• Journal of Practical Agriculture & Fisheries Research
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• v.5 no.1
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• pp.57-63
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• 2003

The bovine rotavirus(BRV), bovine coronavirus(BCV) and bovine viral diarrhea virus(BVDV) are main viruses of bovine viral diarrhea disease. These viruses could be rapidly amplified by the reverse transcriptase polymerase chain reaction(RT-PCR). This study was conducted to develop rapid and accurate diagnostic methods of these viral diseases by multiplex RT-PCR. Specific primers were designed based on the sequences reported by Chang KO et. al. (1997) and Schroeder BA, et. al. (1990), RNA were prepared from the cultured viruses, first-stranded DNAs were synthesised by reverse transcriptase. PCR were conducted to amplify specific regions of the viruses by multiplex. Three bands such as 1,062bp for BRV, 458bp for BCV, and 300bp for BVDV were successfully produced by multiplex RT-PCR. In conclusion, this result suggested that these viruses could be diagnosed rapidly and accurately by multiplex RT-PCR.

• Microbiology and Biotechnology Letters
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• v.40 no.4
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• pp.339-347
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• 2012

Viral safety is an important prerequisite for clinical preparations of mammalian cell culture-derived biopharmaceuticals, because numerous adventitious viruses have been contaminated during the manufacturing process. In particular, Chinese hamster ovary (CHO) cells are highly susceptible to several RNA viruses including reovirus (Reo), bovine viral diarrhea virus (BVDV), and bovine parainfluenza virus (BPIV) and there have been reports of such viral contaminations. Therefore, viral detection during the CHO cell process is necessary to ensure the safety of biopharmaceuticals against viruses. In this study, a multiplex reverse transcription (RT)-PCR assay was developed and subsequently evaluated for its effectiveness as a means to simultaneously detect Reo, BVDV, and BPIV during the manufacture of cell culture-derived biopharmaceuticals. Specific primers for Reo, BVDV, and BPIV were selected, and a multiplex RT-PCR was optimized. The sensitivity of the assay for simultaneous amplification of all viral target RNAs was $7.76{\times}10^2\;TCID_{50}/ml$ for Reo, $7.44{\times}10^1\;TCID_{50}/ml$ for BVDV, and $6.75{\times}10^1\;TCID_{50}/ml$ for BPIV. The multiplex RT-PCR was proven to be very specific to Reo, BVDV, and BPIV and was subsequently applied to the validation of CHO cells artificially infected with each virus. It could detect each viral RNA from CHO cells as well as culture supernatants. Therefore, it was concluded that the multiplex RT-PCR assay can be applied to detection of the adventitious viruses during the manufacture of cell culture-derived biopharmaceuticals.

• Korean Journal of Veterinary Research
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• v.39 no.4
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• pp.743-750
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• 1999

Bovine viral diarrhea viruses (BVDVs) were isolated from cattle with respiratory and diarrhea signs as well as persistently infected cattle. These isolates were analysed serologically to characterize serogroups and to compare serological relationship with reference viruses of type I and II. Most isolates from calf diarrheal cases and persistently infected individuals showed a significant difference in cross-neutralization test with the viruses isolated from nasal discharges showing severe respiratory signs. Serologically most of the commercial vaccine strains could be classified into classical BVDV (type I) such as NADL strain. This serological difference among BVDV isolates suggested the need for new vaccines to protect cattle from both respiratory and enteric BVDV infections in field. The immunogenicity of BVDVs which showed a good propagation capability in MDBK cells and high rates of neutralizing activity (isolate : KD26-1, PHG, B5 and 95002) against all viruses used in this study, was confirmed in guinea pig when treated as single or combined groups.

• Korean Journal of Veterinary Service
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• v.31 no.3
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• pp.265-272
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• 2008

Among calves' diseases, digestive diseases most frequently occur in Korea, and especially viral diarrhea is one of the most important diseases. This study was conducted to get some basic information for the control of the viral diarrhea in calves. The samples were obtained from 319 calves with clinical diarrhea from 195 farms in Gyeongnam south area (Gosung, Tongyung, Hadong) from June 2005 to August 2006, Viral detection was examined by polymerase chain reaction (PCR), Etiological viruses were detected from 171(53,6%) of 319 calves, and bovine rotavirus (BRV) were 130 (40,8%) and bovine viral diarrhea virus (BVDV) 41 (12,9%), and no coronavirus was confirmed, Statistical difference was found in BRV detection between summer (32.6%) and winter (57.7%). However there was no seasonal difference in BVDV. In detection rate of the calves under 19days, BRV was highest (55.1%), but BVDV lowest (5.1%). No big difference was in rate among herd size. However, BRV was lowest (26.8%) in the group over 51 heads, but BVDV was highest (19.5%) in the same.

