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Sample records for bovine viral diarrhea

  1. Molecular biology of bovine viral diarrhea virus

    Science.gov (United States)

    Bovine viral diarrhea viruses (BVDV) are arguably the most important viral pathogen of ruminants worldwide and can cause severe economic loss. Clinical symptoms of the disease caused by BVDV range from subclinical to severe acute hemorrhagic syndrome, with the severity of disease being strain depend...

  2. The evolution of bovine viral diarrhea: a review

    OpenAIRE

    Goens, Denise

    2002-01-01

    The economic importance of bovine viral diarrhea is increasing with the emergence of seemingly more virulent viruses, as evidenced by outbreaks of hemorrhagic syndrome and severe acute bovine viral diarrhea beginning in the 1980s and 1990s. It appears that evolutionary changes in bovine viral diarrhea virus were responsible for these outbreaks. The genetic properties of the classical bovine viral diarrhea virus that contribute to the basis of current diagnostic tests, vaccines, and our unders...

  3. Bovine viral diarrhea virus: biosecurity and control

    Science.gov (United States)

    This paper discusses the recommended procedures involved in setting up biosecurity and control programs designed to limit bovine viral diarrhea virus infections in beef cattle operations. For the purpose of these discussions, a working definition of a biosecurity plan was considered to be an organiz...

  4. DETECTION OF THE BOVINE VIRAL DIARRHEA ANTIBODIES

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    I. V. Goraichuk

    2013-06-01

    Full Text Available Bovine viral diarrhea is a widespread infection of cattle that has a wide range of clinical symptoms in domestic and wild ruminants. It is a major problem in cattle and causes significant economic losses in the cattle industry. The virus infects bovines of all ages and causes both immunosuppression and reproductive, respiratory and digestive disorders. Persistently infected cattle are the main factor in transmission of the disease between and among herds. Comparative results of antibodies presence received by two methods of enzymoimmunoassay and virus neutralization test are given in the paper. During the work, 1010 samples of blood serum of cattle from three farms in the Kharkiv region were selected and analyzed. Bovine viral diarrhea virus concerning antibodies were found by enzymoimmunoassay in 704 samples (69.7% using commercial kit and in 690 samples (68.3% using in house method. After results clarification by virus neutralization test, bovine viral diarrhea antibodies were found in 712 samples (70.5%. Immunoenzyme analysis is recommended for mass screening of cattle for viral diarrhea occurrence. The results confirm that the sensitivity immunoenzyme analysis satisfies the requirements of the diagnostic methods. Using the neutralization reaction of viruses as the «gold standard» of serological methods, it is appropriate to clarify the results of immunoenzyme analysis. Since the results contain a signi ficant number of false positive results, it is necessary to carry out comprehensive studies using both serological and molecular genetics methods.

  5. [Bovine viral diarrhea control in Russian Federation].

    Science.gov (United States)

    Guliukin, M I; Iurov, K P; Glotov, A G; Donchenko, N A

    2013-01-01

    Bovine viral diarrhea (BVD) is one of the greatest challenges for breeding and commercial livestock. It is characterized by lesions of the respiratory and gastrointestinal tract, abortion, infertility, immune deficiency, and persistence of the pathogen. In this work, a set of measures for the rehabilitation and prevention of BVD in cattle is described. It includes the data of the literature, guidance documents for the diagnosis and control of BVD adopted by OIE, EU countries, USA, as well as the results of this research.

  6. Diagnosis and Control of Viral Diseases of Reproductive Importance: Infectious Bovine Rhinotracheitis and Bovine Viral Diarrhea.

    Science.gov (United States)

    Newcomer, Benjamin W; Givens, Daniel

    2016-07-01

    Both bovine viral diarrhea virus and bovine herpesvirus 1 can have significant negative reproductive impacts on cattle health. Vaccination is the primary control method for the viral pathogens in US cattle herds. Polyvalent, modified-live vaccines are recommended to provide optimal protection against various viral field strains. Of particular importance to bovine viral diarrhea control is the limitation of contact of pregnant cattle with potential viral reservoirs during the critical first 125 days of gestation.

  7. Experimental infection of reindeer with bovine viral diarrhea virus

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    J.K. Morton

    1990-08-01

    Full Text Available Two 8-month reindeer (Rangifer tarandus and a 1-month-old Hereford-Holstein calf (Bos taurus were inoculated intranasally with the Singer (cytopathogenic strain of bovine viral diarrhea (BVD virus. Clinical signs in reindeer included loose stools containing blood and mucus, and transient laminitis or coronitis. Signs in the calf were limited to bloody mucus in the stool and lesions in the nasal mucosa. Antibody titers to BVD virus in the reindeer were intermittent, and titers in the calf persisted from days 14 to 63 post-inoculation (PI. Viremia was detected on PI day 4 in one reindeer, days 3-7 in the other, and days 2-7 in the calf. Bovine viral diarrhea virus was isolated from the lung of the calf at necropsy (PI day 63.

  8. Bovine Viral Diarrhea Virus-Associated Disease in Feedlot Cattle.

    Science.gov (United States)

    Larson, Robert L

    2015-11-01

    Bovine viral diarrhea virus (BVDv) is associated with bovine respiratory disease complex and other diseases of feedlot cattle. Although occasionally a primary pathogen, BVDv's impact on cattle health is through the immunosuppressive effects of the virus and its synergism with other pathogens. The simple presence or absence of BVDv does not result in consistent health outcomes because BVDv is only one of many risk factors that contribute to disease syndromes. Current interventions have limitations and the optimum strategy for their uses to limit the health, production, and economic costs associated with BVDv have to be carefully considered for optimum cost-effectiveness.

  9. Bovine respiratory disease model based on dual infections with infection with bovine viral diarrhea virus and bovine corona virus

    Science.gov (United States)

    Bovine respiratory disease complex (BRDC) is the leading cause of economic loss in the U.S. cattle industry. BRDC likely results from simultaneous or sequential infections with multiple pathogens including both viruses and bacteria. Bovine viral diarrhea virus (BVDV) and bovine corona virus (BoCV...

  10. Determining bovine viral diarrhea virus genotypes and biotypes circulating in cattle populations in Mexico

    Science.gov (United States)

    Bovine viral diarrhea (BVD) is the disease in cattle that results from infection with bovine viral diarrhea viruses (BVDV). BVDV is found in cattle populations throughout the world. While the term BVD encompasses a wide range of clinical manifestations, including severe respiratory disease, gastroe...

  11. Diagnosis of natural exposure to bovine viral diarrhea in a vaccinated herd by measuring extended antibody titers against bovine viral diarrhea virus

    OpenAIRE

    Ross, Jeremy

    2003-01-01

    Two abortions occurred in a 150-head commercial cow-calf herd. Bovine viral diarrhea was suspected and confirmed by measuring extended titers against bovine viral diarrhea virus (BVDV) in a sample of 15 breeding females. Fifteen were sero-positive and 11 had significantly high titers (1:972–1:8748), likely due to natural exposure to cattle persistently infected with BVDV.

  12. Molecular diversity of bovine viral diarrhea virus in uruguay.

    Science.gov (United States)

    Maya, L; Puentes, R; Reolón, E; Acuña, P; Riet, F; Rivero, R; Cristina, J; Colina, R

    2016-03-01

    Bovine viral diarrhea (BVD) affects bovine production and reproduction causing significant economic losses all over the world. Two viral species has been recognized: BVDV-1 and BVDV-2, both distributed worldwide. Recently, novel specie of BVDV named HoBi-like pestivirus was discovered. The presence of BVDV was confirmed in 1996 in Uruguay, however, does not exist until today a schedule of compulsory vaccination along the country. Serological studies with samples from all Uruguayan herds were performed during 2000 and 2001 demonstrating that all of them were seropositive to BVDV with a mean prevalence of 69%. In addition, there have been no new studies done since those previously described and it is important to mention that the genetic diversity of BVD has never been described in Uruguay. Nowadays, there is strongly suspect that BVDV is one of the most important causes of reproductive failures in our herds. The aim of this study was to describe for the first time in Uruguay the genetic diversity of BVDV with samples collected from different regions along the country. Serological status of 390 non-vaccinated animals against BVDV with reproductive problems from farms of Rivera, Tacuarembó and Florida departments of Uruguay were studied. All herds were seropositive to BVDV and high proportion of animals were positive (298/390), while 4.1% (16/390) of the animals were positive to Antigen Capture ELISA test and Real Time PCR. Phylogenetic analysis performed with concatenated sequences from the 5'UTR and Npro genomic regions revealed that BVDV-1 and BVDV-2 are infecting our herds, being BVDV-1 the most frequently found. The major subtype was BVDV-1a, followed by BVDV-1i and BVDV-2b. This is the first study that describes the genetic diversity of BVDV in Uruguay and it will contribute to the elaboration of sanitization programs.

  13. Bovine viral diarrhea (BVD: A review emphasizing on Iran perspective

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    Mohammad Khezri

    2015-09-01

    Full Text Available Bovine viral diarrhea (BVD is one of the most important diseases of cattle responsible for major economic losses in dairy industries of Iran. So far, no nationwide program has been taken in Iran to control and eradicate the disease. Moreover, until now, no vaccination program has been practiced against BVD in Iran, although the disease is prevailing in the country. For effective controlling of BVD, it is necessary to cull the affected animals, and new entry of BVD in the farm should be prevented. Focusing on biosecurity in systematic control programs of BVD can also reduce the risks of introduction and spread of other epizootic and zoonotic diseases, thereby improving both cattle health and welfare in general. In this review paper, an overview on BVD emphasizing on Iran perspective has been discussed focusing on clinical manifestations of BVD, routes of transmission of BVD virus (BVDV, its diagnostic methods and possible prevention strategies. [J Adv Vet Anim Res 2015; 2(3.000: 240-251

  14. Molecular detection and characterization of bovine viral diarrhea virus in Mongolian cattle and yaks.

    Science.gov (United States)

    Ochirkhuu, Nyamsuren; Konnai, Satoru; Odbileg, Raadan; Odzaya, Battogtokh; Gansukh, Shura; Murata, Shiro; Ohashi, Kazuhiko

    2016-08-01

    Bovine viral diarrhea virus (BVDV) is classified into two species, namely, Bovine viral diarrhea virus 1 and Bovine viral diarrhea virus 2, and affects cattle worldwide, resulting in significant economic loss. The prevalence of BVDV-1 and BVDV-2 infections and its genotypes in Mongolian animals has not been studied. In this study, we surveyed BVDV infection in dairy cattle and yaks from Bornuur and Bulgan counties by RT-PCR, and the average infection rate in the sampling sites was 15.8 % and 20.0 %, respectively. In addition, molecular features of the 5'-UTR region of the BVDV genome in Mongolian cattle and yaks were identified as belonging to the subtypes BVDV-1a and BVDV-2a, respectively. Determining the prevalence, geographical distribution, and molecular diversity of BVDV-1 and BVDV-2 in various host species in Mongolia is important for further studies and process control programs.

  15. Genetic characterization of a noncytopathic bovine viral diarrhea virus 2b isolated from cattle in China.

    Science.gov (United States)

    Wang, Wei; Shi, Xinchuan; Chen, Chaoyang; Wu, Hua

    2014-10-01

    In January 2013, several clinical signs of cattle with diarrhea, cough, nasal discharge, and fever were reported in Jilin province, China. One virus named SD1301 was isolated and identified. Complete genome of the virus is 12258nt in length and contains a 5'UTR, one open reading frame encoding a polyprotein of 3,897 amino acids and a 3'UTR. Phylogenetic analysis of 5'UTR, N(pro), E1 and E2 gene demonstrated the virus belonged to BVDV 2b, and genetically related to the BVDV strain Hokudai-Lab/09 from Japan in 2010. This bovine viral diarrhea virus displays a unique genetic signature with 27-nucleotide deletion in the 5'UTR, which is similar to the bovine viral diarrhea virus C413 (AF002227). This was the first confirmed isolation of ncp BVDV2b circulating in bovine herd of China.

  16. Case Report: Emergence of bovine viral diarrhea virus persistently infected calves in a closed herd

    Science.gov (United States)

    Bovine viral diarrhea virus (BVDV) continues to have significant economic impact on the cattle industry worldwide. The virus is primarily maintained in the cattle population due to persistently infected animals. Herd surveillance along with good vaccination programs and biosecurity practices are the...

  17. Stochastic simulation modeling to determine time to detect Bovine Viral Diarrhea antibodies in bulk tank milk

    DEFF Research Database (Denmark)

    Foddai, Alessandro; Enøe, Claes; Krogh, Kaspar;

    2014-01-01

    A stochastic simulation model was developed to estimate the time from introduction ofBovine Viral Diarrhea Virus (BVDV) in a herd to detection of antibodies in bulk tank milk(BTM) samples using three ELISAs. We assumed that antibodies could be detected, after afixed threshold prevalence...

  18. Bovine viral diarrhea virus outbreak in a beef cow herd in South Dakota

    Science.gov (United States)

    The objective of this study was to describe the outcome of natural bovine viral diarrhea virus (BVDV) infection in a herd of 136 bred heifers. This outbreak was notable in that a total of 36 PI calves were generated. Of the 136 bred heifers, 8 failed to deliver a calf. Eight calves died shortly a...

  19. Complete Genome Sequence of a Bovine Viral Diarrhea Virus Subgenotype 1h Strain Isolated in Italy

    Science.gov (United States)

    Bazzucchi, Moira; Bertolotti, Luigi; Casciari, Cristina; Rossi, Elisabetta; Rosati, Sergio; De Mia, Gian Mario

    2017-01-01

    ABSTRACT We sequenced the complete genome of bovine viral diarrhea virus (BVDV) strain UM/126/07. It belongs to subgenotype 1h. The complete genome is composed of 12,196 nucleotides organized as one open reading frame encoding 3,898 amino acids. This is the first report of a full-length sequence of BVDV-1h. PMID:28232427

  20. Resolving bovine viral diarrhea virus subtypes from persistently infected US beef calves with complete genome sequence

    Science.gov (United States)

    Bovine viral diarrhea virus (BVDV) is classified into 2 genotypes, BVDV-1 and BVDV-2, each of which contains distinct subtypes with genetic and antigenic differences. Currently, three major subtypes circulate in the United States: BVDV-1a, 1b, and 2a. In addition, a single case of BVDV-2b infection ...

  1. Studies on genetic diversity of bovine viral diarrhea viruses in Danish cattle herds

    DEFF Research Database (Denmark)

    Nagy, Abdou; Fahnøe, Ulrik; Rasmussen, Thomas Bruun;

    2014-01-01

    Scandinavian countries have successfully pursued bovine viral diarrhea virus (BVDV) eradication without the use of vaccines. In Denmark, control and eradication of BVDV were achieved during the last two decades, but occasionally new BVDV infections are detected in some Danish cattle herds. The aim...

  2. Experimental infection of pregnant goats with bovine viral diarrhea virus (BVDV)1 or 2

    Science.gov (United States)

    Infections with bovine viral diarrhea virus (BVDV) of the genus pestivirus, family Flaviviridae, are not limited to cattle but occur in various artiodactyls. Persistently infected (PI) cattle are the main source of BVDV. Persistent infections also occur in heterologous hosts such as sheep and deer. ...

  3. Genetic diversity of bovine viral diarrhea virus in cattle from Mexico

    Science.gov (United States)

    Bovine viral diarrhea virus (BVDV) infects cattle populations worldwide causing significant economic losses though its impact in animal health. Previous studies have reported the prevalence of BVDV species and subgenotypes in cattle from the United States and Canada. In this study, we investigated t...

  4. Complete Genome Sequence of a Bovine Viral Diarrhea Virus Subgenotype 1h Strain Isolated in Italy.

    Science.gov (United States)

    Bazzucchi, Moira; Bertolotti, Luigi; Giammarioli, Monica; Casciari, Cristina; Rossi, Elisabetta; Rosati, Sergio; De Mia, Gian Mario

    2017-02-23

    We sequenced the complete genome of bovine viral diarrhea virus (BVDV) strain UM/126/07. It belongs to subgenotype 1h. The complete genome is composed of 12,196 nucleotides organized as one open reading frame encoding 3,898 amino acids. This is the first report of a full-length sequence of BVDV-1h.

  5. Anti-viral effect of interferon-alpha on bovine viral diarrhea virus.

    Science.gov (United States)

    Sentsui, H; Takami, R; Nishimori, T; Murakami, K; Yokoyama, T; Yokomizo, Y

    1998-12-01

    To get basic information to control persistent virus infection among domestic animals by cytokines, the antiviral activity of four natural human cytokines against bovine viral diarrhea virus (BVDV) was evaluated. Normal bovine peripheral blood mononuclear leukocytes (PBML) and fetal bovine muscular cells (FBMC) were treated with varying doses of human interferon (IFN)-alpha, IFN-gamma, tumor necrosis factor (TNF)-alpha and TNF-beta. The antiviral activity in treated cells was measured by the titration of virus infectivity in comparison with non-treated controls. IFN-alpha significantly suppressed virus growth in both PBML and FBMC. The growth of two cytopathogenic and two noncytopathogenic strains was suppressed in the presence of more than 10(3) u/ml of IFN-alpha. Addition of either TNF-alpha or TNF-beta to IFN-alpha did not potentiate the suppressive effect. IFN-alpha also suppressed the replication of BVDV in PBML from cattle persistently infected with BVDV.

  6. Identification and Characterization of Bovine Viral Diarrhea Virus from Indonesian Cattle (IDENTIFIKASI DAN KARAKTERISASI VIRUS BOVINE VIRAL DIARRHEA DARI SAPI INDONESIA

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    Muharam Saepulloh

    2015-05-01

    Full Text Available Bovine viral diarrhea virus (BVDV is an important viral disease, which a ubiquitous pathogen ofcattle with worldwide economic importance and due to its misdiagnose with other viruses. The goal of thecurrent study was to identify and characterize of BVDV by reverse transcriptase polymerase chainreaction (RT-PCR and followed by sequence genome analyses. Blood, feces, and semen samples werecollected from 588 selected cattle from animals suffering from diarrhea and respiratory manifestation. RTPCRresults showed that the 69 (11.74% samples were positive to BVDV. Further molecularcharacterization was conducted only with 17 PCR positive samples. The results indicated the 17 IndonesianBVD virus isolates were belonging to the genotype-1 of BVDV (BVDV-1 based on sequence analysis anda phylogenetic relationship between Indonesian BVDV isolates and BVDV in the world. This finding is thefirst report of BVD-1 circulated in Indonesian cattle.

  7. Characterization of a bovine viral diarrhea virus originated from cattle in Gansu Province, China.

    Science.gov (United States)

    Gao, Shandian; Shao, Junjun; Du, Junzheng; Lin, Tong; Cong, Guozheng; Zhao, Furong; Chang, Huiyun; Yin, Hong

    2013-08-01

    A bovine viral diarrhea disease virus (BVDV) GS-4 was isolated in Western China form dairy cattle with respiratory disease. Genomic comparison analysis with the 5' half genome sequence encompassing the coding region of N(pro), capsid, and envelope glycoproteins showed that the GS-4 should be classified into BVDV-1b1, which is considered as one of the predominant subgenotypes found in China. This classification was confirmed by phylogenetic analysis based on E2 coding region.

  8. Complete Genome Sequencing of Bovine Viral Diarrhea Virus 1, Subgenotypes 1n and 1o.

    Science.gov (United States)

    Sato, Asuka; Tateishi, Kentaro; Shinohara, Minami; Naoi, Yuki; Shiokawa, Mai; Aoki, Hiroshi; Ohmori, Keitaro; Mizutani, Tetsuya; Shirai, Junsuke; Nagai, Makoto

    2016-02-18

    To gain further insight into the genomic features of bovine viral diarrhea virus 1 (BVDV-1) subgenotypes, we sequenced the complete genome of BVDV-1n Shitara/02/06 and BVDV-1o IS26NCP/01. The complete genome of Shitara/02/06 and IS26NCP/01 shared 77.7 to 79.3% and 78.0 to 85.7% sequence identities with other BVDV-1 subgenotype strains, respectively.

  9. Isolation and identification of a bovine viral diarrhea virus from sika deer in china

    OpenAIRE

    Wang Nan; Sun Changjiang; Wang Quankai; Du Rui; Wang Shijie; Gao Yugang; Zhang Pengju; Zhang Lianxue

    2011-01-01

    Abstract Background Bovine viral diarrhea virus (BVDV) infections continue to cause significantly losses in the deer population. Better isolation and identification of BVDV from sika deer may contribute significantly to the development of prophylactic therapeutic, and diagnostic reagents as well as help in prevention and control of BVDV. However, isolation and identification of BVDV from sika deer is seldom reported in literature. In this study, we collected some samples according to clinical...

  10. Production of a highly immunogenic subunit ISCOM vaccine against Bovine Viral Diarrhea Virus

    DEFF Research Database (Denmark)

    Kamstrup, Søren; Roensholt, L.; Jensen, M.Holm;

    1999-01-01

    Bovine Viral Diarrhea Virus (BVDV) is a major pathogen of cattle in most countries. The main reservoir of virus in herds are BVDV persistently infected animals, which arise as a result of infection of the bovine fetus early in gestation. The spread of virus to the unborn fetus may be prevented......, concentration, and insertion of antigens into immune stimulating complexes (ISCOMs). Vaccines based on two different Danish strains of BVDV were injected into calves and the antisera produced were tested for neutralising activity against a panel of Danish BVDV strains. The two vaccines induced different...

  11. Production effects of pathogens causing bovine leukosis, bovine viral diarrhea, paratuberculosis, and neosporosis.

    Science.gov (United States)

    Tiwari, A; Vanleeuwen, J A; Dohoo, I R; Keefe, G P; Haddad, J P; Tremblay, R; Scott, H M; Whiting, T

    2007-02-01

    The primary purpose of this research was to determine associations among seropositivity for bovine leukemia virus (BLV), bovine viral diarrhea virus (BVDV), Mycobacterium avium ssp. paratuberculosis (MAP), and Neospora caninum (NC) and each of 3 outcome variables (305-d milk, fat, and protein production) in Canadian dairy cattle. Serum samples from up to 30 randomly selected cows from 342 herds on monthly milk testing were tested for antibodies against BLV (IDEXX ELISA; IDEXX Corporation, Westbrook, ME), MAP (IDEXX or Biocor ELISA; Biocor Animal Health, Inc., Omaha, NE), and NC (IDEXX or Biovet ELISA; Biovet Inc., St. Hyacinthe, Quebec, Canada). Up to 5 unvaccinated cattle over 6 mo of age were tested for virus-neutralizing antibodies to the Singer strain of type 1 BVDV. Dairy Herd Improvement records were obtained electronically for all sampled cows. Linear mixed models with herd and cow as random variables were fit, with significant restricted maximum likelihood estimates of outcome effects being obtained, while controlling for potential confounding variables. Bovine leukemia virus seropositivity was not associated with 305-d milk, 305-d fat, or 305-d protein production. Cows in BVDV-seropositive herds (at least one unvaccinated animal with a titer > or =1:64) had reductions in 305-d milk, fat, and protein of 368, 10.2, and 9.5 kg, respectively, compared with cows in BVDV-seronegative herds. Mycobacterium avium ssp. paratuberculosis seropositivity was associated with lower 305-d milk of 212 kg in 4+-lactation cows compared with MAP-seronegative 4+-lactation cows. Neospora caninum seropositivity in primiparous cows was associated with lower 305-d milk, fat, and protein of 158, 5.5, and 3.3 kg, respectively, compared with NC-seronegative primiparous cows. There were no interactions among seropositivity for any of the pathogens and their effects on any of the outcomes examined, although the low MAP seroprevalence limited this analysis. Results from this research

  12. Microarray chip based identification of a mixed infection of bovine herpesvirus 1 and bovine viral diarrhea 2 from Indian cattle.

    Science.gov (United States)

    Ratta, Barkha; Yadav, Brijesh Singh; Pokhriyal, Mayank; Saxena, Meeta; Sharma, Bhaskar

    2014-01-01

    Bovine herpesvirus 1 (BHV1) and bovine viral diarrhea virus 2 (BVD2) are endemic in India although no mixed infection with these viruses has been reported from India. We report first mixed infection of these viruses in cattle during routine screening with a microarray chip. 62 of the 69 probes of BHV1 and 42 of the 57 BVD2 probes in the chip gave positive signals for the virus. The virus infections were subsequently confirmed by RT-PCR. We also discuss the implications of these findings.

  13. Seroprevalence of Bovine Herpes Virus-1, Bovine Herpes Virus-4 and Bovine Viral Diarrhea Virus in Dairy Cattle in Sudan

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    Amira M. Elhassan*, M.A Fadol and A.M. El-Hussein

    2011-10-01

    Full Text Available A survey was conducted to determine prevalence of antibodies against Bovine herpes virus-1 (BoHv-1, Bovine herpes virus-4 (BoHv-4 and Bovine viral diarrhea (BVD in dairy cattle in farms with reproductive problems in two areas in Sudan. Sera samples were collected from Khartoum state and central Sudan during 2005-2008 and analyzed using direct ELISA. The prevalence of antibodies was discussed with respect to age, season, sex, breed and locality BoHv-1 and BVD antibodies were highly prevalent in Khartoum state (51.7 and 50.4%, respectively while in central Sudan BoHv-1 (32.7% antibodies were the most prevalent followed by, BVD (25.7% and BoHv-4 (19.3%. The highest prevalence of antibodies against the three viruses in both areas was found during the rainy season (July to October. The prevalence of antibodies to viruses studied was significantly associated with female sex except for BoHv-1. Prevalence of antibodies to BoHv-4 was significantly associated with breed while those of BoHv-1 and BVD were not. The present results indicated that older cattle were more likely to be seropositive in case of BoHv-4 but to BoHv-1 or BVD viruses. Furthermore, it was found that BoHv-1 and BVD antibodies were highly prevalent in aborted dams. While, infertility problems were highly associated with BoHv-1 antibodies. BVD antibodies showed the highest prevalence in case of death after birth. The results of this study provide better understanding of viral epidemics of reproductive disorders and represent the first report of BoHv-4 antibodies in cattle in Sudan.

  14. Diverse outcomes of bovine viral diarrhea virus infections in a herd naturally infected during pregnancy - a case study

    Science.gov (United States)

    A beef producer purchased Angus crossbred cattle that were pregnant with nursing calves. The purchased cattle, their nursing calves, and subsequent born calves were not initially tested for BVDV. Bovine viral diarrhea virus subtype 2a (BVDV2a) was isolated from an aborted bovine fetus, 6.5 months,...

  15. Short communication. Genotyping and phylogenetic analysis of bovine viral diarrhea virus (BVDV isolates in Kosovo

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    Izedin Goga

    2014-03-01

    Full Text Available Three serum samples positive in Antigen ELISA BVDV have been tested to characterise genetic diversity of bovine viral diarrhea virus (BVDV in Kosovo. Samples were obtained in 2011 from heifers and were amplified by reverse transcription-polymerase chain reaction, sequenced and analysed by computer-assisted phylogenetic analysis. Amplified products and nucleotide sequence showed that all 3 isolates belonged to BVDV 1 genotype and 1b sub genotype. These results enrich the extant knowledge of BVDV and represent the first documented data about Kosovo BVDV isolates.

  16. Genotypic characteristics of bovine viral diarrhea virus 2 strains isolated in northern Italy.

    Science.gov (United States)

    Giangaspero, M; Harasawa, R; Zecconi, A; Luzzago, C

    2001-09-01

    Two strains of Bovine viral diarrhea virus 2 (BVDV-2) were isolated from calves in northern Italy. Variations in the 5'-untranslated region (UTR) of the genome were studied by primary structure alignment and neighbor-joining method based phylogenetic tree analyses and by palindromic nucleotide substitutions at the three variable loci in the 5'-UTR. Genetic analysis indicated their appurtenance to genovar BVDV-2a. Nucleotide sequence at the 5'-UTR of strain BS-95-II, one of the Italian isolates from healthy calves, showed 98% homology to that of the Japanese isolate OY89, a cytopathic strain derived from cattle with mucosal disease.

  17. Immune response to bovine viral diarrhea virus--looking at newly defined targets.

    Science.gov (United States)

    Chase, Christopher C L; Thakur, Neelu; Darweesh, Mahmoud F; Morarie-Kane, Susan E; Rajput, Mrigendra K

    2015-06-01

    Bovine viral diarrhea virus (BVDV) has long been associated with a wide variety of clinical syndromes and immune dysregulation, many which result in secondary bacterial infections. Current understanding of immune cell interactions that result in activation and tolerance are explored in light of BVDV infection including: depletion of lymphocytes, effects on neutrophils, natural killer cells, and the role of receptors and cytokines. In addition, we review some new information on the effect of BVDV on immune development in the fetal liver, the role of resident macrophages, and greater implications for persistent infection.

  18. Latex immunoagglutination assay for bovine viral diarrhea virus utilizing forward light scattering in a microfluidic device

    Science.gov (United States)

    Heinze, Brian C.; Song, Jae-Young; Han, Jin-Hee; Yoon, Jeong-Yeol

    2008-02-01

    We have investigated the utilization of particle agglutination assays using forward light scattering measurements in a microfluidic device towards detecting viral particles. The model viral target was bovine viral diarrhea virus (BVDV). Highly carboxylated polystyrene microspheres (510 nm) were coated with anti-BVDV monoclonal antibodies. This solution was in turn used to detect live modified BVDV. This assay was first performed in a two well slide for proof of concept and then in a simple y-channel microfluidic device with optical fibers arranged in a close proximity setup. Particle immunoagglutination was detected through static light scattering measurements taken at 45° to incident light. In the microfluidic device, modified live BVDV was detected with a detection limit of 0.5 TCID 50 mL -1.

  19. Occurrence of Pseudocowpox virus associated to Bovine viral diarrhea virus-1, Brazilian Amazon.

    Science.gov (United States)

    Alves, Pedro A; Figueiredo, Poliana O; de Oliveira, Cairo H S; Barbosa, José D; Lima, Danillo H S; Bomjardim, Henrique A; Silva, Natália S; Campos, Karinny F; Oliveira, Carlos Magno C; Barbosa-Stancioli, Edel Figueiredo; Abrahão, Jônatas S; Kroon, Erna G; de Souza Trindade, Giliane

    2016-12-01

    In 2011, an outbreak of severe vesicular disease occurred in the state of Pará, Amazon region. Besides proliferative or verrucous lesions, cattle showed atypical clinical signs such as diarrhea and leading to death. The animals were submitted to clinical, pathological and molecular diagnosis, and laboratory tests have confirmed the presence of Pseudocowpox virus (PCPV), a Parapoxvirus genus member, and have also found Bovine viral diarrhea virus-1 (BVDV-1), probably causing persistent infection. The results of molecular diagnostics, followed by sequencing data demonstrated the circulation of both viruses (PCPV and BVDV-1) in an area previously affected by another poxvirus, as Vaccinia virus.The cocirculation between PCPV and BVDV-1 indicates a major concern for animal health because the clinical presentation can be a severe disease. This is the first detection of PCPV in the Brazilian Amazon.

  20. Effects of Preinfection With Bovine Viral Diarrhea Virus on Immune Cells From the Lungs of Calves Inoculated With Bovine Herpesvirus 1.1.

    Science.gov (United States)

    Risalde, M A; Molina, V; Sánchez-Cordón, P J; Romero-Palomo, F; Pedrera, M; Gómez-Villamandos, J C

    2015-07-01

    The aim of this work was to study the interstitial aggregates of immune cells observed in pulmonary parenchyma of calves preinfected with bovine viral diarrhea virus and challenged later with bovine herpesvirus 1. In addition, the intent of this research was to clarify the role of bovine viral diarrhea virus in local cell-mediated immunity and potentially in predisposing animals to bovine respiratory disease complex. Twelve Friesian calves, aged 8 to 9 months, were inoculated with noncytopathic bovine viral diarrhea virus genotype 1. Ten were subsequently challenged with bovine herpesvirus 1 and euthanized at 1, 2, 4, 7, or 14 days postinoculation. The other 2 calves were euthanized prior to the second inoculation. Another cohort of 10 calves was inoculated only with bovine herpesvirus 1 and then were euthanized at the same time points. Two calves were not inoculated with any agent and were used as negative controls. Pulmonary lesions were evaluated in all animals, while quantitative and biosynthetic changes in immune cells were concurrently examined immunohistochemically to compare coinfected calves and calves challenged only with bovine herpesvirus 1. Calves preinfected with bovine viral diarrhea virus demonstrated moderate respiratory clinical signs and histopathologic evidence of interstitial pneumonia with aggregates of mononuclear cells, which predominated at 4 days postinoculation. Furthermore, this group of animals was noted to have a suppression of interleukin-10 and associated alterations in the Th1-driven cytokine response in the lungs, as well as inhibition of the response of CD8+ and CD4+ T lymphocytes against bovine herpesvirus 1. These findings suggest that bovine viral diarrhea virus preinfection could affect the regulation of the immune response as modulated by regulatory T cells, as well as impair local cell-mediated immunity to secondary respiratory pathogens.

  1. Generation of the Bovine Viral Diarrhea Virus E0 Protein in Transgenic Astragalus and Its Immunogenicity in Sika Deer

    OpenAIRE

    Yugang Gao; Xueliang Zhao; Pu Zang; Qun Liu; Gongqing Wei; Lianxue Zhang

    2014-01-01

    The bovine viral diarrhea virus (BVDV), a single-stranded RNA virus, can cause fatal diarrhea syndrome, respiratory problems, and reproductive disorders in herds. Over the past few years, it has become clear that the BVDV infection rates are increasing and it is likely that an effective vaccine for BVDV will be needed. In this study, transgenic Astragalus was used as an alternative productive platform for the expression of glycoprotein E0. The immunogenicity of glycoprotein E0 expressed in tr...

  2. Bovine viral diarrhea virus (BVDV) infection in dairy cattle herds in northeast Thailand.

    Science.gov (United States)

    Nilnont, Theerakul; Aiumlamai, Suneerat; Kanistanont, Kwankate; Inchaisri, Chaidate; Kampa, Jaruwan

    2016-08-01

    Bovine viral diarrhea virus causes a wide range of clinical manifestation with subsequent economic losses in dairy production worldwide. Our study of a population of dairy cattle in Thailand based on 933 bulk tank milk samples from nine public milk collection centers aimed to monitor infective status and to evaluate the effect of the infection in cows as well as to examine the reproductive performance of heifers to provide effective recommendations for disease control in Thailand. The results showed a moderate antibody-positive prevalence in the herd (62.5 %), with the proportion of class-3 herd, actively infected stage, being 17.3 %. Fourteen persistently infected (PI) animals were identified among 1196 young animals from the class-3 herds. Most of the identified PI animals, 11/14, were born in one sub-area where bovine viral diarrhea virus (BVDV) investigation has not been performed to date. With respect to reproductive performance, class-3 herds also showed higher median values of reproductive indices than those of class-0 herds. Cows and heifers in class-3 herds had higher odds ratio of calving interval (CI) and age at first service (AFS) above the median, respectively, compared to class-0 herds (OR = 1.29; P = 0.02 and OR = 1.63; P = 0.02). Our study showed that PI animals were still in the area that was previously studied. Furthermore, a newly studied area had a high prevalence of BVDV infection and the infection affected the reproductive performance of cows and heifers. Although 37.5 % of the population was free of BVDV, the lack of official disease prevention and less awareness of herd biosecurity may have resulted in continuing viral spread and silent economic losses have potentially occurred due to BVDV. We found that BVDV is still circulating in the region and, hence, a national control program is required.

  3. Virulent Properties of Russian Bovine Viral Diarrhea Virus Strains in Experimentally Infected Calves

    Directory of Open Access Journals (Sweden)

    Alexander G. Glotov

    2016-01-01

    Full Text Available The results of experimental study of three noncytopathic and two cytopathic bovine viral diarrhea virus (BVDV strains isolated from cattle in the Siberian region and belonging to the type 1 (subtypes 1a, 1b, and 1d have been presented. All investigated strains caused the development of infectious process in the seronegative 4–6-month-old calves after aerosol challenge with the dose of 6 log10 TCID50. The greatest virulence had noncytopathic strain and cytopathic strain related to the subtypes 1d and 1b, respectively. All strains in infected calves caused some signs of moderate acute respiratory disease and diarrhea: depression 3–5 days postinfection (p.i., refusal to food, severe hyperthermia to 41.9°С, serous exudate discharges from the nasal cavity and eyes, transient diarrhea with blood, leukopenia (up to 2700 cells/mm3, and macroscopic changes in the respiratory organs and intestine. The infected animals recovered from 12 to 15 days p.i. and in 90% cases formed humoral immune response 25 days p.i. (antibody titers to BVDV: 1 : 4–1 : 16. Our results confirmed the presence of virulent BVDV1 strains and showed the need for researches on the molecular epidemiology of the disease, development of more effective diagnostic systems, and optimization of control programs with use of vaccines.

  4. It takes a combination of biosecurity, testing, and vaccination to keep bovine viral diarrhea (BVD) under control

    Science.gov (United States)

    This is the third installment of a 3 part series on bovine viral diarrhea (BVD), written for a lay publication whose core audience in dairy producers. Control of BVD in any dairy operation must rely on the implementation of an organized strategy combining biosecurity, surveillance and increased herd...

  5. Evaluation of temporal surveillance system sensitivity and freedom from bovine viral diarrhea in Danish dairy herds using scenario tree modelling

    DEFF Research Database (Denmark)

    Foddai, Alessandro; Stockmarr, Anders; Boklund, Anette

    2016-01-01

    The temporal sensitivity of the surveillance system (TemSSe) for Bovine Viral Diarrhea (BVD) in Danish dairy herds was evaluated. Currently, the Danish antibody blocking ELISA is used to test quarterly bulk tank milk (BTM). To optimize the surveillance system as an early warning system, we...

  6. Induction of interferon-gamma and downstream pathways during establishment of fetal persistent infection with bovine viral diarrhea virus

    Science.gov (United States)

    Development of transplacental infection depends on the ability of the virus to cross the placenta and replicate within the fetus while counteracting maternal and fetal immune responses.Unfortunately, little is known about this complex process. Non-cytopathic (ncp) strains of bovine viral diarrhea vi...

