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Bovine respiratory disease (BRD) is a complex, multifactorial syndrome and one of the major welfare and economical concerns for the cattle industry. This 1-year cross-sectional study was aimed at documenting the prevalence of BRD-related pathogens and clinical signs before and after a long journey and at identifying possible predisposition factors. Male Limousine beef steers (n = 169) traveling from France to Italy were health checked and sampled with Deep Nasopharyngeal Swabs (DNS) at loading (T0) and 4 days after arrival (T1). Real-time quantitative PCR was used to quantify the presence of bovine viral diarrhea virus (BVDV), bovine respiratory syncytial virus (BRSV), bovine alphaherpesvirus 1 (BoHV-1), bovine coronavirus (BCoV), bovine adenovirus (BAdV), bovine parainfluenza virus 3 (BPIV-3), Histophilus somni, Mannheimia haemolytica, Mycoplasma bovis, and Pasteurella multocida. Weather conditions at departure and arrival were recorded, and the travel conditions were taken from the travel documentation. At T0, even if no animals displayed clinical signs, some of them were already positive for one or more pathogens. At T1, the number of animals displaying clinical signs and positive for BCoV, BAdV, BRSV, H. somni, M. haemolytica, M. bovis, and P. multocida increased dramatically (p < 0.001). Transport also significantly increased co-infection passing from 16.0% at T0 to 82.8% at T1 (p < 0.001). An extra stop during the journey seemed to favor BRSV, M. haemolytica, and P. multocida (p < 0.05). Weather conditions, in particular sudden climate changes from departure to arrival and daily temperature variance, were found to be predisposing factors for many of the pathogens. The farm of arrival also played a role for BRSV, BAdV, and H. somni (p < 0.05). BCoV increased dramatically, but no associations were found confirming that it spreads easily during transport phases. Our findings increased our understanding of factors increasing the likelihood of BRD-related pathogens shedding and can be useful to minimize the incidence of BRD and to implement animal transport regulations.

Aim: The study was designed to measure the seroprevalence of viral and bacterial diseases: Infectious bovine rhinotracheitis, bovine viral diarrhea, bovine leukemia, bovine parainfluenza, bovine respiratory syncytial disease, brucellosis, and paratuberculosis among bovine of Himachal Pradesh during the year 2013-2015. Materials and Methods: The serum samples were collected from seven districts of state, namely, Bilaspur, Kangra, Kinnaur, Lahul and Spiti, Mandi, Sirmour, and Solan. The samples were screened using indirect ELISA kits to measure the seroprevalence of viral and bacterial diseases. Results: The overall seroprevalence of infectious bovine rhinotracheitis was 24.24%, bovine viral diarrhea 1.52%, bovine leukemia 9.09%, bovine parainfluenza 57.58%, bovine respiratory syncytial disease 50%, brucellosis 19.69%, and paratuberculosis 9.09% in Himachal Pradesh. The seroprevalence of bovine rhinotracheitis, bovine leukemia, bovine parainfluenza, bovine respiratory syncytial disease, and paratuberculosis in the state varied significantly (p0.01). Multiple seropositivity has been observed in this study. Bovine parainfluenza virus 3 was observed commonly in mixed infection with almost all viruses and bacteria under study. Conclusion: The viral and bacterial diseases are prevalent in the seven districts of Himachal Pradesh investigated in the study. Therefore, appropriate management practices and routine vaccination programs should be adopted to reduce the prevalence of these diseases.

Bovine parainfluenza virus type 5 (bPIV5) was isolated from cattle with downer cow syndrome in 2012,and included both respiratory and neurotropic pathogens from a variety of animals. In the current study, we conductedserosurveillance using sera obtained from seven Korean farms and optimized a reverse transcription-polymerase chainreaction (RT-PCR) assay to detect bPIV5. The overall seropositive rate for Korean cattle was 21.4% (163/760). Afarm located near the city of Milyang in Gyeoungnam province had a markedly elevated seropositive rate for bPIV5compared to that of the other six farms. The regional seropositive rates were 4.2% (8/192) for Haman, 19.5% (18/55) for Hwasung, 73.9% (65/88) for Milyang, 26.0% (50/192) for Namwon, 1.0% (1/96) for Uljin, 13.5% (13/96)for Yeongju, and 32.7% (8/41) for Yongin. The sensitivity and specificity of three RT-PCR primer sets used to amplifythe conserved fusion gene of bPIV5 were also evaluated. An RT-PCR assay using the bPIVFR3 primer set was 10-fold more sensitive than the assays using the two other primer sets and did not result in non-specific amplification.These results demonstrated that the bPIFR3 primer set can be used to detect bPIV5.Keywords : bovine, parainfluenza virus 5, reverse transcription-polymerase chain reaction, serosurveillance

Vaccine adjuvants are typically designed to stimulate both systemic and mucosal immune responses. Polymeric nanoparticles have been used as adjuvants in the development of vaccines against a number of viral pathogens and tested in laboratory animals. The objective of the study was to assess if synthetic bovine parainfluenza virus type-3 (BPI3V) peptide motifs and solubilised BPI3V proteins encapsulated in poly (dl-lactic-co-glycolide) (PLGA) nanoparticles (NPs) induce specific humoral immune responses in a mouse model following intranasal administration. BPI3V-specific and peptide specific IgG ELISAs were used to measure serum IgG levels to BPI3V. Intranasal delivery of PLGA nanoparticles encapsulating BPI3V proteins elicited an early, gradually increasing BPI3V-specific IgG response that persisted over the subsequent 6 weeks, suggesting slow, persistent release of antigen. PLGA-BPI3V particles administered intranasally induced a stronger IgG antibody response at an earlier time point compared with solubilised BPI3V antigen alone. Such an approach could be deployed in the development of new generation vaccines.