• Journal of Embryo Transfer
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• v.1 no.1
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• pp.16-27
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• 1986

Based on the current importing and exporing regulations for disease control of embryo transfer, some important microorganisms and their control possibilities are reviewed. The results reviewed were sumrnarized as follows: 1. Regulations regarding to the import of embryos vary between importing and exporting countries, but exporting countries examine the donor and embryos for the heaith certification by the requirements of importing countries. 2. Organisms that infect the gametes are 5 kinds of viruses and the diseases caused by them could not be controlled or eradicated using embryo transfer. 3. Organisms that do not infect the gametes are 4 kinds of viruses and the causal organisms are potential candidates for control or eradication by embryo transfer. 4. Organisms that penetrate the zona pellucida and infect the embryo are 6 kinds of viruses including bovine viral diarrhea virus. 5. Organisms that cannot penetrate the zona pellucida or do not infect the embryo are 15 kinds of viruses and the removal from their contaminations are recommended by proper washing procedure and antisera treatment. Bovine and porcine parvovirus, porcine pseudorabies virus and vesicular stomatitis virus are included in these organisms. 6. Bovine embryos that artificially exposed to various pathogenic organisms such as bovine herpes virus, IBR virus, bluetongue virus, bovine viral diarrhea virus and Brucella abortus in vitro are discussed about their infection by several treatments.

• Journal of Animal Science and Technology
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• v.66 no.4
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• pp.781-791
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• 2024

Bovine viral diarrhea (BVD) is a single-stranded, positive-sense ribonucleic acid (RNA) virus belonging to the genus Pestivirus of the Flaviviridae family. BVD frequently causes economic losses to farmers. Among bovine viral diarrhea virus (BVDV) strains, BVDV-1b is predominant and widespread in Hanwoo calves. Reverse-transcription polymerase chain reaction (RT-PCR) is an essential method for diagnosing BVDV-1b and has become the gold standard for diagnosis in the Republic of Korea. However, this diagnostic method is time-consuming and requires expensive equipment. Therefore, Clustered regularly interspaced short palindromic repeats-Cas (CRISPR-Cas) systems have been used for point-of-care (POC) testing of viruses. Developing a sensitive and specific method for POC testing of BVDV-1b would be advantageous for controlling the spread of infection. Thus, this study aimed to develop a novel nucleic acid detection method using the CRISPR-Cas13 system for POC testing of BVDV-1b. The sequence of the BVD virus was extracted from National Center for Biotechnology Information (NC_001461.1), and the 5' untranslated region, commonly used for detection, was selected. CRISPR RNA (crRNA) was designed using the Cas13 design program and optimized for the expression and purification of the LwCas13a protein. Madin Darby bovine kidney (MDBK) cells were infected with BVDV-1b, incubated, and the viral RNA was extracted. To enable POC viral detection, the compatibility of the CRISPR-Cas13 system was verified with a paper-based strip through collateral cleavage activity. Finally, a colorimetric assay was used to evaluate the detection of BVDV-1b by combining the previously obtained crRNA and Cas13a protein on a paper strip. In conclusion, the CRISPR-Cas13 system is highly sensitive, specific, and capable of nucleic acid detection, making it an optimal system for the early point-of-care testing of BVDV-1b.

• Journal of Veterinary Clinics
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• v.11 no.2
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• pp.585-592
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• 1994

A consecutive Jiao Chio acupuncture therapy was performed for 3 days in the 45-90 days old 11 calves of which have been shown severe watery diarrhea. The discharge of the infected calves was yellowish brown in color. Two calves of these patients were infected wi pulmonary disease as well as diarrhea. Thus, Su Qi and Fei Yu acupuncture therapy was carried out additionally after dosing with antibiotics twice for The two infected calves. Blood chemical values and serum neutralizing antibody titers were checked, and total blood cell count was also carried out to know the therapeutic effect before and after(21 days) acupuncture therapeutics. The results are as follows ; 1, The diarrhea has ceased one day after begining of the acupuncture therapy in 5 calves, and the cessation of the diarrhea in remaining calves occurred in 1 calf each on 3rd and 4th day, and 2 calves on 6th day, respectively. Two calves infected with pulmonary disease as well diarrhea were cured 8 days after the begining of the therapcutics. 2. Rotaviruses wire detected in the feces of 2 calves, and bovine diarrhea viruses were detected in the 8 calves by the test for serum neutralizing antibody titers, and bovine coronaviruses were also detected in 5 calves. Four calves of the 5 bovine coronavirus infected calves were also infected with bovine diarrhea viruses. 3. Total leucocyte number, total amount of serum protein, and amount of fibringen were slightly increased, while total erythrocyte number, and erythrocyte packed cell volume were slightly decreased. These valucs were statistically not significant. Electrolytes of Na/sup +/, K/sup +/ and Cl/sup -/ were slightly decreased but these values also were not significant. These results indicate that the acupuncture therapeutics arc significantly effective to remove the viral diarrhea in the young calves.