  7. Bovine viral diarrhea (BVD) can open the door to other problems, including reproductive, respiratory, and enteric disease

    Science.gov (United States)

    This is a review, written for a lay publication whose core audience in dairy producers. A brief history of bovine viral diarrhea (BVD) research is given as well as a review of recent research discoveries. National efforts to reduce antibiotic use have led to a greater emphasis on disease prevention ...

  8. A genome-wide association study for the incidence of persistent bovine viral diarrhea virus infection in cattle

    Science.gov (United States)

    Bovine Viral Diarrhea Viruses (BVDV) comprises a diverse group of viruses that causes disease in cattle. BVDV may establish both, transient and persistent infections depending on the developmental stage of the animal at exposure. The objective was to determine if genomic regions harboring single nuc...

  9. Comparison of the Immune Response Between a Pair of NCP and CP Bovine Viral Diarrhea Virus (BVDV) Type 1 Isolates

    Science.gov (United States)

    Aim: Bovine viral diarrhea virus (BVDV) is a major pathogen of cattle causing severe respiratory and reproductive disease. BVDV vaccines remain an important part of the control strategy. Previous work has described higher antibody responses in animals infected with a noncytopathic (NCP) BVDV when ...

  10. Differential expression of miRNA-423-5p in serum from cattle challenged with bovine viral diarrhea virus

    Science.gov (United States)

    Bovine viral diarrhea virus (BVDV) is an RNA virus that causes respiratory disease in cattle. MicroRNAs have been proposed as indicators of exposure to respiratory pathogens. However, microRNA profiles in cattle exposed to BVDV are currently nonexistent and few studies have been reported; therefore,...

  11. Isolation and Genetic Analysis of Bovine Viral Diarrhea Virus from Infected Cattle in Indiana

    Directory of Open Access Journals (Sweden)

    Roman M. Pogranichniy

    2011-01-01

    Full Text Available Species and biotype distribution was determined in 44 bovine viral diarrhea virus- (BVDV- positive samples submitted to the Animal Disease Diagnostic Laboratory (ADDL in Indiana during 2006–2008. BVDV RNA was detected in the 5′-untranslated region and Npro region using reverse transcriptase PCR followed by sequencing analysis of the PCR product. Additionally, cases were classified into one of six categories according to history and/or lesions: acute symptomatic, hemorrhagic, respiratory distress, reproductive, persistent infection (PI, and mucosal disease (MD. Of 44 BVDV-positive samples, 33 were noncytopathic (ncp, 10 were cytopathic (cp, and one presented both ncp and cp biotypes. Sequencing analysis demonstrated that all samples belonged to BVDV-1a, BVDV-1b, or BVDV-2. The most common isolate was ncp BVDV-1b, (44% followed by ncp BVDV-2a (24%. Among the six categories, respiratory clinical signs were the most common (36% followed by PI (25% and MD (16%.

  12. Enterocytozoon bieneusi in Bovine Viral Diarrhea Virus (BVDV) infected and noninfected cattle herds.

    Science.gov (United States)

    Juránková, J; Kamler, M; Kovařčík, K; Koudela, B

    2013-02-01

    Enterocytozoon bieneusi known as a causative agent of opportunistic infections instigating diarrhoea in AIDS patients was identified also in a number of immunocompetent patients and in a wide range of animals, including cattle. In the present study we tested if the Bovine Viral Diarrhea Virus (BVDV), the most common pathogen underlying immunosuppressive Bovine Viral Diarrhoea (BVD), can enhance the occurrence of opportunistic infections with E. bieneusi in cattle. Six dairy farms were investigated using ELISA to detect antibodies against or antigens arising from BVDV in collected sera. A total of 240 individual faecal samples from four age groups were examined for the presence of E. bieneusi by nested PCR. Sequence analysis of six E. bieneusi positive samples revealed the presence of the genotype I of E. bieneusi, previously described in cattle. The hypothesis expecting higher prevalence of E. bieneusi in BVDV positive cattle herds was not confirmed in this study; however this is the first description about E. bieneusi in cattle in the Czech Republic.

  13. Effects of interferon-tau on cattle persistently infected with bovine viral diarrhea virus.

    Science.gov (United States)

    Kohara, Junko; Nishikura, Yumiko; Konnai, Satoru; Tajima, Motoshi; Onuma, Misao

    2012-08-01

    In this study, the antiviral effects of bovine interferon-tau (boIFN-tau) on bovine viral diarrhea virus (BVDV) were examined in vitro and in vivo. In the in vitro experiments, the replication of cytopathic and non-cytopathic BVDV was inhibited in the bovine cells treated with boIFN-tau. The replication of BVDV was completely suppressed by boIFN-tau at a concentration higher than 10(2) U/ml. In order to examine the effect of boIFN-tau on virus propagation in cattle persistently infected (PI) with non-cytopathic BVDV, boIFN-tau was subcutaneously administered to PI cattle at 10(5) U/kg or 10(6) U/kg body weight 5 times per week for 2 weeks. No physical abnormality such as depression was observed in the cattle during the experiment. The mean BVDV titers in the serum of the PI cattle decreased slightly during the boIFN-tau administration period with the dose of 10(6) U/kg. However, the BVDV titers in the serum returned to the pre-administration level after the final boIFN-tau administration. These results suggest that boIFN-tau demonstrates an anti-BVDV effect, reducing the BVDV level in serum transiently when injected into PI cattle.

  14. Identification of bovine viral diarrhea virus infection in Saanen goats in the Republic of Korea.

    Science.gov (United States)

    Han, Yu-Jung; Chae, Jeong-Byoung; Chae, Joon-Seok; Yu, Do-Hyeon; Park, Jinho; Park, Bae-Keun; Kim, Hyeon-Cheol; Yoo, Jae-Gyu; Choi, Kyoung-Seong

    2016-06-01

    Bovine viral diarrhea virus (BVDV) is one of the most important viral pathogens of livestock and causes substantial economic losses to the livestock industry worldwide. BVDV is not necessarily species specific and is known to infect domesticated and wild ruminants. In the present study, BVDV infection was identified in two Saanen goats from one farm, and two different viral subtypes were found, BVDV-1a and BVDV-2a. Each isolate was closely related to cattle isolates identified in the Republic of Korea. The two sequences obtained in this study were not consistent with border disease virus (BDV). The incidence of BVDV in this farm apparently occurred in the absence of contact with cattle and may be associated with grazing. This study demonstrates that BVDV infection may be possible to transmit among goats without exposure to cattle. Therefore, this result indicates that Saanen goats may act as natural reservoirs for BVDV. This is the first report of BVDV-1a infection in a Saanen goat.

  15. Polymorphic genetic characterization of E2 gene of bovine viral diarrhea virus in China.

    Science.gov (United States)

    Lang, Yifei; Gao, Shandian; Du, Junzheng; Shao, Junjun; Cong, Guozheng; Lin, Tong; Zhao, Furong; Liu, Lihong; Chang, Huiyun

    2014-12-05

    Bovine viral diarrhea virus (BVDV) is one of the wide distributed pathogenic viruses of livestock and wild animals worldwide. E2 glycoprotein is a major structural component of the BVDV virion and plays a key role in viral attachment to host cells and inducing immune responses against viral infection. In order to gain detailed information of the E2 coding region of BVDV circulating in China, 46 positive samples were tested by RT-PCR for the E2 coding region. The 1122 nt nucleotide sequences of full-length E2 were harvested and analyzed. The results suggested that full-length E2 was an ideal target for BVDV genotyping and divided the domestic BVDV isolates into 9 subgenotypes, namely BVDV-1a, -1b1, -1c, -1d, -1o, -1m, -1p, -1q and BVDV-2a, showing great diversity. The difference of nonsynonymous and synonymous substitution rates (dN-dS) inferred both positive and purifying selection of the E2. However, combination of positive and purifying selection at different points indicated purifying selection within the complete E2. Protein properties analysis based on glycosylation sites and epitope prediction demonstrated that the biological character of E2 among individual BVDV subgenotype was similar, but may alter due to amino acid changes. For the first time, the comprehensive collection of E2 sequences of Chinese BVDV isolates was elucidated, which would provide information for future vaccine design and BVD control in China.

  16. Antigenic variability in bovine viral diarrhea virus (BVDV) isolates from alpaca (Vicugna pacos), llama (Lama glama) and bovines in Chile.

    Science.gov (United States)

    Aguirre, I M; Quezada, M P; Celedón, M O

    2014-01-31

    Llamas and alpacas are domesticated South American camelids (SACs) important to ancestral population in the Altiplano region, and to different communities where they have been introduced worldwide. These ungulates have shown to be susceptible to several livestock viral pathogens such as members of the Pestivirus genus and mainly to bovine viral diarrhea virus (BVDV). Seventeen Chilean BVDV isolates were analyzed by serum cross neutralization with samples obtained from five llama, six alpacas, three bovines, plus three reference strains belonging to different subgroups and genotypes. The objective was to describe antigenic differences and similarities among them. Antigenic comparison showed significant differences between different subgroups. Consequently, antigenic similarities were observed among isolates belonging to the same subgroup and also between isolates from different animal species belonging the same subgroup. Among the analyzed samples, one pair of 1b subgroup isolates showed significant antigenic differences. On the other hand, one pair of isolates from different subgroups (1b and 1j) shared antigenic similarities indicating antigenic relatedness. This study shows for the first time the presence of antigenic differences within BVDV 1b subgroup and antigenic similarities within 1j subgroup isolates, demonstrating that genetic differences within BVDV subgroups do not necessary corresponds to differences on antigenicity.

  17. Bovine viral diarrhea virus: molecular cloning of genomic RNA and its diagnostic application

    Energy Technology Data Exchange (ETDEWEB)

    Brock, K.V.

    1987-01-01

    Molecular cloning of a field isolate of bovine viral diarrhea virus (BVDV) strain 72 RNA was done in this study. The sensitivity and specificity of cloned cDNA sequences in hybridization assays with various BVDV strains were determined. cDNA was synthesized from polyadenylated BVDV RNA templates with oligo-dT primers, reverse transcriptase, and DNA polymerase I. The newly synthesized double-stranded BVDV cDNA was C-tailed with terminal deoxytransferase and annealed into G-tailed, Pst-1-cut pUC9 plasmid. Escherichia coli was transformed with the recombinant plasmids and a library of approximately 200 BVDV specific cDNA clones varying in length from 0.5 to 2.6 kilobases were isolated. The sensitivity and specificity of hybridization between the labelled cDNA and BVDV target sequences were determined. Cloned BVDV sequences were isolated from pUC9 plasmid DNA and labelled with /sup 32/P by nick translation. The detection limit by dot blot hybridization assay was 20 pg of purified genomic BVDV RNA. cDNA hybridization probes were specific for all strains of BVDV tested, regardless of whether they were noncytopathic and cytopathic, but did not hybridize with heterologous bovine viruses tested. Probes did not hybridize with uninfected cell culture or cellular RNA. Hybridization probes were at least as sensitive as infectivity assays in detecting homologous virus.

  18. Innate and adaptive immune responses to in utero infection with bovine viral diarrhea virus.

    Science.gov (United States)

    Hansen, Thomas R; Smirnova, Natalia P; Webb, Brett T; Bielefeldt-Ohmann, Helle; Sacco, Randy E; Van Campen, Hana

    2015-06-01

    Infection of pregnant cows with noncytopathic (ncp) bovine viral diarrhea virus (BVDV) induces rapid innate and adaptive immune responses, resulting in clearance of the virus in less than 3 weeks. Seven to 14 days after inoculation of the cow, ncpBVDV crosses the placenta and induces a fetal viremia. Establishment of persistent infection with ncpBVDV in the fetus has been attributed to the inability to mount an immune response before 90-150 days of gestational age. The result is 'immune tolerance', persistent viral replication and shedding of ncpBVDV. In contrast, we describe the chronic upregulation of fetal Type I interferon (IFN) pathway genes and the induction of IFN-γ pathways in fetuses of cows infected on day 75 of gestation. Persistently infected (PI) fetal IFN-γ concentrations also increased at day 97 at the peak of fetal viremia and IFN-γ mRNA was significantly elevated in fetal thymus, liver and spleen 14-22 days post maternal inoculation. PI fetuses respond to ncpBVDV infection through induction of Type I IFN and IFN-γ activated genes leading to a reduction in ncpBVDV titer. We hypothesize that fetal infection with BVDV persists because of impaired induction of IFN-γ in the face of activated Type I IFN responses. Clarification of the mechanisms involved in the IFN-associated pathways during BVDV fetal infection may lead to better detection methods, antiviral compounds and selection of genetically resistant breeding animals.

  19. Bovine viral diarrhea virus antigen detection across whole cattle hides using two antigen-capture enzyme-linked immunosorbent assays.

    Science.gov (United States)

    Vander Ley, Brian L; Ridpath, Julia F; Sweiger, Shaun H

    2012-05-01

    Bovine viral diarrhea virus is a costly disease of cattle that can be controlled by vaccination, biosecurity, and removal of persistently infected cattle. Development and proficiency testing of assays to identify persistently infected cattle requires substantial quantities of known positive- and negative-sample material. The objective of this study was to determine what sections of bovine skin contained Bovine viral diarrhea virus antigen. Two commercially available antigen-capture enzyme-linked immunoassays were used to test subsamples representing the entire skin of 3 persistently infected calves. Both assays detected Bovine viral diarrhea virus antigen in the samples indicated for use by assay protocol. However, one assay identified all subsamples as positive, while the second assay identified 64.4% of subsamples as positive. These results show that use of samples other than those specified by the assay protocol must be validated for each individual assay. In this study, alternative sample sites and use of the entire hide for proficiency testing would be acceptable for only one of the assays tested.

  20. Genetic and antigenic characterization of bovine viral diarrhea viruses isolated from cattle in Hokkaido, Japan.

    Science.gov (United States)

    Abe, Yuri; Tamura, Tomokazu; Torii, Shiho; Wakamori, Shiho; Nagai, Makoto; Mitsuhashi, Kazuya; Mine, Junki; Fujimoto, Yuri; Nagashima, Naofumi; Yoshino, Fumi; Sugita, Yukihiko; Nomura, Takushi; Okamatsu, Masatoshi; Kida, Hiroshi; Sakoda, Yoshihiro

    2016-01-01

    In our previous study, we genetically analyzed bovine viral diarrhea viruses (BVDVs) isolated from 2000 to 2006 in Japan and reported that subgenotype 1b viruses were predominant. In the present study, 766 BVDVs isolated from 2006 to 2014 in Hokkaido, Japan, were genetically analyzed to understand recent epidemics. Phylogenetic analysis based on nucleotide sequences of the 5'-untranslated region of viral genome revealed that 766 isolates were classified as genotype 1 (BVDV-1; 544 isolates) and genotype 2 (BVDV-2; 222). BVDV-1 isolates were further divided into BVDV-1a (93), 1b (371) and 1c (80) subgenotypes, and all BVDV-2 isolates were grouped into BVDV-2a subgenotype (222). Further comparative analysis was performed with BVDV-1a, 1b and 2a viruses isolated from 2001 to 2014. Phylogenetic analysis based on nucleotide sequences of the viral glycoprotein E2 gene, a major target of neutralizing antibodies, revealed that BVDV-1a, 1b and 2a isolates were further classified into several clusters. Cross-neutralization tests showed that BVDV-1b isolates were antigenically different from BVDV-1a isolates, and almost BVDV-1a, 1b and 2a isolates were antigenically similar among each subgenotype and each E2 cluster. Taken together, BVDV-1b viruses are still predominant, and BVDV-2a viruses have increased recently in Hokkaido, Japan. Field isolates of BVDV-1a, 1b and 2a show genetic diversity on the E2 gene with antigenic conservation among each subgenotype during the last 14 years.

  1. Biochemical analysis of bovine viral diarrhea virus polypeptides and studies of strain variation

    Energy Technology Data Exchange (ETDEWEB)

    Raisch, K.P.

    1989-01-01

    Intracellular viral-specific polypeptides from the National Animal Disease Laboratory (NADL) strain of bovine viral diarrhea virus were studied by biosynthesis labelling, radioimmunoprecipitation (RIP), hypertonic initiation block (HIB) and polyacrylamide gel electrophoresis (PAGE). Eighteen virus-specific proteins were identified; thirteen were glycosylated (gp170, p135, p130, gp118, gp82, p80, gp74, gp63, gp60, p59, gp53, gp50, gp45, gp42, p37, gp32, gp25 and p22). When glycosylation was inhibited by tunicamycin, five {sup 35}S-methionine labelled proteins displayed increased electrophoretic mobility (gp170 to p165, gp74 to p66, gp53 to p45, gp50 to p42 and gp25 to p20) and four could not be identified. Similar shifts in mobility were observed following in vitro deglycosylation with endoglycosidases H and F indicating that the nine glycoproteins contained N-linked simple or high mannose containing moieties. Biosynthetic labelling in the presence of the ionophore, monensin, or in vitro deglycosylation with the endoglycosidase, O-glycanase, had no effect, which is consistent with the absence of O-linked carbohydrates in BVDV-specific proteins. N-linked glycosylation of BVDV proteins is critical for infectivity, because the virus from cells treated with tunicamycin was devoid of infectivity, whereas the virus from monensin-treated cells was fully infective. Partitioning of p130, p59, gp53-50, and p37 into solutions of Triton X-114 tentatively identified these molecules as partially hydrophobic transmembrane proteins. Biosynthesis in the presence of {sup 3}H-myristate and {sup 3}H-palmitate did not result in specifically labelled viral proteins indicating predominantly noncovalent nature of putative interactions of these proteins with membranes. Partial proteolytic peptide mapping revealed similarities among gp170, p130 and p80 and between gp53 and gp50.

  2. Morphology and Molecular Composition of Purified Bovine Viral Diarrhea Virus Envelope.

    Directory of Open Access Journals (Sweden)

    Nathalie Callens

    2016-03-01

    Full Text Available The family Flaviviridae includes viruses that have different virion structures and morphogenesis mechanisms. Most cellular and molecular studies have been so far performed with viruses of the Hepacivirus and Flavivirus genera. Here, we studied bovine viral diarrhea virus (BVDV, a member of the Pestivirus genus. We set up a method to purify BVDV virions and analyzed their morphology by electron microscopy and their protein and lipid composition by mass spectrometry. Cryo-electron microscopy showed near spherical viral particles displaying an electron-dense capsid surrounded by a phospholipid bilayer with no visible spikes. Most particles had a diameter of 50 nm and about 2% were larger with a diameter of up to 65 nm, suggesting some size flexibility during BVDV morphogenesis. Morphological and biochemical data suggested a low envelope glycoprotein content of BVDV particles, E1 and E2 being apparently less abundant than Erns. Lipid content of BVDV particles displayed a ~2.3 to 3.5-fold enrichment in cholesterol, sphingomyelin and hexosyl-ceramide, concomitant with a 1.5 to 5-fold reduction of all glycerophospholipid classes, as compared to lipid content of MDBK cells. Although BVDV buds in the endoplasmic reticulum, its lipid content differs from a typical endoplasmic reticulum membrane composition. This suggests that BVDV morphogenesis includes a mechanism of lipid sorting. Functional analyses confirmed the importance of cholesterol and sphingomyelin for BVDV entry. Surprisingly, despite a high cholesterol and sphingolipid content of BVDV envelope, E2 was not found in detergent-resistant membranes. Our results indicate that there are differences between the structure and molecular composition of viral particles of Flaviviruses, Pestiviruses and Hepaciviruses within the Flaviviridae family.

  3. Seroprevalence and risk factors associated with bovine herpesvirus 1 and bovine viral diarrhea virus in North-Eastern Mexico

    Science.gov (United States)

    Segura-Correa, J.C.; Zapata-Campos, C.C.; Jasso-Obregón, J.O.; Martinez-Burnes, J.; López-Zavala, R.

    2016-01-01

    Bovine herpesvirus 1 (BoHV-1) and bovine viral diarrhea virus (BVDV) are well known etiological agents of cattle that produce important economic losses due to reproductive failures and calf mortality, as well as enteric and respiratory disease. Tamaulipas is located northeast of Mexico, an important cattle production and the principal exporter of calf and heifer to the United States. The objectives of this study were to estimate the seroprevalence of BoHV-1 and of BVDV, and to determine the effects of risk factors on these infections. Blood samples of cattle from 57 farms from rural districts of Tamaulipas were collected. The samples were tested for antibodies against BoHV-1 and BVDV using commercial ELISA kits. Data on potential risk factors were obtained using a questionnaire administered to the farmer at the time the blood samples were taken. The seroprevalences for BoHV-1 and BVDV were 64.4% and 47.8%, respectively. In the logistic regression analysis, the significant risk factors were rural district, herd size and cattle introduced to the farm. This study confirms the high seroprevalence of BoHV-1 and BVDV in unvaccinated cattle in Tamaulipas, Mexico. The results of this study could be used for the development of BoHV-1 and BVDV prevention and control program in North-Eastern, Mexico. PMID:27622156

  4. Serological relationships among subgroups in bovine viral diarrhea virus genotype 1 (BVDV-1).

    Science.gov (United States)

    Alpay, Gizem; Yeşilbağ, Kadir

    2015-01-30

    Bovine viral diarrhea virus (BVDV) has various economic impacts associated with diarrhea, poor performance, an increase in the frequency of other infections and lethal outcomes. Both genotypes, namely BVDV-1 and BVDV-2, as well as different subgroups within these genotypes have been reported worldwide. Understanding the serological differences among the BVDV subgroups is important for disease epidemiology and prevention as well as vaccination programs. The aim of this study was to determine the serological relatedness among the subgroups in BVDV-1. For that purpose, sheep hyperimmune sera were collected against representative strains from 6 of the subgroups of BVDV-1 (BVDV-1a, -1b, -1d, -1f, -1h and -1l). The serum samples that gave the peak antibody titer to the homologous strains were used to perform cross neutralization assays. The highest homologous antibody titer (1:5160) was obtained against BVDV-1h. Regarding the cross neutralizing (heterologous) antibodies, the lowest titer (1:20) was produced by the BVDV-1f antiserum against the BVDV-1a and BVDV1-b viruses. The highest cross neutralizing titer (1:2580) achieved by the BVDV-1h antiserum was against the BVDV-1b strain. The cross neutralization results indicated particular serological differences between the recently described subgroup (BVDV-1l) and BVDV-1a/-1b, which are widely used in commercial vaccines. Considering the cross neutralization titers, it is concluded that selected BVDV-1l and BVDV-1h strains can be used for the development of diagnostic and control tools.

  5. Stability of Bovine viral diarrhea virus 1 nucleic acid in fetal bovine samples stored under different conditions.

    Science.gov (United States)

    Ridpath, Julia F; Neill, John D; Chiang, Yu-Wei; Waldbillig, Jill

    2014-01-01

    Infection of pregnant cattle with both species of Bovine viral diarrhea virus (BVDV) can result in reproductive disease that includes fetal reabsorption, mummification, abortion, stillbirths, congenital defects affecting structural, neural, reproductive, and immune systems, and the birth of calves persistently infected with BVDV. Accurate diagnosis of BVDV-associated reproductive disease is important to control BVDV at the production unit level and assessment of the cost of BVDV infections in support of BVDV control programs. The purpose of the current study was to examine the stability of viral nucleic acid in fetal tissues exposed to different conditions, as measured by detection by polymerase chain reaction. Five different types of fetal tissue, including brain, skin and muscle, ear, and 2 different pooled organ samples, were subjected to conditions that mimicked those that might exist for samples collected after abortions in production settings or possible storage conditions after collection and prior to testing. In addition, tissues were archived for 36 months at -20°C and then retested, to mimic conditions that might occur in the case of retrospective surveillance studies. Brain tissue showed the highest stability under the conditions tested. The impact of fecal contamination was increased following archiving in all tissue types suggesting that, for long-term storage, effort should be made to reduce environmental contaminants before archiving.

  6. Implementation of immunohistochemistry on frozen ear notch tissue samples in diagnosis of bovine viral diarrhea virus in persistently infected cattle

    Directory of Open Access Journals (Sweden)

    Bedeković Tomislav

    2011-12-01

    Full Text Available Abstract Background Bovine viral diarrhea is a contagious disease of domestic and wild ruminants and one of the most economically important diseases in cattle. Bovine viral diarrhea virus belongs to the genus Pestivirus, within the family Flaviviridae. The identification and elimination of the persistently infected animals from herds is the initial step in the control and eradication programs. It is therefore necessary to have reliable methods for diagnosis of bovine viral diarrhea virus. One of those methods is immunohistochemistry. Immunohistochemistry on formalin fixed, paraffin embedded tissue is a routine technique in diagnosis of persistently infected cattle from ear notch tissue samples. However, such technique is inappropriate due to complicated tissue fixation process and it requires more days for preparation. On the contrary, immunohistochemistry on frozen tissue was usually applied on organs from dead animals. In this paper, for the first time, the imunohistochemistry on frozen ear notch tissue samples was described. Findings Seventeen ear notch tissue samples were obtained during the period 2008-2009 from persistently infected cattle. Samples were fixed in liquid nitrogen and stored on -20°C until testing. Ear notch tissue samples from all persistently infected cattle showed positive results with good section quality and possibility to determinate type of infected cells. Conclusions Although the number of samples was limited, this study indicated that immunohistochemistry on formalin fixed paraffin embedded tissue can be successfully replaced with immunohistochemistry on frozen ear notch tissue samples in diagnosis of persistently infected cattle.

  7. First report of Bovine Viral Diarrhea Virus antigen from pneumonic cattle in Sudan

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    Intisar Kamil Saeed

    2015-06-01

    Full Text Available To explore the expected role of Bovine Viral Diarrhea Virus (BVDV in pneumonia in cattle, cattle lungs (n=242 showing signs of pneumonia were collected from slaughter houses of three different localities located at Northern, Central and Western Sudan during 2010–2013. The collected samples were tested for the presence of BVDV antigen using Enzyme-Linked Immunosorbent Assay (ELISA, and Fluorescent Antibody Test (FAT. Twenty six (10.7% out of 242 samples were found to be positive for BVDV. Positive results were seen in all the three studied areas, with the highest prevalence (16.7%; n=4/24 at Gezira State in Central Sudan. BVDV genome could be detected in all ELISA positive samples. The results indicated the existence of BVDV infection in cattle in different areas in Sudan, and its possible association with respiratory infections in cattle. Analysis using BLAST indicated that the sequence was identical to the previously reported BVDV-1 (GenBank accession AF220247.1.; nucleotide A was found in our study at position 9 of our sequence, whereas T was present instead in the reference virus. This is the first report of detecting BVDV antigen, genome, and its sequence analysis collected from cattle lungs in Sudan.

  8. Marine natural seaweed products as potential antiviral drugs against Bovine viral diarrhea virus

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    Ana Maria Viana Pinto

    2012-08-01

    Full Text Available Bovine viral diarrhea virus (BVDV is an etiologic agent that causes important economic losses in the world. It is endemic in cattle herds in most parts of the world. The purpose of this study was to evaluate the in vitro cytotoxic effect and antiviral properties of several marine natural products obtained from seaweeds: the indole alkaloid caulerpin (CAV, 1 and three diterpenes: 6-hydroxydichotoma-3,14-diene-1,17-dial (DA, 2, 10,18-diacetoxy-8-hydroxy-2,6-dolabelladiene (DB1, 3 and 8,10,18-trihydroxy-2,6-dolabelladiene (DB3, 4. The screening to evaluate the cytotoxicity of compounds did not show toxic effects to MDBK cells. The antiviral activity of the compounds was measured by the inhibition of the cytopathic effect on infected cells by plaque assay (PA and EC50 values were calculated for CAV (EC=2,0± 5.8, DA (EC 2,8± 7.7, DB1 (EC 2,0±9.7, and DB3 (EC 2,3±7.4. Acyclovir (EC50 322± 5.9 was used in all experiments as the control standard. Although the results of the antiviral activity suggest that all compounds are promising as antiviral agents against BVDV, the Selectivity Index suggests that DB1 is the safest of the compounds tested.

  9. Genomic characterization of three bovine viral diarrhea virus isolates from cattle.

    Science.gov (United States)

    Cai, Dongjie; Song, Quanjiang; Wang, Jiufeng; Zhu, Yaohong

    2016-12-01

    Three strains of the bovine viral diarrhea virus (BVDV) were isolated from cattle in Beijing, China. To investigate their genomic features, we sequenced and characterized the complete genome of each of the isolates. Each of the three virus genomes is about 12,220 bp in length, containing a 5' untranslated region (UTR), one open reading frame (ORF) encoding a 3897-amino-acid polypeptide, and a 3' UTR. The nucleotide sequence of the three isolates were 99.0 % identical to each and other shared nucleotide sequence identities of 73.4 % to 98.3 % with other BVDV-1 strains, about 70.0 % with BVDV-2 strains, about 67.0 % with BVDV-3, and less than 67.0 % with other pestiviruses. Phylogenetic analysis of the full-length genome, 3' UTR, and the N(pro) gene demonstrated that the three viruses were BVDV-1 isolates. This is the first report of complete genome sequences of BVDV 1d isolates from China and might have implications for vaccine development.

  10. Bovine viral diarrhea virus infections: manifestations of infection and recent advances in understanding pathogenesis and control.

    Science.gov (United States)

    Brodersen, B W

    2014-03-01

    Bovine viral diarrhea virus (BVDV) continues to be of economic significance to the livestock industry in terms of acute disease and fetal loss. Many of the lesions relating to BVDV infection have been well described previously. The virus is perpetuated in herds through the presence of calves that are persistently infected. Relationships between various species and biotypes of BVDV and host defenses are increasingly understood. Understanding of the host defense mechanisms of innate immunity and adaptive immunity continues to improve, and the effects of the virus on these immune mechanisms are being used to explain how persistent infection develops. The noncytopathic biotype of BVDV plays the major role in its effects on the host defenses by inhibiting various aspects of the innate immune system and creation of immunotolerance in the fetus during early gestation. Recent advances have allowed for development of affordable test strategies to identify and remove persistently infected animals. With these improved tests and removal strategies, the livestock industry can begin more widespread effective control programs.

  11. Detection of Bovine viral diarrhea virus from three water buffalo fetuses (Bubalus bubalis) in southern Italy.

    Science.gov (United States)

    Martucciello, Alessandra; De Mia, Gian Mario; Giammarioli, Monica; De Donato, Immacolata; Iovane, Giuseppe; Galiero, Giorgio

    2009-01-01

    Bovine viral diarrhea virus (BVDV) is an important pathogen that primarily infects ruminants, leading to several clinical problems including abortion. BVDV-specific antibodies were reported in a wide range of hosts within domestic and wildlife animal populations, and serological studies also indicated BVDV infection in buffaloes. The purpose of this study was to analyze the presence of BVDV in 2 water buffalo (Bubalus bubalis) herds with a history of abortion. Virus isolation from aborted fetuses and from maternal buffy coat and the molecular characterization of the isolates confirmed the presence of BVDV in these animals. The sequence analysis based on the 5' UTR and N(pro) coding regions of the Pestivirus genome revealed that the isolates belong to subgenotype 1b of BVDV. The findings of this study also suggest a possible role of BVDV in causing congenital infection in water buffalo. Its presence in fetal tissues as well as in maternal blood raises questions about the possible development of clinical disease or its influence in abortions in water buffalo.

  12. Spatial and temporal reconstruction of bovine viral diarrhea virus genotype 1 dispersion in Italy.

    Science.gov (United States)

    Luzzago, Camilla; Ebranati, Erika; Sassera, Davide; Lo Presti, Alessandra; Lauzi, Stefania; Gabanelli, Elena; Ciccozzi, Massimo; Zehender, Gianguglielmo

    2012-03-01

    Bovine viral diarrhea virus (BVDV) is a widespread and economically important pathogen of cattle; genetic typing of BVDV isolates distinguished two species, namely BVDV-1 and BVDV-2. BVDV-1 is the most widespread worldwide and it includes at least 11 subtypes. With the aim of clarifying the routes of circulation of BVDV-1 subtypes in an endemic area and in order to investigate the relationships between the genetic diversity of BVDV and its geographic distribution, a phylogenetic analysis of 5' untranslated region of Italian sequences was performed using a new Bayesian framework allowing the spatial-temporal reconstruction of the evolutionary dynamics of highly variable viruses. Our analyses suggested that different BVDV subtypes entered the North-Eastern part of Italy at different times within a time span between 23 and 7 years ago. The largest virus dispersion occurred between the mid 1990s and the early 2000s. A possible gravity-like dynamic of the infection, originating in larger animal population then following patterns of national commercial-flow, should be hypothesized.

  13. Proteins of bovine viral diarrhea virus: characterization, biotype-specific differences, and immunological properties

    Energy Technology Data Exchange (ETDEWEB)

    Donis, R.O.

    1987-01-01

    Virus-specific polypeptides in bovine viral diarrhea-mucosal disease (BVD) virus-infected bovine cells were studied by radiolabeling. A total of 12 polypeptides with apparent Mr of 165, 135, 118, 80, 75, 62, 56-58, 48, 37, 32, 25 and 19 kilodaltons (k) were identified in infected cells. Five glycoproteins were detected in infected cells. Two abundant species had apparent Mr of 48 k and 56-58 k while the minor species had masses of 118, 75 and 65 k. When cells were radiolabeled with L-(/sup 35/S)-methionine in the presence of tunicamycin the 56-58 k migrated with apparent masses of 54 k and 48-50 K in PAGE. Endoglycosidase F digestion of virus-induced polypeptides caused a 4-6 K reduction in the apparent molecular mass of the 56-58 k yielding a 52 k digested product. Tunicamycin caused a drastic reduction in the yield of infectious virus indicating that the carbohydrate moieties serve a vital role in the infection cycle of BVD virus. The noncytopathic biotype BVD (NCB-BVD) virus isolates can be consistently differentiated from cytopathic biotype BVD (CB-BVD) isolates on the basis of unique polypeptide profiles they induce in the infected cell: the most abundant polypeptide in CB-BVD infected cells is the 80 kD polypeptide while NCB-BVD lack this polypeptide and induce a predominant 118 k polypeptide. A panel of 25 murine monoclonal antibodies (Mabs) against the two major glycoproteins of BVD virus was produced. Based on their viral polypeptide specificity and on their ability to neutralize viral infectivity the Mabs in the panel were divided into 3 classes: Class 1 Mabs reacted with the 56-58 k glycoprotein and neutralized the virus, Class 2 Mabs recognized the 56-58 k glycoprotein but were not neutralizing and Class 3 Mabs reacted with the 48 k glycoprotein and did not neutralize the virus. These results identify the 56-58 k as one of the envelope glycoproteins of BVD virus.

  14. Genetic diversity and frequency of bovine viral diarrhea virus (BVDV) detected in cattle in Turkey.

    Science.gov (United States)

    Yilmaz, Huseyin; Altan, Eda; Ridpath, Julia; Turan, Nuri

    2012-09-01

    The aim of this study was to investigate the frequency and diversity of bovine viral diarrhea viruses (BVDV) infecting cattle in Turkey. A total of 1124 bovine blood samples from 19 farms in 4 different Turkish regions were tested by antigen capture ELISA (ACE). BVDV antigen was found in 26 samples from 13 farms. Only 20 of the 26 initial test positive cattle were available for retesting. Of these, 6 of 20 tested positive for BVDV, by ACE and real-time RT-PCR, one month after initial testing. Phylogenetic analysis, based on comparison of the E2 or the 5'UTR coding regions, from 19 of the 26 initial positive samples, indicated that 17 belonged to the BVDV-1 genotype and 2 to the BVDV-2 genotype. Comparison of 5'UTR sequences segregated 8 BVDV-1 strains (strains 5, 6, 10, 11, 12, 13, 17, and 19) to the BVDV1f, 1 strain (strain 8) to the BVDV1i and 1 strain (strain 14) to the BVDV1d subgenotypes. One strain (strain 4) did not group with other subgenotypes but was closer to the BVDV1f. The remaining 6 BVDV-1 strains (strains 1, 2, 3, 7, 9, and 18) segregated to a novel subgenotype. The E2 sequence comparison results were similar, with the exception that strain 5 grouped with the novel subgenotype rather than BVDV1f subgenotype. It appears that among the diverse BVDV strains in circulation there may be a subgenotype that is unique to Turkey. This should be considered in the design of diagnostics and vaccines to be used in Turkey.

  15. Antigenic characterization of Brazilian bovine viral diarrhea virus isolates by monoclonal antibodies and cross-neutralization

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    Botton S.A.

    1998-01-01

    Full Text Available Nineteen Brazilian isolates of bovine viral diarrhea virus (BVDV were characterized antigenically with a panel of 19 monoclonal antibodies (mAbs (Corapi WV, Donis RO and Dubovi EJ (1990 American Journal of Veterinary Research, 55: 1388-1394. Eight isolates were further characterized by cross-neutralization using sheep monospecific antisera. Analysis of mAb binding to viral antigens by indirect immunofluorescence revealed distinct patterns of reactivity among the native viruses. Local isolates differed from the prototype Singer strain in recognition by up to 14 mAbs. Only two mAbs - one to the non-structural protein NS23/p125 and another to the envelope glycoprotein E0/gp48 - recognized 100% of the isolates. No isolate was recognized by more than 14 mAbs and twelve viruses reacted with 10 or less mAbs. mAbs to the major envelope glycoprotein E2/gp53 revealed a particularly high degree of antigenic variability in this glycoprotein. Nine isolates (47.3% reacted with three or less of 10 E2/gp53 mAbs, and one isolate was not recognized by any of these mAbs. Virus-specific antisera to eight isolates plus three standard BVDV strains raised in lambs had virus-neutralizing titers ranging from 400 to 3200 against the homologous virus. Nonetheless, many antisera showed significantly reduced neutralizing activity when tested against heterologous viruses. Up to 128-fold differences in cross-neutralization titers were observed for some pairs of viruses. When the coefficient of antigenic similarity (R was calculated, 49 of 66 comparisons (74.24% between viruses resulted in R values that antigenically distinguish strains. Moreover, one isolate had R values suggesting that it belongs to a distinct serologic group. The marked antigenic diversity observed among Brazilian BVDV isolates should be considered when planning diagnostic and immunization strategies.

