with higher values in the adult cohort compared with the pup cohort. There were no sex-specific differences in prevalence of antibodies for any of the five disease agents.     

Neospora caninum infection in a free-ranging raccoon (Procyon lotor) with concurrent canine distemper virus infection

During a canine distemper virus (CDV) outbreak in raccoons (Procyon lotor) from Cook County, Illinois, a juvenile female suffering from seizures was killed and necropsied. Gross and histologic findings of necrotizing encephalitis and proliferative bronchopneumonia were attributed to CDV infection and considered the cause of clinical signs. A section of cerebellum stained immunohistochemically for Neospora caninum revealed an approximately 40 microm diameter, round to oval cyst with a 2- to 3-microm-thick wall and filled with 1-2 microm diameter, round to oval bradyzoites. Polymerase chain reaction (PCR) results were positive for N. caninum using DNA extracted from the brain. Specific PCR for the closely related organisms Toxoplasma gondii and Hammondia heydorni yielded negative results. This case report provides histologic, immunohistochemical, and molecular evidence that raccoons are a naturally occurring intermediate host of N. caninum.     

Prevalence of selected pathogenic microbial agents in the red fox (Vulpes fulva) and gray fox (Urocyon cinereoargenteus) of southwestern Wisconsin

Free-ranging red foxes (Vulpes fulva) and gray foxes (Urocyon cinereoargenteus) were trapped in southwestern Wisconsin. Fox sera were tested to determine the prevalence of antibody for five different Leptospira interrogans serovars, canine distemper virus (CDV), infectious canine hepatitis virus (ICHV), and Franciscella tularensis infections. Grippotyphosa was the most prevalent leptospiral serovar antibody observed. Twenty-five of 53 (47%) red foxes and 11 of 36 (31%) gray foxes had specific antibodies to grippotyphosa. Juvenile foxes had geometric mean antibody titers to grippotyphosa significantly higher (P less than 0.05) than those of the adults of both species. CDV antibody was detected in sera of red foxes only. Six of 57 (11%) red foxes had CDV antibody. ICHV antibody was detected in 2 of 57 (3%) red foxes and 3 of 32 (9%) gray foxes. Antibody to F. tularensis was not detected in any fox sera.     

Does human proximity affect antibody prevalence in marine-foraging river otters (Lontra canadensis)?

The investigation of diseases of free-ranging river otters (Lontra canadensis) is a primary conservation priority for this species; however, very little is known about diseases of river otters that forage in marine environments. To identify and better understand pathogens that could be important to marine-foraging river otters, other wildlife species, domestic animals, and humans and to determine if proximity to human population could be a factor in disease exposure, serum samples from 55 free-ranging marine-foraging river otters were tested for antibodies to selected pathogens. Thirty-five animals were captured in Prince William Sound, Alaska (USA), an area of low human density, and 20 were captured in the San Juan Islands, Washington State (USA), an area characterized by higher human density. Of 40 river otters tested by indirect immunofluorescent antibody test, 17.5% were seropositive (titer > or =320) for Toxoplasma gondii. All positive animals came from Washington. Of 35 river otters tested for antibodies to Leptospira interrogans using the microscopic agglutination test, 10 of 20 (50%) from Washington were seropositive (titer > or =200). None of the 15 tested animals from Alaska were positive. Antibodies to Neospora caninum (n=40), Sarcocystis neurona (n=40), Brucella abortus (n=55), avian influenza (n=40), canine distemper virus (n=55), phocine distemper virus (n=55), dolphin morbillivirus (n=55), porpoise morbillivirus (n=55), and Aleutian disease parvovirus (n=46) were not detected. Identifying exposure to T. gondii and L. interrogans in otters from Washington State but not in otters from Alaska suggests that living proximal to higher human density and its associated agricultural activities, domestic animals, and rodent populations could enhance river otter exposure to these pathogens.     

Genetic variability and viral seroconversion in an outcrossing vertebrate population

Inverse correlations between genetic variability and parasitism are important concerns for conservation biologists. We examined correlations between neutral genetic variability and the presence of antibodies to canine distemper virus (CDV) and feline parvovirus (FPV) in a free-ranging population of raccoons. Over 3 years there was a strong relationship between age and seroprevalence rates. Most young animals were seronegative to CDV and FPV, but the oldest age class was greater than 80 per cent seropositive to both viruses. CDV-seropositive animals had greater heterozygosity and lower measures of inbreeding compared with CDV-seronegative animals. This relationship was strongest among the youngest animals and did not occur during a 1 year CDV epidemic. In contrast, FPV-seropositive animals only had significantly lower measures of inbreeding in 1 year, perhaps because FPV-associated mortality is relatively low or primarily occurs among very young individuals that were under-represented in our sampling. These results suggest that even in large outcrossing populations, animals with lower heterozygosity and higher measures of inbreeding are less likely to successfully mount an immune response when challenged by highly pathogenic parasites.     