동물세포배양 유래 생물의약품 생산 공정에서 다양한 외래성 바이러스가 오염된 사례가 있기 때문에 바이러스 안전성 보증을 위한 바이러스 검출시험이 필수적이다. Reovirus (Reo), bovine viral diarrhea virus (BVDV), bovine parainfluenza virus (BPIV)는 동물 세포주와 동물 세포 배양 공정에 오염되는 대표적인 RNA 바이러스이다. 세포배양 유래 생물의약품의 안전성을 확보하기 위해, 세포주, 원료물질, 제조공정, 완제품에서 Reo, BVDV, BPIV를 동시에 검출할 수 있는 Multiplex Reverse Transcription (RT)-PCR 시험법을 확립하였다. Reo, BVDV, BPIV에 특이적인 primer를 선별하였으며, multiplex RT-PCR 시험법을 최적화하였다. Reo, BVDV, BPIV를 동시에 검출할 수 있는 multiplex RT-PCR 시험법의 민감도는 각각 $7.76{\times}10^2\;TCID_{50}/ml$ , $7.44{\times}10^1\;TCID_{50}/ml$ , $6.75{\times}10^1\;TCID_{50}/ml$ 이었다. 확립된 multiplex RT-PCR을 생물의약품 제조공정 검증에 적용할 수 있는지 확인하기 위하여 인위적으로 각 바이러스를 오염시킨 CHO 세포에서 검출 시험을 실시한 결과 각 바이러스를 감염시킨 CHO 세포와 세포배양 상청액에서 각 바이러스를 검출할 수 있었다. 위와 같은 결과에서 확립된 multiplex RT-PCR시험법은 세포주, 원료물질, 제조공정, 완제품에서 Reo, BVDV, BPIV를 동시에 검출할 수 있는 특이성과 민감성이 우수한 시험법임을 확인하였다. 【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.】

Viral infections are ubiquitous in humans, animals, and plants. Real-time methods to identify viral infections are limited and do not exist for use in harsh or resource-constrained environments. Previous research identified that tissues produce unique volatile organic compounds (VOC) and demonstrated that VOC concentrations change during pathologic states, including infection, neoplasia, or metabolic disease. Patterns of VOC expression may be pathogen specific and may be associated with an odor that could be used for disease detection. We investigated the ability of two trained dogs to detect cell cultures infected with bovine viral diarrhea virus (BVDV) and to discriminate BVDV-infected cell cultures from uninfected cell cultures and from cell cultures infected with bovine herpes virus 1 (BHV 1) and bovine parainfluenza virus 3 (BPIV 3). Dogs were trained to recognize cell cultures infected with two different biotypes of BVDV propagated in Madin–Darby bovine kidney cells using one of three culture media. For detection trials, one target and seven distractors were presented on a scent wheel by a dog handler unaware of the location of targets and distractors. Detection of BVDV-infected cell cultures by Dog 1 had a diagnostic sensitivity of 0.850 (95% CI: 0.701–0.942), which was lower than Dog 2 (0.967, 95% CI: 0.837–0.994). Both dogs exhibited very high diagnostic specificity (0.981, 95% CI: 0.960–0.993) and (0.993, 95% CI: 0.975–0.999), respectively. These findings demonstrate that trained dogs can differentiate between cultured cells infected with BVDV, BHV1, and BPIV3 and are a realistic real-time mobile pathogen sensing technology for viral pathogens. The ability to discriminate between target and distractor samples plausibly results from expression of unique VOC patterns in virus-infected and -uninfected cells.

Bovine parainfluenza virus 3 (BPIV3) is one of the most important respiratory pathogens in cattle. One BPIV3, named NM09, was isolated from cattle suffering from severe respiratory diseases in 2009. BPIV3 is a potential recombinant vaccine vector. To investigate whether NM09 can infect pigs and determine BPIV3 defense in these animals, BPIV3 antibody-free pigs were inoculated intramuscularly with the BPIV3 NM09 strain in a continuous passage. Clinical signs were observed each day after inoculation. Viral nucleic acid was detected in nasal and anal secretions. Results showed that virus-inoculated pigs displayed few observable clinical signs related to respiratory diseases. The antibody was identified, but the virus could not be detected in the second continuous passage in pigs. Thus, BPIV3 is a potential vaccine vector for genetic engineering.

Objective: This study was aimed at elucidating the association between Para influenza virus 3 (PIV3) and respiratory infections in domestic ruminants in different areas of Sudan. Materials and methods: During 2010-2013, five hundred sixty five lung samples with signs of pneumonia were collected from cattle (n=226), sheep (n=316) and goats (n=23) from slaughter houses in different areas in Sudan. The existence of PIV3 antigen was screened in the collected samples using ELISA and Fluorescent antibody technique. PIV3 genome was detected by PCR, and sequence analysis was conducted. Results: Positive results were found in 29 (12.8%) cattle, 31 (9.8%) sheep and 11 (47.8%) goat samples. All the studied areas showed positive results. Highest prevalence (66.7%) was detected in the sheep and goats in Khartoum, followed by in goats in Nyala (33.3%) at western Sudan. Sequence analyses of PIV3 of different regions of Sudan indicated that these were similar in sequence and length. The BLAST analysis indicated that the test sequences were closely related to the available annotated sequences at the GenBank. All these sequences matched with Bovine parainfluenza virus 3 except two those were matching with Swine parainfluenza virus 3. Conclusion: The results prove the existence of PIV3 infection in cattle, sheep and goats in the studied areas in Sudan and suggest its possible role in the respiratory infections. Genetic analysis indicate that the virus is mostly similar with bovine PIV3. [J Adv Vet Anim Res 2016; 3(3.000): 236-241]

The Bovine Practitioner, No. 12 (1977 November)