  16. Noncytopathic bovine viral diarrhea virus 2 impairs virus control in a mouse model.

    Science.gov (United States)

    Seong, Giyong; Lee, Jin-Sol; Lee, Kyung-Hyun; Shin, Seung-Uk; Yoon, Ji Young; Choi, Kyoung-Seong

    2016-02-01

    Bovine viral diarrhea virus (BVDV) is an economically important pathogen that causes development of mild to severe clinical signs in wild and domesticated ruminants. We previously showed that mice could be infected by BVDV. In the present study, we infected mice intraperitoneally with non-cytopathic (ncp) BVDV1 or ncp BVDV2, harvested the blood and organs of the infected mice at days 4, 7, 10 and 14 postinfection (pi), and performed immunohistochemical analyses to confirm BVDV infection. Viral antigens were detected in the spleens of all infected mice from days 4 through 14 and were also found in the mesenteric lymph nodes, gut-associated lymphoid tissue (GALT), heart, kidney, intestine, and bronchus-associated lymphoid tissue (BALT) of some infected mice. In ncp BVDV2-infected mice, flow cytometric analysis revealed markedly fewer CD4(+) and CD8(+) T lymphocytes and lower expression of costimulatory molecules CD80 (B7-1) and CD86 (B7-2) and major histocompatibility complex (MHC) class II (I-A/I-E) than those in ncp BVDV1-infected mice. Production of the cytokines interleukin (IL)-6 and monocyte chemotactic protein (MCP)-1 was higher in the plasma of ncp BVDV2-infected mice than that in that of ncp BVDV1-infected mice. Our results demonstrate that ncp BVDV1 and ncp BVDV2 interact differently with the host innate immune response in vivo. These findings highlight an important distinction between ncp BVDV1 and ncp BVDV2 and suggest that ncp BVDV2 impairs the host's ability to control the infection and enhances virus dissemination.

  17. Economic effects of exposure to bovine viral diarrhea virus on dairy herds in New Zealand.

    Science.gov (United States)

    Heuer, C; Healy, A; Zerbini, C

    2007-12-01

    The economic loss to dairy farmers associated with bovine viral diarrhea virus (BVDV) is believed to be high in New Zealand, but no estimates are yet available. The aim was therefore to estimate the economic loss associated with BVDV in dairy herds in New Zealand. Bulk tank milk (BTM) from a random sample of 590 herds from the Northland, Bay of Plenty, and Waikato regions was tested for antibody against BVDV. The inhibition percentage (sample to positive ratio), based on a threshold validated in an earlier study, was used to indicate herd-level infection. Herd reproductive indices, herd lactation-average somatic cell counts, and herd average production of milk solids were regressed on BTM inhibition percentage. Herd averages of the overall annual culling rate, the rate of culling because of failure to conceive, the proportion of physiological inter-service intervals, the first-service conception rate, the pregnancy rate at the end of mating, and somatic cell counts were not associated with BVDV antibody in BTM. Abortion rates, rates of calving induction, the time from calving to conception, and the number of services per conception increased, however, whereas milk production decreased with increasing BVDV antibody in BTM. The results indicated significant reproductive and production loss associated with the amount of BVDV antibody in BTM. Total loss attributable to infection with BVDV was similar to reports from other countries and estimated as NZ$87 per cow and year in affected herds, and NZ$44.5 million per year for the New Zealand dairy industry based on an estimated 14.6% affected herds. The loss estimate excludes added cost and negative consequences with respect to animal welfare attributable to increased induction rates, and a greater incidence of production disease because of BVD-induced immune suppression.

  18. Diagnostic gap in Bovine viral diarrhea virus serology during the periparturient period in cattle.

    Science.gov (United States)

    Bachofen, Claudia; Bollinger, Barbara; Peterhans, Ernst; Stalder, Hanspeter; Schweizer, Matthias

    2013-09-01

    Detection of antibodies against Bovine viral diarrhea virus (BVDV) in serum and milk by enzyme-linked immunosorbent assay (ELISA) is a crucial part of all ongoing national schemes to eradicate this important cattle pathogen. Serum and milk are regarded as equally suited for antibody measurement. However, when retesting a seropositive cow 1 day after calving, the serum was negative in 6 out of 9 different ELISAs. To further investigate this diagnostic gap around parturition, pre- and postcalving serum and milk samples of 5 cows were analyzed by BVDV antibody ELISA and serum neutralization test (SNT). By ELISA, 3 out of the 5 animals showed a diagnostic gap in the serum for up to 12 days around calving but all animals remained positive in SNT. In milk, the ELISA was strongly positive after birth but antibody levels decreased considerably within the next few days. Because of the immunoglobulin G (IgG)1-specific transport of serum antibodies into the mammary gland for colostrum production, the IgG subclass specificity of the total and the BVDV-specific antibodies were determined. Although all 5 animals showed a clear decrease in the total and BVDV-specific IgG1 antibody levels at parturition, the precalving IgG1-to-IgG2 ratios of the BVDV-specific antibodies were considerably lower in animals that showed the diagnostic gap. Results showed that BVDV seropositive cows may become "false" negative in several ELISAs in the periparturient period and suggest that the occurrence of this diagnostic gap is influenced by the BVDV-specific IgG subclass response of the individual animal.

  19. Production and characterization of monoclonal antibodies to Brazilian isolates of bovine viral diarrhea virus

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    L.C. Kreutz

    2000-12-01

    Full Text Available Three Brazilian isolates of bovine viral diarrhea virus (BVDV, antigenically distinct from the standard North American isolates, were selected to immunize BALB/c mice in order to obtain hybridoma cells secreting anti-BVDV monoclonal antibodies (mAbs. Two hybridoma clones secreting mAbs, reacting specifically with BVDV-infected cells (mAbs 3.1C4 and 6.F11, were selected after five fusions and screening of 1001 hypoxanthine-aminopterin-thymidine-resistant clones. These mAbs reacted in an indirect fluorescent antibody (IFA assay with all 39 South and North American BVDV field isolates and reference strains available in our laboratory, yet failed to recognize other pestiviruses, namely the hog cholera virus. The mAbs reacted at dilutions up to 1:25,600 (ascitic fluid and 1:100 (hybridoma culture supernatant in IFA and immunoperoxidase (IPX staining of BVDV-infected cells but only mAb 3.1C4 neutralized virus infectivity. Furthermore, both mAbs failed to recognize BVDV proteins by IPX in formalin-fixed paraffin-embedded tissues and following SDS-PAGE and immunoblot analysis of virus-infected cells, suggesting they are probably directed to conformational-type epitopes. The protein specificity of these mAbs was then determined by IFA staining of CV-1 cells transiently expressing each of the BVDV proteins: mAb 3.1C4 reacted with the structural protein E2/gp53 and mAb 6.F11 reacted with the structural protein E1/gp25. Both mAbs were shown to be of the IgG2a isotype. To our knowledge, these are the first mAbs produced against South American BVDV isolates and will certainly be useful for research and diagnostic purposes.

  20. Phylogeography, phylodynamics and transmission chains of bovine viral diarrhea virus subtype 1f in Northern Italy.

    Science.gov (United States)

    Cerutti, Francesco; Luzzago, Camilla; Lauzi, Stefania; Ebranati, Erika; Caruso, Claudio; Masoero, Loretta; Moreno, Ana; Acutis, Pier Luigi; Zehender, Gianguglielmo; Peletto, Simone

    2016-11-01

    Bovine viral diarrhea virus (BVDV) type 1 in Italy is characterized by high genetic diversity, with at least 20 subtypes. Subtype 1f is endemic in a restricted geographic area, meaning that it has local distribution. We investigated the population dynamics of BVDV-1f in Northern Italy and characterized the transmission chains of a subset of samples from Piedmont and Aosta Valley regions. A total of 51 samples from 1966 to 2013 were considered and 5' UTR sequences were used for phylogeography. A subset of 12 samples was selected for Npro gene sequencing and further characterization of the transmission chains using both molecular and epidemiological data. Phylogeography estimated the root of BVDV-1f tree in Veneto in 1965. Four significant subclades included sequences clustering by region: Lombardy (n=3), Lombardy and Emilia-Romagna (n=7), Piedmont (n=17), Piedmont and Aosta Valley (n=21). The Piedmont-only subclade has a ladder-like branching structure, while the Piedmont and Aosta Valley subclade has a nearly complete binary structure. In the subset, the outbreak reconstruction identified one sample from Piedmont as the most probable source of infection for the Aosta Valley cases. An ad hoc questionnaire submitted to public veterinarians revealed connections between sampled and non-sampled farms by means of trades, exhibitions and markets. According to the phylogeography, BVDV-1f moved westward, entering from Veneto, and spreading to Lombardy and Emilia-Romagna in the early 1990s, and finally to Piedmont and Aosta Valley in the first decade of 2000s. Both phylogeographic analyses on the whole dataset and on the selection of Npro dataset pointed out that subtype 1f entered Aosta Valley from Piedmont. The integration of molecular and epidemiological data revealed connections between farms, and such approach should be considered in any control plan. In Aosta Valley, the study showed that BVDV1f can be controlled only monitoring the introduction of cattle from Piedmont

  1. Genetic diversity of bovine viral diarrhea virus 1: Italian isolates clustered in at least seven subgenotypes.

    Science.gov (United States)

    Giammarioli, Monica; Pellegrini, Claudia; Casciari, Cristina; Rossi, Elisabetta; De Mia, Gian Mario

    2008-11-01

    Bovine viral diarrhea virus (BVDV) is an economically important pathogen of cattle. Two approved species are recognized, namely BVDV-1 and BVDV-2. To date, only 4 subgenotypes of BVDV-2 are known, and at least 11 distinct subgenotypes have been detected for BVDV-1. In a previous study, the genetic characteristics of 38 field isolates of BVDV from northern Italy were investigated, and all 38 isolates were classified as BVDV-1 and could be assigned to 5 different subgenotypes, namely BVDV-1b, BVDV-1d, BVDV-1e, BVDV-1h, and BVDV-1f. However, the circulation of BVDV-2 has been reported in Italy as well. The aim of the current study was to type 88 BVD viruses found throughout Italy. Genetic study was based on the 5'-UTR, supported by select comparison within the N(pro) coding region. Phylogenetic analysis showed that 5 isolates could be typed as BVDV-2a. The remaining 83 isolates were typed as BVDV-1 and were found to belong to 7 distinct subgenotypes, namely BVDV-1a (n = 8), BVDV-1b (n = 37), BVDV-1d (n = 3), BVDV-1e (n = 22), BVDV-1f (n = 4), BVDV-1g (n = 4), and BVDV-1h (n = 5). The majority of cattle farms in the current study were predominantly infected by BVDV-1b and BVDV-1e isolates, whereas the other BVDV subgenotypes occurred only sporadically. The results also provided evidence for circulation of additional subgenotypes BVDV-1a and BVDV-1g. The occurrence of BVDV-2 was also reconfirmed.

  2. Bovine Viral Diarrhea Virus in Zoos: A Perspective from the Veterinary Team

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    Jack J Kottwitz

    2016-01-01

    Full Text Available The many different species in close proximity make zoological collections a unique environment for disease transmission. Bovine Viral Diarrhea Virus (BVDV is of special concern with zoos due to the numerous exotic ruminant species that this virus can infect. BVDV occurs as both a non-cytopathic and a cytopathic strain both of which are capable of infecting exotic ruminants. The cytopathic strain causes mucosal disease and death. Infection with the non-cytopathic strain may produce persistently infected (PI animals. PI individuals may show vague clinical signs, including abortion. Management of BVDV in zoos should focus on identification of PI individuals and prevention of infection of other animals of the collection. Variability makes serological testing as the sole method of screening for BVDV infection undesirable in exotic ruminants. Combination testing provides a definitive answer, especially in sensitive wildlife. Use of a combination of antigen-capture ELISA (ACE with haired skin, Real Time-PCR (RT-PCR on whole blood, and antibody detection via serum neutralization has the greatest potential to identify PI animals. An animal that is positive on both ACE and RT-PCR, but is negative on serology should be considered highly suspicious of being a PI, and should be isolated and undergo repeat testing 4 to 6 weeks later to confirm positive status. This testing methodology also allows screening of pregnant and newborn animals. Isolation or culling may need to be considered in animals determined to be positive via combination testing. These decisions should only be made after careful consideration and evaluation, especially with endangered species.

  3. Isolation and identification of a bovine viral diarrhea virus from sika deer in china

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    Wang Nan

    2011-02-01

    Full Text Available Abstract Background Bovine viral diarrhea virus (BVDV infections continue to cause significantly losses in the deer population. Better isolation and identification of BVDV from sika deer may contribute significantly to the development of prophylactic therapeutic, and diagnostic reagents as well as help in prevention and control of BVDV. However, isolation and identification of BVDV from sika deer is seldom reported in literature. In this study, we collected some samples according to clinical sign of BVDV to isolation and identification of BVDV from sika deer. Results we isolated a suspected BVDV strain from livers of an aborted fetus from sika deer in Changchun (China using MDBK cell lines, named as CCSYD strain, and identified it by cytopathic effect (CPE, indirect immunoperoxidase test (IPX and electron microscopy(EM. The results indicated that this virus was BVDV by a series of identification. The structural proteins E0 gene was cloned and sequenced. The obtained E0 gene sequence has been submitted to GenBank with the accession number: FJ555203. Alignment with other 9 strains of BVDV, 7 strains of classical swine fever virus (CSFV and 3 strains of border disease virus(BDV in the world, showed that the homology were 98.6%-84.8%, 76.0%-74.7%, 76.6%-77.0% for nucleotide sequence, respectively. The phylogenetic analysis indicated that new isolation and identification CCSYD strain belonged to BVDV1b. Conclusion To the best of our knowledge, this is the first report that BVDV was isolated and identified in sika deer. This current research contributes development new BVDV vaccine to prevent and control of BVD in sika deer.

  4. Innate immune responses of calves during transient infection with a noncytopathic strain of bovine viral diarrhea virus

    DEFF Research Database (Denmark)

    Muller-Doblies, D.; Arquint, A.; Schaller, P.

    2004-01-01

    with the temporal activation of the alpha/beta interferon-regulated Mx gene in white blood cells (WBC) and skin as well as the upregulation of the acute-phase serum proteins haptoglobin (Hp) and serum amyloid A (SAA). The viral strain used did provoke transient health impairment, namely, fever and leukopenia...... that were associated with viremia, viral shedding with nasal secretions, and antiviral seroconversion. Complete recovery was observed within 3 weeks. Elevated levels of SAA and Hp were apparent from days 4 to 13 and 8 to 11, respectively. In WBC, the levels of Mx mRNA and Mx protein were elevated from days......In this study, six immunocompetent calves were experimentally infected with a noncytopathic strain of bovine viral diarrhea virus (BVDV), and the effects of the viral infection on parameters of the innate immune response of the host were analyzed. Clinical and virological data were compared...

  5. Molecular analyses detect natural coinfection of water buffaloes (Bubalus bubalis) with bovine viral diarrhea viruses (BVDV) in serologically negative animals.

    Science.gov (United States)

    Craig, María I; König, Guido A; Benitez, Daniel F; Draghi, María G

    2015-01-01

    Infection of water buffaloes (Bubalus bubalis) with bovine viral diarrhea viruses (BVDV) has been confirmed in several studies by serological and molecular techniques. In order to determine the presence of persistently infected animals and circulating species and subtypes of BVDV we conducted this study on a buffalo herd, whose habitat was shared with bovine cattle (Bossp.). Our serological results showed a high level of positivity for BVDV-1 and BVDV-2 within the buffalo herd. The molecular analyses of blood samples in serologically negative animals revealed the presence of viral nucleic acid, confirming the existence of persistent infection in the buffaloes. Cloning and sequencing of the 5' UTR of some of these samples revealed the presence of naturally mix-infected buffaloes with at least two different subtypes (1a and 1b), and also with both BVDV species (BVDV-1 and BVDV-2).

  6. Mutations induced in the NS5B gene of bovine viral diarrhea virus by antiviral treatment convey resistance to the compound

    Science.gov (United States)

    Bovine viral diarrhea virus (BVDV) is a widespread bovine pathogen for which there is no specific therapeutic agent. A previous study using 2-(2-benzimidazolyl)-5-[4-(2-imidazolino)phenyl]furan dihydrochloride (DB772) to treat calves persistently infected with BVDV resulted in a decrease in the vira...

  7. Impact of species and subgenotypes of bovine viral diarrhea virus on control by vaccination.

    Science.gov (United States)

    Fulton, Robert W

    2015-06-01

    Bovine viral diarrhea viruses (BVDV) are diverse genetically and antigenically. This diversity impacts both diagnostic testing and vaccination. In North America, there are two BVDV species, 1 and 2 with 3 subgenotypes, BVDV1a, BVDV1b and BVDV2a. Initially, US vaccines contained BVDV1a cytopathic strains. With the reporting of BVDV2 severe disease in Canada and the USA there was focus on protection by BVDV1a vaccines on BVDV2 disease. There was also emphasis of controlling persistently infected (PI) cattle resulted in studies for fetal protection afforded by BVDV1a vaccines. Initially, studies indicated that some BVDV1a vaccines gave less than 100% protection against BVDV2 challenge for fetal infection. Eventually vaccines in North America added BVDV2a to modified live virus (MLV) and killed BVDV1a vaccines. Ideally, vaccines should stimulate complete immunity providing 100% protection against disease, viremias, shedding, and 100% fetal protection in vaccinates when challenged with a range of diverse antigenic viruses (subgenotypes). There should be a long duration of immunity stimulated by vaccines, especially for fetal protection. MLV vaccines should be safe when given according to the label and free of other pathogens. While vaccines have now included BVDV1a and BVDV2a, with the discovery of the predominate subgenotype of BVDV in the USA to be BVDV1b, approximately 75% or greater in prevalence, protection in acute challenge and fetal protection studies became more apparent for BVDV1b. Thus many published studies examined protection by BVDV1a and BVDV2a vaccines against BVDV1b in acute challenge and fetal protection studies. There are no current BVDV1b vaccines in the USA. There are now more regulations on BVDV reproductive effects by the USDA Center for Veterinary Biologics (CVB) regarding label claims for protection against abortion, PI calves, and fetal infections, including expectations for studies regarding those claims. Also, the USDA CVB has a memorandum

  8. Isolation of a mutant MDBK cell line resistant to bovine viral diarrhea virus infection due to a block in viral entry.

    Science.gov (United States)

    Flores, E F; Donis, R O

    1995-04-20

    A cell line, termed CRIB, resistant to infection with bovine viral diarrhea virus (BVDV) has been derived from the MDBK bovine kidney cell line. CRIB cells were obtained by selection and cloning of cells surviving infection with a highly cytolytic BVDV strain. CRIB cells contain no detectable infectious or defective BVDV as ascertained by cocultivation, animal inoculation, indirect immunofluorescence, Western immunoblot, Northern hybridization, and RNA PCR. Inoculation of CRIB cells with 24 cytopathic and noncytopathic BVDV strains does not result in expression of viral genes or amplification of input virus. Karyotype and isoenzyme analyses demonstrated that CRIB are genuine bovine cells. CRIB cells are as susceptible as the parental MDBK cells to 10 other bovine viruses, indicating that these cells do not have a broad defect blocking viral replication. Transfection of CRIB cells with BVDV RNA or virus inoculation in the presence of polyethylene-glycol results in productive infection, indicating that the defect of CRIB cells is at the level of virus entry. CRIB cells are the first bovine cells reported to be resistant to BVDV infection in vitro and may be a useful tool for studying the early interactions of pestiviruses with host cells.

  9. Development of a sandwich Dot-ELISA for detecting bovine viral diarrhea virus antigen with E2 recombinant protein

    Institute of Scientific and Technical Information of China (English)

    Yuelan ZHAO; Yuzhu ZUO; Lei ZHANG; Jinghui FAN; Hanchun YANG; Jianhua QIN

    2009-01-01

    The IgG antibodies of rabbit anti-E2 protein of the bovine viral diarrhea virus were prepared by a general method from high efficiency serum immunized by E2 recombinant protein antigen expressed in E. coli prokaryotic expression system and were labeled to make enzymelabeled antibody with the method of NaIO4. A sandwich Dot enzyme-linked immunosorbent assay (Dot-ELISA) for the detection of BVDV was developed. The optimal reaction conditions of Dot-ELISAwere determined. The results show that optimal coating antibody was 300 μg·mL-1, the working concentration of HRP-labeled antibody was 1:50. The optimal blocking reagent and time were 5% bovine serum and 45 rain. The minimum detection of the content of antigen reached 1.35μg·mL-1. Compared with the routine IDEXX ELISA test kit with the whole virus, its specificity, sensitivity and coincidence rate were 90.48%, 96.55% and 95.24%, respectively. Compared with the sandwich Dot-ELISA with the negative staining electron microscope and RT-PCR, the coincidence rates were 90.9% and 93.1%, respectively. In addition, Bovine viral diarrhea virus (BVDV) antigen of 178 samples collected from cow farms in the Hebei Province, China, were detected by the developed Dot-ELISA and the IDEXX BVDV antigen Test Kit simultaneously, BVDV antigen positive rate was 39.89%-41.01%. The result of detecting clinical samples demonstrated that the established method showed its specificity, sensitivity and repeatability, whereas the results were easily interpreted without an ELISA reader.

  10. Evaluation of the epidemiological and economic consequences of control scenarios for bovine viral diarrhea virus in dairy herds.

    Science.gov (United States)

    Santman-Berends, I M G A; Mars, M H; van Duijn, L; van Schaik, G

    2015-11-01

    Bovine viral diarrhea virus (BVDV) is an important endemic infection. However, no information was available on whether it would be economically beneficial to implement a national control program in the Netherlands. Therefore, a stochastic simulation model was developed in which control scenarios were added to compare the epidemiological and economic consequences of BVDV control in Dutch dairy herds in the next 10 yr. In the epidemiological part of the model, herds could be classified as susceptible, infectious, recovered, or vaccinated. The outputs of the epidemiological module served as input for the economic module. Net costs that could be attributed to bovine viral diarrhea consisted of production losses, costs for testing, and culling persistently infected cattle in the present voluntary Dutch BVDV control program and costs for vaccination. Four different control scenarios were simulated, involving testing and culling of persistently infected (based on serum or ear-notch testing), and monitoring BVDV statuses and vaccination and were derived from BVDV control programs that are currently executed in Europe. The costs and benefits of BVDV control in the current situation and in each of the simulated control scenarios were evaluated assuming an annual discount rate of 2%. The model estimated a mean BVDV herd prevalence of 18.0% in 2014 and showed a slightly decreasing prevalence over time. The outputs seemed realistic for the present situation in the Netherlands when compared with actual survey data. The average annual net costs associated with bovine viral diarrhea were estimated at €27.8 million for the dairy industry. Two control scenarios were beneficial in controlling BVDV during the study period (between 2015 and 2025). In the scenario where tracing and removing of PI animals and monitoring of the subsequent status was obligatory, the benefit to cost (B/C) ratio was 1.5 (€1.5 benefit for each invested euro). In the scenario in which the BVDV status of

  11. Fine mapping of Loci on BTA2 and BTA26 Associated with Bovine Viral Diarrhea Persistent Infection and Linked with Bovine Respiratory Disease in Cattle

    Directory of Open Access Journals (Sweden)

    Ricardo eZanella

    2011-11-01

    Full Text Available Bovine respiratory disease (BRD is considered to be the most costly infectious disease in the cattle industry. Bovine viral diarrhea virus (BVDV is one of the pathogens involved with the BRD complex of disease. Bovine viral diarrhea virus infection also negatively impacts cow reproduction and calf performance. Loci associated with persistently infected animals (BVD-PI and linked with BRD have previously been identified near 14 Mb on bovine chromosome 2 (BTA2 and 15.3 Mb on bovine chromosome 26 (BTA26. The objective of this study was to refine the loci associated with BVD-PIand linked with BRD. Association testing for BVD-PI was performed on a population of 65 BVD-PI calves, 51 of their dams, and 60 unaffected calves (controls with 175 single nucleotide polymorphisms (SNPs on BTA2 and 209 SNPs on BTA26. Comparisons were made between BVD-PI calves and controls calves and the dams of BVD-PI calves and controls calves. For the linkage analysis of BRD, the same markers were used to genotype 2 half sib-families consisting of the sires and 72 BRD positive and 148 BRD negative offspring. Using an allelic chi-square test, 11 loci on BTA2 and 8 loci on BTA26 were associated with the dams of the BVD-PI calves (P < 0.05 and 5 loci on BTA2 and 10 loci on BTA26 were associated with BVD-PI calves. One locus on BTA2 and two loci on BTA26 were found to be linked (P < 0.05 with BRD. These results further refined the loci associated and linked with BVD-PI and BRD, respectively.

  12. Bovine viral diarrhea viruses (BVDV) and their cousins the HoBi-like viruses: Multi symptom, multi host, multi tasking pathogens

    Science.gov (United States)

    The term bovine viral diarrhea (BVD) has come to refer to a diverse collection of clinical presentations that include respiratory, enteric and reproductive symptoms accompanied by immunosuppression. While the majority of cases are subclinical in nature two forms exist, mucosal disease and hemorrhag...

  13. Identification of amino acid changes in the envelope glycoproteins of bovine viral diarrhea viruses isolated from alpaca that may be involved in host adaptation

    Science.gov (United States)

    Bovine viral diarrhea viruses (BVDV) are most commonly associated with infections of cattle. However, BVDV is often isolated from closely related ruminants with a number of BVDV-1b viruses being isolated from alpacas that were both acutely and persistently infected (PI). The complete nucleotide se...

  14. Full-length coding sequence for 12 bovine viral diarrhea virus isolates from persistently infected cattle in a feedyard in Kansas

    Science.gov (United States)

    We report here the full-length coding sequence of 12 bovine viral diarrhea virus (BVDV) isolates from persistently infected cattle from a feedyard in southwest Kansas, USA. These 12 genomes represent the three major genotypes (BVDV 1a, 1b, and 2a) of BVDV currently circulating in the United States....

  15. HoBi-like virus challenge of pregnant cows that had previously given birth to calves persistently infected with bovine viral diarrhea virus

    Science.gov (United States)

    The ability of bovine viral diarrhea viruses (BVDV) to establish persistent infection (PI) following fetal infection is central to keeping these viruses circulating. Similarly, an emerging species of pestivirus, HoBi-like viruses, is also able to establish PIs. Dams that are not PI, but carrying PI ...

  16. Bovine viral diarrhea virus type 2 impairs macrophage responsiveness to toll-like receptor ligation with the exception of toll-like receptor 7

    Science.gov (United States)

    Bovine viral diarrhea virus (BVDV) is a member of the Flaviviradae family. BVDV isolates are classified into two biotypes based on the development of cytopathic (cp) or non-cytopathic (ncp) effects in epithelial cell culture. In addition, BVDV isolates are further separated into species, BVDV1 and 2...

  17. Evidence of bovine viral diarrhea virus infection in three species of sympatric wild ungulates in Nevada: Life history strategies may maintain endemic infections in wild populations

    Science.gov (United States)

    Evidence for bovine viral diarrhea virus (BVDV) infection was detected in 2009-10 during a pneumonia die-off in Rocky Mountain bighorn sheep (Ovis canadensis canadensis), and sympatric mountain goats (Oreamnos americanum) in adjacent mountain ranges in Elko County, Nevada. Seroprevalence to BVDV-1 ...

  18. Preliminary mapping of non-conserved epitopes on envelope glycoprotein E2 of bovine viral diarrhea virus type 1 and 2

    NARCIS (Netherlands)

    Jelsma, H.; Loeffen, W.L.A.; Beuningen, van A.R.; Rijn, van P.A.

    2013-01-01

    Bovine viral diarrhea virus (BVDV) belongs together with Classical swine fever virus (CSFV) and Border disease virus (BDV) to the genus Pestivirus in the Flaviviridae family. BVDV has been subdivided into two different species, BVDV1 and BVDV2 based on phylogenetic analysis. Subsequent characterizat

  19. Comparison of levels and duration of detection of antibodies to bovine viral diarrhea virus 1, bovine viral diarrhea virus 2, bovine respiratory syncytial virus, bovine herpesvirus 1, and bovine parainfluenza virus 3 in calves fed maternal colostrum or a colostrum-replacement product.

    Science.gov (United States)

    Chamorro, Manuel F; Walz, Paul H; Haines, Deborah M; Passler, Thomas; Earleywine, Thomas; Palomares, Roberto A; Riddell, Kay P; Galik, Patricia; Zhang, Yijing; Givens, M Daniel

    2014-04-01

    Colostrum-replacement products are an alternative to provide passive immunity to neonatal calves; however, their ability to provide adequate levels of antibodies recognizing respiratory viruses has not been described. The objective of this study was to compare the serum levels of IgG at 2 d of age and the duration of detection of antibodies to bovine viral diarrhea virus 1 (BVDV-1), bovine viral diarrhea virus 2 (BVDV-2), bovine respiratory syncytial virus (BRSV), bovine herpesvirus 1 (BHV-1), and bovine parainfluenza virus 3 (BPIV-3) in calves fed maternal colostrum (MC) or a colostrum replacement (CR) at birth. Forty newborn male Holstein calves were assigned to the CR or the MC group. Group CR (n = 20) received 2 packets of colostrum replacement (100 g of IgG per 470-g packet), while group MC (n = 20) received 3.8 L of maternal colostrum. Blood samples for detection of IgG and virus antibodies were collected from each calf at birth, at 2 and 7 d, and monthly until the calves became seronegative. Calves in the MC group had greater IgG concentrations at 2 d of age. The apparent efficiency of absorption of IgG was greater in the MC group than in the CR group, although the difference was not significant. Calves in the CR group had greater concentrations of BVDV neutralizing antibodies during the first 4 mo of life. The levels of antibodies to BRSV, BHV-1, and BPIV-3 were similar in the 2 groups. The mean time to seronegativity was similar for each virus in the 2 groups; however, greater variation was observed in the antibody levels and in the duration of detection of immunity in the MC group than in the CR group. Thus, the CR product provided calves with more uniform levels and duration of antibodies to common bovine respiratory viruses.

  20. Evaluation of reproductive protection against bovine viral diarrhea virus and bovine herpesvirus-1 afforded by annual revaccination with modified-live viral or combination modified-live/killed viral vaccines after primary vaccination with modified-live viral vaccine.

    Science.gov (United States)

    Walz, Paul H; Givens, M Daniel; Rodning, Soren P; Riddell, Kay P; Brodersen, Bruce W; Scruggs, Daniel; Short, Thomas; Grotelueschen, Dale

    2017-02-15

    The objective of this study was to compare reproductive protection in cattle against bovine viral diarrhea virus (BVDV) and bovine herpesvirus 1 (BoHV-1) provided by annual revaccination with multivalent modified-live viral (MLV) vaccine or multivalent combination viral (CV) vaccine containing temperature-sensitive modified-live BoHV-1 and killed BVDV when MLV vaccines were given pre-breeding to nulliparous heifers. Seventy-five beef heifers were allocated into treatment groups A (n=30; two MLV doses pre-breeding, annual revaccination with MLV vaccine), B (n=30; two MLV doses pre-breeding, annual revaccination with CV vaccine) and C (n=15; saline in lieu of vaccine). Heifers were administered treatments on days 0 (weaning), 183 (pre-breeding), 366 (first gestation), and 738 (second gestation). After first calving, primiparous cows were bred, with pregnancy assessment on day 715. At that time, 24 group A heifers (23 pregnancies), 23 group B heifers (22 pregnancies), and 15 group C heifers (15 pregnancies) were commingled with six persistently infected (PI) cattle for 16days. Ninety-nine days after PI removal, cows were intravenously inoculated with BoHV-1. All fetuses and live offspring were assessed for BVDV and BoHV-1. Abortions occurred in 3/23 group A cows, 1/22 group B cows, and 11/15 group C cows. Fetal infection with BVDV or BoHV-1 occurred in 4/23 group A offspring, 0/22 group B offspring, and 15/15 group C offspring. This research demonstrates efficacy of administering two pre-breeding doses of MLV vaccine with annual revaccination using CV vaccine to prevent fetal loss due to exposure to BVDV and BoHV-1.

  1. Comparison of humoral immune responses in dairy heifers vaccinated with 3 different commercial vaccines against bovine viral diarrhea virus and bovine herpesvirus-1.

    Science.gov (United States)

    DesCôteaux, Luc; Cécyre, Dominique; Elsener, Johanne; Beauchamp, Guy

    2003-10-01

    A randomized clinical trial was conducted to compare the humoral immune response to 3 different commercial vaccines in dairy heifers housed in 3 different dairy farms in Quebec. All heifers were seronegative to type 1 bovine viral diarrhea virus (BVDV) (Singer strain), type 2 BVDV (NVSL 125c strain), and bovine herpesvirus-1 (BHV-1) at the beginning of the trial. In addition, control heifers in group 1 remained seronegative to the 2 viruses till the end of the trial. Significant differences in humoral immune responses occurred among the 3 commercial vaccines at 4 weeks and 6 months following vaccination. The vaccine in group 2 elicited higher mean antibody titers and seroconversion rates to both type 1 and type 2 BVDV than that in groups 3 or 4. Vaccines in groups 2 and 3 induced higher mean antibody titers to BHV-1 than did the vaccine in group 4.

  2. Experimental infection with cytopathic bovine viral diarrhea virus in mice induces megakaryopoiesis in the spleen and bone marrow.

    Science.gov (United States)

    Seong, Giyong; Lee, Jin-Sol; Lee, Kyung-Hyun; Choi, Kyoung-Seong

    2016-02-01

    Here, we infected mice with cytopathic bovine viral diarrhea virus 1 (cp BVDV1) by oral inoculation and investigated the effects of infection by histopathological, immunohistochemical (IHC), hematological methods. Twelve mice were infected, and samples were obtained at day 2, 5, and 9 postinfection (pi). Most of the infected mice exhibited clinical signs of illness such as reduced movement, crouching, loose feces, loss of appetite, and reduced water intake. Blood samples from six mice were positive for BVDV based on reverse transcription polymerase chain reaction (RT-PCR). Blood analysis also revealed thrombocytopenia and lymphopenia. Viral antigens were detected in the spleen (12/12), bone marrow (12/12), and/or mesenteric lymph nodes (4/12) of all infected mice by IHC analysis. The spleens showed significant histopathological changes including (i) substantially increased numbers of megakaryocytes, (ii) lymphocyte depletion, and (iii) hemorrhages. The bone marrow also had an increased number of megakaryocytes, although this increase was not as strong as it was in the spleen. Severe lymphoid depletion was observed in the mesenteric lymph nodes. Viral infections were present in the lymphocytes but not detected in megakaryocytes of the spleen, bone marrow, or mesenteric lymph nodes. These results suggest that the increased numbers of megakaryocytes may be a direct result of BVDV infection. BVDV infection in mice following oral inoculation of cp BVDV1 leads to megakaryopoiesis in the spleen and bone marrow to replenish the platelets.

  3. Peripheral blood mononuclear cells from field cattle immune to bovine viral diarrhea virus (BVDV) are permissive in vitro to BVDV.

    Science.gov (United States)

    Gupta, V; Mishra, N; Pateriya, A; Behera, S P; Rajukumar, K

    2014-01-01

    The aim of this study was to determine the in vitro permissivity of peripheral blood mononuclear cells (PBMCs) from bovine viral diarrhea virus (BVDV)-immune field cattle to homologous and heterologous BVDVs. PBMCs from seventeen BVDV-naïve and sixteen BVDV-immune animals were infected with noncytopathic BVDV-1 or BVDV-2. The immune status of cattle was indicated by the presence of virus neutralizing antibodies, while viral load of PBMCs was determined by real-time RT-PCR. The results revealed that the PBMCs from naïve or immune animals were permissive to either BVDV-1 or BVDV-2, but the viral load was significantly higher for the naïve than for the immune animals. Furthermore, the load of homologous virus in PBMCs from immune animals was lower than that of heterologous virus. Our results provide evidence that the PBMCs from BVDV-immune cattle in field are susceptible to reinfection with homologous or heterologous BVDV, albeit to a lower extent in the former case.

  4. Genetic diversity of bovine viral diarrhea viruses from the Galicia region of Spain.

    Science.gov (United States)

    Factor, C; Yus, E; Eiras, C; Sanjuan, M L; Cerviño, M; Arnaiz, I; Diéguez, F J

    2016-01-01

    This study examined the frequency and diversity of bovine viral diarrhoea viruses (BVDVs) infecting cattle in Galicia (northwestern Spain). A total of 86 BVDV strains were typed in samples of serum from 79 persistently infected animals and 3 viraemic animals and of abomasal fluid from 4 fetuses. Samples came from 73 farms participating in a voluntary BVDV control programme. Typing was based on a 288-bp sequence from the 5' untranslated region amplified using primers 324 and 326. Of the 86 strains, 85 (98.8 per cent) belonged to species BVDV-1 and 1 (1.2 per cent) belonged to BVDV-2; 73 strains (84.9 per cent) were typed as BVDV-1b, 2 as BVDV-1e and 6 as BVDV-1d. One strain each was typed as belonging to 1a, 1h, 1k and 1l. The sole BVDV-2 strain was classified as 2a. These results identify BVDV-1b as the predominant species, and they indicate the presence of viral types not previously described anywhere in Spain. This is also the first report of BVDV-2 in Galicia and only the second report of BVDV-2 in Spain.

  5. Genetic diversity of bovine viral diarrhea viruses from the Galicia region of Spain

    Science.gov (United States)

    Factor, C.; Yus, E.; Eiras, C.; Sanjuan, M. L.; Cerviño, M.; Arnaiz, I.; Diéguez, F. J.

    2016-01-01

    This study examined the frequency and diversity of bovine viral diarrhoea viruses (BVDVs) infecting cattle in Galicia (northwestern Spain). A total of 86 BVDV strains were typed in samples of serum from 79 persistently infected animals and 3 viraemic animals and of abomasal fluid from 4 fetuses. Samples came from 73 farms participating in a voluntary BVDV control programme. Typing was based on a 288-bp sequence from the 5′ untranslated region amplified using primers 324 and 326. Of the 86 strains, 85 (98.8 per cent) belonged to species BVDV-1 and 1 (1.2 per cent) belonged to BVDV-2; 73 strains (84.9 per cent) were typed as BVDV-1b, 2 as BVDV-1e and 6 as BVDV-1d. One strain each was typed as belonging to 1a, 1h, 1k and 1l. The sole BVDV-2 strain was classified as 2a. These results identify BVDV-1b as the predominant species, and they indicate the presence of viral types not previously described anywhere in Spain. This is also the first report of BVDV-2 in Galicia and only the second report of BVDV-2 in Spain. PMID:27843559

  6. Generation of the bovine viral diarrhea virus e0 protein in transgenic astragalus and its immunogenicity in sika deer.

