Serological investigations of red foxes (Vulpes vulpes L.) for determination of the spread of tick-borne encephalitis in Northrhine-Westphalia

Serum samples from 786 red foxes shot between January 1995 and August 1996 in the southern half of Northrhine-Westphalia, located in western Germany, were tested for the presence of antibodies against tick-borne encephalitis (TBE) virus using the Immunozym FSME IgG All Species-ELISA (Immuno, Heidelberg, Germany) as a screening test: 759 sera were negative, 23 (2.9%) were borderline, and four (0.5%) were positive. Nine of the 27 ELISA reactive sera were confirmed by the TBE Western-Blot (Immuno, Heidelberg, Germany). Furthermore these 27 sera were tested for neutralizing antibodies by means of a plaque reduction neutralization test (PRNT) against TBE and West Nile viruses. Only one single serum was found to have a neutralization titre (+1:800 PRNT80) against TBE virus. All other 26 sera were negative for neutralizing antibodies against TBE or West Nile virus. Since the titre of the single serum is low, it can be interpreted that if TBE virus is present, its prevalence is extremely low. Northrhine-Westphalia is not classified as a TBE-endemic area. Further calculated serological testing of game and virological investigation of collected ticks in the affected area seem to be meaningful and necessary.     

Arbovirus antibodies in wild game caught in Moravia]

The HIT method was used to examine blood serums of the game in Moravia (roebuck, red deer, fallow deer, mouflon, wild boar, brown hare) for the presence of antibodies to arboviruses of these groups: alphavirus (Sindbis-SIN), flavivirus (West Nile-WN), tick-borne encephalitis (TBE) and Bunyamwera (Tahyna-TAH, Calovo-CVO). Antibodies to all viruses were detected, and namely in these frequencies: SIN 0.9%, WN 16.9%, TAH 41.5%, CVO 23.1% and TBE 8.5%.     

Tick-borne encephalitis (TBE) with special emphasis on infection in horses

The tick-borne encephalitis (TBE), also known as early summer meningo-encephalitis, is a geographically limited virus infection transmitted mainly by ticks. The importance of TBE is largely underestimated. The causative agent TBE-Virus (TBEV) is grouped into the genus Flavivirus of the virus family Flaviviridae. Clinical disease including fatal outcomes has been described for men and dogs. With regard to horses only a limited number of case reports is available. In a study performed at the Institute of Virology, Justus-Liebig-University Giessen serum samples from the German endemic region of Marburg-Biedenkopf were tested for antibodies against TBEV. From 240 sera tested 7 (2.9%) were regarded as positive in a serum neutralization test (SNT). In an ELISA, performed in parallel to confirm the SNT results, 5 out of 7 positive sera from the SNT were also positive. The remaining two samples with low SNT-titres and all sera from horses negative in the SNT were also negative in the ELISA. This article is focussed on TBE of horses. In this context different aspects of TBE are included such as properties of the causative agent, interactions between causative agent, host animals and environment, spread of TBEV, pathogenesis, clinical symptoms, diagnosis and control.     

Acute encephalitis and hydrocephalus in dogs caused by canine parainfluenza virus

Gnotobiotic puppies were inoculated intracerebrally with a strain of canine parainfluenza virus (CPI-78-238). Four of eight dogs developed histological evidence of acute encephalitis. Clinical signs of encephalitis were seen in two of these four dogs; one had signs and lesions of interstitial pneumonia. Of six inoculated dogs observed for six months after infection, five developed internal hydrocephalus. Virus was reisolated from two dogs with acute encephalitis but not from dogs with hydrocephalus. Hemagglutination-inhibition antibodies persisted throughout the observation period of six months at high levels in the serum and cerebrospinal fluid of hydrocephalic dogs.     

