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Elaboration of triplex PCR for detection of selected viral infections in waterfowl
2019
Kozdruń, Wojciech | Czekaj, Hanna | Styś-Fijoł, Natalia | Piekarska, Karolina | Samanta Niczyporuk, Jowita
Viral infections are the greatest threat to waterfowl and cause significant economic losses. Diagnosis and differentiation of three goose viruses is difficult in the field and often requires laboratory confirmation. Therefore, the aim of the study was to develop a triplex PCR and optimise its parameters for simultaneous detection of DNA of goose parvovirus (GPV), goose polyomavirus (GHPV), and goose circovirus (GoCV). The DNA of viruses isolated from field cases from the National Veterinary Research Institute’s own collection was used for the study. The primer attachment temperature, the number of reaction cycles, and the Taq DNA polymerase and Mg²⁺ concentrations were optimised. The sensitivity and specificity of this triplex PCR was also determined. Based on the obtained results, triplex PCR parameters were optimised for simultaneous detection of DNA of GPV, GHPV, and GoCV in one sample. The following PCR products of the expected size were obtained: GPV DNA of 806 bp, GoCV DNA of 571 bp, and GHPV DNA of 180 bp. The developed triplex PCR method proved to be useful for simultaneous detection of infections with three waterfowl viruses and will be used in relevant laboratory diagnostics.
Mostrar más [+] Menos [-]Routes and prevalence of shedding of psittacine beak and feather disease virus
1991
Ritchie, B.W. | Niagro, F.D. | Latimer, K.S. | Steffens, W.L. | Pesti, D. | Ancona, J. | Lukert, P.D.
Psittacine beak and feather disease (PBFD) virus was recovered from the feces and crop washings from various species of psittacine birds diagnosed with PBFD. High concentrations of the virus also could be demonstrated in feather dust collection from a room where 22 birds with active cases of PBFD were being housed. The virions recovered from the feces, crop, and feather dust were confirmed to be PBFD virus by ultrastructural, physical, or antigenic characteristics. Virus recovered from the feather dust and feces hemagglutinated cockatoo erythrocytes. The specificity of the agglutination was confirmed by hemagglutination inhibition, using rabbit antibodies against PBFD virus. During the test period, 26% (8 of 31) of the birds screened were found to be excreting PBFD virus in their feces, and 21% (3 of 14) of crop washings were positive for PBFD virus. Some birds in the sample group had active cases of diarrhea, whereas others had normal-appearing feces. Diarrhea was found to be the only significant indicator of whether a bird was likely to be excreting virus from the digestive tract. These findings suggest that exposure of susceptible birds to PBFD virus may occur from contact with contaminated feather dust, feces, or crop secretions. Viral particles that were morphologically similar to parvovirus (2- to 24 nm-icosahedral nonenveloped virions) also were recovered from feces of some of the birds.
Mostrar más [+] Menos [-]Organ distribution of avian polyomavirus DNA and virus-neutralizing antibody titers in healthy adult budgerigars
1993
Phalen, D.N. | Wilson, V.G. | Graham, D.L.
Tissue specimens and serum samples obtained from adult budgerigars in various stages of reproduction housed in an aviary with enzootic avian polyomavirus (APV) disease were examined by means of polymerase chain reaction techniques for APV DNA. Although the birds were apparently healthy, APV DNA could be detected in all 40 birds examined (inapparent infection rate, 100%). Viral DNA was found in most organ systems examined. Analysis of data suggested that organ virus concentrations were lower in breeding than in nonbreeding birds. Serum samples from 144 birds were examined for virus-neutralizing (VN) antibody. All serum samples had detectable VN antibody titers. Determining VN titer had a sensitivity of 100% for detection of APV infection in birds and was more sensitive than analysis of droppings by use of polymerase chain reaction techniques to detect APV infection in 6-month-old birds. Analysis of the data suggested that lower VN antibody titers were associated with longer duration of continuous breeding.
Mostrar más [+] Menos [-]Hemagglutination by psittacine beak and feather disease virus and use of hemagglutination inhibition for detection of antibodies against the virus
1991
Ritchie, B.W. | Niagro, F.D. | Latimer, K.S. | Steffens, W.L. | Pesti, D. | Lukert, P.D.
Conditions for psittacine beak and feather disease (PBFD) virus hemagglutination and hemagglutination-inhibition (HI) test reactions are defined. The PBFD virus was found to hemagglutinate cockatoo and some guinea pig erythrocytes. The HI test was used to assay serum antibody titer in birds with active PBFD virus infections and in others that had been exposed to diseased birds. On the basis of HI antibody titers in psittacine birds that had been exposed to PBFD virus, but remained clinically normal, we suggest that some birds exposed to the virus are able to mount an effective immune response. Birds with active PBFD virus infections had lower antibody values than did birds that had been exposed to the virus, but remained clinically normal. On the basis of these findings, the ability to develop a suitable HI antibody response may be crucial in determining the disease status of susceptible birds exposed to the PBFD virus. If HI antibodies are found to have neutralizing activity, then the fact that a high HI titer was induced in birds inoculated with purified PBFD virus might suggest that an immunization program would be effective in preventing PBFD virus infections.
Mostrar más [+] Menos [-]Improved method for recovering ascarid and other helminth eggs from soil associated with epizootics and during survey studies
1983
Kazacos, K.R.
improved method for recovery of ascarid and other nematode eggs from soil, emphasis on Baylisascaris procyonis and Toxocara cati
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