    Science.gov (United States)

    Gao, Yugang; Zhao, Xueliang; Zang, Pu; Liu, Qun; Wei, Gongqing; Zhang, Lianxue

    2014-01-01

    The bovine viral diarrhea virus (BVDV), a single-stranded RNA virus, can cause fatal diarrhea syndrome, respiratory problems, and reproductive disorders in herds. Over the past few years, it has become clear that the BVDV infection rates are increasing and it is likely that an effective vaccine for BVDV will be needed. In this study, transgenic Astragalus was used as an alternative productive platform for the expression of glycoprotein E0. The immunogenicity of glycoprotein E0 expressed in transgenic Astragalus was detected in deer. The presence of pBI121-E0 was confirmed by polymerase chain reaction (PCR), transcription was verified by reverse transcription- (RT-) PCR, and recombinant protein expression was confirmed by ELISA and Western blot analyses. Deer that were immunized subcutaneously with the transgenic plant vaccine developed specific humoral and cell-mediated immune responses against BVDV. This study provides a new method for a protein with weak immunogenicity to be used as part of a transgenic plant vaccine.

  7. Generation of the Bovine Viral Diarrhea Virus E0 Protein in Transgenic Astragalus and Its Immunogenicity in Sika Deer

    Directory of Open Access Journals (Sweden)

    Yugang Gao

    2014-01-01

    Full Text Available The bovine viral diarrhea virus (BVDV, a single-stranded RNA virus, can cause fatal diarrhea syndrome, respiratory problems, and reproductive disorders in herds. Over the past few years, it has become clear that the BVDV infection rates are increasing and it is likely that an effective vaccine for BVDV will be needed. In this study, transgenic Astragalus was used as an alternative productive platform for the expression of glycoprotein E0. The immunogenicity of glycoprotein E0 expressed in transgenic Astragalus was detected in deer. The presence of pBI121-E0 was confirmed by polymerase chain reaction (PCR, transcription was verified by reverse transcription- (RT- PCR, and recombinant protein expression was confirmed by ELISA and Western blot analyses. Deer that were immunized subcutaneously with the transgenic plant vaccine developed specific humoral and cell-mediated immune responses against BVDV. This study provides a new method for a protein with weak immunogenicity to be used as part of a transgenic plant vaccine.

  8. Clinical and virological characteristics of calves experimentally infected with a Brazilian isolate of bovine viral diarrhea virus type 1a

    Directory of Open Access Journals (Sweden)

    Luana Marchi Quadros

    Full Text Available ABSTRACT: To study the pathogenicity of the Brazilian bovine viral diarrhea virus (BVDV type 1a 241.10 isolate, four calves were intranasally inoculated with a viral suspension containing 107.2 TCID50 mL-1. One calf was left uninoculated and kept in contact with the other calves to investigate viral transmissibility. After inoculation, the animals were monitored daily for clinical signs of infection. The presence of the virus in the blood and nasal secretions was confirmed by virus isolation in cell culture. White blood cells were quantified prior to and every 3 days after infection, and the presence of antibodies was checked every 7 days, starting at day 0 until day 42 post-inoculation (pi. After infection, nasal and ocular serous secretions were observed between days 1 and 5 pi, along with a mild cough from days 2 to 4 pi; however, no severe clinical signs were present. Body temperature was slightly elevated between days 4 and 6 pi. The control calf did not develop any of the signs observed in the infected animals. Cell culture-mediated virus isolation confirmed viremia between days 4 and 8 pi and the presence of the virus in the nasal secretions between days 1 and 10 pi. All infected animals showed a decrease in white blood cell count. Antibodies could be detected from day 14 pi, and these levels remained high until day 35 pi. The control calf had no viremia, viral presence in nasal secretions, or positive serology, indicating the absence of viral transmission. Thus, isolate BVDV 1a 241.10 has low pathogenicity and transmissibility but retains immunosuppressive capacity.

  9. Experimental infection with non-cytopathic bovine viral diarrhea virus 1 in mice induces inflammatory cell infiltration in the spleen.

    Science.gov (United States)

    Han, Yu-Jung; Kwon, Young-Je; Lee, Kyung-Hyun; Choi, Eun-Jin; Choi, Kyoung-Seong

    2016-09-01

    Previously, our study showed that oral inoculation of mice with cytopathic (cp) bovine viral diarrhea virus (BVDV) led to lymphocyte depletion and increased numbers of megakaryocytes in the spleen as well as thrombocytopenia and lymphopenia. In the present study, to investigate the possible differences in the detection of viral antigen, histopathological lesions, and hematologic changes between non-cytopathic (ncp) BVDV1 and cp BVDV1, mice were orally administered low and high doses of ncp BVDV1 and were necropsied at days 0, 2, 5, and 9 postinfection (pi). None of the ncp BVDV1-infected mice exhibited clinical signs of illness, unlike those infected with cp BVDV1. Statistically significant thrombocytopenia was observed during ncp BVDV1 infection, and lymphopenia was found only in mice infected with a high dose at day 9 pi. Interestingly, ncp BVDV1 infection increased the numbers of basophils, eosinophils, neutrophils, and monocytes in some infected mice. Viral antigen was detected in the lymphocytes of the spleen, mesenteric lymph nodes, Peyer's patches, and bone marrow by immunohistochemistry. Lymphoid depletion was evident in the mesenteric lymph nodes of mice infected with a high dose and also found in the Peyer's patches of some infected mice. Infiltration of inflammatory cells, including neutrophils and monocytes, and an increased number of megakaryocytes were seen in the spleen. These results suggest that the distribution of viral antigens is not associated with the presence of histopathological lesions. Inflammatory cell infiltration was observed in the spleens as a result of viral replication and may be attributable to the host reaction to ncp BVDV1 infection. Together, these findings support the possibility that mice can be used as an animal model for BVDV infection.

  10. Priming Cross-Protective Bovine Viral Diarrhea Virus-Specific Immunity Using Live-Vectored Mosaic Antigens

    Science.gov (United States)

    Fang, Xin; Waghela, Suryakant D.; Bray, Jocelyn; Njongmeta, Leo M.; Herring, Andy; Abdelsalam, Karim W.; Chase, Christopher; Mwangi, Waithaka

    2017-01-01

    Bovine viral diarrhea virus (BVDV) plays a key role in bovine respiratory disease complex, which can lead to pneumonia, diarrhea and death of calves. Current vaccines are not very effective due, in part, to immunosuppressive traits and failure to induce broad protection. There are diverse BVDV strains and thus, current vaccines contain representative genotype 1 and 2 viruses (BVDV-1 & 2) to broaden coverage. BVDV modified live virus (MLV) vaccines are superior to killed virus vaccines, but they are susceptible to neutralization and complement-mediated destruction triggered by passively acquired antibodies, thus limiting their efficacy. We generated three novel mosaic polypeptide chimeras, designated NproE2123; NS231; and NS232, which incorporate protective determinants that are highly conserved among BVDV-1a, 1b, and BVDV-2 genotypes. In addition, strain-specific protective antigens from disparate BVDV strains were included to broaden coverage. We confirmed that adenovirus constructs expressing these antigens were strongly recognized by monoclonal antibodies, polyclonal sera, and IFN-γ-secreting T cells generated against diverse BVDV strains. In a proof-of-concept efficacy study, the multi-antigen proto-type vaccine induced higher, but not significantly different, IFN-γ spot forming cells and T-cell proliferation compared to a commercial MLV vaccine. In regards to the humoral response, the prototype vaccine induced higher BVDV-1 specific neutralizing antibody titers, whereas the MLV vaccine induced higher BVDV-2 specific neutralizing antibody titers. Following BVDV type 2a (1373) challenge, calves immunized with the proto-type or the MLV vaccine had lower clinical scores compared to naïve controls. These results support the hypothesis that a broadly protective subunit vaccine can be generated using mosaic polypeptides that incorporate rationally selected and validated protective determinants from diverse BVDV strains. Furthermore, regarding biosafety of using a

  11. Predominance of bovine viral diarrhea virus 1b and 1d subtypes during eight years of survey in Poland.

    Science.gov (United States)

    Kuta, A; Polak, M P; Larska, M; Żmudziński, J F

    2013-10-25

    The genetic diversity of bovine viral diarrhea virus (BVDV) was determined from 65 animals persistently infected with BVDV and diagnosed between 2004 and 2011 in Poland. The samples originated from 28 herds in 12 provinces, where over 90% of the whole cattle population of Poland is reared. Phylogenetic analysis based on the fragments of two genomic regions of BVDV namely, 5'-untranslated region (5'-UTR) and N(pro) was performed. All the BVDV isolates belonged to BVDV-1 species and were further divided into four subtypes. There were 31 viruses of BVDV-1b subtype (47.6%) present in 12 herds, 24 of BVDV-1d subtype (36.9%) in 9 herds, 8 of BVDV-1f subtype (12.3%) in 5 herds and 2 BVDV-1g subtype (3.0%) in 2 herds. Neither BVDV-1a subtype, nor BVDV-2 species or any atypical bovine pestivirus were found among isolates tested. Despite increasing import of live cattle in the recent years, genetic diversity of Polish BVDV isolates was rather low.

  12. Two doses of bovine viral diarrhea virus DNA vaccine delivered by electroporation induce long-term protective immune responses.

    Science.gov (United States)

    van Drunen Littel-van den Hurk, Sylvia; Lawman, Zoe; Snider, Marlene; Wilson, Don; van den Hurk, Jan V; Ellefsen, Barry; Hannaman, Drew

    2013-02-01

    Bovine viral diarrhea virus (BVDV) is a pathogen of major importance in cattle, so there is a need for new effective vaccines. DNA vaccines induce balanced immune responses and are relatively inexpensive and thus promising for both human and veterinary applications. In this study, newborn calves with maternal antibodies were vaccinated intramuscularly (i.m.) with a BVDV E2 DNA vaccine with the TriGrid Delivery System for i.m. delivery (TDS-IM). Two doses of this vaccine spaced 6 or 12 weeks apart were sufficient to induce significant virus-neutralizing antibody titers, numbers of activated T cells, and reduction in viral shedding and clinical presentations after BVDV-2 challenge. In contrast to the placebo-treated animals, the vaccinated calves did not lose any weight, which is an excellent indicator of the well-being of an animal and has a significant economic impact. Furthermore, the interval between the two vaccinations did not influence the magnitude of the immune responses or degree of clinical protection, and a third immunization was not necessary or beneficial. Since electroporation may enhance not only the magnitude but also the duration of immunity after DNA immunization, the interval between vaccination and challenge was extended in a second trial, which showed that two doses of this E2 DNA vaccine again significantly reduced clinical disease against BVDV for several months. These results are promising and support this technology for use against infectious diseases in cattle and large species, including humans, in general.

  13. Prevalence study and genetic typing of bovine viral diarrhea virus (BVDV in four bovine species in China.

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    Mingliang Deng

    Full Text Available To determine the nationwide status of persistent BVDV infection in different bovine species in China and compare different test methods, a total of 1379 serum samples from clinical healthy dairy cattle, beef cattle, yaks (Bos grunniens, and water buffalo (Bubalus bubalis were collected in eight provinces of China from 2010 to 2013. The samples were analyzed using commercial antibody (Ab and antigen (Ag detection kits, and RT-PCR based on the 5'-UTR and Npro gene sequencing. Results showed that the overall positive rates for BVDV Ab, Ag and RT-PCR detection were 58.09% (801/1379, 1.39% (14/1010, and 22.64% (146/645, respectively, while the individual positive rates varied among regions, species, and farms. The average Ab-positive rates for dairy cattle, beef cattle, yaks, and water buffalo were 89.49% (298/333, 63.27% (248/392, 45.38% (236/520, and 14.18% (19/134, respectively, while the Ag-positive rates were 0.00% (0/116, 0.77% (3/392, 0.82% (3/368, and 5.97% (8/134, respectively, and the nucleic acid-positive rates detected by RT-PCR were 32.06% (42/131, 13.00% (26/200, 28.89% (52/180, and 19.40% (26/134, respectively. In addition, the RT-PCR products were sequenced and 124 5'-UTR sequences were obtained. Phylogenetic analysis of the 5'-UTR sequences indicated that all of the 124 BVDV-positive samples were BVDV-1 and subtyped into either BVDV-1b (33.06%, BVDV-1m (49.19%, or a new cluster, designated as BVDV-1u (17.74%. Phylogenetic analysis based on Npro sequences confirmed this novel subtype. In conclusion, this study revealed the prevalence of BVDV-1 in bovine species in China and the dominant subtypes. The high proportion of bovines with detectable viral nucleic acids in the sera, even in the presence of high Ab levels, revealed a serious threat to bovine health.

  14. Viral causes of diarrhea.

    Science.gov (United States)

    Goodgame, R W

    2001-09-01

    Viruses are important causes of diarrhea. In healthy adults, the main clinical manifestation is acute, self-limited gastroenteritis. Advances in molecular diagnostics have shown that epidemics of acute gastroenteritis most frequently are due to caliciviruses spread through contaminated food or through person-to-person contact. Application of similar technology is needed to make a definitive statement about the role of such candidate viruses as rotavirus, astrovirus, and adenovirus as the cause of nonepidemic acute gastroenteritis in adults. Rarely a previously healthy adult gets acute CMV colitis. CMV and EBV mainly cause diarrhea in immunocompromised patients, however. Advances in prophylaxis and treatment have reduced the frequency and severity of these diseases. Acute infantile gastroenteritis is caused by rotavirus, calcivirus, astrovirus, and adenovirus. These viral diseases of the gut are seen by the physician as routine and rare clinical problems.

  15. Extended Genetic Diversity of Bovine Viral Diarrhea Virus and Frequency of Genotypes and Subtypes in Cattle in Italy between 1995 and 2013

    OpenAIRE

    Camilla Luzzago; Stefania Lauzi; Erika Ebranati; Monica Giammarioli; Ana Moreno; Vincenza Cannella; Loretta Masoero; Elena Canelli; Annalisa Guercio; Claudio Caruso; Massimo Ciccozzi; Gian Mario De Mia; Pier Luigi Acutis; Gianguglielmo Zehender; Simone Peletto

    2014-01-01

    Genetic typing of bovine viral diarrhea virus (BVDV) has distinguished BVDV-1 and BVDV-2 species and an emerging putative third species (HoBi-like virus), recently detected in southern Italy, signaling the occurrence of natural infection in Europe. Recognizing the need to update the data on BVDV genetic variability in Italy for mounting local and European alerts, a wide collection of 5′ UTR sequences (n = 371) was selected to identify the frequency of genotypes and subtypes at the herd level....

  16. 牛病毒性腹泻病毒RT-PCR诊断方法的建立%The Foundation of PCR Diagnosis Method For Bovine Viral Diarrhea Virus

    Institute of Scientific and Technical Information of China (English)

    魏澍; 刘金玲

    2012-01-01

    A pair of primers were designed according to the sequences of bovine viral diarrhea virus strain in public. Using these primers approximately 190 bp-long DNA products were amplified by RT-PCR from genetype I and genetype II products of bovine viral diarrhea virus, but not from other control samples. This method can detect as little as 0.10 ng bovine viral diarrhea virus RNA.%根据已发表的牛病毒性腹泻病毒株的基因序列,分析合成了一对扩增跨幅为190bp左右的引物,对牛病毒性腹泻病基因I型或基因II型的毒株进行RT—PCR扩增,结果是取得了与预期大小一致的RT-PCR产物,而对照样品的扩增全为阴性;该方法最低可检测到0.10遗的牛病毒性腹泻病毒RNA。

  17. Modelling the spread of bovine viral diarrhea virus (BVDV) in a beef cattle herd and its impact on herd productivity.

    Science.gov (United States)

    Damman, Alix; Viet, Anne-France; Arnoux, Sandie; Guerrier-Chatellet, Marie-Claude; Petit, Etienne; Ezanno, Pauline

    2015-02-24

    Bovine viral diarrhea virus (BVDV) is a common pathogen of cattle herds that causes economic losses due to reproductive disorders in breeding cattle and increased morbidity and mortality amongst infected calves. Our objective was to evaluate the impact of BVDV spread on the productivity of a beef cow-calf herd using a stochastic model in discrete time that accounted for (1) the difference in transmission rates when animals are housed indoors versus grazing on pasture, (2) the external risk of disease introductions through fenceline contact with neighboring herds and the purchase of infected cattle, and (3) the risk of individual pregnant cattle generating persistently infected (PI) calves based on their stage in gestation. The model predicted the highest losses from BVDV during the first 3 years after disease was introduced into a naive herd. During the endemic phase, the impact of BVDV on the yearly herd productivity was much lower due to herd immunity. However, cumulative losses over 10 years in an endemic situation greatly surpassed the losses that occurred during the acute phase. A sensitivity analysis of key model parameters revealed that herd size, the duration of breeding, grazing, and selling periods, renewal rate of breeding females, and the level of numerical productivity expected by the farmer had a significant influence on the predicted losses. This model provides a valuable framework for evaluating the impact of BVDV and the efficacy of different control strategies in beef cow-calf herds.

  18. Actinobacteria from Termite Mounds Show Antiviral Activity against Bovine Viral Diarrhea Virus, a Surrogate Model for Hepatitis C Virus

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    Marina Aiello Padilla

    2015-01-01

    Full Text Available Extracts from termite-associated bacteria were evaluated for in vitro antiviral activity against bovine viral diarrhea virus (BVDV. Two bacterial strains were identified as active, with percentages of inhibition (IP equal to 98%. Both strains were subjected to functional analysis via the addition of virus and extract at different time points in cell culture; the results showed that they were effective as posttreatments. Moreover, we performed MTT colorimetric assays to identify the CC50, IC50, and SI values of these strains, and strain CDPA27 was considered the most promising. In parallel, the isolates were identified as Streptomyces through 16S rRNA gene sequencing analysis. Specifically, CDPA27 was identified as S. chartreusis. The CDPA27 extract was fractionated on a C18-E SPE cartridge, and the fractions were reevaluated. A 100% methanol fraction was identified to contain the compound(s responsible for antiviral activity, which had an SI of 262.41. GC-MS analysis showed that this activity was likely associated with the compound(s that had a peak retention time of 5 min. Taken together, the results of the present study provide new information for antiviral research using natural sources, demonstrate the antiviral potential of Streptomyces chartreusis compounds isolated from termite mounds against BVDV, and lay the foundation for further studies on the treatment of HCV infection.

  19. Use of homologous recombination in yeast to create chimeric bovine viral diarrhea virus cDNA clones

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    Sandra Arenhart

    Full Text Available Abstract The open reading frame of a Brazilian bovine viral diarrhea virus (BVDV strain, IBSP4ncp, was recombined with the untranslated regions of the reference NADL strain by homologous recombination in Saccharomyces cerevisiae, resulting in chimeric full-length cDNA clones of BVDV (chi-NADL/IBSP4ncp#2 and chi-NADL/IBSP4ncp#3. The recombinant clones were successfully recovered, resulting in viable viruses, having the kinetics of replication, focus size, and morphology similar to those of the parental virus, IBSP4ncp. In addition, the chimeric viruses remained stable for at least 10 passages in cell culture, maintaining their replication efficiency unaltered. Nucleotide sequencing revealed a few point mutations; nevertheless, the phenotype of the rescued viruses was nearly identical to that of the parental virus in all experiments. Thus, genetic stability of the chimeric clones and their phenotypic similarity to the parental virus confirm the ability of the yeast-based homologous recombination to maintain characteristics of the parental virus from which the recombinant viruses were derived. The data also support possible use of the yeast system for the manipulation of the BVDV genome.

  20. [Genome sequencing and analysis of the bovine viral diarrhea virus-2 strain JZ05-1 isolated in China].

    Science.gov (United States)

    Li, Qing-chao; Miao, Li-guang; Li, Hai-tao; Liu, Yan-huan; Zhang, Guang-lei; Xiao, Jia-mei

    2010-05-01

    Bovine viral diarrhea virus (BVDV) is a member of the genus Pestivirus, which is a widespread problem for beef and dairy herds, and has given rise to a significant loss in the livestock industry all over the world. The BVDV strain JZ05-1 isolated from cattle in Jilin, China generated cytopathic effect (CPE) in MDBK cells. Eight overlapped gene fragments were amplified by RT-PCR and sequenced, the complete genom sequence of BVDV strain JZ05-1 was assembled. According to the results, the JZ05-1 genome was composed of 12285 nucleotides in length (GenBank accession No. GQ888686), which could be divided into three regions: a 387 nt 5'-untranslated region (UTR), a 11694 nt single large open reading frame encoding a polyprotein, and a 204 nt 3'-UTR. The 5'-UTR and genome sequences were analyzed by sequence alignment and construction of phylogenetic trees. The strain JZ05-1 was classified as BVDV type 2a. The BVDV-2 strain JZ05-1 genome showed high similarity to the p11Q isolated in Canada and the XJ-04 isolated in China, with 90% and 91% identity in nucleotide sequence, respectively. Compared with the similarity within the BVDV-2 genotype (96%), the JZ05-1 had low sequence similarity to other BVDV-2 strains.

  1. Computational Study Exploring the Interaction Mechanism of Benzimidazole Derivatives as Potent Cattle Bovine Viral Diarrhea Virus Inhibitors.

    Science.gov (United States)

    Wang, Jinghui; Yang, Yinfeng; Li, Yan; Wang, Yonghua

    2016-07-27

    Bovine viral diarrhea virus (BVDV) infections are prevailing in cattle populations on a worldwide scale. The BVDV RNA-dependent RNA polymerase (RdRp), as a promising target for new anti-BVDV drug development, has attracted increasing attention. To explore the interaction mechanism of 65 benzimidazole scaffold-based derivatives as BVDV inhibitors, presently, a computational study was performed based on a combination of 3D-QSAR, molecular docking, and molecular dynamics (MD) simulations. The resultant optimum CoMFA and CoMSIA models present proper reliabilities and strong predictive abilities (with Q(2) = 0. 64, R(2)ncv = 0.93, R(2)pred = 0.80 and Q(2) = 0. 65, R(2)ncv = 0.98, R(2)pred = 0.86, respectively). In addition, there was good concordance between these models, molecular docking, and MD results. Moreover, the MM-PBSA energy analysis reveals that the major driving force for ligand binding is the polar solvation contribution term. Hopefully, these models and the obtained findings could offer better understanding of the interaction mechanism of BVDV inhibitors as well as benefit the new discovery of more potent BVDV inhibitors.

  2. Development of fetal and placental innate immune responses during establishment of persistent infection with bovine viral diarrhea virus.

    Science.gov (United States)

    Smirnova, Natalia P; Webb, Brett T; Bielefeldt-Ohmann, Helle; Van Campen, Hana; Antoniazzi, Alfredo Q; Morarie, Susan E; Hansen, Thomas R

    2012-08-01

    Transplacental viral infections are dependent upon complex interactions between feto-placental and maternal immune responses and the stage of fetal development at which the infection occurs. Bovine viral diarrhea virus (BVDV) has the ability to cross the placenta and infect the fetus. Infection early in gestation with non-cytopathic (ncp) BVDV leads to persistent infection. Establishment of fetal persistent infection results in life-long viremia, virus-specific immunotolerance, and may have detrimental developmental consequences. We have previously shown that heifers infected experimentally with ncp BVDV type 2 on d. 75 of gestation had transient robust up-regulation of the type I interferon (IFN) stimulated genes (ISGs) 3-15 days after viral inoculation. Blood from persistently infected (PI) fetuses, collected 115 days post maternal infection, demonstrated moderate chronic up-regulation of ISGs. This infection model was used to delineate timing of the development of innate immune responses in the fetus and placenta during establishment of persistent infection. It was hypothesized that: (i) chronic stimulation of innate immune responses occurs following infection of the fetus and (ii) placental production of the type I IFN contributes to up-regulation of ISGs in PI fetuses. PI fetuses, generated by intranasal inoculation of pregnant heifers with ncp BVDV, and control fetuses from uninfected heifers, were collected via Cesarean sections on d. 82, 89, 97, 192, and 245 of gestation. Fetal viremia was confirmed starting on d. 89. Significant up-regulation of mRNA encoding cytosolic dsRNA sensors -RIG-I and MDA5 - was detected on d. 82-192. Detection of viral dsRNA by cytosolic sensors leads to the stimulation of ISGs, which was reflected in significant up-regulation of ISG15 mRNA in fetal blood on d. 89, 97, and 192. No difference in IFN-α and IFN-β mRNA concentration was found in fetal blood or caruncular tissue, while a significant increase in both IFN-α and IFN

  3. Development and evaluation of a replicon particle vaccine expressing the E2 glycoprotein of bovine viral diarrhea virus (BVDV in cattle

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    Loy John Dustin

    2013-01-01

    Full Text Available Abstract Background Bovine viral diarrhea virus is one of the most significant and costly viral pathogens of cattle worldwide. Alphavirus-derived replicon particles have been shown to be safe and highly effective vaccine vectors against a variety of human and veterinary pathogens. Replicon particles are non-propagating, DIVA compatible, and can induce both humoral and cell mediated immune responses. This is the first experiment to demonstrate that Alphavirus-based replicon particles can be utilized in a standard prime/boost vaccination strategy in calves against a commercially significant bovine pathogen. Findings Replicon particles that express bovine viral diarrhea virus sub-genotype 1b E2 glycoprotein were generated and expression was confirmed in vitro using polyclonal and monoclonal antibodies specific to E2. Vaccine made from particles was generated in Vero cells and administered to BVDV free calves in a prime/boost regimen at two dosage levels. Vaccination resulted in neutralizing antibody titers that cross-neutralized both type 1 and type 2 BVD genotypes following booster vaccination. Additionally, high dose vaccine administration demonstrated some protection from clinical disease and significantly reduced the degree of leukopenia caused by viral infection. Conclusions Replicon particle vaccines administered in a prime/boost regimen expressing BVDV E2 glycoprotein can induce cross-neutralizing titers, reduce leukopenia post challenge, and mitigate clinical disease in calves. This strategy holds promise for a safe and effective vaccine to BVDV.

  4. Comparison of the breadth and complexity of bovine viral diarrhea (BVDV) populations circulating in 34 persistently infected cattle generated in one outbreak.

    Science.gov (United States)

    Ridpath, J F; Bayles, D O; Neill, J D; Falkenberg, S M; Bauermann, F V; Holler, L; Braun, L J; Young, D B; Kane, S E; Chase, C C L

    2015-11-01

    Exposure to bovine viral diarrhea viruses (BVDV) results in acute and persistent infections. Persistent infections result from in utero exposure during the first trimester of gestation. Clinical presentation, in persistently infected cattle (PI), is highly variable. The reasons for this variation is largely unknown. The BVDV circulating in PI exist as quasispecies (swarms of individual viruses). An outbreak resulting in 34 PI cattle presented an opportunity to compare a large number of PI׳s. Methods were developed to compare the circulating viral populations within PI animals. It was found that PI animals generated in the same outbreak carry circulating viral populations that differ widely in size and diversity. Further, it was demonstrated that variation in PI viral populations could be used as a quantifiable phenotype. This observation makes it possible to test the correlation of this phenotype to other phenotypes such as growth rate, congenital defects, viral shed and cytokine expression.

  5. Global transcriptomic profiling of bovine endometrial immune response in vitro. II. Effect of bovine viral diarrhea virus on the endometrial response to lipopolysaccharide.

    Science.gov (United States)

    Oguejiofor, Chike F; Cheng, Zhangrui; Abudureyimu, Ayimuguli; Anstaett, Olivia L; Brownlie, Joe; Fouladi-Nashta, Ali A; Wathes, D Claire

    2015-10-01

    Infection with noncytopathic bovine viral diarrhea virus (ncpBVDV) is associated with uterine disease and infertility. This study investigated the influence of ncpBVDV on immune functions of the bovine endometrium by testing the response to bacterial lipopolysaccharide (LPS). Primary cultures of mixed epithelial and stromal cells were divided into four treatment groups (control [CONT], BVDV, CONT+LPS, and BVDV+LPS) and infected with ncpBVDV for 4 days followed by treatment with LPS for 6 h. Whole-transcriptomic gene expression was measured followed by Ingenuity Pathway Analysis. Differential expression of 184 genes was found between CONT and BVDV treatments, showing interplay between induction and inhibition of responses. Up-regulation of TLR3, complement, and chemotactic and TRIM factors by ncpBVDV all suggested an ongoing immune response to viral infection. Down-regulation of inflammatory cytokines, chemokines, CXCR4, and serine proteinase inhibitors suggested mechanisms by which ncpBVDV may simultaneously counter the host response. Comparison between BVDV+LPS and CONT+LPS treatments showed 218 differentially expressed genes. Canonical pathway analysis identified the key importance of interferon signaling. Top down-regulated genes were RSAD2, ISG15, BST2, MX2, OAS1, USP18, IFIT3, IFI27, SAMD9, IFIT1, and DDX58, whereas TRIM56, C3, and OLFML1 were most up-regulated. Many of these genes are also regulated by IFNT during maternal recognition of pregnancy. Many innate immune genes that typically respond to LPS were inhibited by ncpBVDV, including those involved in pathogen recognition, inflammation, interferon response, chemokines, tissue remodeling, cell migration, and cell death/survival. Infection with ncpBVDV can thus compromise immune function and pregnancy recognition, thereby potentially predisposing infected cows to postpartum bacterial endometritis and reduced fertility.

  6. Use of three-dimensional accelerometers to evaluate behavioral changes in cattle experimentally infected with bovine viral diarrhea virus.

    Science.gov (United States)

    Bayne, Jenna E; Walz, Paul H; Passler, Thomas; White, Brad J; Theurer, Miles E; van Santen, Edzard

    2016-06-01

    OBJECTIVE To assess the use of 3-D accelerometers to evaluate behavioral changes in cattle experimentally infected with a low-virulent strain of bovine viral diarrhea virus (BVDV). ANIMALS 20 beef steers (mean weight, 238 kg). PROCEDURES Calves were allocated to a BVDV (n = 10) or control (10) group. On day 0, calves in the BVDV group were inoculated with a low-virulent strain of BVDV (4 × 10(6) TCID50, intranasally), and calves in the control group were sham inoculated with BVDV-free medium (4 mL; intranasally). An accelerometer was affixed to the right hind limb of each calf on day -7 to record activity (lying, walking, and standing) continuously until 35 days after inoculation. Baseline was defined as days -7 to -1. Blood samples were collected at predetermined times for CBC, serum biochemical analysis, virus isolation, and determination of anti-BVDV antibody titers. RESULTS All calves in the BVDV group developed viremia and anti-BVDV antibodies but developed only subclinical or mild disease. Calves in the control group did not develop viremia or anti-BVDV antibodies. Mean time allocated to each activity did not differ significantly between the BVDV and control groups on any day except day 8, when calves in the BVDV group spent less time standing than the calves in the control group. Following inoculation, calves in both groups tended to spend more time lying and less time walking and standing than they did during baseline. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that behavioral data obtained by accelerometers could not distinguish calves subclinically infected with BVDV from healthy control calves. However, subtle changes in the behavior of the BVDV-infected calves were detected and warrant further investigation.

  7. Increase in proto-oncogene mRNA transcript levels in bovine lymphoid cells infected with a cytopathic type 2 bovine viral diarrhea virus.

    Science.gov (United States)

    Neill, John D; Ridpath, Julia F

    2008-08-01

    Infection of susceptible animals with bovine viral diarrhea viruses (BVDV) can result in an array of disease symptoms that are dependent in part on the strain of infecting virus and the physiological status of the host. BVDV are lymphotrophic and exist as two biotypes. Cytopathic BVDV kill cells outright while noncytopathic strains can readily establish persistent infections. The molecular mechanisms behind these different affects are unknown. To gain a better understanding of the mechanisms of disease, serial analysis of gene expression (SAGE), a powerful method for global gene expression analysis, was employed to examine gene expression changes in BVDV-infected BL3 cells, a bovine B-cell lymphosarcoma cell line. SAGE libraries were constructed from mRNA derived from BL3 cells that were noninfected or infected with the cytopathic BVDV2 strain 296c. Annotation of the SAGE data showed the expression of many genes that are characteristic of B cells and integral to their function. Comparison of the SAGE databases also revealed a number of genes that were differentially expressed. Of particular interest was the increased numbers of transcripts encoding proto-oncogenes (c-fos, c-jun, junB, junD) in 296c-infected cells, all of which are constituents of the AP-1 transcriptional activation complex. Real-time RT-PCR confirmed these results and indicated that the actual increases were larger than that predicted by SAGE. In contrast, there was no corresponding increase in protein levels, but instead a significant decrease of c-jun and junB protein levels in the infected BL3 cells was observed. Rather than an increase in transcription of these genes, it appeared that these proto-oncogenes transcripts accumulated in the BVDV2-infected cells.

  8. The bovine viral diarrhea virus E2 protein formulated with a novel adjuvant induces strong, balanced immune responses and provides protection from viral challenge in cattle.

    Science.gov (United States)

    Snider, Marlene; Garg, Ravendra; Brownlie, Robert; van den Hurk, Jan V; van Drunen Littel-van den Hurk, Sylvia

    2014-11-28

    Bovine viral diarrhea virus (BVDV) is still one of the most serious pathogens in cattle, meriting the development of improved vaccines. Recently, we developed a new adjuvant consisting of poly[di(sodium carboxylatoethylphenoxy)]-phosphazene (PCEP), either CpG ODN or poly(I:C), and an immune defense regulator (IDR) peptide. As this adjuvant has been shown to mediate the induction of robust, balanced immune responses, it was evaluated in an E2 subunit vaccine against BVDV in lambs and calves. The BVDV type 2 E2 protein was produced at high levels in a mammalian expression system and purified. When formulated with either CpG ODN or poly(I:C), together with IDR and PCEP, the E2 protein elicited high antibody titers and production of IFN-γ secreting cells in lambs. As the immune responses were stronger when poly(I:C) was used, the E2 protein with poly(I:C), IDR and PCEP was subsequently tested in cattle. Robust virus neutralizing antibodies as well as cell-mediated immune responses, including CD8(+) cytotoxic T cell (CTL) responses, were induced. The fact that CTL responses were demonstrated in calves vaccinated with an E2 protein subunit vaccine indicates that this adjuvant formulation promotes cross-presentation. Furthermore, upon challenge with a high dose of virulent BVDV-2, the vaccinated calves showed almost no temperature response, weight loss, leukopenia or virus replication, in contrast to the control animals, which had severe clinical disease. These data suggest that this E2 subunit formulation induces significant protection from BVDV-2 challenge, and thus is a promising BVDV vaccine candidate; in addition, the adjuvant platform has applications in bovine vaccines in general.

  9. Contribution of Leptospira, Neospora caninum and bovine viral diarrhea virus to fetal loss of beef cattle in New Zealand.

    Science.gov (United States)

    Sanhueza, J M; Heuer, C; West, D

    2013-10-01

    The profitability of beef breeding farms in New Zealand depends principally on optimal reproductive performance. The aim of this study was to estimate the impact of four major pathogens, bovine viral diarrhea virus (BVDV), Neospora caninum (N. caninum), Leptospira borgpetersenii serovar Hardjo (Hardjo), and Leptospira interrogans serovar Pomona (Pomona), on rates of fetal loss in commercial beef breeding herds. Farms reporting fetal loss were recruited, and a blood sample from aborting cows (cases) was collected. Controls were normally calving cows from the same farm. At least four controls were selected from each farm contributing cases. Samples were tested using ELISA for detection of antibodies against BVDV and N. caninum, and microscopic agglutination test (MAT) for detection of antibody against Hardjo and Pomona. A selection of titer cut-offs was conducted to evaluate the relationship between fetal loss and seropositivity to each pathogen using conditional logistic regression. The cut-off titer with the strongest association with fetal loss was included in the multivariate model. A significant increased risk of fetal loss was found for animals seropositive to N. caninum (odds ratio (OR)=3.36; 95% confidence interval (95% CI)=1.27-8.89), Hardjo (OR=1.84; 95% CI=1.01-3.33), and Pomona in non-vaccinated cows (OR=14.91, 95% CI=1.73-128.84) at the ELISA titer ≥ 30, and MAT titers of ≥ 1:384 and ≥ 1:768 for a positive sample, respectively. A marginally non-significant increased risk of fetal loss was found for animals exposed to BVDV (OR=2.01; 95% CI=0.99-4.11) at the ELISA titer of ≤ 1. Vaccination did not affect ORs for Hardjo or BVDV and no herd vaccinated against N. caninum. Approximately 14.0% of all fetal loss in the beef breeding cattle population in New Zealand may be attributable to BVDV (3.5%), N. caninum (3.0%), Hardjo (4.7%), and Pomona (3.6%).

  10. Induction of interferon-gamma and downstream pathways during establishment of fetal persistent infection with bovine viral diarrhea virus.

    Science.gov (United States)

    Smirnova, Natalia P; Webb, Brett T; McGill, Jodi L; Schaut, Robert G; Bielefeldt-Ohmann, Helle; Van Campen, Hana; Sacco, Randy E; Hansen, Thomas R

    2014-04-01

    Development of transplacental infection depends on the ability of the virus to cross the placenta and replicate within the fetus while counteracting maternal and fetal immune responses. Unfortunately, little is known about this complex process. Non-cytopathic (ncp) strains of bovine viral diarrhea virus (BVDV), a pestivirus in the Flaviviridae family, cause persistent infection in early gestational fetuses (infected, PI), but are cleared by immunocompetent animals and late gestational fetuses (>150 days; transiently infected, TI). Evasion of innate immune response and development of immunotolerance to ncp BVDV have been suggested as possible mechanisms for the establishment of the persistent infection. Previously we have observed a robust temporal induction of interferon (IFN) type I (innate immune response) and upregulation of IFN stimulated genes (ISGs) in BVDV TI fetuses. Modest chronic upregulation of ISGs in PI fetuses and calves reflects a stimulated innate immune response during persistent BVDV infection. We hypothesized that establishing persistent fetal BVDV infection is also accompanied by the induction of IFN-gamma (IFN-γ). The aims of the present study were to determine IFN-γ concentration in blood and amniotic fluid from control, TI and PI fetuses during BVDV infection and analyze induction of the IFN-γ downstream pathways in fetal lymphoid tissues. Two experiments with in vivo BVDV infections were completed. In Experiment 1, pregnant heifers were infected with ncp BVDV type 2 on day 75 or 175 of gestation or kept naïve to generate PI, TI and control fetuses, respectively. Fetuses were collected by Cesarean section on day 190. In Experiment 2, fetuses were collected on days 82, 89, 97, 192 and 245 following infection of pregnant heifers on day 75 of gestation. The results were consistent with the hypothesis that ncp BVDV infection induces IFN-γ secretion during acute infection in both TI and PI fetuses and that lymphoid tissues such as spleen

  11. Evaluation of transmission of bovine viral diarrhea virus (BVDV) between persistently infected and naive cattle by the horn fly (Haematobia irritans).