Serological Investigations of Red Foxes (Vulpes vulpes L.) for Determination of the Spread of Tick‐borne Encephalitis in Northrhine‐Westphalia

Serum samples from 786 red foxes shot between January 1995 and August 1996 in the southern half of Northrhine‐Westphalia, located in western Germany, were tested for the presence of antibodies against tick‐borne encephalitis (TBE) virus using the Immunozym FSME IgG All Species‐ELISA^® (Immuno, Heidelberg, Germany) as a screening test: 759 sera were negative, 23 (2.9 %) were borderline, and four (0.5 %) were positive. Nine of the 27 ELISA reactive sera were confirmed by the TBE Western‐Blot (Immuno, Heidelberg, Germany). Furthermore these 27 sera were tested for neutralizing antibodies by means of a plaque reduction neutralization test (PRNT) against TBE and West Nile viruses. Only one single serum was found to have a neutralization titre (+ 1 : 800 PRNT₈₀) against TBE virus. All other 26 sera were negative for neutralizing antibodies against TBE or West Nile virus. Since the titre of the single serum is low, it can be interpreted that if TBE virus is present, its prevalence is extremely low. Northrhine‐Westphalia is not classified as a TBE‐endemic area. Further calculated serological testing of game and virological investigation of collected ticks in the affected area seem to be meaningful and necessary.     

Cervids as sentinel‐species for tick‐borne encephalitis virus in Norway ‐ A serological study

Tick‐borne encephalitis virus (TBEV) is the causative agent of tick‐borne encephalitis (TBE). TBEV is one of the most important neurological pathogens transmitted by tick bites in Europe. The objectives of this study were to investigate the seroprevalence of TBE antibodies in cervids in Norway and the possible emergence of new foci, and furthermore to evaluate if cervids can function as sentinel animals for the distribution of TBEV in the country. Serum samples from 286 moose, 148 roe deer, 140 red deer and 83 reindeer from all over Norway were collected and screened for TBE immunoglobulin G (IgG) antibodies with a modified commercial enzyme‐linked immunosorbent assay (ELISA) and confirmed by TBEV serum neutralisation test (SNT). The overall seroprevalence against the TBEV complex in the cervid specimens from Norway was 4.6%. The highest number of seropositive cervids was found in south‐eastern Norway, but seropositive cervids were also detected in southern‐ and central Norway. Antibodies against TBEV detected by SNT were present in 9.4% of the moose samples, 1.4% in red deer, 0.7% in roe deer, and nil in reindeer. The majority of the positive samples in our study originated from areas where human cases of TBE have been reported in Norway. The study is the first comprehensive screening of cervid species in Norway for antibodies to TBEV, and shows that cervids are useful sentinel animals to indicate TBEV occurrence, as supplement to studies in ticks. Furthermore, the results indicate that TBEV might be spreading northwards in Norway. This information may be of relevance for public health considerations and supports previous findings of TBEV in ticks in Norway.     

Experimental infection of Japanese encephalitis virus in dogs

A previous serosurvey of Japanese encephalitis virus (JEV) among dogs suggested that dogs are well suited for use as sentinels for assessing the risk of JEV transmission to humans. To examine the clinical symptoms and duration of anti-JEV antibodies in dogs, three dogs were experimentally challenged with JEV. All JEV-infected dogs did not show any clinical signs or abnormal blood tests, except for C-reactive protein. Virus-neutralization titers rapidly increased and were maintained until 70 days postinfection, and neither the virus nor the viral genome was detected in blood. Thus, since dogs live in close proximity to humans as companion animals, they are well suited for use as sentinels for surveying the human risk of JEV infection.     

Evaluation of serological tests for detecting tick-borne encephalitis virus (TBEV) antibodies in animals

Tick-borne encephalitis (TBE) in animals is not well understood yet. TBE virus (TBEV) serology in several host species could be valuable for epidemiological analyses in the field as well as for the detection of clinical cases. However, performance and suitability of the available test systems are not well assessed. Therefore, we evaluated two commercial TBEV-ELISA kits in a pilot study and compared them for their suitability in veterinary applications. For this purpose, we tested 163 field collected goat sera and evaluated the results by serum neutralization test (SNT) as "gold standard". Twenty-eight SNT positive sera (17.2%) were detected. The best suited ELISA kit was used for determination of a species-specific cutoff for horses, cattle, sheep, goats, pigs, mice, dogs, rabbits and monkeys with defined sera from animals without known or with improbable contact to TBEV. The level of non-specific ELISA results does not only differ between animal species but may also be influenced by the age of the tested animals. The number of sera which tested false positive by ELISA was higher in older than in young sheep. In order to obtain defined polyclonal sera as references, two dogs, cattle, goats, sheep, rabbits and pigs each, as well as one horse and 90 mice were immunized four times with a commercially available TBEV vaccine. In conclusion, our results demonstrated that commercial TBEV-ELISA kits are suitable for application in veterinary medicine for both, verification of clinical TBE cases and epidemiological screening. However, positive ELISA results should be verified by SNT. Only a very low number of false negative ELISA-results were found.     

