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J Vet Sci. 2018 May 31;19(3):350-357. doi: 10.4142/jvs.2018.19.3.350.

Dembo polymerase chain reaction technique for detection of bovine abortion, diarrhea, and respiratory disease complex infectious agents in potential vectors and reservoirs.

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Research and Education Center for Prevention of Global Infectious Diseases of Animals, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Tokyo 183-0045, Japan.
United Graduate School of Veterinary Science, Gifu University, Gifu 501-1193, Japan.
Faculty of Veterinary Science, Paraclinic Department, Kabul University, Kabul 1006, Afghanistan.
Laboratory of Comparative Pathology, Department of Clinical Science, Faculty of Veterinary Medicine, Hokkaido University, Sapporo 060-0808, Japan.
Center for Animal Disease Control, University of Miyazaki, Miyazaki 889-2192, Japan.
Faculty of Veterinary Science, Nippon Veterinary and Life Science University, Tokyo 180-8602, Japan.
Education and Research Center for Food Animal Health (GeFAH), Gifu University, Gifu 501-1193, Japan.
Canine-Lab. Inc., Tokyo 184-0012, Japan.
Department of Microbiology and Immunology, University of Texas Medical Branch at Galveston, TX 77555-1019, USA.


Bovine abortion, diarrhea, and respiratory disease complexes, caused by infectious agents, result in high and significant economic losses for the cattle industry. These pathogens are likely transmitted by various vectors and reservoirs including insects, birds, and rodents. However, experimental data supporting this possibility are scarce. We collected 117 samples and screened them for 44 bovine abortive, diarrheal, and respiratory disease complex pathogens by using Dembo polymerase chain reaction (PCR), which is based on TaqMan real-time PCR. Fifty-seven samples were positive for at least one pathogen, including bovine viral diarrhea virus, bovine enterovirus, Salmonella enterica ser. Dublin, Salmonella enterica ser. Typhimurium, and Neospora caninum; some samples were positive for multiple pathogens. Bovine viral diarrhea virus and bovine enterovirus were the most frequently detected pathogens, especially in flies, suggesting an important role of flies in the transmission of these viruses. Additionally, we detected the N. caninum genome from a cockroach sample for the first time. Our data suggest that insects (particularly flies), birds, and rodents are potential vectors and reservoirs of abortion, diarrhea, and respiratory infectious agents, and that they may transmit more than one pathogen at the same time.


Dembo polymerase chain reaction; cattle; disease reservoirs; disease vectors; virulence factors

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