    Science.gov (United States)

    Chamorro, Manuel F; Passler, Thomas; Givens, M Daniel; Edmondson, Misty A; Wolfe, Dwight F; Walz, Paul H

    2011-02-01

    Identifying reservoirs and transmission routes for bovine viral diarrhea virus (BVDV) are important in developing biosecurity programs. The aim of this study was to evaluate BVDV transmission by the hematophagous horn fly (Haematobia irritans). Flies collected from four persistently infected cattle were placed in fly cages attached to principal (n = 4) and control (n = 4) BVDV-naïve calves housed individually in isolation rooms. Flies were able to feed on principal calves, but a barrier prevented fly feeding from control calves. Flies were tested for BVDV by RT-PCR and virus isolation at time of collection from PI cattle and after 48 h of exposure on BVDV-naïve calves. Blood samples were collected from calves and tested for BVDV infection. Virus was isolated from fly homogenates at collection from PI animals and at removal from control and principal calves. All calves remained negative for BVDV by virus isolation and serology throughout the study. Bovine viral diarrhea virus may be detected in horn flies collected from PI cattle, but horn flies do not appear to be an important vector for BVDV transmission.

  12. Detecção de ácidos nucléicos de Brucella spp., Leptospira spp., herpesvirus bovino e vírus da diarréia viral bovina, em fetos bovinos abortados e em animais mortos no perinatal Detection of Brucella spp., Leptospira spp., bovine herpesvirus and bovine viral diarrhea virus nucleic acids in aborted fetuses and bovines dead perinatal

    Directory of Open Access Journals (Sweden)

    A. Cortez

    2006-12-01

    Full Text Available Samples of 114 bovine fetuses and 10 calves, which dead in perinatal period, were examined for detection of DNA. The most common detected agent was Brucella spp. in 17 samples (13.7% followed by Leptospira spp. in 4 cases (3.2%,bovine herpesvirus (BHV and bovine viral diarrhea (BVDV in 3 animals (2.4% each, and 1 for the association of BVDV and BHV. In 77.4 % (96/124 of the samples it was not possible to detect any agent.

  13. Antiviral Activity of Bacillus sp. Isolated from the Marine Sponge Petromica citrina against Bovine Viral Diarrhea Virus, a Surrogate Model of the Hepatitis C Virus

    Directory of Open Access Journals (Sweden)

    Clarice Weis Arns

    2013-04-01

    Full Text Available The Hepatitis C virus causes chronic infections in humans, which can develop to liver cirrhosis and hepatocellular carcinoma. The Bovine viral diarrhea virus is used as a surrogate model for antiviral assays for the HCV. From marine invertebrates and microorganisms isolated from them, extracts were prepared for assessment of their possible antiviral activity. Of the 128 tested, 2 were considered active and 1 was considered promising. The best result was obtained from the extracts produced from the Bacillus sp. isolated from the sponge Petromica citrina. The extracts 555 (500 µg/mL, SI>18 and 584 (150 µg/mL, SI 27 showed a percentage of protection of 98% against BVDV, and the extract 616, 90% of protection. All of them showed activity during the viral adsorption. Thus, various substances are active on these studied organisms and may lead to the development of drugs which ensure an alternative therapy for the treatment of hepatitis C.

  14. DETECTION OF ANTIBODIES AGAINST BOVINE HERPES VIRUS 1, BOVINE VIRAL DIARRHEA VIRUS AND BOVINE RESPIRATORY SYNCYTIAL VIRUS IN EARLY AND ULTRA-EARLY WEANED BEEF CALVES

    Directory of Open Access Journals (Sweden)

    Diego Daniel Gonzalez

    2013-01-01

    Full Text Available Bovine respiratory disease is the leading cause of morbidity and mortality in weaned calves. In Argentina, two weaning practices have been implemented. In the early weaning, the calf is removed from the cow at 60-70 days of age while in ultra-early weaning the calf is weaned at 30-45 days of age. The purposes of both systems is to improve cow body condition, calf performance, conception rates and forage availability for the cow. In this study we evaluated the antibody response against BVDV and BoHV1 in early and ultra-early weaned calves that had received a conventional vaccination schedule (first dose at weaning and a booster 21 days post-weaning. Passively acquired immunity may provide protection against disease caused by these viruses. The presence of antibodies against BRSV, a virus that was not present in the vaccines used, was also evaluated as an indirect indicator of viral circulation in the herd. At the time of vaccination, calves presented a wide range of maternally-derived antibody titers. Vaccination against BoHV-1 did not evoke seroconvertion and antibody titers continued to decay throughout the experience. After vaccination, seroconversion to BVDV could be detected in calves with low antibody titers, while higher antibody titers exerted an inhibitory effect of the active humoral response.

  15. Genetic Variability of Bovine Viral Diarrhea Virus and Evidence for a Possible Genetic Bottleneck during Vertical Transmission in Persistently Infected Cattle.

    Directory of Open Access Journals (Sweden)

    Natalie Dow

    Full Text Available Bovine viral diarrhea virus (BVDV, a Pestivirus in the family Flaviviridae, is an economically important pathogen of cattle worldwide. The primary propagators of the virus are immunotolerant persistently infected (PI cattle, which shed large quantities of virus throughout life. Despite the absence of an acquired immunity against BVDV in these PI cattle there are strong indications of viral variability that are of clinical and epidemiological importance. In this study the variability of E2 and NS5B sequences in multiple body compartments of PI cattle were characterized using clonal sequencing. Phylogenetic analyses revealed that BVDV exists as a quasispecies within PI cattle. Viral variants were clustered by tissue compartment significantly more often than expected by chance alone with the central nervous system appearing to be a particularly important viral reservoir. We also found strong indications for a genetic bottleneck during vertical transmission from PI animals to their offspring. These quasispecies analyses within PI cattle exemplify the role of the PI host in viral propagation and highlight the complex dynamics of BVDV pathogenesis, transmission and evolution.

  16. Genetic Variability of Bovine Viral Diarrhea Virus and Evidence for a Possible Genetic Bottleneck during Vertical Transmission in Persistently Infected Cattle.

    Science.gov (United States)

    Dow, Natalie; Chernick, Adam; Orsel, Karin; van Marle, Guido; van der Meer, Frank

    2015-01-01

    Bovine viral diarrhea virus (BVDV), a Pestivirus in the family Flaviviridae, is an economically important pathogen of cattle worldwide. The primary propagators of the virus are immunotolerant persistently infected (PI) cattle, which shed large quantities of virus throughout life. Despite the absence of an acquired immunity against BVDV in these PI cattle there are strong indications of viral variability that are of clinical and epidemiological importance. In this study the variability of E2 and NS5B sequences in multiple body compartments of PI cattle were characterized using clonal sequencing. Phylogenetic analyses revealed that BVDV exists as a quasispecies within PI cattle. Viral variants were clustered by tissue compartment significantly more often than expected by chance alone with the central nervous system appearing to be a particularly important viral reservoir. We also found strong indications for a genetic bottleneck during vertical transmission from PI animals to their offspring. These quasispecies analyses within PI cattle exemplify the role of the PI host in viral propagation and highlight the complex dynamics of BVDV pathogenesis, transmission and evolution.

  17. 内蒙古牛病毒性腹泻病毒血清学调查%Seroepidemiological Survey on Bovine Viral Diarrhea Virus Infection in Inner Mongolia

    Institute of Scientific and Technical Information of China (English)

    童钦; 吕晓妍; 王炜; 项键

    2013-01-01

    To survey the prevalence of bovine viral diarrhea virus on dairy in Inner Mongolia. Cytopathic biotype bovine viral diarrhea virus was used as neutralizing test antigen. 509 sera collected from 6 dairy farms in Ulanhot, Banyan Nur, Hohhot Areas were detected by neutralization test. The results showed that: 297 sera were positive, 6 dairy farms sera positive rate was 55.10%, 38.80%, 45.50%, 50.00%, 77.80%, 33.33% respectively. The average positive rate of Ulanhot, Banyan Nur, Hohhot was 77.80%, 48.70%, 33.33%, total positive rate was 58.35%. The results suggested that bovine viral diarrhea virus exist in dairy farms in Inner Mongolia.%为了解内蒙古自治区集约化养牛场病毒性腹泻的感染情况.用致细胞病变型牛病毒性腹泻病毒毒株制备中和抗原,通过中和实验对采自乌兰浩特、巴彦淖尔和呼和浩特地区6个集约化奶牛场的509份血清进行了牛病毒性腹泻病毒抗体检测.结果表明:共检出阳性血清297份,6个牛场的牛病毒性腹泻病毒血清阳性率分别为55.10%、38.80%、45.50%、50.00%、77.80%、33.33%;乌兰浩特、巴彦淖尔、呼和浩特这3个地区的平均阳性率分别为77.80%、48.70%、33.33%,总血清阳性率为58.35%.说明内蒙古地区存在牛病毒性腹泻病毒的感染,应采取相应的措施净化牛场.

  18. Weaning management of newly received beef calves with or without continuous exposure to a persistently infected bovine viral diarrhea virus pen mate: Effects on rectal temperature and serum proinflammatory cytokine and haptog

    Science.gov (United States)

    Exposure to animals persistently infected (PI) with bovine viral diarrhea virus (BVDV) results in immunomodulation in cohorts. It is hypothesized that the extent of modulation differs for low-risk, preconditioned (PC) vs. high-risk, auction market (AM) beef cattle. Our objective was to compare immun...

  19. Weaning management of newly received beef calves with or without continuous exposure to a persistently infected bovine viral diarrhea virus pen mate: Effects on rectal temperature, peripheral blood leukocytes and serum

    Science.gov (United States)

    Exposure to animals persistently infected (PI) with bovine viral diarrhea virus (BVDV) results in immunomodulation in cohorts. It is hypothesized that the extent of modulation differs for preconditioned (PC) vs. auction market (AM) cattle. Our objective was to compare immune responses of PC or AM ca...

  20. The Progress of the Detection Methods of Bovine Viral Diarrhea Virus%牛病毒性腹泻病毒检测方法的研究进展

    Institute of Scientific and Technical Information of China (English)

    童钦; 霍志云; 胡桂学; 王炜; 武华

    2012-01-01

    In order to quickly and exactly detect bovine viral diarrhea virus, reduce the economic loss of cattle industry. This paper reviewed the method of detecting bovine viral diarrhea virus, and summarized the advantages and disadvantages of these methods. The results showed that according to the farmers on the virus rapid accurate diagnosis needs, prospects for a new detection of bovine viral diarrhea virus method and immune colloidal gold test strip method. Immune colloidal gold test strip method improvement of bovine viral diarrhea virus detection methods, contribute to the rapid diagnosis of the virus.%为了快速准确地诊断出牛病毒性腹泻病毒,减少养牛业的经济损失.综述了近年来检测牛病毒性腹泻病毒的方法,总结出这些方法的优缺点.结果表明:根据养殖户对该病毒快速准确诊断的实际需要,展望了一种新的检测牛病毒性腹泻病毒的方法——免疫胶体金试纸条法.表明免疫胶体金试纸条法完善了牛病毒性腹泻病毒的检测方法,有助于在现场快速诊断该病毒.

  1. Lesions and distribution of viral antigen following an experimental infection of young seronegative calves with virulent bovine virus diarrhea virus-type II.

    Science.gov (United States)

    Ellis, J A; West, K H; Cortese, V S; Myers, S L; Carman, S; Martin, K M; Haines, D M

    1998-07-01

    During the past several years, acute infections with bovine viral diarrhea virus (BVDV) have been causally linked to hemorrhagic and acute mucosal disease-like syndromes with high mortality. The majority of BVDVs isolated in such cases have been classified as type II on the basis of genetic and antigenic characteristics. It was our objective to examine clinical disease, lesions and potential sites of viral replication, following experimental BVDV type II infection in young calves. On approximately day 35 after birth, calves that had received BVDV-antibody-negative colostrum were infected by intranasal inoculation of 5 x 10(5) TCID50 of BVDV type II isolate 24,515 in 5 mL of tissue culture fluid (2.5 mL/nostril). Calves were monitored twice daily for signs of clinical disease. Approximately 48-72 h after infection, all calves developed transient pyrexia (39.4-40.5 degrees C) and leukopenia. Beginning on approximately day 7 after infection, all calves developed watery diarrhea, pyrexia (40.5-41.6 degrees C), marked leukopenia (> or = 75% drop from preinoculation values), variable thrombocytopenia, and moderate to severe depression. Calves were euthanized on days 10, 11, or 12 after infection due to severe disease. Gross and histological lesions consisted of multifocal bronchointerstitial pneumonia (involving 10%-25% of affected lungs), bone marrow hypoplasia and necrosis, and minimal erosive lesions in the alimentary tract. Immunohistochemical staining for BVDV revealed widespread viral antigen usually within epithelial cells, smooth muscle cells and mononuclear phagocytes in multiple organs, including lung, Peyer's patches, gastric mucosa, thymus, adrenal gland, spleen, lymph nodes, bone marrow, and skin. This BVDV type II isolate caused rapidly progressive, severe multisystemic disease in seronegative calves that was associated with widespread distribution of viral antigen and few gross or histological inflammatory lesions.

  2. Bovine viral diarrhea virus type 2 in vivo infection modulates TLR4 responsiveness in differentiated myeloid cells which is associated with decreased MyD88 expression.

    Science.gov (United States)

    Schaut, Robert G; McGill, Jodi L; Neill, John D; Ridpath, Julia F; Sacco, Randy E

    2015-10-02

    Symptoms of bovine viral diarrhea virus (BVDV) infection range from subclinical to severe, depending on strain virulence. Several in vitro studies showed BVDV infection impaired leukocyte function. Fewer studies have examined the effects of in vivo BVDV infection on monocyte/macrophage function, especially with strains of differing virulence. We characterized cytokine production by bovine myeloid cells isolated early or late in high (HV) or low virulence (LV) BVDV2 infection. Given BVDV infection may enhance susceptibility to secondary bacterial infection, LPS responses were examined as well. Monocytes from HV and LV infected calves produced higher levels of cytokines compared to cells from controls. In contrast, monocyte-derived macrophage cytokine levels were generally reduced. Modulated cytokine expression in HV BVDV2 macrophages was associated with decreased MyD88 expression, likely due to its interaction with viral NS5A. These data and those of others, suggest that certain Flaviviridae may have evolved strategies for subverting receptor signaling pathways involving MyD88.

  3. Effect of timing of challenge following short-term natural exposure to bovine viral diarrhea virus type 1b on animal performance and immune response in beef steers.

    Science.gov (United States)

    Carlos-Valdez, L; Wilson, B K; Burciaga-Robles, L O; Step, D L; Holland, B P; Richards, C J; Montelongo, M A; Confer, A W; Fulton, R W; Krehbiel, C R

    2016-11-01

    Bovine respiratory disease (BRD) is the most common and economically detrimental disease of beef cattle during the postweaning period, causing the majority of morbidity and mortality in feedlots. The pathogenesis of this disease often includes an initial viral infection, which can predispose cattle to a secondary bacterial infection. The objective of this experiment was to determine the effects of timing of an intratracheal (MH) challenge relative to 72 h of natural exposure to bovine viral diarrhea virus (BVDV) type 1b persistently infected (PI) calves on performance, serum antibody production, total and differential white blood cell (WBC) count, rectal temperature, clinical severity score (CS), and haptoglobin (Hp). Steers ( = 24; 276 ± 31 kg initial BW) were randomly allocated to 1 of 3 treatments (8 steers/treatment) in a randomized complete block design. Treatments were steers not exposed to calves PI with BVDV 1b and not challenged with MH (CON), steers intratracheally challenged with MH 84 h after being exposed to calves PI with BVDV 1b for 72 h (LateCh), and steers intratracheally challenged with MH 12 h after being exposed to calves PI with BVDV 1b for 72 h (EarlyCh). Performance (ADG, DMI, and G:F) was decreased ( marketing channels allow for variation in the timing of respiratory pathogen exposure, understanding the physiological changes in morbid cattle will lead to improved management of BRD.

  4. Platelet aggregation responses and virus isolation from platelets in calves experimentally infected with type I or type II bovine viral diarrhea virus.

    Science.gov (United States)

    Walz, P H; Bell, T G; Grooms, D L; Kaiser, L; Maes, R K; Baker, J C

    2001-10-01

    Altered platelet function has been reported in calves experimentally infected with type II bovine viral diarrhea virus (BVDV). The purpose of the present study was to further evaluate the ability of BVDV isolates to alter platelet function and to examine for the presence of a virus-platelet interaction during BVDV infection. Colostrum-deprived Holstein calves were obtained immediately after birth, housed in isolation, and assigned to 1 of 4 groups (1 control and 3 treatment groups). Control calves (n = 4) were sham inoculated, while calves in the infected groups (n = 4 for each group) were inoculated by intranasal instillation with 10(7) TCID50 of either BVDV 890 (type II), BVDV 7937 (type II), or BVDV TGAN (type I). Whole blood was collected prior to inoculation (day 0) and on days 4, 6, 8, 10, and 12 after inoculation for platelet function testing by optical aggregometry by using adenosine diphosphate and platelet activating factor. The maximum percentage aggregation and the slope of the aggregation curve decreased over time in BVDV-infected calves; however, statistically significant differences (Freidman repeated measures ANOVA on ranks, P infected with the type II BVDV isolates. Bovine viral diarrhea virus was not isolated from control calves, but was isolated from all calves infected with both type II BVDV isolates from days 4 through 12 after inoculation. In calves infected with type I BVDV, virus was isolated from 1 of 4 calves on days 4 and 12 after inoculation and from all calves on days 6 and 8 after inoculation. Altered platelet function was observed in calves infected with both type II BVDV isolates, but was not observed in calves infected with type I BVDV. Altered platelet function may be important as a difference in virulence between type I and type II BVDV infection.

  5. 牛病毒性腹泻分子及血清流行病学研究进展%Advances on Serological and Molecular Epidemiology of Bovine Viral Diarrhea

    Institute of Scientific and Technical Information of China (English)

    王建领; 付彤; 刘杰; 张龙现; 菅复春

    2012-01-01

    牛病毒性腹泻病毒主要侵害牛,尤其是犊牛,引起以腹泻(急性感染症状)和黏膜病(慢性持续性感染症状)为临床特征的疾病,给养牛业造成了巨大损失.为此,主要就牛病毒性腹泻的病原特征、分子及血清流行病学概况进行了综述.%Bovine viral diarrhea virus mainly infects cow,especially calves,and causes the disease with clinical features of diarrhea (acute symptoms) and mucosal disease (persistent infection with chronic symptoms). With the purpose to provide a reference for its prevention and control, the new progresses of bovine viral diarrhea pathogen characteristics,molecular and serological epidemiology were summarized in this review.

  6. In vitro replication activity of bovine viral diarrhea virus in an epithelial cell line and in bovine peripheral blood mononuclear cells.

    Science.gov (United States)

    Turin, Lauretta; Lucchini, Barbara; Bronzo, Valerio; Luzzago, Camilla

    2012-11-01

    The present study focused on the in vitro infection of Madin-Darby bovine kidney (MDBK) cells and bovine peripheral blood mononuclear cells (PBMCs) from naÏve animals with non-cytopathic (ncp, BVDV-1b NY-1) and cytopathic (cp, BVDV-1a NADL) strains. Infections with 0.1 and 1 multiplicity of infections (MOI) and incubation times of 18 and 36 hr were compared. Twelve BVDV naÏve heifers were enrolled to collect PBMCs. The viral loads in MDBK cells and in PBMCs after in vitro infections were measured by real-time polymerase chain reaction (PCR) assays. The highest viral loads were measured at 1 MOI and 36 hr post infection in both cell systems and the lowest at 0.1 MOI and 18 hr with the exception of the cp strain NADL in PBMCs, for which the highest viral load was observed at 0.1 MOI and 36 hr. Viral load mean values were higher for the cp strain than the ncp strain irrespective of the extent of the infection period and MOI. The models of infection studied uncovered different replication activities respectively according to the biotype of virus, the cell substrate and the duration of infection. Replication tends to be higher in PBMCs, particularly at low MOIs and for the ncp strain.

  7. Both foot-and-mouth disease virus and bovine viral diarrhea virus replication are inhibited by Mx1 protein originated from porcine.

    Science.gov (United States)

    Shi, Huijun; Fu, Qiang; Ren, Yan; Wang, Dawei; Qiao, Jun; Wang, Pengyan; Zhang, Hui; Chen, Chuangfu

    2015-01-01

    Mx1 protein is I type interferons (IFNs)-induced 76-kDa guanosine triphosphatases (GTPases) that belong to the dynamin superfamily of large GTPases. Mx1 proteins have attracted attention because some display antiviral activity against pathogenic RNA and DNA viruses. Meanwhile, Mx1 gene generally exists in organisms or cells of mammalian, fish and chicken. Blocking a wide range of RNA virus replication by inhibiting nuclear viral mRNA synthesis is a unique property of Mx1 protein. In order to investigate a novel prevention measure against foot-and-mouth disease virus (FMDV) and bovine viral diarrhea virus (BVDV), which frequently break out in Xinjiang Uygur Autonomous Region of China, we investigated the effects of porcine Mx1 protein on FMDV and BVDV replication by measuring viral reverse transcriptase activity at various time intervals. In our study, Mx1 protein was overexpressed in BHK-21 and MDBK cells mediated by lentivirus prior to infect with FMDV and BVDV. FMDV and BVDV replication levels were monitored by quantitative real-Time PCR. The results showed porcine Mx1 overexpression significantly inhibited both FMDV and BVDV replication within 12 and 36 hours post-infection (pi). The finding may provide a new therapeutic approach for preventing from FDMV and BVDV infection.

  8. Isolation and Identification of Bovine Viral Diarrhea Virus Shandong Strain%牛病毒性腹泻病毒山东株的分离与鉴定

    Institute of Scientific and Technical Information of China (English)

    侯佩莉; 杨宏军; 宋玲玲; 王洪梅; 刘来兴; 何洪彬

    2013-01-01

    牛病毒性腹泻病毒是牛病毒性腹泻-黏膜病的重要病原体,该病毒病是极为复杂、呈多临床类型表现的传染病,给世界养牛业造成了巨大的经济损失.本研究从疑似牛病毒性腹泻-黏膜病的粪便中分离得到一株病毒能使MDBK细胞产生细胞病变,经RT-PCR检测及扩增产物测序进一步证实,该病毒为牛病毒性腹泻-黏膜病病毒,TCID50测定该病毒的滴度为107.15TCID50/ml.该病毒株的分离鉴定为牛病毒性腹泻病毒疫苗的研制奠定了基础.%Bovine viral diarrhea virus ( BVDV) is the causative agent of bovine viral diarrhoea, and is u-biquitous in cattle. It is very complex and presents a wide range of clinical manifestations, which causes large economic loss to cattle industry around the world. In this study, a virus strain causing cytopathic effect of ma-din -darby bovinekidney (MDBK) was isolated from the bovine excrement and identified as BVDV by RT -PCR and sequencing analysis. The virus titer was 10 7.15 TCID50/ml. The isolation and identification of this virus strain laid foundation for exploring its vaccine.

  9. Bovine Viral Diarrhea Virus Type 2 Impairs Macrophage Responsiveness to Toll-Like Receptor Ligation with the Exception of Toll-Like Receptor 7.

    Directory of Open Access Journals (Sweden)

    Robert G Schaut

    Full Text Available Bovine viral diarrhea virus (BVDV is a member of the Flaviviridae family. BVDV isolates are classified into two biotypes based on the development of cytopathic (cp or non-cytopathic (ncp effects in epithelial cell culture. BVDV isolates are further separated into species, BVDV1 and 2, based on genetic differences. Symptoms of BVDV infection range from subclinical to severe, depending on strain virulence, and may involve multiple organ systems and induction of a generalized immunosuppression. During BVDV-induced immune suppression, macrophages, critical to innate immunity, may have altered pathogen recognition receptor (PRR signaling, including signaling through toll-like receptors (TLRs. Comparison of BVDV 2 strains with different biotypes and virulence levels is valuable to determining if there are differences in host macrophage cellular responses between viral phenotypes. The current study demonstrates that cytopathic (cp, noncytopathic (ncp, high (hv or low virulence (lv BVDV2 infection of bovine monocyte-derived macrophages (MDMΦ result in differential expression of pro-inflammatory cytokines compared to uninfected MDMΦ. A hallmark of cp BVDV2 infection is IL-6 production. In response to TLR2 or 4 ligation, as might be observed during secondary bacterial infection, cytokine secretion was markedly decreased in BVDV2-infected MDMΦ, compared to non-infected MDMΦ. Macrophages were hyporesponsive to viral TLR3 or TLR8 ligation. However, TLR7 stimulation of BVDV2-infected MDMΦ induced cytokine secretion, unlike results observed for other TLRs. Together, these data suggest that BVDV2 infection modulated mRNA responses and induced a suppression of proinflammatory cytokine protein responses to TLR ligation in MDMΦ with the exception of TLR7 ligation. It is likely that there are distinct differences in TLR pathways modulated following BVDV2 infection, which have implications for macrophage responses to secondary infections.

  10. A nationwide database linking information on the hosts with sequence data of their virus strains: A useful tool for the eradication of bovine viral diarrhea (BVD) in Switzerland.

    Science.gov (United States)

    Stalder, Hanspeter; Hug, Corinne; Zanoni, Reto; Vogt, Hans-Rudolf; Peterhans, Ernst; Schweizer, Matthias; Bachofen, Claudia

    2016-06-15

    Pestiviruses infect a wide variety of animals of the order Artiodactyla, with bovine viral diarrhea virus (BVDV) being an economically important pathogen of livestock globally. BVDV is maintained in the cattle population by infecting fetuses early in gestation and, thus, by generating persistently infected (PI) animals that efficiently transmit the virus throughout their lifetime. In 2008, Switzerland started a national control campaign with the aim to eradicate BVDV from all bovines in the country by searching for and eliminating every PI cattle. Different from previous eradication programs, all animals of the entire population were tested for virus within one year, followed by testing each newborn calf in the subsequent four years. Overall, 3,855,814 animals were tested from 2008 through 2011, 20,553 of which returned an initial BVDV-positive result. We were able to obtain samples from at least 36% of all initially positive tested animals. We sequenced the 5' untranslated region (UTR) of more than 7400 pestiviral strains and compiled the sequence data in a database together with an array of information on the PI animals, among others, the location of the farm in which they were born, their dams, and the locations where the animals had lived. To our knowledge, this is the largest database combining viral sequences with animal data of an endemic viral disease. Using unique identification tags, the different datasets within the database were connected to run diverse molecular epidemiological analyses. The large sets of animal and sequence data made it possible to run analyses in both directions, i.e., starting from a likely epidemiological link, or starting from related sequences. We present the results of three epidemiological investigations in detail and a compilation of 122 individual investigations that show the usefulness of such a database in a country-wide BVD eradication program.

  11. Introduction to Bovine Viral Diarrhea-Mucosal Disease%牛病毒性腹泻-黏膜病研究概述

    Institute of Scientific and Technical Information of China (English)

    王国超; 白鸽; 王选; 吴奇强

    2016-01-01

    As an acute and febrile infectious disease caused by bovine viral diarrhea virus(BVDV),bovine vi-ral diarrhea-mucosal disease (BVD-MD)is characterized by diarrhea,mucosa exfoliative,pregnant cow a-bortion,dead fetus and abnormal fetus,persistent infection and immunotolerance,immunosupression.The epidemiological studies showed that BVD is widely prevalent in many countries and regions with developed cattle industry.The disease has been prevalent in more than 20 provinces with a high infection rates.Fur-thermore,cattle with BVDV are the potential source of contamination to bioproducts and bioactive materi-als,cause severe economic loss to animal husbandry production and related industry.This paper mainly in-troduced the pathogen classification and typing,infection and epidemic states at home and abroad,preven-tive strategies.%牛病毒性腹泻-黏膜病是由牛病毒性腹泻病毒(BVDV)引起的牛的一种急性、热性传染病,主要表现为腹泻、黏膜脱落、母畜流产、产死胎和畸胎,持续性感染、免疫耐受与免疫抑制等。流行病学调查发现,该病在世界范围内广泛流行,特别是在许多畜牧业发达的国家和地区普遍存在。我国有20多个省、市、自治区有该病流行,并且有较高的感染率。携带 BVDV 的牛是牛源生物制品及生物活性材料的潜在污染源,给畜牧业生产及相关领域造成严重的经济损失。论文主要对牛病毒性腹泻-黏膜病的病原学分类与分型、感染与流行情况、国内外现状及防控策略进行了综述。

  12. Aislamiento y caracterización del virus de la diarrea viral bovina en un ternero con síndrome purpúrico Characterization of bovine viral diarrhea virus isolated from cattle with hemorrhagic syndrome

    Directory of Open Access Journals (Sweden)

    A Gollán

    2006-01-01

    Full Text Available El Virus de la Diarrea Viral Bovina (VDVB produce en el ganado bovino numerosas patologías que van desde pérdidas reproductivas hasta afecciones de poca significación clínica en el aparato digestivo. Se ha reportado variabilidad entre las cepas de VDVB, que se manifiesta por la existencia de los biotipos citopatogénicos (CP y no citopatogénicos (NCP, y los tipos virales I y II. El presente trabajo describe los hallazgos patológicos y virológicos en un ternero que clínicamente exhibió trombocitopenia y diarrea. La cepa viral aislada (334/3 fue caracterizada molecularmente por secuenciación de la región 5' no-codificante (5' RNC. Los análisis realizados revelaron un 90-98% de homología con las cepas de referencia tipo I, no encontrándose cambios asociados a las cepas VDVB tipo II.The Bovine Viral Diarrhea (BVDV virus causes numerous pathologies that range from reproductive losses to infections of little clinical significance in the bovine digestive tract. Variation have been reported among the strains of the BVDV, which are classified into two biotypes; cytopathogenic (CP and non-cytopathogenic (NCP, and the viral types I and II. This work describes the pathological findings in a calf with diarrhea and severe thrombocytopenia. The strain isolated (334/3 was molecularly characterized by sequencing of the 5’non-coding region (5’ NCR. These analyses revealed 90-98% homologies with reference strains type I strains and the changes associated with BVDV type II, were not found.

  13. Isolation and identification of bovine viral diarrhea virus from swine%猪源牛病毒性腹泻病毒的分离鉴定

    Institute of Scientific and Technical Information of China (English)

    杨小燕; 魏春华; 刘建奎; 戴爱玲; 李晓华

    2011-01-01

    从福建省某猪场疑似猪瘟的发病仔猪采集病料,处理后接种于牛肾细胞(MDBK),分离到1株病毒,并对其进行了系统鉴定.结果显示,病料接种MDBK细胞72 h后产生了明显的细胞病变;该分离毒株可以被牛病毒性腹泻病毒(BVDV)阳性血清中和,中和效价为1:106;该分离毒株对氯仿、乙醚、胰酶、酸敏感,不耐热,56℃30 min可被灭活,PCR检测该分离毒株的细胞培养液,结果可扩增出325 bp的片段,该片段的核苷酸序列与BVDV NADL株标准种毒的同源性为89.2%.结果表明,该分离毒株为猪源牛病毒性腹泻病毒.%One strain of virus was isolated from the infected swine in Fujian Province by cultivating it in Madin-Darby bovine kidney(MDBK) cells, then it was systematically identified by a series of methods. In result,the isolated virus caused typical cytopathogenic effect in the MDBK cells at 72 hours after infection.The isolated virus could be neutralized by the positive sera with bovine viral diarrhea virus at titer 1: 106.The isolate was sensitive to chloroform, ether, trypsin, acid and temperature, and could be inactivated at 56 ℃ for 30 minutes. A DNA fragment of 325 bp was amplified by PCR from the MDBK cells infected with the isolated virus. The identity of this gene sequence to the isolated virus and NADL strain from GenBank was 89.2 %. It was concluded from the results that the isolated virus from the infected swine is a bovine viral diarrhea virus.

  14. Development of an enhanced bovine viral diarrhea virus subunit vaccine based on E2 glycoprotein fused to a single chain antibody which targets to antigen-presenting cells.

    Science.gov (United States)

    Pecora, Andrea; Malacari, Darío A; Pérez Aguirreburualde, María S; Bellido, Demian; Escribano, José M; Dus Santos, María J; Wigdorovitz, Andrés

    2015-01-01

    Bovine viral diarrhea virus (BVDV) is an important cause of economic losses worldwide. E2 is an immunodominant protein and a promising candidate to develop subunit vaccines. To improve its immunogenicity, a truncated E2 (tE2) was fused to a single chain antibody named APCH, which targets to antigen-presenting cells. APCH-tE2 and tE2 proteins were expressed in the baculovirus system and their immunogenicity was firstly compared in guinea pigs. APCH-tE2 vaccine was the best one to evoke a humoral response, and for this reason, it was selected for a cattle vaccination experiment. All the bovines immunized with 1.5 μg of APCH-tE2 developed high levels of neutralizing antibodies against BVDV up to a year post-immunization, demonstrating its significant potential as a subunit vaccine. This novel vaccine is undergoing scale-up and was transferred to the private sector. Nowadays, it is being evaluated for registration as the first Argentinean subunit vaccine for cattle.

  15. Evidence of Bovine viral diarrhea virus Infection in Three Species of Sympatric Wild Ungulates in Nevada: Life History Strategies May Maintain Endemic Infections in Wild Populations.

    Science.gov (United States)

    Wolff, Peregrine L; Schroeder, Cody; McAdoo, Caleb; Cox, Mike; Nelson, Danielle D; Evermann, James F; Ridpath, Julia F

    2016-01-01

    Evidence for bovine viral diarrhea virus (BVDV) infection was detected in 2009-2010 while investigating a pneumonia die-off in Rocky Mountain bighorn sheep (Ovis canadensis, canadensis), and sympatric mountain goats (Oreamnos americanum) in adjacent mountain ranges in Elko County, Nevada. Seroprevalence to BVDV-1 was 81% (N = 32) in the bighorns and 100% (N = 3) in the mountain goats. Serosurveillance from 2011 to 2015 of surviving bighorns and mountain goats as well as sympatric mule deer (Odocoileus hemionus), indicated a prevalence of 72% (N = 45), 45% (N = 51), and 51% (N = 342) respectively. All species had antibody titers to BVDV1 and BVDV2. BVDV1 was isolated in cell culture from three bighorn sheep and a mountain goat kid. BVDV2 was isolated from two mule deer. Six deer (N = 96) sampled in 2013 were positive for BVDV by antigen-capture ELISA on a single ear notch. Wild ungulates and cattle concurrently graze public and private lands in these two mountain ranges, thus providing potential for interspecies viral transmission. Like cattle, mule deer, mountain goats, and bighorn sheep can be infected with BVDV and can develop clinical disease including immunosuppression. Winter migration patterns that increase densities and species interaction during the first and second trimester of gestation may contribute to the long term maintenance of the virus in these wild ungulates. More studies are needed to determine the population level impacts of BVDV infection on these three species.

  16. Risk and prevention of bovine viral diarrhea virus (BVDV) transmission through embryo production via somatic cell nuclear transfer (SCNT) using oocytes from persistently infected donors.

    Science.gov (United States)

    Gregg, K; Riddell, K P; Chen, S H; Galik, P K; Xiang, T; Guerra, T; Marley, M S; Polejaeva, I; Givens, M D

    2010-07-01

    The objective was to assess the risk of transmission of bovine viral diarrhea virus (BVDV) through embryo production via somatic cell nuclear transfer (SCNT), with oocytes obtained from persistently infected (PI) donors. Using ultrasound-guided follicular aspiration following superstimulation, oocytes were obtained from five female beef cattle, including three that were PI and two that were negative for BVDV. In the three PI cattle, seven aspirations yielded 32 oocytes (PI-1: three aspirations yielding six oocytes; PI-2: two aspirations yielding 14 oocytes; and PI-3: two aspirations yielding 12 oocytes). The oocyte recovery rate was better in negative control cattle, with 32 oocytes obtained from the two cattle in a single superstimulation and aspiration session. Oocytes were processed individually for SCNT, evaluated, and tested for BVDV. Nearly all (31/32) oocytes from the three PI donors were positive for BVDV by PCR, with detected viral RNA copy number ranging from 1 to 1.1 x 10(5). The proportion of oocytes acceptable for SCNT embryo production (based on oocyte quality and maturation status) was only 16 to 35% from PI donors, but was 81% from control donors. Therefore, routine testing of unacceptable (discarded) oocytes could be an effective approach to identify batches that might contain infected oocytes from PI donors. Identification and removal of high-risk batches of oocytes would minimize the risk of BVDV transmission through SCNT embryo production.