Prevalence of antibodies against Saint Louis encephalitis and Jamestown Canyon viruses in California horses

Jamestown Canyon (JC) and Saint Louis encephalitis (SLE) viruses are mosquito-transmitted viruses that have long been present in California. The objective of this study was to determine the seroprevalence of these two viruses in horses prior to the introduction of West Nile (WN) virus. Approximately 15% of serum samples collected in 1998 from 425 horses on 44 equine operations horses throughout California had serum antibodies to JC virus, whereas antibodies were not detected to SLE virus. The results indicate that horses in California were commonly infected prior to 1998 with mosquito-transmitted Bunyaviruses that are identical or closely related to JC virus, but not with SLE virus. The different seroprevalence of SLE and JC viruses in horses likely reflects the unique ecology of each virus, and it is predicted that WN virus will have a wider distribution in California than closely related SLE virus.     

Nonsuppurative encephalitis in a dog

A 4-year-old male German Hunting Terrier presented with tremor, dyspnea, trismus, spasms of the musculature of the larynx and pharynx, and hypothermia and subsequently died despite intensive clinical care. Prior clinical signs included vomitus and diarrhea. Microscopic examination of the brain revealed a multifocal nonsuppurative brain stem encephalitis; a few intralesional neurons contained intranuclear inclusions. By immunohistochemistry, Aujeszky disease virus (Suid herpesvirus 1) antigen was detected in neurons in the brain and in ganglion cells of the trigeminal ganglia. Viral culture of brain tissue confirmed the presence of Aujeszky disease virus. Histopathologic findings in the brain with the identification of Aujeszky disease virus by immunohistochemistry and polymerase chain reaction are consistent with Aujeszky disease virus-induced encephalitis. Sequencing revealed a 100% homology of the isolated Aujeszky disease virus with Aujeszky disease virus isolates of wild boar from Eastern Germany.     

Clinical encephalitis following avian encephalomyelitis vaccination in broiler breeder pullets

Twelve-to-fourteen-week-old broiler breeder pullets and cockerels from two different companies were found to have encephalitis 2 weeks after vaccination with avian encephalomyelitis (AE) vaccine. Histologic examination and virus isolation indicated that affected chickens had AE. All chickens had been vaccinated with either one of two serials of AE vaccine approximately 2 weeks before the onset of clinical signs.     

Post-vaccinal distemper encephalitis in two Border Collie cross littermates

Abstract

CASE HISTORY: One 4.5-month-old male Border Collie cross presented with aggression and seizures in October 2006. A 16-month-old, female, spayed Border Collie cross presented with hypersalivation and a dropped jaw and rapidly became stuporous in September 2007. The dogs were littermates and developed acute neurological signs 5 and 27 days, respectively, after vaccination with different modified live vaccines containing canine distemper virus.

HISTOPATHOLOGICAL FINDINGS: Sections of brain in both dogs showed evidence of encephalitis mainly centred on the grey matter of brainstem nuclei, where there was extensive and intense parenchymal and perivascular infiltration of histiocytes and lymphocytes. Intra-nuclear and intra-cytoplasmic inclusions typical of distemper were plentiful and there was abundant labelling for canine distemper virus using immunohistochemistry.

DIAGNOSIS: Post-vaccinal canine distemper.

CLINCIAL RELEVANCE: Post-vaccinal canine distemper has mainly been attributed to virulent vaccine virus, but it may also occur in dogs whose immunologic nature makes them susceptible to disease induced by a modified-live vaccine virus that is safe and protective for most dogs.     

