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The infection of dogs with distemper virus (CDV), adenovirus type 1 (CAV-1) and parvovirus (CPV) is still being diagnosed by practitioners, even in vaccinated dogs. Laboratory techniques used for the isolation and identification of these viruses are usually time consuming and often their accuracy is not satisfactory. The presented studies concentrated on the adaptation of PLA (Peroxidase Linked Assay) for the evaluation of CDV, CAV-1 and CPV replication in tissue cultures. Comparable studies on titration of viruses tested on the basis of cytopathic effects and results of PLA revealed that the titre of CDV and CPV evaluated by CPE was lower than calculated using PLA. The difference was equal to 0.6 log for CDV and 0.7 log CPV. These results confirm four to five-fold higher sensitivity of PLA. Titre of adenovirus type 1 measured by both methods did not demonstrate significant differences which can be explained by production of regular and characteristic CPE and rapid spread of cell destruction. Results of PLA revealed that the expression of viral antigens takes place between 18 and 24 hours after cell inoculation, whereas the first cytopathic effect was visible after 48-96 hours. The repeatability of results obtained by PLA was significantly higher with p<0.05 than titration of viruses on the bases of CPE in the case of CAV-1 and CDV. The presented results confirm the high sensitivity and specificity of PLA, which could be widely used in studies on CDV, CAV-1 and CPV.
Canine distemper (febris catarrhalis contagiosa et nervosa, vulpium et mustelarium) is one of the most dangerous viral diseases in animals, affecting mostly foxes and minks. The disease is caused by the virus member of the genus Morbilliviruses of the family Paramyxoviridae. The following animal families are susceptible to the virus: Canidae, Mustelidae, Procyonidae, Hyaenidae, Viverridae, Felidae, Ursidae and Phocidae. The symptoms of the disease may develop in the digestive, respiratory, nervous or urinary system. The disease may also manifest itself in conjunctivitis, skin lesions, and abortion. Infected animals become the reservoir of the virus, dangerous to other susceptible individuals and species. Diagnosis is based on the epizootiological history, clinical symptoms and laboratory examinations: IF and PCR. Post mortem diagnosis is based on the histopathological examination of the samples of the lungs, brain, stomach and urinary bladder mucous membrane. A direct virus isolation is of no practical importance, because the procedure is too time-consuming and the cytopathic effect occurs after several or a dozen days. In uncertain cases a very useful method, though rarely used nowadays, is a biological test on young animals susceptible to the distemper virus. Fur-bearing carnivores kept on farms and non-domestic animals living in national parks must not be treated, euthanasia should be performed instead. In exceptional cases, if the infected animal is very valuable and does not pose a threat to other animals, individual treatment may be administered. The most efficient way of avoiding the disease in animals is a specific prophylaxis based on vaccination. Controlling the distemper in the marine and non-domestic mammals is very difficult and practically not established. There are some clinical tests on using oral vaccines, similar to those used in wild foxes against rabies. However, the vaccines based on the living, modified distemper virus can be dangerous for zoological animals, especially for large cats and wild carnivores.
The aim of the study was to to detect and characterize the canine distemper virus isolated from 4 foxes and 6 dogs, naturally infected in Poland, as well as from two vaccines. A 200-bp fragment of the CDV H gene was amplified and sequenced. Sequencing of the PCR products from the isolates led to the identification of 4 groups. The most numerous group No. 1 was represented by virus strains isolated from 4 foxes and 4 dogs. In group No. 2 there were two sequences of CDV isolated from two dogs. These sequences varied from group 1 by transitions in positions 133 and 135. The isolates from groups 1 and 2 showed significant differences of nucleotide as well as aminoacid sequences in comparison with the isolates obtained from vaccines and with various CDV sequences obtained from GenBank. The results may indicate the appearance of new CDV variants in eastern Poland.
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