  17. Evidence of bovine viral diarrhea virus infection in three species of sympatric wild ungulates in Nevada: life history strategies may maintain endemic infections in wild populations

    Directory of Open Access Journals (Sweden)

    Peregrine Lee Wolff

    2016-03-01

    Full Text Available Evidence for bovine viral diarrhea virus (BVDV infection was detected in 2009-10 while investigating a pneumonia die-off in Rocky Mountain bighorn sheep (Ovis canadensis canadensis, and sympatric mountain goats (Oreamnos americanum in adjacent mountain ranges in Elko County, Nevada. Seroprevalence to BVDV-1 was 81% (N=32 in the bighorns and 100% (N=3 in the mountain goats. Serosurveillance from 2011 to 2015 of surviving bighorns and mountain goats as well as sympatric mule deer (Odocoileus hemionus, indicated a prevalence of 72% (N=45, 45% (N=51, and 51% (N=342 respectively. All species had antibody titers to BVDV1 and BVDV2. BVDV1 was isolated in cell culture from three bighorn sheep and a mountain goat kid. BVDV2 was isolated from two mule deer. Six deer (N=96 sampled in 2013 were positive for BVDV by antigen-capture ELISA on ear notch. Wild ungulates and cattle concurrently graze public and private lands in these two mountain ranges, thus providing potential for interspecies viral transmission. Like cattle, mule deer, mountain goats, and bighorn sheep can be infected with BVDV and can develop clinical disease including immunosuppression. Winter migration patterns that increase densities and species interaction during the first and second trimester of gestation may contribute to the long term maintenance of the virus in these wild ungulates. More studies are needed to determine the population level impacts of BVDV infection on these three species.

  18. Recombinant E2 glycoprotein of bovine viral diarrhea virus induces a solid humoral neutralizing immune response but fails to confer total protection in cattle

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    S. Chimeno Zoth

    2007-06-01

    Full Text Available Two recombinant baculoviruses were produced in order to obtain a bovine viral diarrhea virus (BVDV immunogen: AcNPV/E2 expressing E2 glycoprotein, and AcNPV/E0E1E2 expressing the polyprotein region coding for the three structural proteins of BVDV (E0, E1, and E2. Mice were immunized with Sf9 cells infected with the recombinant baculoviruses in a water in oil formulation and the production of neutralizing antibodies was evaluated. Since E2 elicited higher neutralizing antibody titers than E0-E1-E2 polyprotein, it was selected to immunize cattle. Calves received two doses of recombinant E2 vaccine and were challenged with homologous BVDV 37 days later. The recombinant immunogen induced neutralizing titers which showed a mean value of 1.5 ± 0.27 on the day of challenge and reached a top value of 3.36 ± 0.36, 47 days later (84 days post-vaccination. On the other hand, sera from animals which received mock-infected Sf9 cells did not show neutralizing activity until 25 days post-challenge (62 days post-vaccination, suggesting that these antibodies were produced as a consequence of BVDV challenge. Even when no total protection was observed in cattle, in vitro viral neutralization assays revealed that the recombinant immunogen was able to induce neutralizing antibody synthesis against the homologous strain as well as against heterologous strains in a very efficient way.

  19. 河南省奶牛病毒性腹泻-黏膜病血清学调查%Serological Survey of Bovine Viral Diarrhea-mucosal Disease in Henan Province

    Institute of Scientific and Technical Information of China (English)

    石冬梅; 张华; 邓红雨; 刘太宇; 王金合

    2011-01-01

    为了解河南省奶牛病毒性腹泻一黏膜病的发生情况,2009年5月对河南省的豫东、豫西、豫南、豫北和郑州市五区域20个奶牛小区的355头奶牛随机采血,采用ELISA方法,检测奶牛病毒性腹泻-黏膜病血清抗体,结果显示20个奶牛小区有17个奶牛小区感染有病毒性腹泻-黏膜病病毒,场感染率为85%,各小区奶牛病毒性腹泻-黏膜病检测阳性率在0%~88.24%,血清阳性率平均为53.8%,表明该病在河南省奶牛中感染率较高.%The aim of this study was to investigate the prevalence of Bovine viral diarrhea-mucosal disease in Henan Province. 355 bovine sera from twenty cattle breeding farms in different parts of Henan province was collected and antibodies to Bovine viral diarrhea virus (BVDV) were detected by ELISA. The result showed that 85 percent of twenty cattle breeding farms were infected by Bovine viral diarrhea virus. The highest positive rate in cattle breeding farms was 88.24 percent, and the average rate also arrived at 53.8 percent. It indicated that the Bovine viral diarrhea-mucosal disease was widespread in the cattle breeding farms of Henan province.

  20. Construction and immunogenicity of the recombinant Lactobacillus acidophilus pMG36e-E0-LA-5 of bovine viral diarrhea virus.

    Science.gov (United States)

    Zhao, Yuelan; Jiang, Lufeng; Liu, Teng; Wang, Min; Cao, Wenbo; Bao, Yongzhan; Qin, Jianhua

    2015-12-01

    Bovine viral diarrhea/mucosal disease (BVD/MD) is an infectious disease of cattle with a worldwide distribution, creating a substantial economic impact. It is caused by bovine viral diarrhea virus (BVDV). This research was conducted to construct the recombinant Lactobacillus acidophilus (L. acidophilus) pMG36e-E0-LA-5 of BVDV E0 gene and to test its immunogenicity and protective efficacy against BVDV infection in the mice model. The BVDV E0 gene was sub-cloned into the expression vector and then transformed into the L. acidophilus LA-5 strain by electroporation. The recombinant L. acidophilus pMG36e-E0-LA-5 was confirmed by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. The mice were immunized orally with the recombinant L. acidophilus pMG36e-E0-LA-5. The serum IgG antibody and fecal sIgA antibody responses, expression levels of interleukin (IL)-12 (IL-12) and interferon gamma (IFN-γ) were detected respectively. On the 7th day after the last-immunization, the mice were inoculated with BVDV to evaluate the protective efficiency of the recombinant L. acidophilus pMG36e-E0-LA-5. The results showed that the expressed products protein E0 in the L. acidophilus LA-5 resulted in single band of 27kDa by SDS-PAGE and its strong reactivity with BVDV antibody was confirmed by Western blotting. The IgG and sIgA antibodies responses, IL-12 and IFN-γ expression levels in the vaccinated mice with recombinant L. acidophilus pMG36e-E0-LA-5 were significantly higher than those in the control mice. The protective rate of the vaccinated mice against BVDV increased significantly, and a 90.00% protection rate in virulent challenge was observed. These results indicated that the recombinant L. acidophilus pMG36e-E0-LA-5 strain was successfully constructed and it could effectively improve the immune response in mice and might provide protection against BVDV.

  1. Coinfections of Sudanese dairy cattle with bovine herpes virus 1, bovine viral diarrhea virus, bluetongue virus and bovine herpes virus 4 and their relation to reproductive disorders

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    Amira M. Elhassan

    2016-12-01

    Reults: The meta-analysis of the data indicated high seroprevalence of coinfections with various combinations of these agents; only few animals were singly infected. An infection with BHV-1 was observed to be higher than the prevalence of associations between BHV-1 and the other three viral agents. Prevalence of seropositivities to coinfection with BHV-1/BTV; BHV-1/BVD; BHV-1/BTV/BVD were the highest while seropositivities prevalences that involved BHV-4 were much lower. The highest abortion rates were encountered in coinfections with BHV-1/BVD/BTV (31% and BHV-1/BVD/BTV/BHV-4 (30% while most infertility cases were noticed in coinfection with BHV-1/BVD/BTV (44% and BHV-1/BVD/BTV/BHV-4 (21%, and coinfections with the four viruses were encountered in most of the death after birth cases (25%. Overall mixed infections with BHV-1/BVD/BTV (34% and BHV-1/BVD/BTV/BHV-4 (22.5% were involved in the majority of reproductive problems studied. Conclusion: Mixed infections constitutes the vast majority of cases and are involved in the majority of reproductive disorders investigated. The high prevalence of seropositivity to all of the four viruses should call for an intervention strategy to reduce the impact of these viruses. [J Adv Vet Anim Res 2016; 3(4.000: 332-337

  2. Extended Genetic Diversity of Bovine Viral Diarrhea Virus and Frequency of Genotypes and Subtypes in Cattle in Italy between 1995 and 2013

    Directory of Open Access Journals (Sweden)

    Camilla Luzzago

    2014-01-01

    Full Text Available Genetic typing of bovine viral diarrhea virus (BVDV has distinguished BVDV-1 and BVDV-2 species and an emerging putative third species (HoBi-like virus, recently detected in southern Italy, signaling the occurrence of natural infection in Europe. Recognizing the need to update the data on BVDV genetic variability in Italy for mounting local and European alerts, a wide collection of 5′ UTR sequences (n = 371 was selected to identify the frequency of genotypes and subtypes at the herd level. BVDV-1 had the highest frequency, followed by sporadic BVDV-2. No novel HoBi-like viruses were identified. Four distribution patterns of BVDV-1 subtypes were observed: highly prevalent subtypes with a wide temporal-spatial distribution (1b and 1e, low prevalent subtypes with a widespread geographic distribution (1a, 1d, 1g, 1h, and 1k or a restricted geographic distribution (1f, and sporadic subtypes detected only in single herds (1c, 1j, and 1l. BVDV-1c, k, and l are reported for the first time in Italy. A unique genetic variant was detected in the majority of herds, but cocirculation of genetic variants was also observed. Northern Italy ranked first for BVDV introduction, prevalence, and dispersion. Nevertheless, the presence of sporadic variants in other restricted areas suggests the risk of different routes of BVDV introduction.

  3. Extended genetic diversity of bovine viral diarrhea virus and frequency of genotypes and subtypes in cattle in Italy between 1995 and 2013.

    Science.gov (United States)

    Luzzago, Camilla; Lauzi, Stefania; Ebranati, Erika; Giammarioli, Monica; Moreno, Ana; Cannella, Vincenza; Masoero, Loretta; Canelli, Elena; Guercio, Annalisa; Caruso, Claudio; Ciccozzi, Massimo; De Mia, Gian Mario; Acutis, Pier Luigi; Zehender, Gianguglielmo; Peletto, Simone

    2014-01-01

    Genetic typing of bovine viral diarrhea virus (BVDV) has distinguished BVDV-1 and BVDV-2 species and an emerging putative third species (HoBi-like virus), recently detected in southern Italy, signaling the occurrence of natural infection in Europe. Recognizing the need to update the data on BVDV genetic variability in Italy for mounting local and European alerts, a wide collection of 5' UTR sequences (n = 371) was selected to identify the frequency of genotypes and subtypes at the herd level. BVDV-1 had the highest frequency, followed by sporadic BVDV-2. No novel HoBi-like viruses were identified. Four distribution patterns of BVDV-1 subtypes were observed: highly prevalent subtypes with a wide temporal-spatial distribution (1b and 1e), low prevalent subtypes with a widespread geographic distribution (1a, 1d, 1g, 1h, and 1k) or a restricted geographic distribution (1f), and sporadic subtypes detected only in single herds (1c, 1j, and 1l). BVDV-1c, k, and l are reported for the first time in Italy. A unique genetic variant was detected in the majority of herds, but cocirculation of genetic variants was also observed. Northern Italy ranked first for BVDV introduction, prevalence, and dispersion. Nevertheless, the presence of sporadic variants in other restricted areas suggests the risk of different routes of BVDV introduction.

  4. Expression of Bovine Viral Diarrhea Virus Envelope Glycoprotein E2 in Yeast Pichia pastoris and its Application to an ELISA for Detection of BVDV Neutralizing Antibodies in Cattle.

    Science.gov (United States)

    Behera, Sthita Pragnya; Mishra, Niranjan; Nema, Ram Kumar; Pandey, Pooja Dubey; Kalaiyarasu, Semmannan; Rajukumar, Katherukamem; Prakash, Anil

    2015-01-01

    The aim of this article is to express envelope glycoprotein E2 of bovine viral diarrhea virus (BVDV) in yeast Pichia pastoris and its utility as a diagnostic antigen in ELISA. The BVDV E2 gene was cloned into the pPICZαA vector followed by integration into the Pichia pastoris strain X-33 genome for methanol-induced expression. SDS-PAGE and Western blot results showed that the recombinant BVDV E2 protein (72 kDa) was expressed and secreted into the medium at a concentration of 40 mg/L of culture under optimized conditions. An indirect ELISA was then developed by using the yeast-expressed E2 protein. Preliminary testing of 300 field cattle serum samples showed that the E2 ELISA showed a sensitivity of 91.07% and a specificity of 92.02% compared to the reference virus neutralization test. The concordance between the E2 ELISA and VNT was 91.67%. This study demonstrates feasibility of BVDV E2 protein expression in yeast Pichia pastoris for the first time and its efficacy as an antigen in ELISA for detecting BVDV neutralizing antibodies in cattle.

  5. Rapid genome detection of Schmallenberg virus and bovine viral diarrhea virus by use of isothermal amplification methods and high-speed real-time reverse transcriptase PCR.

    Science.gov (United States)

    Aebischer, Andrea; Wernike, Kerstin; Hoffmann, Bernd; Beer, Martin

    2014-06-01

    Over the past few years, there has been an increasing demand for rapid and simple diagnostic tools that can be applied outside centralized laboratories by using transportable devices. In veterinary medicine, such mobile test systems would circumvent barriers associated with the transportation of samples and significantly reduce the time to diagnose important infectious animal diseases. Among a wide range of available technologies, high-speed real-time reverse transcriptase quantitative PCR (RT-qPCR) and the two isothermal amplification techniques loop-mediated isothermal amplification (LAMP) and recombinase polymerase amplification (RPA) represent three promising candidates for integration into mobile pen-side tests. The aim of this study was to investigate the performance of these amplification strategies and to evaluate their suitability for field application. In order to enable a valid comparison, novel pathogen-specific assays have been developed for the detection of Schmallenberg virus and bovine viral diarrhea virus. The newly developed assays were evaluated in comparison with established standard RT-qPCR using samples from experimentally or field-infected animals. Even though all assays allowed detection of the target virus in less than 30 min, major differences were revealed concerning sensitivity, specificity, robustness, testing time, and complexity of assay design. These findings indicated that the success of an assay will depend on the integrated amplification technology. Therefore, the application-specific pros and cons of each method that were identified during this study provide very valuable insights for future development and optimization of pen-side tests.

  6. Application of restriction fragment length polymorphism analysis to simple and rapid genotyping of bovine viral diarrhea virus strains isolated in Japan.

    Science.gov (United States)

    Seki, Yoshihisa; Seimiya, Yukio M; Motokawa, Masato; Yaegashi, Gakuji; Nagai, Makoto; Hayashi, Michiko

    2008-04-01

    The E2 regions of 177 bovine viral diarrhea virus (BVDV) strains isolated in Japan between 1957 and 2006 were analyzed for genotyping. The strains were classified into 8 genotypes (1a, 1b, 1c, 1d, 1e, 1f, So and 2a) based on the phylogenetic analysis. The restriction fragment length polymorphism (RFLP) analysis of the RT-PCR products using 6 selected enzymes (Apo I, Mly I, BstAP I, Pvu II, Ear I, EcoR V) disclosed the cutting patterns classified into 11 groups (I-XI), each of that consisted of strains belonging to a single genotype. Namely, groups-I and -II were composed by genotype-1a strains, groups-III and -IV by 1b strains, and groups-V and -VI by 1c strains. Other groups-VII, -VIII, -IX, -X and -XI comprised genotypes-1d, -1e, -1f, -So and -2a strains, respectively. The results suggest that the RFLP analysis can simply and rapidly differentiate the 8 genotypes of BVDV strains.

  7. Genetic characterization of bovine viral diarrhea virus strains in Beijing, China and innate immune responses of peripheral blood mononuclear cells in persistently infected dairy cattle.

    Science.gov (United States)

    Weng, Xiao Gang; Song, Quan Jiang; Wu, Qiong; Liu, Ming Chao; Wang, Meng Ling; Wang, Jiu Feng

    2015-01-01

    To acquire epidemiological data on the bovine viral diarrhea virus (BVDV) and identify cattle persistently infected (PI) with this virus, 4,327 samples from Holstein dairy cows were screened over a four-year period in Beijing, China. Eighteen BVD viruses were isolated, 12 from PI cattle. Based on genetic analysis of their 5'-untranslated region (5'-UTR), the 18 isolates were assigned to subgenotype BVDV-1m, 1a, 1d, 1q, and 1b. To investigate the innate immune responses in the peripheral-blood mononuclear cells of PI cattle, the expression of Toll-like receptors (TLRs), RIG-I-like receptors, interferon-α (IFN-α), IFN-β, myxovirus (influenza virus) resistance 1 (MX1), and interferon stimulatory gene 15 (ISG15) was assessed by qPCR. When compared with healthy cattle, the expression of TLR-7, IFN-α, and IFN-β mRNA was downregulated, but the expression of MX1 and ISG-15 mRNA was upregulated in PI cattle. Immunoblotting analysis revealed that the expression of interferon regulatory factor 3 (IRF-3) and IRF-7 was lower in PI cattle than in healthy cattle. Thus, BVDV-1m and 1a are the predominant subgenotypes in the Beijing region, and the strains are highly divergent. Our findings also suggest that the TLR-7/IRF-7 signaling pathway plays a role in evasion of host restriction by BVDV.

  8. One year duration of immunity of the modified live bovine viral diarrhea virus type 1 and type 2 and bovine herpesvirus-1 fractions of Vista® Once SQ vaccine.

    Science.gov (United States)

    Purtle, Lisa; Mattick, Debra; Schneider, Corey; Smith, Linda; Xue, Wenzhi; Trigo, Emilio

    2016-03-18

    Three studies were performed to determine the duration of immunity of the bovine viral diarrhea virus type 1 and type 2 (BVDV-1 and BVDV-2) and bovine herpesvirus-1 (BHV-1) fractions of a commercially prepared modified-live vaccine. Vista® Once SQ (Vista®) vaccine contains five modified-live viruses, BVDV-1, BVDV-2, BHV-1, bovine respiratory syncytial virus, and bovine parainfluenza 3 virus, and two modified-live bacteria, Pasteurella multocida and Mannheimia haemolytica. For all three studies, calves were administered a single dose of vaccine or placebo vaccine subcutaneously, and were challenged with one of the three virulent viruses at least one year following vaccination. Calves were evaluated daily following challenge for clinical signs of disease associated with viral infection, nasal swab samples were evaluated for virus shedding, and serum was tested for neutralizing antibodies. Following the BVDV-1 and BVDV-2 challenges, whole blood was evaluated for white blood cell counts, and for the BVDV-2 study, whole blood was also evaluated for platelet counts. Calves vaccinated with BVDV type 1a, were protected from challenge with BVDV type 1b, and had significant reductions in clinical disease, fever, leukopenia, and virus shedding compared to control calves. Vaccinated calves in the BVDV-2 study were protected from clinical disease, mortality, fever, leukopenia, thrombocytopenia, and virus shedding compared to controls. Vaccinated calves in the BHV-1 study were protected from clinical disease and fever, and had significantly reduced duration of nasal virus shedding. These three studies demonstrated that a single administration of the Vista® vaccine to healthy calves induces protective immunity against BVDV-1, BVDV-2 and BHV-1 that lasts at least one year following vaccination.

  9. Immunogenicity of an inactivated Chinese bovine viral diarrhea virus 1a (BVDV 1a) vaccine cross protects from BVDV 1b infection in young calves.

    Science.gov (United States)

    Wang, Wei; Shi, Xinchuan; Wu, Yongwang; Li, Xiaoxin; Ji, Ye; Meng, Qingsen; Zhang, Shucheng; Wu, Hua

    2014-08-15

    Bovine viral diarrhea virus (BVDV) 1a and 1b strains are the predominant subgenotypes in China. Because of the genetic and antigenic variability among different BVDV strains, a vaccine effective in one region may fail to protect against infections caused by different virus strains in another region. No BVDV vaccine developed with the predominant strains in China are available. In this study, the immunogenicity of an inactivated Chinese BVDV 1a NM01 vaccine strain was evaluated by challenging with a Chinese BVDV 1b JL strain. Ten 2-4-month-old calves were intramuscularly vaccinated with a single dose of the vaccine strain and boosted with same dose three weeks after the first vaccination, with five mock immunized calves serving as a control group. The average titer of neutralization antibody to BVDV 1a and BVDV 1b of immunized calves reached 1:410 and 1:96, respectively, at 21 days post the second vaccination. Twenty-one days post the second vaccination, all calves were challenged with strain JL. The clinical signs, such as the temperature and leukopenia of the immunized calves and viral shedding, were significantly less than the mock immunized calves after challenging with the virulent BVDV 1b strain, indicating that the BVDV 1a vaccine strain elicited efficacious protection against the endemic BVDV 1b strain in China. To the best of our knowledge, this is the first report of an inactivated BVDV vaccine which demonstrated effective cross-protection against BVDV type 1b infection in China.

  10. Stability of the resistance to the thiosemicarbazone derived from 5,6-dimethoxy-1-indanone, a non-nucleoside polymerase inhibitor of bovine viral diarrhea virus.

    Directory of Open Access Journals (Sweden)

    Eliana F Castro

    Full Text Available Bovine viral diarrhea virus (BVDV is the prototype Pestivirus. BVDV infection is distributed worldwide and causes serious problems for the livestock industry. The thiosemicarbazone of 5,6-dimethoxy-1-indanone (TSC is a non-nucleoside polymerase inhibitor (NNI of BVDV. All TSC-resistant BVDV variants (BVDV-TSCr T1-5 present an N264D mutation in the NS5B gene (RdRp whereas the variant BVDV-TSCr T1 also presents an NS5B A392E mutation. In the present study, we carried out twenty passages of BVDV-TSCr T1-5 in MDBK cells in the absence of TSC to evaluate the stability of the resistance. The viral populations obtained (BVDV R1-5 remained resistant to the antiviral compound and conserved the mutations in NS5B associated with this phenotype. Along the passages, BVDV R2, R3 and R5 presented a delay in the production of cytopathic effect that correlated with a decrease in cell apoptosis and intracellular accumulation of viral RNA. The complete genome sequences that encode for NS2 to NS5B, Npro and Erns were analyzed. Additional mutations were detected in the NS5B of BVDV R1, R3 and R4. In both BVDV R2 and R3, most of the mutations found were localized in NS5A, whereas in BVDV R5, the only mutation fixed was NS5A V177A. These results suggest that mutations in NS5A could alter BVDV cytopathogenicity. In conclusion, the stability of the resistance to TSC may be due to the fixation of different compensatory mutations in each BVDV-TSCr. During their replication in a TSC-free medium, some virus populations presented a kind of interaction with the host cell that resembled a persistent infection: decreased cytopathogenicity and viral genome synthesis. This is the first report on the stability of antiviral resistance and on the evolution of NNI-resistant BVDV variants. The results obtained for BVDV-TSCr could also be applied for other NNIs.

  11. Cloning and sequence analysis of genetic variation on NS2-3 of bovine viral diarrhea virus (HB-DCZ) strain in Hebei Province, China

    Institute of Scientific and Technical Information of China (English)

    ZHAO Yuelan; QIN Jianhua; GUO Hongbin; ZUO Yuzhu; ZHANG Baoning; ZHANG Lei

    2007-01-01

    The objective of this research is to analyze the genetic characterization of a bovine viral diarrhea virus (BVDV) strain (HB-DCZ strain) isolated from China and describe its relationship with other BVDV strains.Special primers (forward:5'- gagatctcgggaggtac -3',reverse:5'-cctctcggcatgatcccgaaa -3) are used to amplify partial NS2-3 sequence of HB-DCZ strain by reverse transcription polymerase chain reaction (RT-PCR).The product of PCR is cloned into pMD18-T vector,and then transfected into JM109.The recombinant plasmids are extracted and identified by EcoR Ⅰand Hind Ⅲ enzyme digestion.The NS 2-3 nucleotide fragment of the isolated virus is sequenced,and then amino acid sequence is deduced.Nucleotide sequence and deduced amino acid sequence of the strain are analyzed and compared with other BVDV strains from Genebank with the aid of DNAstar software.The results show that the obtained fragment of HB-DCZ strain contains 665 bp nucleotides,which indicate that there is no insertion in the isolated virus genome.The homologies of nucleotide sequences show that HB-DCZ strain has 99.1%,97.4%,92.3%,77%,76.9%,76.4% and 76.2%,sequence similarity with 184,ZM195,Osloss,Oregon C24V,Singer,NADL,and S D-I,respectively.According to the nucleotide sequences of the obtained fragments,the 208 corresponding amino acids are deduced.The homologies of amino acid sequences show that HB-DCZ strain had 100%,93.3%,91.3% and 83.2% sequence similarity with VEDEVAC,Osloss,ILLC and Oregon C24V,respectively.In conclusion,HB-DCZ strain has no exogenous sequence insertion,no gene recombination,no gene rearrangement and no gene deficiency.HB-DCZ strain is closely related to BVDV Osloss strain,and belongs to subtype Ib.

  12. 牛病毒性腹泻病毒的分子生物学研究进展%Advance in molecular biology of bovine viral diarrhea virus

    Institute of Scientific and Technical Information of China (English)

    孙宏进; 陶洁; 朱礼倩; 朱国强

    2011-01-01

    牛病毒性腹泻病(Bovine Viral Diarrhea, BVD)是由牛病毒性腹泻病毒(BVDV)引起的,主要侵害牛的一种重要传染病,临床主要表现为发热、黏膜糜烂、溃疡、白细胞减少、持续感染(Persistently Infected, PI)、咳嗽及怀孕母牛流产或产生畸形胎儿等等.羊、猪、鹿和多种野生动物等也能感染和传播.该病呈世界性分布,从世界范围内讲已有60多年的历史,在我国也已存在20多年之久,给各国畜牧业造成了巨大的经济损失,但目前为止,还没有有效的预防和控制BVDV的措施.本文阐述了BVDV的基因组结构、编码的蛋白质、分子流行病学、生物型及生物型之间转化机制以及国外BVDV新型疫苗研制等方面的进展情况,以便人们进一步了解该病分子生物学方面的信息.

  13. Suitability of vaccinia virus and bovine viral diarrhea virus (BVDV for determining activities of three commonly-used alcohol-based hand rubs against enveloped viruses

    Directory of Open Access Journals (Sweden)

    Steinmann Jochen

    2007-02-01

    Full Text Available Abstract Background A procedure for including activity against enveloped viruses in the post-contamination treatment of hands has been recommended, but so far no European standard is available to implement it. In 2004, the German Robert Koch-Institute (RKI and the German Association for the Control of Virus Disease (DVV suggested that vaccinia virus and bovine viral diarrhea virus (BVDV should be used as test viruses in a quantitative suspension test to determine the activity of a disinfectant against all enveloped viruses. Methods We have studied the activities of three commonly-used alcohol-based hand rubs (hand rub A, based on 45% propan-2-ol, 30% propan-1-ol and 0.2% mecetronium etilsulfate; hand rub B, based on 80% ethanol; hand rub C, based on 95% ethanol against vaccinia virus and BVDV, and in addition against four other clinically relevant enveloped viruses: herpes simplex virus (HSV types 1 and 2, and human and avian influenza A virus. The hand rubs were challenged with different organic loads at exposure time of 15, 30 and 60 s. According to the guidelines of both BGA/RKI and DVV, and EN 14476:2005, the reduction of infectivity of each test virus was measured on appropriate cell lines using a quantitative suspension test. Results All three alcohol-based hand rubs reduced the infectivity of vaccinia virus and BVDV by ≥ 4 log10-steps within 15 s, irrespective of the type of organic load. Similar reductions of infectivity were seen against the other four enveloped viruses within 15 s in the presence of different types of organic load. Conclusion Commonly used alcohol-based hand rubs with a total alcohol concentration ≥ 75% can be assumed to be active against clinically relevant enveloped viruses if they effectively reduce the infectivities of vaccinia virus and BVDV in a quantitative suspension test.

  14. A search for RNA insertions and NS3 gene duplication in the genome of cytopathic isolates of bovine viral diarrhea virus

    Directory of Open Access Journals (Sweden)

    V.L. Quadros

    2006-07-01

    Full Text Available Calves born persistently infected with non-cytopathic bovine viral diarrhea virus (ncpBVDV frequently develop a fatal gastroenteric illness called mucosal disease. Both the original virus (ncpBVDV and an antigenically identical but cytopathic virus (cpBVDV can be isolated from animals affected by mucosal disease. Cytopathic BVDVs originate from their ncp counterparts by diverse genetic mechanisms, all leading to the expression of the non-structural polypeptide NS3 as a discrete protein. In contrast, ncpBVDVs express only the large precursor polypeptide, NS2-3, which contains the NS3 sequence within its carboxy-terminal half. We report here the investigation of the mechanism leading to NS3 expression in 41 cpBVDV isolates. An RT-PCR strategy was employed to detect RNA insertions within the NS2-3 gene and/or duplication of the NS3 gene, two common mechanisms of NS3 expression. RT-PCR amplification revealed insertions in the NS2-3 gene of three cp isolates, with the inserts being similar in size to that present in the cpBVDV NADL strain. Sequencing of one such insert revealed a 296-nucleotide sequence with a central core of 270 nucleotides coding for an amino acid sequence highly homologous (98% to the NADL insert, a sequence corresponding to part of the cellular J-Domain gene. One cpBVDV isolate contained a duplication of the NS3 gene downstream from the original locus. In contrast, no detectable NS2-3 insertions or NS3 gene duplications were observed in the genome of 37 cp isolates. These results demonstrate that processing of NS2-3 without bulk mRNA insertions or NS3 gene duplications seems to be a frequent mechanism leading to NS3 expression and BVDV cytopathology.

  15. A simulation model to quantify the value of implementing whole-herd Bovine viral diarrhea virus testing strategies in beef cow-calf herds.

    Science.gov (United States)

    Nickell, Jason S; White, Brad J; Larson, Robert L; Renter, David G; Sanderson, Mike W

    2011-03-01

    Although numerous diagnostic tests are available to identify cattle persistently infected (PI) with Bovine viral diarrhea virus (BVDV) in cow-calf herds, data are sparse when evaluating the economic viability of individual tests or diagnostic strategies. Multiple factors influence BVDV testing in determining if testing should be performed and which strategy to use. A stochastic model was constructed to estimate the value of implementing various whole-herd BVDV cow-calf testing protocols. Three common BVDV tests (immunohistochemistry, antigen-capture enzyme-linked immunosorbent assay, and polymerase chain reaction) performed on skin tissue were evaluated as single- or two-test strategies. The estimated testing value was calculated for each strategy at 3 herd sizes that reflect typical farm sizes in the United States (50, 100, and 500 cows) and 3 probabilities of BVDV-positive herd status (0.077, 0.19, 0.47) based upon the literature. The economic value of testing was the difference in estimated gross revenue between simulated cow-calf herds that either did or did not apply the specific testing strategy. Beneficial economic outcomes were more frequently observed when the probability of a herd being BVDV positive was 0.47. Although the relative value ranking of many testing strategies varied by each scenario, the two-test strategy composed of immunohistochemistry had the highest estimated value in all but one herd size-herd prevalence permutation. These data indicate that the estimated value of applying BVDV whole-herd testing strategies is influenced by the selected strategy, herd size, and the probability of herd BVDV-positive status; therefore, these factors should be considered when designing optimum testing strategies for cow-calf herds.

  16. Bovine immunoglobulin G does not have an inhibitory effect on diagnostic polymerase chain reaction utilizing magnetic bead extraction methods as demonstrated on the detection of Bovine viral diarrhea virus in dairy calves.

    Science.gov (United States)

    Chigerwe, Munashe; Crossley, Beate M

    2013-07-01

    The objective of the current study was to investigate if the presence of colostral-derived immunoglobulin G (IgG) in blood is an inhibitor of diagnostic polymerase chain reaction (PCR) for detection of Bovine viral diarrhea virus (BVDV). Eleven precolostral and 11 postcolostral blood samples in ethylenediamine tetra-acetic acid (EDTA) anticoagulant as well as serum samples were collected from 11 Holstein bull calves. Calves were fed 3 liters of colostrum once, by oroesophageal tubing. Postcolostral, blood, and serum samples were collected at 48 hr of age. Serum IgG concentrations were determined in the precolostral and postcolostral serum samples using radial immunodiffusion. The blood samples (precolostral and postcolostral) were spiked with BVDV, and 2 diagnostic PCR extraction methods were applied to each sample. The extraction and amplification efficiencies of the 2 PCR methods on the precolostral and postcolostral EDTA blood samples were evaluated. Two of the 11 calves had inadequate passive transfer of colostral immunoglobulins at 48 hr of age based on the serum IgG concentrations. All blood samples from calves were negative for BVDV prior to the spiking with the virus. Evaluation of the 2 different methods among 3 different virus concentrations demonstrated that there was no difference in extraction or amplification efficiency in precolostral and postcolostral samples. The results of this study suggest that bovine IgG is not an inhibitor of PCR used for detection of BVDV in cattle. The methods used in the current study are acceptable for PCR detection of BVDV in cattle.

  17. Development and immune efficacy assessment of an inactivated oil-emulsion vaccine against bovine viral diarrhea disease%牛病毒性腹泻油乳剂灭活苗的研制与免疫效果试验

    Institute of Scientific and Technical Information of China (English)

    赵月兰; 张磊; 王安忠; 左玉柱; 秦建华

    2009-01-01

    牛病毒性腹泻-黏膜病(bovine viral diarrheamucosal disease, BVD-MD) 是由牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)引起的牛传染病,其特征是消化道黏膜糜烂、坏死、胃肠炎和腹泻、免疫耐受与持续性感染、免疫抑制、先天性缺陷、母畜流产、产死胎和畸形胎等。1946年Olafson等[1]在美国首次报道本病,之后世界各国均有该病报道。

  18. Possibility for use of RT-PCR technique in establishing presence of bovine viral diarrhea virus in sperm of breeding bulls

    Directory of Open Access Journals (Sweden)

    Petrović Tamaš

    2005-01-01

    Full Text Available The bovine viral diarrhea (BVD virus is a significant health-economic pathogen in cattle which can be excreted and spread also through sperm of persistently or acutely infected bulls. Native sperm of 6 bulls, found to be negative to the BVD virus by isolating the virus and using the RT-PCR method, was experimentally infected with a tenfold dilution of the non-cytopathogen 22146 strain of the BVD virus with a titer of 105,5. This way, dilutions of the BVD virus from 10-1 to 10-6 (5 x 104 TCID/50 do 0,5 TCID/50 in 0.1 ml native sperm were obtained. From sperm infected in this way, the virus was reisolated on FTB cell culture in a microtiter plate with 96 pools in which each sample of the infected sperm was set up in three samples, and each of them was titrated to a dilution of 1:2 to 1:256. The presence of the BVD virus was proven using the technique of fluorescent antibodies in a second blind passage on FTB culture cells. For cell culture an extremely toxic effect of native sperm to a dilution of 1:64 was established. The BVD virus was reisolated from sperm in all three sperm samples with 5 x 104, 5 x 103 i 5 x 102 TCID/50, and it was not reisolated from sperm with 50, with 5, and with 0.5 TCID/50 BVD virus in 0.1 ml native sperm. At the same time, the presence of the BVD viral genome was proved using the RT-PCR method in the same samples of artificially infected native sperm of bulls. A positive re suit was established in native sperm with 5 x 104, 5 x 103, 5 x 102 and 50 TCID/50 BVD virus n 0.1 ml native sperm. The experiment proved that the RT-PCR method has advantages over the isolation of the BVD virus from samples of native sperm of bulls. These are: shortterm investigations (1 to 2 days and greater sensitivity (10 times bigger than the isolation of the virus. The isolation of the virus takes at least 10 days, and its greater sensitivity is primarily a result of the cyrotoxic effect of native sperm of bulls on cell culture.

  19. The internal initiation of translation in bovine viral diarrhea virus RNA depends on the presence of an RNA pseudoknot upstream of the initiation codon

    Directory of Open Access Journals (Sweden)

    Moes Lorin

    2007-11-01

    Full Text Available Abstract Background Bovine viral diarrhea virus (BVDV is the prototype representative of the pestivirus genus in the Flaviviridae family. It has been shown that the initiation of translation of BVDV RNA occurs by an internal ribosome entry mechanism mediated by the 5' untranslated region of the viral RNA 1. The 5' and 3' boundaries of the IRES of the cytopathic BVDV NADL have been mapped and it has been suggested that the IRES extends into the coding of the BVDV polyprotein 2. A putative pseudoknot structure has been recognized in the BVDV 5'UTR in close proximity to the AUG start codon. A pseudoknot structure is characteristic for flavivirus IRESes and in the case of the closely related classical swine fever virus (CSFV and the more distantly related Hepatitis C virus (HCV pseudoknot function in translation has been demonstrated. Results To characterize the BVDV IRESes in detail, we studied the BVDV translational initiation by transfection of dicistronic expression plasmids into mammalian cells. A region coding for the amino terminus of the BVDV SD-1 polyprotein contributes considerably to efficient initiation of translation. The translation efficiency mediated by the IRES of BVDV strains NADL and SD-1 approximates the poliovirus type I IRES directed translation in BHK cells. Compared to the poliovirus IRES increased expression levels are mediated by the BVDV IRES of strain SD-1 in murine cell lines, while lower levels are observed in human cell lines. Site directed mutagenesis revealed that a RNA pseudoknot upstream of the initiator AUG is an important structural element for IRES function. Mutants with impaired ability to base pair in stem I or II lost their translational activity. In mutants with repaired base pairing either in stem 1 or in stem 2 full translational activity was restored. Thus, the BVDV IRES translation is dependent on the pseudoknot integrity. These features of the pestivirus IRES are reminiscent of those of the classical

  20. Bovine viral diarrhea virus in free-ranging wild ruminants in Switzerland: low prevalence of infection despite regular interactions with domestic livestock

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    Casaubon Julien

    2012-10-01

    Full Text Available Abstract Background In the frame of an eradication program for bovine viral diarrhea (BVD in Swiss livestock, the question was raised whether free-ranging wildlife could threaten the success of this sanitary measure. Therefore, we conducted serological and virological investigations on BVD virus (BVDV infections in the four indigenous wild ruminant species (roe deer, red deer, Alpine chamois and Alpine ibex from 2009 to 2011, and gathered information on interactions between wild and domestic ruminants in an alpine environment by questionnaire survey. Results Thirty-two sera out of 1’877 (1.7%, 95% confidence interval [CI] 1.2-2.4 were seropositive for BVDV, and a BVDV1 sub genotype h virus was found in a seropositive chamois (0.05%, 95% CI 0.001-0.3. The seropositive animals originated from sub-alpine or alpine regions and significantly more seropositive red deer, chamois and ibex than roe deer were found. There were no statistically significant differences between sampling units, age classes, genders, and sampling years. The obtained prevalences were significantly lower than those documented in livestock, and most positive wild ruminants were found in proximity of domestic outbreaks. Additionally, BVDV seroprevalence in ibex was significantly lower than previously reported from Switzerland. The survey on interspecific interactions revealed that interactions expected to allow BVDV transmission, from physical contacts to non-simultaneous use of the same areas, regularly occur on pastures among all investigated ruminant species. Interactions involving cervids were more often observed with cattle than with small ruminants, chamois were observed with all three domestic species, and ibex interacted mostly with small ruminants. Interactions related to the use of anthropogenic food sources were frequently observed, especially between red deer and cattle in wintertime. Conclusions To our knowledge, this is the first report of BVDV RNA isolated from an

  1. Herd-level prevalence and risk factors for bovine viral diarrhea virus infection in cattle in the State of Paraíba, Northeastern Brazil.