Serosurvey of viral infections in free-ranging Namibian cheetahs (Acinonyx jubatus)

Cheetahs (Acinonyx jubatus) in captivity have unusually high morbidity and mortality from infectious diseases, a trait that could be an outcome of population homogeneity or the immunomodulating effects of chronic stress. Free-ranging Namibian cheetahs share ancestry with captive cheetahs, but their susceptibility to infectious diseases has not been investigated. The largest remaining population of free-ranging cheetahs resides on Namibian farmlands, where they share habitat with domestic dogs and cats known to carry viruses that affect cheetah health. To assess the extent to which free-ranging cheetahs are exposed to feline and canine viruses, sera from 81 free-ranging cheetahs sampled between 1992 and 1998 were evaluated for antibodies against canine distemper virus (CDV), feline coronavirus (feline infectious peritonitis virus; FCoV/ FIPV), feline herpesvirus 1 (FHV1), feline panleukopenia virus (FPV), feline immunodeficiency virus (FIV), and feline calicivirus (FCV) and for feline leukemia virus (FeLV) antigens. Antibodies against CDV, FCoV/FIPV, FHV1, FPV, and FCV were detected in 24, 29, 12, 48, and 65% of the free-ranging population, respectively, although no evidence of viral disease was present in any animal at the time of sample collection. Neither FIV antibodies nor FeLV antigens were present in any free-ranging cheetah tested. Temporal variation in FCoV/FIPV seroprevalence during the study period suggested that this virus is not endemic in the free-ranging population. Antibodies against CDV were detected in cheetahs of all ages sampled between 1995 and 1998, suggesting the occurrence of an epidemic in Namibia during the time when CDV swept through other parts of sub-Saharan Africa. This evidence in free-ranging Namibian cheetahs of exposure to viruses that cause severe disease in captive cheetahs should direct future guidelines for translocations, including quarantine of seropositive cheetahs and preventing contact between cheetahs and domestic pets.     

Serosurvey of free-ranging Amur tigers in the Russian Far East

Wild Amur tigers (Panthera tigris altaica, n=44) from the Russian Far East were tested for antibodies to feline leukemia virus, feline corona virus (FCoV), feline immunodeficiency virus, feline parvovirus (FPV), canine distemper virus (CDV), Toxoplasma gondii, and Bartonella henselae. Antibodies to FCoV, CDV, FPV, and T. gondii were detected in 43, 15, 68, and 42% of tigers, respectively. No differences were detected in antibody prevalence estimates between tigers captured as part of a research program and those captured to mitigate human-tiger conflicts. Domestic dogs (Canis familiaris) were tested as a potential source for CDV; 16% were vaccinated against CDV and 58% of unvaccinated dogs were antibody positive for CDV. A high percentage of tigers were exposed to potential pathogens that could affect the survival of this species. We recommend continued monitoring of wild tigers throughout Asia, development of standardized sampling and postmortem examination procedures, and additional research to better understand potential domestic and wild animal sources for these pathogens.     

Morbidity-mortality factors and survival of an urban coyote population in Arizona

The health of coyotes (Canis latrans) in urban areas has not been studied. Our objectives were to assess the health of coyotes in Tucson (Arizona, USA) by determining the prevalence of antibodies to selected pathogens, estimating survival rates, and identifying sources of mortality. We drew blood from 22 coyotes to evaluate the prevalence of heartworm (Dirofilaria immitis) antigens, and antibodies to canine distemper virus (CDV), infectious canine hepatitis (ICH), canine parvovirus (CPV), and seven serovars of Leptospira interrogans. We trapped and radiocollared 19 coyotes to determine survival rates. We performed necropsies on 19 coyotes to quantify their general health, the presence of internal and external parasites, and causes of mortality. No coyotes tested positive for heartworm antigens. The prevalence of antibody to CDV, ICH, and CPV was 27, 50, and 100%, respectively. Twenty-seven percent of coyotes tested positive for one of five serovars of L. interrogans. The diseases for which coyotes in Tucson possessed antibodies appear to be enzootic in the population. The annual survival rate of coyotes was 0.72. Eleven necropsied coyotes were killed by cars, five coyotes were hit by cars, two were killed by a trapper, and the cause of death for one coyote was unknown. Coyotes in Tucson appear to be exposed to the viral, bacterial, and parasitic infections common in many coyote populations, but humans are the major source of mortality.     