Necrotizing encephalitis of unknown cause in Fennoscandian Arctic foxes (Alopex lagopus).

A neurologic disease affected a colony of endangered Fennoscandian arctic foxes (Alopex lagopus) kept in captivity for breeding purposes. Several outbreaks of disease occurred between 1994 and 2004. The clinical signs included ataxia, indications of anosmia, blindness, and abnormal behavior. The disease was characterized by severe necrotizing encephalitis affecting mostly the cranial cerebrum, basal ganglia, and olfactory bulbs. Investigations to identify the etiology of the disease included testing for several infectious agents known to cause encephalitis in carnivores. Tests for Toxoplasma gondii, Encephalitozoon cuniculi, Neospora caninum, canine distemper virus, rabies, adenovirus type 1, Borna disease virus, and Listeria monocytogenes were negative. The colony was closed, and the cause of the disease remains undetermined.     

Prevalence and risk factors of Japanese encephalitis virus (JEV) in livestock and companion animal in high-risk areas in Malaysia

Japanese encephalitis (JE) is vector-borne zoonotic disease which causes encephalitis in humans and horses. Clinical signs for Japanese encephalitis virus (JEV) infection are not clearly evident in the majority of affected animals. In Malaysia, information on the prevalence of JEV infection has not been established. Thus, a cross-sectional study was conducted during two periods, December 2015 to January 2016 and March to August in 2016, to determine the prevalence and risk factors in JEV infections among animals and birds in Peninsular Malaysia. Serum samples were harvested from the 416 samples which were collected from the dogs, cats, water birds, village chicken, jungle fowls, long-tailed macaques, domestic pigs, and cattle in the states of Selangor, Perak, Perlis, Kelantan, and Pahang. The serum samples were screened for JEV antibodies by commercial IgG ELISA kits. A questionnaire was also distributed to obtain information on the animals, birds, and the environmental factors of sampling areas. The results showed that dogs had the highest seropositive rate of 80% (95% CI: ±?11.69) followed by pigs at 44.4% (95% CI: ±?1.715), cattle at 32.2% (95% CI: ±?1.058), birds at 28.9% (95% CI: ±?5.757), cats at 15.6% (95% CI: ±?7.38), and monkeys at 14.3% (95% CI: ±?1.882). The study also showed that JEV seropositivity was high in young animals and in areas where mosquito vectors and migrating birds were prevalent.     

Comparison of Immunoperoxidase Plaque Staining and Neutralizing Tests for Canine Distemper Virus

The use of immunoperoxidase plaque staining (IP) in the antibody test for canine distemper virus (CDV) and its diagnostic value were investigated. Antibody titres by the IP test and neutralizing test (NT) against CDV were compared in 114 specific pathogen-free (SPF) dogs, including 84 CDV-vaccinated and 30 unimmunized dogs. All of the unimmunized dogs, which had negative NT titres, also showed negative IP titres, and all of the CDV-vaccinated dogs, which had positive NT titres, showed positive IP titres. Furthermore, the IP titres showed similar changes with time after vaccination to the NT titres in five CDV-vaccinated SPF dogs. These findings indicate that the IP test has similar specificity to the NT.Next, the IP and NT titres were compared in 190 clinically suspected cases of canine distemper (CD), including 107 dogs with neurological signs and 83 dogs without neurological signs, and also in 50 clinically healthy dogs. Among the suspected cases of CD, of 46 dogs with neurological signs and 29 dogs without neurological signs, and which had negative NT titres, 33 (72%) and 19 (66%), respectively, showed positive IP titres. Many suspected cases of CD showed a high IP titre despite a low NT titre. In contrast, among healthy dogs, none of 9 NT-negative dogs showed positive IP titres.The difference in the IP and NT titres (log₁₀ [reciprocal of IP titre] – log₁₀ [reciprocal of NT titre]) was measured to investigate the agreement between IP test and NT. The differences were 1.488±0.867, 1.245±0.833, 0.595±0.351 and 0.594±0.372, respectively, in the suspected cases of CD with neurological signs, the suspected cases of CD without neurological signs, the healthy dogs and the SPF dogs. The differences in the suspected cases of CD were significantly high compared to those for the SPF dogs (p<0.001).Furthermore, in 8 dogs diagnosed as cases of CD from their paired sera, whose initial sera showed a markedly low NT titre compared to the IP titre, 5 dogs without neurological signs developed neurological signs later.These findings suggest that the IP test can detect CD during its early phases and in dogs with low antibody levels, undetectable by NT, and so would be useful for serological diagnosis.     