• Clinical and Experimental Vaccine Research
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• v.11 no.1
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• pp.30-42
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• 2022

Purpose: The key objective of this study was to formulate a local combined inactivated gel adjuvanted vaccine containing bovine viral diarrhea virus (BVDV)-1, BVDV-2 viruses and Clostridium perfringens type A toxoid. The study evaluated its ability to enhance protective active immune response in camels' calves against these infectious pathogens under field conditions. Materials and Methods: The local BVDV cytopathic strains and a local strain of toxigenic C. perfringens type A were used in vaccines formulation. Vaccines A and B were monovalent vaccines against C. perfringens and both strains of BVDVs, respectively. While the vaccine C was the combined vaccine used against the three agents. All vaccines were adjuvanted with Montanide gel. Sterility, safety, and potency tests were applied on the formulated vaccines. Virus neutralization and toxin anti-toxin neutralization tests were used to evaluate the immune responses. Results: Both monovalent (vaccine A) and combined vaccines (vaccine C) showed a protective level (4.5 and 3 IU/mL, respectively) against C. perfringens from the 2nd-week post-vaccination. The titer declined to 3 and 2 IU/mL, respectively at the 5th-month post-vaccination. The titer against BVDV, the monovalent vaccine (vaccine B) reached the beak (1.95 IU/mL) at the 1st-month post-vaccination and lasted till 6th-month post-vaccination (0.92 and 0.94 IU/mL) for BVDV-1a and BVDV-2, respectively. Conclusion: Vaccination of camels with the combined vaccine adjuvanted by Montanide gel containing C. perfringens type A toxoid and BVDV strains with 6-month intervals is recommended to protect camels safely and efficiently against such infections in the field.

• Journal of Microbiology and Biotechnology
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• v.18 no.7
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• pp.1317-1325
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• 2008

Viral safety is an important prerequisite for clinical preparations of plasma-derived pharmaceuticals. One potential way to increase the safety of therapeutic biological products is the use of a virus-retentive filter. In order to increase the viral safety of human antihemophilic factor IX, particularly in regard to non-enveloped viruses, a virus removal process using a polyvinylidene fluoride membrane filter (Viresolve NFP) has been optimized. The most critical factor affecting the filtration efficiency was operating pH and the optimum pH was 6 or 7. Flow rate increased with increasing operating pressure and temperature. Recovery yield in the optimized production-scale process was 96%. No substantial changes were observed in the physical and biochemical characteristics of the filtered factor IX in comparison with those before filtration. A 47-mm disk membrane filter was used to simulate the process performance of the production-scale cartridges and to test if it could remove several experimental model viruses for human pathogenic viruses, including human hepatitis A virus (HAV), porcine parvovirus (PPV), murine encephalomyocarditis virus (EMCV), human immunodeficiency virus type 1 (HIV), bovine viral diarrhea virus (BVDV), and bovine herpes virus (BHV). Non-enveloped viruses (HAV, PPV, and EMCV) as well as enveloped viruses (HIV, BVDV, and BHV) were completely removed during filtration. The log reduction factors achieved were $\geq$6.12 for HAV, $\geq$4.28 for PPV, $\geq$5.33 for EMCV, $\geq$5.51 for HIV, $\geq$5.17 for BVDV, and $\geq$5.75 for BHV. These results indicate that the virus filtration process successfully improved the viral safety of factor IX.

• Microbiology and Biotechnology Letters
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• v.37 no.4
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• pp.377-382
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• 2009

Viral safety is an important prerequisite for clinical preparations of all biopharmaceuticals derived from plasma, cell lines, or tissues of human or animal origin. To ensure the safety, implementation of multiple viral clearance (inactivation and/or removal) steps has been highly recommended for manufacturing of biopharmaceuticals. Of the possible viral clearance strategies, Ultraviolet-C (UVC) irradiation has been known as an effective viral inactivating method. However it has been dismissed by biopharmaceutical industry as a result of the potential for protein damage and the difficulty in delivering uniform doses. Recently a continuous flow UVC reactor (UVivatec) was developed to provide highly efficient mixing and maximize virus exposure to the UV light. In order to investigate the effectiveness of UVivatec to inactivate viruses without causing significant protein damage, the feasibility of the UVC irradiation process was studied with a commercial therapeutic protein. Recovery yield in the optimized condition of $3,000\;J/m^2$ irradiation was more than 98%. The efficacy and robustness of the UVC reactor was evaluated with regard to the inactivation of human immunodeficiency virus (HIV), hepatitis A virus (HAV), bovine herpes virus (BHV), bovine viral diarrhea virus (BVDV), porcine parvovirus (PPV), bovine parvovirus (BPV), minute virus of mice (MVM), reovirus type 3 (REO), and bovine parainfluenza virus type 3 (BPIV). Non enveloped viruses (HAV, PPV, BPV, MVM, and REO) were completely inactivated to undetectable levels by $3,000\;J/m^2$ irradiation. Enveloped viruses such as HIV, BVDV, and BPIV were completely inactivated to undetectable levels. However BHV was incompletely inactivated with slight residual infectivity remaining even after $3,000\;J/m^2$ irradiation. The log reduction factors achieved by UVC irradiation were ${\geq}3.89$ for HIV, ${\geq}5.27$ for HAV, 5.29 for BHV, ${\geq}5.96$ for BVDV, ${\geq}4.37$ for PPV, ${\geq}3.55$ for BPV, ${\geq}3.51$ for MVM, ${\geq}4.20$ for REO, and ${\geq}4.15$ for BPIV. These results indicate that UVC irradiation using UVivatec was very effective and robust in inactivating all the viruses tested.