Exposure to feline and canine pathogens in bobcats and gray foxes in urban and rural zones of a national park in California

Exposure of bobcats (Lynx rufus) and gray foxes (Urocyon cinereoargenteus) to a range of common canine and feline pathogens was assessed in urban and rural zones of Golden Gate National Recreation Area, a National Park in the San Francisco Bay Area, (California, USA) from 1992 to 1995. Testing included serology for canine distemper virus, canine parvovirus (CPV), canine adenovirus, Leptospira interrogans, feline calicivirus (FCV), feline panleukopenia virus, feline herpesvirus, feline enteric coronavirus (FECV), feline immunodeficiency virus, feline leukemia virus, Toxoplasma gondii, and Bartonella henselae. Testing was also performed for Dirofilaria immitis. Significantly more gray foxes were seropositive for CPV in the urban zone than in the rural zone. In addition, radio-tracking of gray foxes in the rural zone indicated that all three of the rural CPV-seropositive foxes had traveled into adjoining small towns, whereas only one of the 11 seronegative animals had done so. Significantly more bobcats were seropositive for FCV in the rural zone than in the urban zone. Individual bobcats with positive FCV antibody titers had patterns of movement that intercepted park inholdings where domestic cats lived. Bobcat samples were seronegative for all five of the other viral feline pathogens, with the exception of a FECV-seropositive bobcat. High seroprevalence was detected for B. henselae and T. gondii in both zones. Variation in the seroprevalence for different pathogens might be related to differences in the exposure of bobcats and foxes to domestic animals: in the urban zone, gray foxes were located in residential areas outside the park, whereas bobcats were not. Although for most of the pathogens examined there was no relationship between urbanization and exposure, our results for CPV in foxes and FCV in bobcats indicated that proximity to urban areas or contact with humans can increase the risk of disease exposure for wild carnivore populations. Combining behavioral information from radio-tracking with data on pathogen exposure or disease incidence can provide valuable insights into the ecology of wildlife disease that might be missed with broad-scale, population-level comparisons alone.     

Diseases, parasites and survival of coyotes in south-central Georgia.

Serologic testing, radio-telemetry and post-mortem diagnostic evaluations were used to investigate survival and causes of mortality among 17 coyotes (Canis latrans) in south-central Georgia (USA). Prevalence of canine heartworm (Dirofilaria immitis) microfilariae was lower (P = 0.057) among fall-captured (22%) than among winter-captured (75%) coyotes. Prevalence of heartworm was higher among adults than juveniles in the fall, but no significant difference was detected between animals captured in winter. Antibodies were found against canine parvovirus (65%), canine parainfluenza virus (59%), infectious canine hepatitis virus (41%), and Toxoplasma gondii (18%). Antibodies were not found to Brucella canis, canine coronavirus, five serovars of Leptospira interrogans, or canine distemper virus. Seroprevalence of canine parvovirus was lower (P = 0.009) among fall-captured animals (33%) than winter-captured animals (100%). The Kaplan-Meier estimate of annual survival was 0.500 for all animals. Juvenile survival did not differ (P = 0.79) from adult survival, but male survival (S = 0.217) was lower (P = 0.11) than female survival (S = 0.804). Two of nine (22%) mortalities were human-caused, one was due to concurrent canine parvovirus and canine distemper virus infections, one animal died of trauma, two were considered natural mortalities of unknown cause, and no cause of death could be determined for the remaining three animals. Natural mortality may be significant for coyotes in south-central Georgia, although there was no apparent link between exposure to pathogens and the animals' subsequent fate in our small sample.     

A serosurvey of viral infections in lions (Panthera leo), from Queen Elizabeth National Park, Uganda

Serum samples from 14 lions (Panthera leo) from Queen Elizabeth National Park, Uganda, were collected during 1998 and 1999 to determine infectious disease exposure in this threatened population. Sera were analyzed for antibodies against feline immunodeficiency virus (FIV), feline calicivirus (FCV), feline herpesvirus 1 (feline rhinotracheitis: FHV1), feline/canine parvovirus (FPV/CPV), feline infectious peritonitis virus (feline coronavirus: FIPV), and canine distemper virus (CDV) or for the presence of feline leukemia virus (FeLV) antigens. Ten lions (71%) had antibodies against FIV, 11 (79%) had antibodies against CDV, 11 (79%) had antibodies against FCV, nine (64%) had antibodies against FHV1, and five (36%) had antibodies against FPV. Two of the 11 CDV-seropositive lions were subadults, indicating recent exposure of this population to CDV or a CDV-like virus. No lions had evidence of exposure to FeLV or FIPV. These results indicate that this endangered population has extensive exposure to common feline and canine viruses.     

