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Summary

Rapid and accurate detection of a virus in a population is a critical factor in the eventual treatment and/or control of the virus. In this study, we examined use of the polymerase chain reaction (pcr) to detect swine influenza virus in nasal swab specimens from infected pigs. This approach was first standardized, using viral rna purified by guanidinium/phenol-chloroform extraction and placed in the same transport medium as the swabs. By using highly conserved primers for the swine H1 hemagglutinin, we amplified a 591-base pair fragment that was analyzed by use of agarose gel electrophoresis, Southern blot, and dna sequencing.

To evaluate pcr as a potential diagnostic tool for detection of swine influenza virus infection, we obtained nasal swab specimens from experimentally infected pigs. Amplification by pcr and reamplification of extracted samples with internal primers yielded detectable bands for an amount of virus less than that required to infect embryonating chicken eggs. We also tested swab specimens from pigs involved in 3 separate, natural episodes of swine influenza. These swab specimens were extracted, amplified and reamplified, producing visible bands on the gel and in Southern blots. We performed Southern blot analyses on all pcr products, to confirm that they were from viral H1 rna. We also cloned and sequenced a 591-base pair product from 1 specimen and found that it was 100% identical to the hemagglutinin gene sequence of A/Sw/Ind/1726/88. Results indicate that pcr can be used to detect swine influenza virus, even in nasal swab specimens, the specimen typically collected for diagnosis of virus infection.

Free access
in American Journal of Veterinary Research

Objective

To monitor the prevailing viral respiratory tract infections in cattle after transportation to feedlots.

Animals

100 cattle with signs of respiratory tract disease on arrival at 2 feedlots.

Procedures

Nasal swab samples were obtained from each animal and were used for inoculation of defined cell culture systems that detected bovine viruses known to cause respiratory tract infections, as well as viruses previously not recognized as respiratory pathogens for cattle.

Results

Bovine respiratory coronaviruses were isolated from 38 of the 100 cattle, including 6 of 50 cattle from California, 22 of 31 cattle from Oklahoma, 6 of 11 cattle from Texas, and 4 of 8 cattle of unknown origin. Parainfluenza 3 viruses also were isolated from 4 California cattle, but other bovine viruses were not detected.

Clinical Implications

The high rate of coronavirus isolations from feedlot cattle with signs of respiratory tract disease implied wide distribution and high susceptibility among cattle to this infection, which had not been detected by use of viral isolation systems in previous etiologic evaluations of feedlot cattle affected with bovine respiratory disease complex. (J Am Vet Med Assoc 1996; 208:1452-1455)

Free access
in Journal of the American Veterinary Medical Association

calves with BRD may help veterinarians when designing antimicrobial treatment protocols for dairy operations. Multiple sampling methods are used to identify the various bacterial respiratory pathogens associated with BRD, with the nasal swab (NS), deep

Open access
in American Journal of Veterinary Research

influenza virus by rRT-PCR assay of nasal swab specimens through the University of Wisconsin Shelter Medicine Program and the WVDL. These 300 dogs represented a sampling of dogs with respiratory disease from each shelter. The combined annual intake for dogs

Full access
in Journal of the American Veterinary Medical Association

horses with neurologic signs. Blood, nasal swab specimen, and CSF sampling and processing —Blood samples and nasal swab specimens were obtained on day −1 and 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, and 21 days after infection. Blood was drawn into 10-mL

Full access
in American Journal of Veterinary Research

hours after birth from a small subset (n = 34) of calves for which both birth and the first nursing episode were observed. A blood sample (approx 9 mL at initial vaccination and 25 mL at preconditioning and weaning) for serum acquisition and nasal swab

Full access
in American Journal of Veterinary Research

inoculation in blood tubes without an anticoagulant to obtain serum for VN and VI. Nasal swab specimens for VI were also obtained and body temperature measured by use of a rectal thermometer on days 0, 3, 5, 6, 7, 8, 9, 11, and 14 after BVDV challenge

Full access
in American Journal of Veterinary Research

serum samples and nasal swab specimens obtained from calves vaccinated with a modified-live BVDV vaccine. In 1 study, 11 serum samples obtained from 78% of vaccinated calves between 3 and 10 days after vaccination and tested by use of RT-PCR assay had

Full access
in American Journal of Veterinary Research

and that calves with otitis media would be more likely to have nasal swab specimens with positive M bovis culture results versus calves without otitis media. Materials and Methods Animals —All study procedures were approved by the Animal Care

Full access
in Journal of the American Veterinary Medical Association

miscellaneous illnesses were treated on an individual basis according to clinical signs. Collection of samples —For each group of calves, paired nasal swab specimens and fecal samples were obtained prior to shipping, at arrival at the feedlot research station

Full access
in American Journal of Veterinary Research