    Science.gov (United States)

    Fernandes, Leise Gomes; Nogueira, Adriana Hellmeister de Campos; De Stefano, Eliana; Pituco, Edviges Maristela; Ribeiro, Cláudia Pestana; Alves, Clebert José; Oliveira, Tainara Sombra; Clementino, Inácio José; de Azevedo, Sérgio Santos

    2016-01-01

    Serological surveys based on a planned sampling on bovine viral diarrhea virus (BVDV) infection in Brazilian cattle herds are scarce. A cross-sectional study was carried out to determine herd- and animal-level seroprevalences and to identify risk factors associated with herd-level seroprevalence for BVDV infection in the State of Paraíba, Northeastern Brazil, from September 2012 to January 2013. The state was divided into three sampling strata, and for each stratum, the prevalence of herds infected with BVDV and the prevalence of seropositive animals was estimated by a two-stage sampling survey. In total, 2443 animals were sampled from 478 herds. A virus-neutralization test was used for BVDV antibody detection. A herd was considered positive when at least one seropositive animal was detected. The herd- and animal-level prevalences in the State of Paraíba were 65.5% (95% confidence interval (CI) = 61.1-69.7%) and 39.1% (95% CI = 33.1-45.6%), respectively. The frequency of seropositive animals per herd ranged from 10 to 100% (median of 50%). The risk factors identified were as follows: more than six calves aged ≤12 months (odds ratio (OR) = 3.72; 95% CI = 2.08-6.66), animal purchasing (OR = 1.66; 95% CI = 1.08-2.55), pasture rental (OR = 2.15; 95% CI = 1.35-3.55), and presence of veterinary assistance (OR = 2.04; 95% CI = 1.10-3.79). Our findings suggest that the implementation of control and prevention measures among farmers, with the aim of preventing dissemination of the agent in the herds, is necessary. Special attention should be given to addressing the identified risk factors, such as sanitary control prior to animal purchasing and to discourage the pasture rental, as well as to encourage the vaccination in the herds.

  2. Greater numbers of nucleotide substitutions are introduced into the genomic RNA of bovine viral diarrhea virus during acute infections of pregnant cattle than of non-pregnant cattle

    Directory of Open Access Journals (Sweden)

    Neill John D

    2012-08-01

    Full Text Available Abstract Background Bovine viral diarrhea virus (BVDV strains circulating in livestock herds show significant sequence variation. Conventional wisdom states that most sequence variation arises during acute infections in response to immune or other environmental pressures. A recent study showed that more nucleotide changes were introduced into the BVDV genomic RNA during the establishment of a single fetal persistent infection than following a series of acute infections of naïve cattle. However, it was not known if nucleotide changes were introduce when the virus crossed the placenta and infected the fetus or during the acute infection of the dam. Methods The sequence of the open reading frame (ORF from viruses isolated from four acutely infected pregnant heifers following exposure to persistently infected (PI calves was compared to the sequences of the virus from the progenitor PI calf and the virus from the resulting progeny PI calf to determine when genetic change was introduced. This was compared to genetic change found in viruses isolated from a pregnant PI cow and its PI calf, and in three viruses isolated from acutely infected, non-pregnant cattle exposed to PI calves. Results Most genetic changes previously identified between the progenitor and progeny PI viruses were in place in the acute phase viruses isolated from the dams six days post-exposure to the progenitor PI calf. Additionally, each progeny PI virus had two to three unique nucleotide substitutions that were introduced in crossing the placenta and infection of the fetus. The nucleotide sequence of two acute phase viruses isolated from steers exposed to PI calves revealed that six and seven nucleotide changes were introduced during the acute infection. The sequence of the BVDV-2 virus isolated from an acute infection of a PI calf (BVDV-1a co-housed with a BVDV-2 PI calf had ten nucleotides that were different from the progenitor PI virus. Finally, twenty nucleotide changes were

  3. 9 CFR 113.311 - Bovine Virus Diarrhea Vaccine.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Bovine Virus Diarrhea Vaccine. 113.311... Virus Vaccines § 113.311 Bovine Virus Diarrhea Vaccine. Bovine Virus Diarrhea Vaccine shall be prepared..., and immunogenic shall be used for preparing the production seed virus for vaccine production....

  4. Serologic study on Bovine Viral Diarrhea in dairy cattle from scale dairy farms in Liaoning%辽宁地区规模化奶牛场牛病毒性腹泻-黏膜病血清学调查

    Institute of Scientific and Technical Information of China (English)

    姚伟

    2015-01-01

    牛病毒性腹泻-黏膜病(Bovine Viral Diarrhea,BVD)是由牛病毒性腹泻-黏膜病病毒(Bovine Viral Di-arrhea Virus,BVDV)引起的接触性传染病,对奶牛业造成巨大的经济损失.为了摸清辽宁地区BVDV感染情况,本研究首次对辽宁省14个市27家规模化奶牛场共793份样品进行了BVD血清学检测,结果显示BVD平均阳性率为74.0%.进一步分析结果显示,BVD抗体阳性率存在养殖场规模和地区差异,该研究结果将为辽宁地区BVD的防控和净化提供基础.

  5. The Review of the Detection Technology for Bovine Viral Diarrhea-Mucosal Disease%牛病毒性腹泻-粘膜病的检测技术研究进展

    Institute of Scientific and Technical Information of China (English)

    赵丹; 王建华; 王万骞

    2014-01-01

    对牛病毒性腹泻-粘膜病的病毒分离、琼脂扩散试验、微量中和试验、免疫荧光技术、酶联免疫吸附试验、核酸杂交技术、聚合酶链反应等检测技术进行了综述,对7种技术运用于牛病毒性腹泻-粘膜病的检测作了简要介绍,为进一步研究牛病毒性腹泻病毒提供参考。%This paper gave a review of the diagnostic methods including virus isolation, agar gel immunodiffusion, virus micro-neutralization test, fluorescent antibody test, enzyme-linked immunosorbent assay, nucleic acid hybridization, polymerase chain reaction, introduced seven technology to detect bovine viral diarrhea virus, and provided references for the further research of bovine viral diarrhea virus.

  6. Development and Application of One Step RT-PCR Method for Detection of Bovine Viral Diarrhea Virus%牛病毒性腹泻病毒一步法RT-PCR检测方法的建立与应用

    Institute of Scientific and Technical Information of China (English)

    王春庆

    2013-01-01

    To establish a rapid bovine viral diarrhea virus (BVDV) pathogen detection methods, based on BVDV gene sequence in GenBank, we synthesized one pair of primers, established one step RT-PCR for BVDV detection. The method was used for infectious bovine rhinotracheitis virus (IBRV), classical swine fever virus (CSFV), bovine para influenza virus 3 (BPIV3) , all of the PCR results were negative, the sensitivity was 1 ng RNA. The established one step PCR method had good specificity, sensitivity, reproducibility, it could detect very low levels of BVDV quickly and accurately, it provided a kind of rapid, sensitive, specific and precise molecular biology detection method for pathogen detection and molecular epidemiology of material such as BVDV.%为建立一种快速检测牛病毒性腹泻病毒病原的方法,本研究根据GenBank上登录的牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)基因组序列,设计1对引物,建立了检测BVDV的一步法RT-PCR方法.该方法对牛传染性鼻气管炎病毒、猪瘟病毒、牛副流感病毒3型的扩增结果均为阴性,检测的敏感性达1 ng RNA.该一步法RT-PCR方法具有良好的特异性、敏感性、重复性,可以准确快速检测出极低含量的BVDV,将为BVDV的病原检测及分子流行病学调查等提供一种快速、灵敏、特异、准确的分子生物学检测方法.

  7. 宁夏部分地区牛病毒性腹泻的血清学检测%Serological Investigation of Bovine Viral Diarrhea in Some Regions of Ningxia

    Institute of Scientific and Technical Information of China (English)

    王晓亮; 王玉梅; 张玉玲; 吴亚文; 李知新; 张雯; 胡永浩

    2016-01-01

    [目的]掌握宁夏地区部分牛场犊牛病毒性腹泻病毒感染情况,为犊牛病毒性腹泻的净化和综合防治提供依据。[方法]从宁夏4个市9个县(区)的25个发病牛场(户)采集血清样品191份,采用ELISA方法,进行血清学调查。[结果]宁夏4个地级市犊牛病毒性腹泻的抗体阳性率高达64.92%,并有一定程度的抗原阳性,且主要发生在1月龄以内的犊牛。[结论]牛病毒性腹泻是具有重大经济意义的疾病,是今后宁夏养牛业应重点防范的疾病之一。%Objective]The aim of the study was to understand the infection of bovine viral diarrhea virus(BVDV)on some cattle farms in some parts of Ningxia and to provide the basis for cleaning up and comprehensive prevention and control of bovine viral diarrhea. [Methods] 191 bovine serum samples were collected from 25 cattle farms in 4 cities of Ningxia to detect the infection of BVDV by ELISA method. [Results]Results showed that BVDV antibody positive rate was as high as 64.92% in 4 cities of Ningxia,and antigen positive was detected in a certain degree. And most of BVD occurred in less than one month old calves. [Conclusion]Bovine viral diarrhea is one of the diseases with great eco-nomic signiifcance on cattle farms,and it was also the disease that should be prevented primarily for the cattle industry in Ningxia in the future.

  8. 内蒙古地区奶牛病毒性腹泻/黏膜病血清流行病学调查%Serum Epidemiology Investigation of Bovine Viral Diarrhea-mucosal Disease in Inner Mongolia Area

    Institute of Scientific and Technical Information of China (English)

    李智勇; 石顺利; 王艳杰; 陈德浩; 李平安; 关平原

    2014-01-01

    应用ELLSA试验对来自内蒙古地区17个大﹑中﹑小型奶牛场的2391份牛血清样品进行了牛病毒性腹泻/黏膜病抗体检测,并对其中222份抗体阴性牛应用ELISA试验进行牛病毒性腹泻/黏膜病的抗原检测。结果表明:17个奶牛场均检出BVDV抗体阳性,共检出阳性血清2125份,阳性率最高达100%,最低为46.8%,平均为88.9%。对14个奶牛场进行了BVDV抗原检测,在5个奶牛场检出BVDV抗原阳性,阳性率为3.6%(8/222)。表明内蒙古地区奶牛场普遍存在牛病毒性腹泻/黏膜病感染,感染率较高,并且牛群中存在持续性感染(PI)牛。%2 391 bovine serum samples were collected from 17 dairy farms of Inner Mongolia to detect the antibody level of bovine viral diarrhea-mucosal disease by ELISA. The antigen detection of bovine viral diarrhea-mucosal disease was made on 222 antibody-negative serum samples by ELISA. The results showed that positive BVDV antibody was detected in 17 dairy farms. And there were 2 125 positive serum samples,and the highest positive rate was 100%and the lowest positive rate was 46.8%and the average positive rate was 88.9%. BVDV antigens in 14 dairy farms were detected and that in 5 dairy farms were positive. The positive rate was 3.6%(8/222). It was shown that the bovine viral diarrhea generally existed in dairy farms of Inner Mongolia with higher infection rate. And persistent infection (PI) cattle were existed in cow herd.

  9. 牛病毒性腹泻病研究进展及防控建议%Analysis of Bovine Viral Diarrhea Prevalence and Comprehensive Prevention and Control in China

    Institute of Scientific and Technical Information of China (English)

    郝宝成; 梁剑平; 王学红; 郭文柱; 郭志廷; 杨贤鹏

    2013-01-01

    Bovine viral diarrhea virus (BVDV) is a pestivirus that is enzootic in most cattle populations throughout the world. This virus is present throughout the body of persistently infected cattle. In recent years, as more and more common the formation and development of large-scale farming. The jeopardized of Bovine viral diarrhea / mucosal disease for farming industry has become increasingly apparent. The BVDV's morphology of pathogens, genotype classification, hazards, diagnostic tests, the prevalence and prevention and control status were summarized in order to improve the prevention and control system and purification of eliminating the disease provide a reference for the country as soon as possible.%牛病毒性腹泻病由牛病毒性腹泻病毒引起,主要感染牛并引发疾病,呈世界性分布,可造成严重的经济损失.近年来,随着规模化养殖的发展,牛病毒性腹泻病对养殖业的危害日益显现.本文主要对牛病毒性腹泻病的病原特性、基因型分类、危害、诊断检测方法、国内流行情况和防控现状等进行综述,以期为国家尽快建立完善的防控体系和净化消除该病提供参考.

  10. Establishment and Application of a RT-PCR Assay for Detection of Bovine Viral Diarrhea Virus%牛病毒性腹泻病毒RT-PCR检测方法的建立及应用

    Institute of Scientific and Technical Information of China (English)

    张淑琴; 郭利; 张亭亭; 吴永旺; 武华

    2011-01-01

    根据GenBank中登录的牛病毒性腹泻病毒(BVDV)基因序列,设计合成了1对特异性引物,建立了检测BVDV的RT-PCR方法.通过对该方法的特异性、敏感性和重复性进行试验,结果显示,该方法可从BVDV标准毒株Oregon C24V中扩增出471 bp的特异性片段,而对猪瘟病毒、牛传染性鼻气管炎病毒、牛呼吸道合胞体病毒、牛副流感病毒、MDBK正常细胞的扩增结果均为阴性.经对标准毒株的细胞毒进行检测,其敏感度达10-1TCIDso/mL应用该方法对临床腹泻病牛各脏器样品进行检测,结果比病毒分离方法更为敏感,操作简便.表明建立的RT-PCR方法具有特异、灵敏、高效、快速的特点,可用于BVDV的临床检测及流行病学监测.%A reverse transcription PCR (RT-PCR) assay for detection of bovine viral diarrhea virus (BVDV)was established using a pair of specific primers of BVDV.This method could specifically amplify a 471 bp fragment from BVDV Oregon C24V strain, but not from classical swine fever virus (CSFV) ,infectious bovine rhinotracheitis virus (IBRV), bovine respiratory syncytial virus (BRSV) ,parainfluenza virus-3 (PI3) and normal MDBK cells.The organs were detected by RT-PCR and virus isolated that collected from a sick calf with bovine viral diarrhea (BVD) like syndrome.These results indicated that the established RT-PCR assay was specific, sensitive, efficient and rapid for detection, monitoring and epidemiological investigation of BVDV infection.

  11. Isolation and identification of bovine viral diarrhea virus from imported fetal bovine serum%进口胎牛血清中牛病毒性腹泻病毒的分离及鉴定

    Institute of Scientific and Technical Information of China (English)

    王竞晗; 李素; 何文瑞; 赵权; 仇华吉

    2016-01-01

    目的 对进口胎牛血清中的牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)进行分离及鉴定.方法 Trizol法提取待检胎牛血清中的病毒RNA后,利用套式RT-PCR扩增BVDV,并利用生物信息学Lasergene 7.0软件对其核苷酸序列进行同源性分析.取生长状态良好的MDBK细胞,按1%比例加入待检胎牛血清,收获F1代BVDVFBS2014,反复冻融3次后,再次接种MDBK细胞,连传4代.采用荧光定量RT-PCR法、间接免疫荧光试验及抗原捕获ELISA法对收获的病毒进行鉴定.结果 扩增产物大小均与预期相符,与已报道的BVDV-1NADL株的核苷酸序列一致性为92.2%,属于BVDV-1a亚型毒株;分离的病毒经鉴定为BVDV,命名为BVDV FBS2014,病毒基因拷贝数为104拷贝/μl,感染MDBK细胞可见特异性绿色荧光;各代次的病毒收获液中BVDV的基因拷贝数均大于104.92拷贝/μl,BVDV抗原检测结果均为阳性.结论 购买的进口胎牛血清中成功分离到可致MDBK细胞产生细胞病变的病毒,经鉴定所分离的病毒为BVDV,命名为FBS2014株,该病毒属于BVDV-1a亚型毒株.

  12. Caracterização preliminar de amostras do vírus da Diarréia Viral Bovina (BVDV isoladas no Brasil Preliminary characterization of brazilian isolates of bovine viral diarrhea virus (BVDV

    Directory of Open Access Journals (Sweden)

    Sônia A. Botton

    1998-04-01

    ção contra o vírus.This article reports the preliminary characterization of 19 Brazilian bovine viral diarrhea virus (BVDV isolates, regarding the biological, antigenic and molecular properties. Eleven viruses were isolated from bovine fetuses, six were obtained from blood of animals from herds with reproductive problems, and two were isolated from clinical cases of gastroenteric disease. The clinical cases affected young animals and were characterized by diarrhea, oronasal and digestive erosions and ulceration, and occasional digestive bleeding and vulvar petechial hemorrhage. Sixteen isolates (84.2%, including those obtained from fetuses and clinical cases, were of the non-cytopathic (ncp biotype. Replication of three isolates (15.8% in tissue culture was characterized by appearance of cellular vacuolation and progressive destruction of the monolayers. Analysis of these isolates after cloning revealed a mixed population of cytopathic (cp and non-cytopathic viruses. Analysis of viral polypeptides by SDS-PAGE followed by "Western immunoblot" revealed the production of the non-structural protein NS3/p80 in cells infected with the cp viruses. In contrast, generation of NS3/p80 was not observed in cells infected with the ncp isolates, which only expressed the precursor polypeptide NS23/p125. Analysis of reactivity with a panel of 15 monoclonal antibodies (MAbs revealed a marked antigenic variability among the isolates, mainly in the envelope glycoprotein E2/gp53. Although one MAb to this glycoprotein recognized 18 isolates (94.7%, the other nine E2/gp53 MAbs recognized zero to 57.9% of the isolates. The marked antigenic diversity observed among the brazilian BVDV isolates may have important implications on diagnosis and immunization strategies.

  13. Monitoring bovine viral diarrhea virus (BVDV infection status in dairy herds Monitoramento do estado de infecção pelo vírus da diarrhéia viral bovina (BVDV em rebanhos bovinos leiteiros

    Directory of Open Access Journals (Sweden)

    Francisco J. Diéguez

    2008-12-01

    Full Text Available This study was designed to assess the relationship between antibodies against bovine viral diarrhea virus (BVDV determined in the bulk tank milk (BTM and the within-herd seroprevalence. We also assessed the efficiency of measuring antibody levels in BTM samples to monitor BVDV infection status in a herd. In the 81 farms included in the study, BTM samples were obtained and blood samples withdrawn from all cattle older than one year. The infection status was then determined in serum and milk using a commercial blocking ELISA based on the detection of anti-p80 antibodies. Apart from these baseline serum and milk samples, another BTM sample was collected from each herd 9 months later, and a third BTM sample obtained 9 months after this. In these second and third milk samples, anti-BVDV antibodies were determined using the same ELISA kit. Statistical tests revealed good agreement between herd seroprevalences (% seropositive animals in the herd and the antibody levels detected in the BTM samples. During the 18 months of follow-up, the farms with persistently infected cattle at the study outset (14.8% of the herds showed a significant decrease in BTM antibody titers after virus clearance. Conversely, a significant increase in BTM antibody levels was observed in the herds infected with BVDV during the follow-up period. Our findings indicate that monitoring antibody levels in the BTM is a useful method of identifying changes in the BVDV infection status of a herd.Os objetivos do presente estudo foram avaliar a relação entre os níveis de anticorpos frente ao vírus da diarréia viral bovina (BVDV no tanque de leite e a prevalência de animais seropositivos em cada rebanho; e também avaliar a eficiência da medição dos níveis de anticorpos no tanque de leite como método de monitoramento do status de infecção frente ao BVDV. Nos rebanhos estudados, obtiveram-se amostras de soro de todos os animais com idade superior a um ano, assim como uma

  14. Establishment and Initial Application of RT- PCR for Quick Detectionof Bovine Viral Diarrhea Virus%牛病毒性腹泻病毒反转录PCR快速检测方法的建立与初步应用

    Institute of Scientific and Technical Information of China (English)

    李倬; 田镔; 李明生; 平玲

    2012-01-01

    根据GenBank中登录的牛病毒性腹泻病毒(BVDV)5'非编码区基因序列,设计合成了1对特异性引物,建立了检测BVDV的反转录PCR快速检测方法.通过对该方法的特异性、敏感性和重复性进行试验,结果显示,该方法从BVDV标准毒株Oregon C24V中扩增出267 bp的特异性片段,该方法重复性好,反应批内检测结果相同.与牛轮状病毒、牛冠状病毒、猪瘟病毒和F4新生牛肾传代正常细胞无交叉反应,具有高度的特异性,而且敏感性高,最低检出限为10~1.84 TCID50/mL.利用该方法对42份临床腹泻病牛疑似粪便样品进行了检测,结果检出7份阳性,而同时利用IDEXX公司抗原检测试剂盒检出阳性只有6份.表明,建立的该方法具有快速、敏感、特异等优点,是牛病毒性腹泻病毒病的临床诊断和流行病学调查的有力工具.%A reverse transcription- coupled PCR(RT- PCR) assay for quick detection of bovine viral diarrhea virus(BVDV) was established using a pair of specific primers based on the 5"UTR gene of BVDV published in GenBank. The results showed that this method could specifically amplify a 267 bp fragment from BVDV Oregon C24 V strain, reproducibility of this assay were reliable and the results were fully consistent. The specificity test proved that this assay had a high specificity which had no cross- reaction with bovine rotavirus, bovine coronavirus, classical swine fever virus and normal F4 NBPC cells. The assay also had good sensitivity, and the detection limit was up to 10-- 1.84 TCID50/mL. 7 positive of 42 samples from clinical diarrhea bovine were detected by RT- PCR and only 6 positive were detected by IDEXX diagnostic kit test at the same time. The results revealed that established RT - PCR assay possessed some advantages such as fast, sensitive and specific. It may be used for clinical diagnosis and the epidemiologic survey of bovine viral diarrhea/mucosal disease as a powerful tool.

  15. Excreção e transmissão do vírus da diarréia viral bovina por bezerros persistentemente infectados Shedding and transmission of bovine viral diarrhea virus by persistently infected calves

    Directory of Open Access Journals (Sweden)

    Sandra Arenhart

    2009-09-01

    /48 (16,6% foram soropositivos para anticorpos no dia 10, 26/48 (54,1% no dia 40 e 37/48 (77% haviam soroconvertido no dia 100, quando encerrou-se o monitoramento. Estes resultados demonstram que a viremia e excreção viral contínua em altos títulos por animais PI assegura a transmissão rápida do BVDV a animais mantidos em contato, sendo a transmissão notadamente mais rápida em condições intensivas e de alta densidade animal.Persistently infected (PI calves born to cows infected with non-cytopathic bovine virus diarrhea virus (BVDV represent the main reservoir of the virus in nature. We herein report an investigation on the patterns of virus shedding and transmission by five PI calves produced experimentally through inoculation of pregnant cows with Brazilian BVDV isolates. Five calves that survived intrauterine infection were born healthy, lacking neutralizing antibodies to BVDV and harboring virus in the blood. After weaning - and following the disappearance of colostral antibodies - PI calves were monitored for virus in serum and body secretions (ocular, oral, nasal and genital at weekly intervals for up to 150 days. For each animal, the virus titers in serum showed minor variations throughout the collections (with one exception that presented an increase late in infection, yet the titers varied widely among animals (from 10² to 10(6TCID50/mL. Virus shedding in secretions was detected steadily during all the observation period with minor titer variations for each particular animal. The highest titers were generally detected in nasal and ocular secretions (titers 10(4 to 10(6TCID50mL whereas genital and oral secretions usually contained low amount of virus (10² to 10³TCID50mL. To evaluate the kinetics of virus transmission by these animals, one PI was introduced on a group of 10 seronegative calves maintained with a high animal density simulating the conditions of an intensive management. All 10 contact calves seroconverted to BVDV by day 30. Another PI calf

  16. Analysis of mRNA expression for genes associated with regulatory T lymphocytes (CD25, FoxP3, CTLA4, and IDO) after experimental infection with bovine viral diarrhea virus of low or high virulence in beef calves.

    Science.gov (United States)

    Palomares, Roberto A; Hurley, David J; Woolums, Amelia R; Parrish, Jacqueline E; Brock, Kenny V

    2014-12-01

    Immunosuppression caused by bovine viral diarrhea virus (BVDV) has been associated with lymphocyte depletion, leukopenia and impairment of leukocyte function; however, no work has been done on the relationship between BVDV and regulatory T lymphocytes (Tregs). The objective of this study was to compare the mRNA expression of genes associated with Tregs (CD25, FoxP3, CTLA4, and IDO), after experimental infection of beef calves with low (LV) or high (HV) virulence BVDV. Thirty BVDV-naïve calves were randomly assigned to three groups. Calves were intra-nasally inoculated with LV (n=10, strain SD-1) or HV (n=10, strain 1373) BVDV or BVDV-free cell culture medium (control, n=10). Quantitative RT-PCR was used to determine the expression of target genes in tracheo-bronchial lymph nodes and spleen on day 5 post-infection. The mRNA expression of CD25 was up-regulated in tracheo-bronchial lymph nodes of LV (Pviral strains, or differences in viral infectivity of the host cells.

  17. Advance in Nonstructural Protein NS3 of Bovine Viral Diarrhea Virus%牛病毒性腹泻病毒非结构蛋白NS3的研究进展

    Institute of Scientific and Technical Information of China (English)

    侯佩莉; 孙涛; 何洪彬

    2012-01-01

    非结构蛋白NS3是致细胞病变型牛病毒性腹泻病毒的分子标记蛋白,它具有多种生物学功能.研究结果表明,NS3在病毒非结构蛋白的加工、成熟、基因组复制与转录过程中以及宿主细胞致细胞病变效应的产生中起重要作用.文章总结了NS3的生物学特性及其相关功能,为阐明NS3的致细胞病变作用机理奠定基础.%Nonstructural protein NS3 is considered a molecular marker for the cytopathic biotype of bovine viral diarrhea virus and has a variety of biological functions. It has been reported that NS3 played an important role in the processing of other nonstructural proteins, the release of mature particles, viral RNA replication, transcription and the generation of cytopathic effect in the host cell. This paper summarizes the biological properties of NS3 and its related functions, which lays the foundation for further study on the mechanism of NS3-induced cytopathic effect.

  18. 牛病毒性腹泻病毒EO基因原核表达载体的构建及表达%Construction of Prokaryotic Expression Vector of Bovine Viral Diarrhea Virus EO Gene and Its Expression

    Institute of Scientific and Technical Information of China (English)

    王璐; 郭春娟; 吴星星; 宫玉玲; 季新成; 冉多良

    2012-01-01

    C24V strains of bovine viral diarrhea virus (BVDV) were used to be inoculated into MDBK cells to extract viral RNA.to amplify BVDV-EO gene and the fragments obtained were connected with the pET-28a expression vector to be transformed into E. coli BL-21 to screen out the posetive clones. The results showed that identification of pET-28a-E0 prokaryotic expression vector was successfully constructed. The bacteria were collected after IPTG induction by SDS-PAGE and Western-blot identification, protein i-dentification results show that to be expressed in E. coli.%采用RT-PCR方法,利用牛病毒性腹泻病毒(BVDV)C24V株接种MDBK细胞,提取病毒RNA,扩增出BVDV-E0基因,将所得片段与pET-28a表达载体连接,转化至BL-21大肠杆菌中,筛选阳性克隆,鉴定后证明pET-28a-E0原核表达载体构建成功.经IPTG诱导后收集菌体进行SDS-PAGE和Western-blot鉴定,鉴定结果表明,目的蛋白在大肠杆菌中得以表达.

  19. Comparative study of three nucleic acid amplification assays for the detection of bovine viral diarrhea virus%牛病毒性腹泻病毒三种核酸检测方法的比较

    Institute of Scientific and Technical Information of China (English)

    范晴; 谢芝勋; 刘加波; 庞耀珊; 邓显文; 谢志勤; 谢丽基

    2012-01-01

    利用针对牛病毒性腹泻病毒(BVDV)的常规RT-PCR、实时荧光定量RT-PCR(real-time RT-PCR)和RT-环介导等温扩增技术(RT-LAMP)3种核酸检测方法对不同质粒浓度的样品和417份临床疑似样品进行了检测,以比较3种核酸检测方法的优越性。结果显示,3种核酸检测方法中,real-time RT-PCR和RT-LAMP一样敏感,均能检测到10拷贝/μL,常规RT-PCR只能检测到1×104拷贝/μL,但临床样品检测表明,常规RT-PCR方法敏感度低,会造成一部分漏检,RT-LAMP灵敏度高又会造成错检。综合比较3种方法后,推荐用RT-LAMP结合real-time RT-PCR,不仅节约成本,且结果更为准确可靠,可提高牛病毒性腹泻的检出率。%Ten fold dilution series samples and 417 clinical samples suspected with bovine viral diarrhea were used to evaluate three BVDV(bovine viral diarrhea virus)-specific detection methods:conventional RT-PCR,real-time RT-PCR and RT-LAMP(loop-mediated isothermal amplification).Results showed that the real-time RT-PCR and RT-LAMP had the same high sensitivity,and both of them could detect as lower as 10 copies/μL of samples,but the conventional RT-PCR had a lower sensitivity with its detection limit of 1×104 copies/μL.Clinically,a contrast analysis of these three methods showed that the RT-PCR had a higher undetected rate duo to its low sensitivity to clinical samples;the error rate of RT-LAMP was high due to its high sensitivity.These results suggested that the method of RT-LAMP combining with real-time RT-PCR is the recommended methods in clinical detection of BVDV,which is not only cost efficient but also accurate.

  20. Effects of long- or short-term exposure to a calf identified as persistently infected with bovine viral diarrhea virus on feedlot performance of freshly weaned, transport-stressed beef heifers.

    Science.gov (United States)

    Elam, N A; Thomson, D U; Gleghorn, J F

    2008-08-01

    A single experiment with a completely randomized design was conducted to evaluate the effects of long- or short-term exposure to a calf identified as persistently infected with bovine viral diarrhea virus (PI-BVD) on feedlot performance and carcass characteristics of freshly weaned, transport-stressed beef heifers. Two hundred eighty-eight heifers that had been vaccinated for BVD before weaning and transport were processed and given a metaphylactic antibiotic treatment at arrival and were fed common receiving, growing, and finishing diets for a 215-d period. Treatments were designed to directly or adjacently expose the cattle to a PI-BVD heifer. Directly exposed treatments were 1) negative control with no PI-BVD calf exposure (control), 2) PI-BVD calf commingled in the pen for 60 h and then removed (short-term exposure), and 3) PI-BVD calf commingled in the pen for the duration of the study (long-term exposure); and spatially exposed treatments were 1) negative control with no PI-BVD calf exposure (adjacent pen control), 2) PI-BVD calf commingled in the adjacent pen for 60 h and then removed (adjacent pen short-term exposure), and 3) PI-BVD calf commingled in the adjacent pen for the duration of the study (adjacent pen long-term exposure). Exposure to a PI calf transiently (60 h) or for the duration of the feeding period (215 d) did not affect (P > or = 0.25) final BW compared with heifers that were not exposed. Neither period nor overall DMI was affected (P > or = 0.37) by PI-BVD calf exposure, and no differences (P > or = 0.44) were observed between short- and long-term exposed heifers in the direct or spatially exposed groups. Likewise, total trial ADG was not affected (P > or = 0.36) and overall efficiency of gain (P > or = 0.19) was unaffected by PI-BVD calf exposure in the direct or spatially exposed groups. The results from this study suggest that exposing previously vaccinated, freshly weaned, transport- stressed beef calves to a calf that is persistently

  1. 猪源牛病毒性腹泻病毒SD0803株细胞传代研究%BIOLOGICAL CHARACTERISTICS OF PASSAGED ON MDBK CELLS BOVINE VIRAL DIARRHEA VIRUS OF PIG ORIGIN

    Institute of Scientific and Technical Information of China (English)

    孙春清; 邓宇; 张宏彪; 龙进学; 韦祖樟; 童光志; 袁世山

    2012-01-01

    To develop a Bovine viral diarrhea virus(BVDV) vaccine,the BVDV strain SD0803 of pig origin was passaged in MDBK cells for 40 times.Viral RNA was extracted from the 40th passage virus and 5 segments were amplified in RT-PCR.The complete genomic sequence was megaligned and compared with its parental virus using DNASTAR software.The results showed that the homology between the 40th passage and parental virus was 99.8% in nucleotides and 99.6 % in amino acids.Twenty three nucleotide mutations were identified,of which 15 were sense mutations.Amino acid mutations were mainly located on E2 and NS5B.To compare the growth characteristics between the parent virus and passaged viruses,supernatants were collected from infected MDCK cells at passages 1,10,20,30 and 40,and measured the amounts of released viral RNAs in RT-PCR.The multi-step growth curves showed that the parent virus and passaged viruses had high replication efficiency in MDBK cells,and shared similar growth properties.Some mutations that occurred during virus passages had no effect on the virus titers as determined by titration.%为研究猪源牛病毒性腹泻病毒(Bovine viral diarrhea virus,BVDV)生物学特性,将本实验室分离得到的SD0803毒株在马-达氏牛肾细胞(mardin-darby bovine kidney,MDBK)上连续传40代,提取第40代病毒基因组RNA,设计扩增及测序引物,用RT-PCR方法分5段扩增第40代病毒基因片段,并进行全长测序,利用DNASTAR软件进行序列拼接及分析,与亲代病毒SD0803序列比对分析;用Real-time PCR方法测定第1、10、20、30和40代细胞上清中的病毒复制效率,并绘制多步生长曲线。测序结果表明,第40代病毒与亲代病毒核苷酸序列的同源性为99.8%,氨基酸序列的同源性为99.6%,其中有23处发生核苷酸突变,14处为有义突变,氨基酸变化主要集中在E2和NS5B区域。多步生长曲线显示,传代病毒和亲本病毒均能在MDBK细胞上获得较高的复制效率,并

  2. Análise antigênica e molecular de amostras citopáticas do vírus da diarréia viral bovina Antigenic and molecular analysis of cytopathic isolates of bovine viral diarrhea virus

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    Fernando Luiz Tobias

    2000-03-01

    Mucosas.Seven cytopathic isolates of bovine viral diarrhea virus (BVDV isolated from clinical cases and blood of calves from herds with reproductive problems were analysed. All isolates contained a mixture of cytopathic (cp and noncytopathic (ncp viruses which were biologically cloned yielding pure populations of viruses of either biotype. The cp and ncp viruses obtained by cloning were characterized antigenically with a panel of monoclonal antibodies (MAbs and regarding to the expression of the non-structural polypeptide NS3. The analysis of MAb binding revealed two patterns of reactivity: 1. In five isolates, the cp and ncp viruses from the same isolate were antigenically very similar to each other, suggesting they represent true "pairs"of viruses. 2. Two isolates, however, yielded cp and ncp viruses antigenically different from each other. Western immunoblot analysis of non-structural polypeptides of ncp viruses revealed a unique band of reactivity, with a mass of approximately 125kDa, corresponding to the NS23 of the standard BVDV Singer strain. In addition to NS23, the cp viruses expressed a polypeptide of approximatelly 80kDa, corresponding to the NS3. The NS23 of two cp viruses displayed an altered migration in SDS-PAGE compared to the other viruses. In one virus, the NS23 had a molecular mass lower than expected whereas in other virus, two bands of reactivity were observed: one smaller and another bigger than the standard NS23, respectively. Our findings confirm previous results that cytopathic BVDV field isolates usually contain a mixture of viruses of both biotypes and that cytopathogenicity correlates with the expression of NS3. The isolation of cp viruses from the blood of clinically normal cattle, however, demonstrates that their occurrence is not restricted to cases of mucosal disease.

  3. Research Progress of Interaction Between Bovine Viral Diarrhea Virus and Host Cell%牛病毒性腹泻病毒与宿主细胞的相互作用研究

    Institute of Scientific and Technical Information of China (English)

    韩猛立; 黄新; 宋天增; 杨井泉; 薄新文; 钟发刚

    2012-01-01

    自1946年Olafson等首次在美国纽约州发现牛病毒性腹泻病毒(BVDV)以来,该病毒在世界各地广泛流行和传播,尤其对一些畜牧业发达国家的奶业和牛肉业造成了巨大损失.了解牛病毒性腹泻病的致病机理,研制出更加安全可靠有效的疫苗,对控制该病的发生和蔓延尤为重要.对BVDV的病原学、生物学特性以及对宿主细胞的相互作用等进行了阐述,以期为BVDV的预防及治疗奠定基础.%The bovine viral diarrhea virus (BVDV) has been prevalenced and disseminated worldwidely, since it was confirmed by Olafson in New York State in 1946. This virus caused large losses in dairy and beef industries, especially in the countries with advanced animal husbandry. Understanding the pathogenic mechanism and utilization of the safe and effective vaccine are very important for the control of occurrence and spread of this disease. The etiology, biological characteristics and interaction of host cell BVDV were described so as to provide the basic data for the prevention and therapy of BVDV.

  4. Acute infection with bovine viral diarrhea virus of low or high virulence leads to depletion and redistribution of WC1(+) γδ T cells in lymphoid tissues of beef calves.