A Sarcocystis sp.-like protozoan and concurrent canine distemper virus infection associated with encephalitis in a raccoon (Procyon lotor).

A raccoon (Procyon lotor) with signs of weakness was captured in upstate New York (USA). Despite attempted care in a rehabilitation facility, the animal died and was examined because of suspected infectious neurologic disease. The cerebrum had a marked, locally extensive, neutrophilic, necrotizing encephalitis with numerous associated intralesional protozoal organisms, and a moderate to marked multifocal perivascular nonsuppurative meningoencephalitis. Based on morphology and immunohistochemical staining, the organism was a Sarcocystis sp.-like protozoan. Rabies antigen and canine distemper virus (CDV) inclusions were not detected. However, the animal was positive for canine distemper virus based on peroxidase anti-peroxidase staining.     

First Evidence of Canine Distemper in Brazilian Free-Ranging Felids

Serum samples from 19 jaguars (Panthera onca), nine pumas (Puma concolor), and two ocelots (Leopardus pardalis) were collected between January 1999 and March of 2005 and tested for presence of canine distemper virus (CDV). All cats were free-ranging animals living in two protected areas in the Brazilian Atlantic Forest. In addition, 111 domestic dogs from nearby areas were sampled for CDV. Our results show the first evidence of CDV exposure in Brazilian free-ranging felids. From the 30 samples analyzed, six jaguars and one puma were tested seropositive for CDV. All seropositive large felids were from Ivinhema State Park, resulting in 31.5% of the sampled jaguars or 60% of the total jaguar population in Ivinhema State Park, and 11.28% of the sampled pumas. From the total 111 domestic dogs sampled, 45 were tested seropositive for CDV. At Morro do Diabo State Park, 34.6% of the dogs sampled were positive for CDV, and 100% at Ivinhema State Park. Canine distemper virus in wild felids seems to be related with home range use and in close association with domestic dogs living in nearby areas.     

Tyzzer's disease as a complication of canine distemper in a raccoon

Intercurrent canine distemper and Tyzzer's disease were diagnosed in a mature raccoon (Procyon lotor) submitted for necropsy. Clinical, gross and microscopic findings characteristic of canine distemper virus (CDV), included ataxia, dyspnea, suppurative conjunctivitis, interstitial pneumonitis and generalized lymphocytolysis. Inclusion bodies typical of CDV infection were present in many epithelial tissues. Acute multifocal hepatic necrosis and acute segmental necrotizing enteritis were attributed to the presence of Bacillus piliformis organisms in these lesions, confirmed by special stains and electron microscopy. This is apparently the first reported case of Tyzzer's disease in a raccoon.     

Serologic survey for selected virus infections in polar bears at Svalbard

Polar bears (Ursus maritimus) were chemically immobilized and sampled at Svalbard, Norway, and on the pack ice in the Barents Sea from late March to mid-May between 1990 and 1998. Plasma samples were tested for the presence of antibodies to canine distemper virus (CDV), calicivirus, phocid herpesvirus type 1 (PhHV-1), and rabies virus. A seroprevalence of 8% to CDV and 2% to calicivirus were found, whereas no antibodies were detected against PhHV-1 or rabies virus. This serologic survey indicates that polar bears in this region are exposed to morbillivirus and calicivirus, although the nature of these viruses and infections are unknown. Morbillivirus and calicivirus are potential pathogens in seals, but it is unknown whether they may cause health problems in polar bears.     

Humoral response and protection from experimental challenge following vaccination of raccoon pups with a modified-live canine distemper virus vaccine.