Comparative analyses of canine distemper viral isolates from clinical cases of canine distemper in vaccinated dogs

Sequence and phylogenetic analyses of three isolates of canine distemper virus (CDV) isolated from three dogs with a vaccination history were compared with the same analyses of vaccine virus isolated from a vaccine used for dogs. The three dogs showed clinical signs of a recent major type of CD in Japan, including oculonasal discharge and diarrhea, and pathological findings including non-suppurative encephalitis, pneumonia, mild gastroenteritis and lymphoid depletion. Inclusion bodies were in the stomach without inflammation and encephalitis was without clinical signs. One of the highest titers of CDV in different organs of the three dogs was commonly systemic lymphatic organs, including the spleen, lymph nodes and tonsils. New isolates of CDV joined to the clades of the Asia 1 group that is far from the vaccine group. These results surely indicate that wild strains of CDV from dogs with a vaccination history were not reversed vaccine virus, and that the dogs showed characteristics of recent CD in Japan.     

Rapid and specific serodiagnosis of western equine encephalitis virus infection in horses

Paired sera from 28 nonvaccinated horses with serologically confirmed western equine encephalitis (WEE) virus infections were evaluated for immunoglobulin (Ig)M and IgG directed against WEE virus, by use of enzyme immunoassay. Twenty-one of the horses developed greater than or equal to 4-fold increases or decreases in serum IgM titers in paired serum samples, confirming the diagnosis of WEE in these horses. Of the remaining 7 horses, 1 had stable IgM titers, 1 had a 2-fold increase in IgM titer between paired sera, 2 had 2-fold decreases in IgM titer, and for 3 horses adequate volumes were not available for both sera of the pair. Twenty-nine of 56 blood samples collected from these 28 horses had been collected within the first 3 days after clinical disease was recognized; all 28 horses and 48 of 53 available serum samples had IgM antibody to WEE virus. Immunoglobulin M also was detected in sera of 27 of 45 other nonvaccinated horses that had illnesses clinically compatible with WEE. Sera with IgM did not have cross-reacting IgM against eastern equine encephalitis virus. Therefore, the sensitivity, specificity, and lack of persistence of IgM was useful in the rapid diagnosis of WEE virus infections in horses.     

Magnetic resonance imaging findings in histologically confirmed Pug dog encephalitis

The purpose of the study was to describe magnetic resonance (MR) imaging features of histologically confirmed necrotizing encephalitis in four Pugs and to compare those findings with MR imaging characteristics of necrotizing encephalitis in other breeds. All dogs had the following common findings: lesions restricted to the forebrain, both cerebral hemispheres diffusely but asymmetrically affected, lesions affected gray and white matter resulting in loss of distinction between both, most severe lesions in occipital and parietal lobes, lesions were irregularly T2-hyperintense and T1-isointense to slightly T1-hypointense, and no cavitation. There were various degrees of contrast enhancement of brain and leptomeninges. Asymmetry of lateral ventricles and midline shift was seen in one dog each. Two dogs had brain herniation, which may have contributed to the progression of neurologic signs. Hyperintensity on T2-weighted and fluid attenuated inversion recovery images in the hippocampus and piriform lobe was consistent with excitotoxic edema, whereas similar imaging features in other forebrain areas corresponded to areas of inflammation or liquefaction on histopathology. In comparison with necrotizing encephalitis in other canine breeds, Pug dog encephalitis has some unique MR imaging features. Therefore, these characteristics cannot be applied to other breeds, nor should imaging features of necrotizing encephalitis of other canine breeds be used for interpretation of MR images in Pug dogs.