Eight 8-wk-old raccoon pups (Procyon lotor) with maternal canine distemper virus (CDV) neutralizing antibodies (NAb) and 24 8-wk-old seronegative pups were administered a commercial modified-live CDV vaccine (Galaxy, D, Solvay Animal Health, Inc., Kitchener, Ontario, Canada). All 24 seronegative raccoons had detectable serum CDV NAb titers 14 days after the initial dose. Titers rose to maximum levels 4 wk post-vaccination. Mean titers for groups of vaccinated seronegative pups were maintained between 1:256 and 1:2,048 for the remainder of the 3 mo observation period. Geometric means of the serum CDV NAb titer of eight seronegative pups given a single vaccine dose at 8 wk of age did not differ significantly from those of eight pups that were given serial doses at 8, 12, and 16 wk of age, or from those of eight pups vaccinated once at 16 wk of age. Seven unvaccinated 8-wk-old raccoon pups used as controls remained seronegative throughout the trial. Seven out of eight 8-wk-old pups with maternal antibodies, vaccinated at 8, 12, and 16 wk of age, failed to develop a rise in their CDV NAb titers until at least 18 wk of age, 2 wk after the third vaccination. Titers in eight unvaccinated raccoons with maternal antibodies declined steadily to undetectable levels at 20 wk of age. A half-life of 10.55 days was calculated for maternally-derived CDV NAb in raccoon pups. Sixteen vaccinated raccoons were protected from clinical disease following experimental oronasal challenge with a virulent raccoon strain of CDV, 13 to 23 wk after vaccination. Serum CDV NAb titers at the time of challenge ranged from 1:12 to 1:384 and increased during the period of observation. Three of four unvaccinated seronegative raccoons used as controls failed to mount any detectable CDV NAb and were euthanatized after developing clinical signs of canine distemper 26, 29, and 30 days post-challenge (PC). Necropsies confirmed the diagnosis. The fourth control raccoon exhibited transient equivocal clinical signs, mounted a sluggish humoral response, but was clinically normal when euthanatized 42 days PC. In this raccoon, there was focal non-suppurative encephalitis with intranuclear inclusion bodies typical of CDV infection.     

The removal of viruses during the purification of equine antisera using filtration aids Hyflo Super-Cel and Fulmon Super A.

The manufacturing process in Australia for equine antisera against various venoms/toxins is based primarily on ammonium sulphate precipitation of pepsin-digested IgG, whereby Fc and F(ab')(2)fragments are separated. The capacity of the process to remove non-enveloped and enveloped model viruses was assessed using a scaled-down process. Each virus was added to mid-process samples from equine plasma before the material was applied to Hyflo Super-Celtrade mark filtration followed by Fulmonttrade mark Super A filtration. Samples were analysed pre- and post-filtration and the log clearance of the viruses calculated. The mean clearance factors for viral load of canine adenovirus type II (CAV(2)), poliovirus type 1 (PV1), infectious bovine rhinotracheitis virus (IBR) and canine distemper virus (CDV) were 5.3 logs, 4.2 logs, 5.7 logs and 4. 0 logs respectively. Clearance results as virus is adsorbed to the filtration aids which are removed from the process, thereby demonstrating improved viral safety of equine antisera produced by CSL.     

水貂犬瘟热动物模型的建立

目的建立水貂犬瘟热动物模型,并利用水貂犬瘟热模型评价不同犬瘟热强毒株的毒力,为水貂犬瘟热病毒疫苗的研究奠定基础。方法从猴、藏獒、犬的病料中分离犬瘟热病毒,测定犬瘟热病毒的毒力,并进行传代培养。利用犬的犬瘟热动物模型筛选稳定的犬瘟热强毒株,进行水貂犬瘟热动物模型的建立及其毒力评估。结果筛选出了稳定的犬瘟热强毒株并进行了家犬动物实验,同时表现出了强烈的临床症状,并利用不同的代次毒进行了犬瘟热动物模型的建立。结论成功建立了犬瘟热动物模型并对不同来源的犬瘟热病毒毒力进行了评估。     

Seroprevalence of Toxoplasma gondii in mesocarnivores of the Canadian prairies

The protozoon Toxoplasma gondii has a worldwide distribution and affects many species of warm-blooded animals. In the Canadian prairies, mesocarnivores such as striped skunks (Mephitis mephitis) and raccoons (Procyon lotor) have experienced an increase in density and distribution, and they are in close contact with human dwellings. However, there has been no systematic study on the seroprevalence of T. gondii in these mesocarnivore populations. The objectives of the current project were to determine the serum antibody prevalence of T. gondii in Canadian prairie mesocarnivores and to study the relationship between antibody prevalence and species, sex, age, location, and year of collection. Antibodies to T. gondii were found in 5 of 24 (20.8%) skunks from Saskatchewan trapped in 1999 and 5 of 40 (12.5%) in 2000. Seroprevalences for T. gondii in raccoons and skunks trapped in Manitoba were 2 of 10 (20%) raccoons trapped in 2002, 7 of 44 (15.9%) trapped in 2003, and 16 of 37 (43.2%) trapped in 2004; and in 13 of 99 (13.1%) skunks trapped in 2003, 29 of 131 (22.1%) trapped in 2004, 53 of 165 (32.1%) trapped in 2005, and 30 of 51 (58.8%) trapped in 2006. Age, location, and year, but not the host species, were important variables in the determining the seroprevalence of T. gondii in skunks and raccoons. Results confirm that T. gondii is endemic in the skunk and raccoon populations in the Canadian prairies.