    Science.gov (United States)

    Palomares, Roberto A; Sakamoto, Kaori; Walz, Heather L; Brock, Kenny V; Hurley, David J

    2015-10-15

    The objective of this study was to determine the abundance and distribution of γδ T lymphocytes in lymphoid tissue during acute infection with high (HV) or low virulence (LV) non-cytopathic bovine viral diarrhea virus (BVDV) in beef calves. This study was performed using tissue samples from a previous experiment in which thirty beef calves were randomly assigned to 1 of 3 groups: LV [n=10; animals inoculated intranasally (IN) with LV BVDV-1a (strain SD-1)], HV [n=10; animals inoculated IN with HV BVDV-2 (strain 1373)], and control (n=10; animals inoculated with cell culture medium). On day 5 post inoculation, animals were euthanized, and samples from spleen and mesenteric lymph nodes (MLN) were collected to assess the abundance of WC1(+) γδ T cells. A higher proportion of calves challenged with BVDV showed signs of apoptosis and cytophagy in MLN and spleen samples compared to the control group. A significantly lower number of γδ T cells was observed in spleen and MLN from calves in HV and LV groups than in the control calves (P<0.05). In conclusion, acute infection with HV or LV BVDV resulted in depletion of WC1(+) γδ T cells in mucosal and systemic lymphoid tissues at five days after challenge in beef calves. This reduction in γδ T cells in the studied lymphoid tissues could be also due to lymphocyte trafficking to other tissues.

  5. 牛病毒性腹泻病毒基因2型毒株的分离与鉴定%ISOLATION AND IDENTIFICATION OF BOVINE VIRAL DIARRHEA VIRUS-2 FROM THE ESTABLISED MDCK CELL LINE

    Institute of Scientific and Technical Information of China (English)

    张淑琴; 谭斌; 程世鹏

    2016-01-01

    本研究采用特异性RT-PCR从某实验室送检MDBK细胞中分离到1株牛病毒性腹泻病毒(Bovine viral diarrhea virus,BVDV),命名为BVDV-C1.通过免疫荧光、电镜观察和5'UTR序列测定及分子进化分析,结果表明其在MDBK细胞中无细胞病变产生,应用牛病毒性腹泻病毒2型特异性单克隆抗体检测,在感染细胞胞浆内呈现特异性荧光信号,而牛病毒性腹泻病毒1型特异性单克隆抗体未检测到荧光.电镜观察病毒粒子呈圆形,有囊膜,直径约为50 nm.5'UTR序列分析表明该毒株属于BVDV-2b基因亚型.BVDV-C1株的分离对进一步开展疫苗研发、流行病学调查以及致病机理等方面的研究具有重要意义.

  6. 牛病毒性腹泻病病毒荧光定量PCR检测体系的建立与评价%Establishment and evaluation of real-time PCR for detection of bovine viral diarrhea virus

    Institute of Scientific and Technical Information of China (English)

    史利军; 孙宇; 尹惠琼; 孙卫华; 吕茂民; 章金刚

    2009-01-01

    基于实时荧光定量PCR技术建立了一种有效地检测牛病毒性腹泻病病毒(Bovine viral diarrhea virus,BVDV)核酸的方法.对BVDV基因组进行同源比对,选取5'UTR区作为扩增目的区,经软件分析后设计特异扩增引物,扩增片段长度为203 bp.选用SYBR染料作为扩增时信号指示剂,经扩增曲线分析表明,建立的方法可有效地检测BVDV.检测体系可检测到10~2 copies/μL的样品拷贝数.故本研究建立的BVDV实时定量检测体系可用于易感动物,牛源血液生物制品及其他可能感染或污染BVDV样品的检测.

  7. 牛病毒性腹泻病二价弱毒活疫苗制备工艺研究%Study on Preparation Technique of Divalent Attenuated Virus Live Vaccine of Bovine Viral Diarrhea Virus

    Institute of Scientific and Technical Information of China (English)

    李海涛; 苗利光; 刘艳环; 朱言柱; 安亚雄; 王坤; 冯卓

    2013-01-01

    研究了BVDV2/JZ05-1和BVDV1/JZ05-3二价弱毒活疫苗的制备工艺.通过对BVDV2/JZ05-1和BVDV1/JZ05-3疫苗毒传代培养测定毒价后,试制出5批毒价分别是10.4TCID50/mL和105.6TCID50/mL的BVDV二价弱毒活疫苗,经检验符合试验设计要求.%The aim of this study was to explore the preparation technique of divalent attenuated virus live vaccine of bovine viral diarrhea virus 2/JZ05-1(BVDV2/JZ05-1) and BVDV1/JZ05-3.BVDV2/JZ05-1 and BVDV1/JZ05-3 were continuously cultivated in cell culture.The poison titor was examined.5 groups of divalent attenuated BVDV live vaccine were produced.The poison prices were 105.4TCID50/mL and 105.6TCID50/mL,respectively.The vaccines were examined and was within the range of the test design requirement.

  8. 抗CP型、NCP型牛病毒性腹泻病毒高免卵黄抗体的制备%Preparation of IgY against Cytopathic (CP) and Noncytopathic (NCP) Bovine Viral Diarrhea Virus (BVDV)

    Institute of Scientific and Technical Information of China (English)

    赵玉龙; 吾莫尔; 薄新文; 李岩; 钟发刚; 李娜; 库朝锋

    2009-01-01

    本研究采用致细胞病变(cytopathic,CP)型牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)标准毒和非致细胞病变(noncytopathic,NCP)型新疆优势毒株免疫产蛋鸡,用改良PEG法提取卵黄抗体(IgY),并对提取的IgY采用SDS-PAGE检测纯度,间接ELISA检测免疫后每隔7 d的抗体效价,并测定所得抗体对NCP型BVDV的中和效价.结果表明,用该法提取的IgY纯度较高;间接ELISA结果证明,经过4次免疫后,抗CP型BVDV的效价达到1:32000,抗NCP型BVDV的效价达到1:40000,3个月后再次检测,卵黄抗体效价未见明显下降.最后一次免疫14 d的抗体对NCP型BVDV的中和效价达到1×10-3.

  9. Fast detection and determination of bovine viral diarrhea virus in blood samples%进口奶牛病毒性腹泻病毒快速检测鉴定

    Institute of Scientific and Technical Information of China (English)

    赵秀玲; 闻伟刚; 黄素文; 张吉红; 杜华宏

    2007-01-01

    采用ELISA方法对3 073头奶牛全血进行牛病毒性腹泻病毒(Bovine viral diarrhea virus,BVDV)检测,结果发现编号a889的血样呈阳性反应,另外有3份血样(a126、a803、b1277)呈可疑反应;进一步的白细胞抽提物ELISA反应表明,a889血样呈阳性反应,其余呈阴性反应.对a889血样进行RT-PCR扩增,得到1条310 bp大小的特异性条带,与预期片段大小吻合;经序列同源性比较分析,确定a889血样中存在牛病毒性腹泻病毒.

  10. Failed detection of Bovine viral diarrhea virus 2 subgenotype a (BVDV-2a) by direct fluorescent antibody test on tissue samples due to reduced reactivity of field isolates to raw anti-BVDV antibody.

    Science.gov (United States)

    Yan, Lifang; Pace, Lanny W; Baughman, Brittany; Wilson, Floyd D; Zhang, Shuping; Zhang, Michael Z

    2016-03-01

    Bovine viral diarrhea virus 1 (BVDV-1) is associated with mild or subclinical infections, whereas BVDV-2 is frequently implicated in outbreaks of severe thrombocytopenia and acute fatal disease. In the present study, the carcass of a beef breed cow and tissue samples of a beef calf were received for laboratory diagnosis. Both animals exhibited severe clinical signs compatible with thrombocytopenia or hemorrhagic syndrome. Direct fluorescent antibody test (DFAT) failed to detect BVDV antigen in the tissue specimens of both cases. However, immunohistochemistry (IHC) revealed the presence of BVDV antigen in oral and esophageal mucosa and Peyer patches of the beef breed cow. Real-time reverse transcription polymerase chain reaction (real-time RT-PCR) detected BVDV-2 in selected tissues of both animals. Subsequently, BVDV was isolated from both cases and subjected to genetic and serologic characterizations. Mutations in the 5'-untranslated genomic region (5'-UTR) primer and probe binding sites and the E2 gene were associated with reduced efficiency of an established real-time RT-PCR assay and amino acid alterations in the E2 glycoprotein, respectively. Both viral isolates were classified by real-time RT-PCR and phylogenetic analysis as BVDV-2 subgenotype a. Unlike BVDV reference strains Singer and 125c, the isolates cross-reacted with anti-BVDV-1 and anti-BVDV-2 reference sera, indicating antigenic variations in field isolates. The isolates also showed reduced reactivity to porcine anti-BVDV antiserum (the raw serum used to produce BVDV DFA conjugate). In summary, data from the present investigation indicated that genetic and antigenic variations affected the performance of detection assays, especially DFAT, highlighting the need for regular evaluation and modification of BVDV tests.

  11. Development of one-step SYBR Green real-time RT-PCR for quantifying bovine viral diarrhea virus type-1 and its comparison with conventional RT-PCR

    Directory of Open Access Journals (Sweden)

    Lou Sai

    2011-07-01

    Full Text Available Abstract Background Bovine viral diarrhea virus (BVDV is a worldwide pathogen in cattle and acts as a surrogate model for hepatitis C virus (HCV. One-step real-time fluorogenic quantitative reverse transcription polymerase chain reaction (RT-PCR assay based on SYBR Green I dye has not been established for BVDV detection. This study aims to develop a quantitative one-step RT-PCR assay to detect BVDV type-1 in cell culture. Results One-step quantitative SYBR Green I RT-PCR was developed by amplifying cDNA template from viral RNA and using in vitro transcribed BVDV RNA to establish a standard curve. The assay had a detection limit as low as 100 copies/ml of BVDV RNA, a reaction efficiency of 103.2%, a correlation coefficient (R2 of 0.995, and a maximum intra-assay CV of 2.63%. It was 10-fold more sensitive than conventional RT-PCR and can quantitatively detect BVDV RNA levels from 10-fold serial dilutions of titrated viruses containing a titer from 10-1 to 10-5 TCID50, without non-specific amplification. Melting curve analysis showed no primer-dimers and non-specific products. Conclusions The one-step SYBR Green I RT-PCR is specific, sensitive and reproducible for the quantification of BVDV in cell culture. This one-step SYBR Green I RT-PCR strategy may be further optimized as a reliable assay for diagnosing and monitoring BVDV infection in animals. It may also be applied to evaluate candidate agents against HCV using BVDV cell culture model.

  12. 牛病毒性腹泻病毒BVDV2/JZ05-2毒株牛感染模型建立%Bovine Viral Diarrhea Virus BVDV2/JZ05-2 Strain Infection Modeling

    Institute of Scientific and Technical Information of China (English)

    冯卓; 刘艳环; 朱言柱; 李海涛; 钟宏鹏; 王坤; 张润祥; 苗利光

    2013-01-01

    试验用4~6月龄健康牛22头,共分5组,4个试验组5头/组,对照组2头。各试验组分别鼻内接种强毒107 FAID50/mL、106 FAID50/mL、105 FAID50/mL、104 FAID50/mL ,4mL/头,2mL/鼻孔。对照组鼻内接种MDBK细胞培养液冻融物,4mL/头,2mL/鼻孔。试验动物接种强毒后,每天观察实验动物的反应,包括精神状态、体温、食欲、黏膜颜色、呼吸频率、咳嗽、鼻腔分泌物及腹泻等;每天采集鼻拭子用于病毒分离。攻毒前2d、0d ,攻毒后隔天采血用于白细胞计数和病毒分离;试验于病毒接种后14d结束。试验过程中如有动物死亡,对死亡动物进行病理学检查。结果,106.6 FAID50/头BVDV2/JZ05-2攻击4~9月龄的牛,可引起被攻击牛有规律地体温升高和白细胞数量下降,同时可以从鼻拭子中分离到病毒。以这些指标作为感染模型可以有效地评价BVDV2疫苗免疫效果。%The aim of this experiment was to build bovine viral diarrhea virus BVDV 2/JZ05-2 strain infection model .Twenty two healthy cattle (4~6 months old ) were divided into 5 groups .In four experimental groups ,each group had 5 cattle .The control group had 2 cattle .The con-centrations of the BVDV2/JZ05-2 strain used in the experimental groups were 107 FAID50/mL ,106 FAID50/mL ,105 FAID50/mL ,104 FAID50/mL ,respectively .In control group ,the cattle were given MDBK nutrient solution .The volume of the sample was 4 mL through intranasal in dif-ferent groups .After inoculated the strong virus ,the clinical symptom was recorded everyday .The clinical symptom included psychosis ,tempera-ture ,appetite ,mucosa colour ,respiratory rate ,cough ,nasal cavity secreta and diarrhea .Nose swab was collected to separate the strong virus ev-eryday .The blood was obtained to determine white blood cell count and separate virus in -2 day ,0 day and 2 day .After the virus was inoculat-ed for 14 days ,the experiment was finished .If the dead cattle

  13. Identificação do vírus da Diarréia Viral Bovina tipo 2 (BVDV-2 no sul do Brasil Identification of bovine virus diarrhea virus type-2 (BVDV-2 in southern Brazil

    Directory of Open Access Journals (Sweden)

    Eduardo F. Flores

    2000-06-01

    Full Text Available Amostras do vírus da Diarréia Viral Bovina (BVDV, denominadas de BVDV tipo 2 (BVDV-2, foram inicialmente identificadas em surtos de BVD aguda e enfermidade hemorrágica e têm sido isoladas predominantemente na América do Norte. O presente artigo descreve dois casos de enfermidade gastroentérica/respiratória seguidos de isolamento e identificação de amostras de BVDV tipo 2 no sul do Brasil. Os vírus foram isolados de duas novilhas de diferentes rebanhos. Um dos animais apresentou enfermidade aguda, cursando com anorexia, atonia ruminal, diarréia escura ou muco-sanguinolenta, tenesmo e descarga nasal muco-purulenta. O outro animal desenvolveu enfermidade de longa duração (7 meses, caracterizada por crescimento retardado, anorexia, quadros recorrentes de diarréia, dermatite interdigital, hemorragias digestivas e genitais ocasionais, conjuntivite, artrite e pneumonia crônica. Congestão disseminada das mucosas, ulcerações extensivas e profundas na língua, palato e esôfago, áreas necróticas na mucosa do rúmen, áreas de congestão e ulcerações cobertas com fibrina no intestino delgado foram os achados mais proeminentes. Antígenos do BVDV foram demonstrados por imunohistoquí-mica no epitélio da língua, nos pulmões e em linfonodos mesentéricos. Amostras não-citopáticas do BVDV foram isoladas em cultivo celular a partir de leucócitos e do baço dos animais afetados e identificadas por imunofluorescência. Caracterização antigênica e análise filogenética desses isolados, e de outras duas amostras de BVDV isoladas de fetos coletados em matadouros, revelou tratar-se de BVDV tipo 2. A presença do BVDV tipo 2 na população bovina do Brasil possui um significado epidemiológico importante e pode ter conseqüências para o diagnóstico, estratégias de imunização e produção de vacinas.Highly virulent bovine viral diarrhea virus (BVDV isolates, named BVDV type-2 (BVDV-2, were initially identified in outbreaks of acute

  14. Effects of injectable trace minerals on humoral and cell-mediated immune responses to Bovine viral diarrhea virus, Bovine herpes virus 1 and Bovine respiratory syncytial virus following administration of a modified-live virus vaccine in dairy calves.

    Science.gov (United States)

    Palomares, R A; Hurley, D J; Bittar, J H J; Saliki, J T; Woolums, A R; Moliere, F; Havenga, L J; Norton, N A; Clifton, S J; Sigmund, A B; Barber, C E; Berger, M L; Clark, M J; Fratto, M A

    2016-10-01

    Our objective was to evaluate the effect of an injectable trace mineral (ITM) supplement containing zinc, manganese, selenium, and copper on the humoral and cell mediated immune (CMI) responses to vaccine antigens in dairy calves receiving a modified-live viral (MLV) vaccine containing BVDV, BHV1, PI3V and BRSV. A total of 30 dairy calves (3.5 months of age) were administered a priming dose of the MLV vaccine containing BHV1, BVDV1 & 2, BRSV, PI3V, and an attenuated-live Mannheimia-Pasteurella bacterin subcutaneously (SQ). Calves were randomly assigned to 1 of 2 groups: (1) administration of ITM SQ (ITM, n=15) or (2) injection of sterile saline SQ (Control; n=15). Three weeks later, calves received a booster of the same vaccine combination SQ, and a second administration of ITM, or sterile saline, according to the treatment group. Blood samples were collected on days 0, 7, 14, 21, 28, 42, 56, and 90 post-vaccination for determination of antibody titer, viral recall antigen-induced IFN-γ production, and viral antigen-induced proliferation by peripheral blood mononuclear cells (PBMC). Administration of ITM concurrently with MLV vaccination resulted in higher antibody titers to BVDV1 on day 28 after priming vaccination compared to the control group (P=0.03). Calves treated with ITM showed an earlier enhancement in PBMC proliferation to BVDV1 following vaccination compared to the control group. Proliferation of PBMC after BVDV stimulation tended to be higher on day 14 after priming vaccination in calves treated with ITM than in the control group (P=0.08). Calves that received ITM showed higher PBMC proliferation to BRSV stimulation on day 7 after priming vaccination compared to the control group (P=0.01). Moreover, calves in the ITM group also had an enhanced production IFN-γ by PBMC after stimulation with BRSV on day 21 after priming vaccination compared to day 0 (P<0.01). In conclusion, administration of ITM concurrently with MLV vaccination in dairy calves

  15. Cobaias como modelo para teste de vacinas inativadas contra o herpesvírus bovino tipo 1 e o vírus da diarréia viral bovina Guinea pigs as a model test of bovine herpesvirus type 1 and bovine viral diarrhea virus inactivated vaccines

    Directory of Open Access Journals (Sweden)

    Letícia Frizzo da Silva

    2007-08-01

    Full Text Available O presente trabalho relata a avaliação de cobaias como modelo para testes de imunogenicidade de vacinas inativadas contra o herpesvírus bovino tipo 1 (BoHV-1 e o vírus da diarréia viral bovina (BVDV. Para isso, cobaias (n=60 e bovinos (n=10 foram imunizados duas vezes, com intervalo de 28 dias, com uma vacina experimental contendo antígenos dos dois vírus, e testados para anticorpos neutralizantes 28 dias após a segunda dose. Os bovinos foram vacinados com a dose recomendada para a espécie (5mL; as cobaias foram distribuídas em seis grupos e imunizadas com doses fracionadas (0,005mL a 1,6mL. Os grupos de cobaias imunizadas com doses equivalentes a 1/16 (0,320mL e 1/8 (0,640mL da dose bovina desenvolveram títulos médios geométricos (GMTs de 6,46 e 7,56, respectivamente, estatisticamente semelhantes aos dos bovinos (GMT=8 (P>0,05. Uma alta correlação dose-resposta (R²=0,95 foi observada entre as doses vacinais e os títulos de anticorpos neutralizantes anti-BoHV-1 nos grupos de cobaias. Por outro lado, não foi possível o estabelecimento de uma dose vacinal que induzisse em cobaias uma resposta neutralizante anti-BVDV em níveis semelhantes à induzida em bovinos. Apenas as cobaias imunizadas com as doses maiores (0,640 e 1,6mL desenvolveram títulos neutralizantes de magnitude moderada (GMTs de 8 e 9, respectivamente, porém estatisticamente inferiores ao GMT dos bovinos (GMT=34,9 (PThe present study reports the use of guinea pigs as a model to study the immunogenicity of bovine herpesvirus type 1 (BoHV-1 and bovine viral diarrhea virus (BVDV inactivated vaccines. To this purpose, guinea pigs (60 and calves (10 were immunized twice with a 28 day interval with an experimental vaccine containing antigens of both viruses and tested for virus neutralizing (VN antibodies 28 days after the second dose. Calves were immunized with the recommended dose (5mL, while the guinea pigs were distributed in six groups and immunized with

  16. 牛病毒性腹泻病毒弱毒活疫苗免疫持续期的研究%The Efficacy of the Live Attenuated Bovine Viral Diarrhea Virus SM Vaccine Strain

    Institute of Scientific and Technical Information of China (English)

    张淑琴; 郭利; 王伟; 张亭亭; 吴永旺; 武华

    2012-01-01

    The study was designed to evalutate neutralizing(SN) antibodies against bovine viral diarrhea virus(BVDV) in the calves vaccinated with the live attenuated bovine viral diarrhea virus SM vaccine.A total of 30 healthy calves at weaning were randomly divided into two groups,with 15 calves in vaccinate group and 15 in control group.The attenuated BVDV SM vaccine strain was administrated via intramuscular injection on the neck at a single dose and challenged with a virulent BVDV-JL virus by nasal spray.The blood sample were collected at different time points for monitoring SN antibody titers.The results indicated that the SN antibody titers in the vaccinated calves maintained at a relative high level until 12 month post-vaccination,and the vaccinated calves were effectively protected from the challenge,as detected by the white blood cell count(WBC) and,virus shedding.therefore,the duration of immunity of the vaccine was at lease for 9 months.%为检测牛病毒性腹泻病毒(BVDV)弱毒活疫苗在免疫牛体内抗体产生及其消长规律,评价弱毒疫苗的保护效力,并确定免疫持续期,本试验对免疫试验牛每头颈部肌肉接种BVDV SM株弱毒疫苗104.5TCID50,监测血清抗体效价,进行免疫持续期的确定。在疫苗免疫后的6个月、9个月和12个月,分别抽取5头免疫组和5头对照组牛,采用BVDV-JL强毒株进行攻毒试验,每头牛攻毒剂量为6×107.0TCID50/mL。结果显示疫苗免疫后12个月时血清中和抗体效价仍维持在1∶1048以上。攻毒结果显示,在3个不同时间点进行强毒攻击后,免疫组所有动物白细胞数量都没有下降也没有分离到病毒,而对照组动物白细胞数下降均超过30%,6个月和9个月时动物血清中均能分离到病毒,而12个月对照组动物由于年龄大,没有分离到病毒,因此暂定此疫苗的免疫持续期为9个月。

  17. 牛病毒性腹泻弱毒活疫苗免疫持续期的研究%The Efficacy of the Live Attenuated Bovine Viral Diarrhea Virus SM Vaccine Strain

    Institute of Scientific and Technical Information of China (English)

    张淑琴; 郭利; 王炜; 张亭亭; 吴永旺; 武华

    2012-01-01

    The study was designed to evaluate neutralizing (SN) antibodies against bovine viral diarrhea virus (BVDV) in the calves vaccinated with the live attenuated bovine viral diarrhea virus SM vaccine. A total of 30 healthy calves at weaning were randomly divided into two groups, with 15 calves in vaccinate group and 15 in control group. The attenuated BVDV SM vaccine strain was administrated via intramuscular injection on the neck at a single dose and challenged with a virulent BVDV-JL virus by nasal spray. The blood samples were collected at different time points for monitoring SN antibody titers. The results indicated that the SN antibody titers in the vaccinated calves maintained at a relative high level until 12 month post-vaccination, and the vaccinated calves were effectively protected from the challenge, as detected by the white blood cell count (WBC) and virus shedding. Therefore, the duration of immunity of the vaccine was at lease for 9 months.%为检测牛病毒性腹泻病毒(BVDV)弱毒活疫苗在免疫牛体内抗体产生及其消长规律,评价弱毒疫苗的保护效力,并确定免疫持续期,本试验对免疫试验牛每头颈部肌肉接种BVDV SM株弱毒疫苗104.5TCID50/头,监测血清抗体效价,进行免疫持续期的确定.在疫苗免疫后的6、9和12个月分别抽取5头免疫组和5头对照组牛采用BVDV-JL强毒株进行攻毒试验,每头牛攻毒剂量为6×107.0 TCID50/mL.结果显示疫苗免疫后12个月时血清中和抗体效价仍维持在1∶1048以上,攻毒结果显示3个时间点强毒攻击后,免疫组所有动物白细胞数量都没有下降也没有分离到病毒,而对照组动物白细胞数下降均超过30%,6和9个月动物均分离到病毒,而12个月对照组动物由于年龄大,没有分离到病毒,因此暂定此疫苗的免疫持续期为9个月.

  18. Isolation and identification of a bovine viral diarrhea virus isolate from yak in Qinghai Province%青海牦牛病毒性腹泻病毒的分离与鉴定

    Institute of Scientific and Technical Information of China (English)

    张芳芳; 王光华; 郑福英; 宫晓炜; 周继章; 吴润; 邱昌庆

    2011-01-01

    采集青海省泽库县某牧户疑似牦牛病毒性腹泻病牛的样品,将RT-PCR检测为阳性的样品处理后接种MDBK细胞,并盲传至第10代,检测每一代细胞中牛病毒性腹泻病毒的E0基因。通过分离株接种细胞后产生的病变效应观察、免疫荧光试验、电镜观察、RT-PCR扩增以及序列分析鉴定了该病毒。结果表明,盲传的每代细胞中均可检测到E0基因;接毒后的细胞在盲传至第7代时出现明显的细胞病变;免疫荧光试验中能观察到细胞内发出的特异性荧光;经浓缩、纯化的病毒在电镜下呈直径为40~60nm的病毒粒子,将其命名为QHZK株。对克隆的E0基因测序后提交至GenBank(登录号:JF927789),并将获得的序列与其他国内外分离株的E0序列比对,同源性为73.6%~98.2%;系统进化分析表明该分离株属于牛病毒性腹泻病毒1b亚型。%Samples were collected from yak with suspected bovine viral diarrhea on a yak farm in Zeku County,Qinghai Province.MDBK cell lines were inoculated with RT-PCR positive specimens,followed by blind passage culture for 10 generations,and the E0 gene of bovine viral diarrhea virus(BVDV) was detected from the cell cultures of each generation.The isolates were cultured in MDBK cell lines,and identified by cytopathic effect(CPE),direct immunofluorescence assay(DIFA),electron microscopy(EM),RT-PCR and sequence analysis. The results indicated that the E0 gene was detected from the cell cultures of each generation,and typical CPE was observed from cell cultures of the seventh generation.Specific fluorescence in cell cultures can be see in DIFA.Virions of 40-60nm were observed under EM,and was named QHZK strain.The obtained E0 gene sequence was submitted to GenBank with the accession number:JF927789.Alignment with other 9 strains of BVDV available in GenBank showed a homology of 73.6%-98.2% for nucleotide sequence.The phylogenetic analysis indicated that the isolated

  19. Expression and Characterization of E0 Protein of Bovine Viral Diarrhea Virus%牛病毒性腹泻病毒E0基因的原核表达与活性检测

    Institute of Scientific and Technical Information of China (English)

    严伟行

    2013-01-01

    A fragment of about 650 bp was amplified by RT-PCR technique with specific primers based on BVDV genome sequence. Then the target fragment was directionally cloned into pET32a vector. After identifying with enzyme cut and sequencing, the recombinant plasmid was transformed into E. coli BL21(DE3). The recombinant protein E0 was expressed in inclusion body form in E. coli after induction with IPTG. After purification, the purified protein was analyzed by Western blotting, the results showed that the purified recombinant protein retained better antigenicity.%参考牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)基因组序列设计1对引物,扩增出650 bp的E0基因片段.将目的片段克隆至pMD18 T载体,经酶切鉴定获得阳性重组质粒并对其进行测序.将E0基因定向亚克隆到pET32a表达载体中,酶切及测序鉴定正确后,转化BL21表达菌,经IPTG诱导得到了以包涵体形式表达的重组蛋白.重组蛋白经亲和层析法纯化后,免疫印迹检测证明纯化的重组蛋白具有良好的活性.

  20. 广西牛病毒性腹泻病毒GX4分离株全基因测序及序列分析%Identification and sequence analysis of bovine viral diarrhea virus in Guangxi province

    Institute of Scientific and Technical Information of China (English)

    范晴; 谢芝勋; 谢志勤; 刘加波; 庞耀珊; 邓显文; 谢丽基; 罗思思

    2015-01-01

    牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)属于瘟病毒属,是养牛业中常见的病原体.通过MDBK细胞首次从广西分离获得1株牛源牛病毒性腹泻病毒,命名为GX4.设计引物对其全基因组进行扩增,获得其全基因序列,测序结果表明,GX4全基因组为12 218 bp,编码3 898个氨基酸,GenBank登录号JN704144.1.对全基因序列进行分析,根据其5'-UTR,确定GX4属于BVDV-1b基因亚型;与BVDV其他参考毒株的比对发现,GX4与巴西分离株IBSP4ncp同源性最高,核苷酸同源性为94.4%,推导氨基酸同源性为96.2%,并且P125基因无外源序列插入,属于非细胞病变型.分子流行病学的结果,揭示了目前流行株的差异性,为该病的防控措施的制定提供参考.

  1. 基因2型牛病毒性腹泻病毒新疆及山东分离株的鉴定%IDENTIFICATION OF BOVINE VIRAL DIARRHEA VIRUS GENOTYPE 2 ISOLATES OF XJ-04 AND SD-06

    Institute of Scientific and Technical Information of China (English)

    任敏; 朱礼倩; 焦海宏; 林燕清; 陶洁; 蒋颖; 刘振华; 王传彬; 朱国强

    2009-01-01

    将疑似为基因2型牛病毒性腹泻病毒(Bovine viral diarrhea virus genotyp02,BVDV-2)感染阳性的不同地区源病料样品接种马-达氏牛肾细胞(Mardin-Darby bovine Kidney,MDBK)单层细胞,进行病毒分离培养和传代.通过BVDV-2型特异性RT-PCR检测结果表明,共分离到2株细胞源BVDV-2病毒,命名为XJ-04和SD-06分离株.间接免疫荧光试验表明,上述分离的细胞源BVDV-2病毒能被鼠源BVDV-2重组E2蛋白抗血清或鼠抗BVDV-2单克隆抗体特异性识别,产生特异性胞内荧光.XJ-04和SD-06分离株分别盲传至13代和8代,光镜下观察到病毒感染的MDBK细胞内空泡化、细胞脱落、呈现拉网状的典型细胞病变.2株细胞源BVDV-2病毒感染的MDBK细胞制备超薄切片,在透射电镜下,细胞内质网中观察到约60 nm大小的病毒粒子.经差速离心、20%蔗糖垫底超速离心提纯的病毒粒子,经磷钨酸染色,负染电镜下观察到病毒有囊膜、粒子大小约60 nm的病毒颗粒.

  2. Effects of on-arrival versus delayed clostridial or modified live respiratory vaccinations on health, performance, bovine viral diarrhea virus type I titers, and stress and immune measures of newly received beef calves.

    Science.gov (United States)

    Richeson, J T; Kegley, E B; Gadberry, M S; Beck, P A; Powell, J G; Jones, C A

    2009-07-01

    Stress, commonly associated with weaning, marketing, and shipment of feeder cattle, can compromise immune function, and vaccine administration during immunosuppression may reduce vaccine efficacy and calf growth. Four treatments were compared in a 2 x 2 factorial arrangement to evaluate the effect of on-arrival (d 0) vs. delayed (d 14) administration of clostridial (CLOS) and respiratory (RESP) vaccines on health, performance, bovine viral diarrhea virus (BVDV) antibody titers, and physiological immune measurements of high-risk, newly received calves. Crossbred bull and steer calves (n = 263) were weighed (239 +/- 1.2 kg), stratified by sex, and randomly assigned to vaccination treatment: 1) arrival CLOS, arrival RESP (ACAR); 2) arrival CLOS, delayed RESP (ACDR); 3) delayed CLOS, arrival RESP (DCAR); and 4) delayed CLOS, delayed RESP (DCDR). Body weight and blood samples were collected on d 0, 14, 28, 42, and 56. Average daily gain did not differ (P > or = 0.34), averaging 0.98, 0.93, 0.95, and 0.91 kg/d for ACAR, ACDR, DCAR, and DCDR, respectively, for the entire 56-d trial. Vaccination timing did not affect morbidity (P > or = 0.23); however, there tended to be a CLOS timing effect (P = 0.07) and RESP timing effect (P = 0.09) on days to initial bovine respiratory disease (BRD) treatment. Average days to initial BRD treatment were less for ACAR (6 +/- 0.8 d) compared with DCDR (8 +/- 0.8 d; P = 0.01). Greater white blood cell counts were observed for DCDR than ACDR (P = 0.01), with ACAR and DCAR being intermediate. Serum cortisol concentrations were greater on d 0 than d 14 (P < 0.01) or d 28 (P = 0.01) but no treatment x day interaction (P = 0.21) was observed. Timing of RESP administration affected (P = 0.001) serum BVDV type I titers, with greater (P < 0.01) levels in calves receiving RESP vaccine on arrival. Delaying CLOS or RESP vaccination did not affect BW gain or morbidity in high risk, newly received stocker calves. Calves administered RESP vaccine on d 0

  3. Progress on Bovine Viral Diarrhea Virus and Its Evolution%牛病毒性腹泻病毒及其进化研究进展

    Institute of Scientific and Technical Information of China (English)

    郭燕; 王新华; 钟发刚

    2007-01-01

    牛病毒性腹泻病毒(Boyine viral diarrhoea virus,BVDV)在我国发现有20多年,而在世界范围来讲已有近100年的历史.由于牛病毒性腹泻病毒造成奶牛生产性能下降、繁殖障碍、持续感染等,是导致集约化奶牛场严重亏损的重要原因.且牛病毒性腹泻导致的黏膜病致死率几乎100%,已经严重阻碍养牛业的发展,但目前我国对牛病毒性腹泻还没有好的防控措施.文章对牛病毒性腹泻的发生历史总结,便于在牛病毒性腹泻病毒的诊断和预防中参考.

  4. Bovine viral diarrhea diagnostic: immunohistochemistry standardization for routine Padronização da técnica de imunoistoquímica para o diagnóstico etiológico de rotina da diarréia bovina a vírus

    Directory of Open Access Journals (Sweden)

    G.I. Andrade

    2005-10-01

    Full Text Available The immunohistochemistry standardization for bovine viral diarrhea virus (BVDV diagnostic was described. The formalin-fixed tissue samples from a heifer with mucosal disease were used as positive control. The validation of the first phase results was performed using samples from an aborted fetus and a calf infected with reference strains of BVDV. The best results were seen using monoclonal antibodies and a commercial kit consisting of labelled streptavidin biotin (LSAB reagents and the diaminobenzidine (DAB substrate-chromogen reagent. The immunohistochemistry demonstrated to be an useful method for routine diagnosis for the controll and detection of BVDV infection.

  5. PREVALENCE OF BOVINE VIRAL DIARRHOEA VIRUS IN WEST BENGAL, INDIA

    Directory of Open Access Journals (Sweden)

    Reshmi Ghosh

    2015-12-01

    Full Text Available Bovine Viral Diarrhea (BVD is one of the most economically important diseases in cattle. The present study was undertaken to diagnose the persistently infected (PI animals by AntigenELISA and Reverse Transcriptase PCR using serum samples from organized farms as well as rural areas of West Bengal. The results showed that out of 964 serum samples tested 07 (0.73% was positive for BVDV by Antigen-ELISA. For further confirmation, RNA was extracted from the positive samples and RT-PCR was performed with 5' UTR specific primers which showed 294 bp amplicons. This finding showed circulation of BVDV in cattle in West Bengal, India.

  6. 牦牛病毒性黏膜病病毒P125基因的克隆及序列分析%Cloning and Bioinformatics Analysis of the P125 Gene of Bovine Viral Diarrhea Virus Strain Yak

    Institute of Scientific and Technical Information of China (English)

    刘亚刚; 孙凯; 王研; 王盼盼; 胡炳峰; 王文伯

    2012-01-01

    According to the sequence data of BVDV strain published by GenBank, five set of primers were designed and used to amplify P125 gene of BVDV strain yak by the method of PCR. A specific 3475 bp DNA segment was amplified, which was cloned into pMD19-T vector. The positive recombinant clone was identified by plasmid PCR and restriction enzyme digestion. Compared with BLAST, the P125 gene of BVDV strain yak exhibited the highest homology with strain NADL, but they only shared 63. 4% , showing that the P125 gene of BVDV strain yak had great variation. This might be the virus to adapt to yak and the ecological environment of the plateau,or the virus might also have an independent source of genetic.%试验参考GenBank中发表的牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)毒株基因组序列设计了5对引物,利用PCR扩增出预期的3475 bp目的片段;将扩增产物连接至pMD19-T载体,经质粒PCR鉴定及双酶切鉴定获得阳性重组质粒并进行核苷酸序列测定.经BLAST同源性分析表明,牦牛BVDV P125基因与BVDV-Ⅰ型的NADL毒株P125基因的同源性为63.4%.系统分析结果表明,牦牛P125基因与BVDV标准毒株P125基因在遗传进化与亲缘关系上均较远,这可能是因为环境诱变的结果或该毒株具有新的遗传衍化来源.

  7. Preparation and characterization of antisera against recombinant E2 protein of bovine viral diarrhea virus%牛病毒性腹泻病毒E2蛋白的多克隆抗体制备及鉴定

    Institute of Scientific and Technical Information of China (English)

    高欲燃; 朱远茂; 康健; 史鸿飞; 李娇; 任宪刚; 冯军科; 于作; 薛飞

    2011-01-01

    To prepare the polyclonal antibody against recombinant E2 protein of bovine viral diarrhea virus (BVDV) in rabbits, E. coli BL21 (DE3) was transformed with the recombinant plasmid pET30a-E2. The recombinant E2 protein was expressed in E. coli after cultivation and induction. The purified recombinant E2 protein could be recognized by specific BVDV antisera in western blot. Then the purified recombinant E2 protein was used as antigen for immunizing rabbit to prepare polyclonal antibody against the recombinant E2 protein. The result of virus nentralization test showed that the titer of the polyclonal antibody to neutralize BVDV was 1:2,048. The polyclonal antibody against the recombinant E2 protein of BVDV also had highly reactivity and specialty in immunofluorescence analysis and western blot. The polyclonal antibody against recombinant E2 protein of BVDV developed in rabbits could be used in detection of BVDV in China and provided a good basis for establishing an ELISA for detecting of E2 protein of BVDV.%为制备牛病毒性腹泻病毒(BVDV)重组E2蛋白的兔源多克隆抗体,本研究利用表达BVDV E2蛋白的重组质粒pET30a-E2转化E.coli BL21(DE3),经诱导表达获得重组E2蛋白.Western blot检测显示纯化蛋白能够与BVDV参考阳性血清反应.以纯化的重组E2蛋白免疫新西兰白兔制备多克隆抗体,病毒中和试验测定其中和效价为1:2 048,间接免疫荧光和western blot试验表明其具有良好的反应性和特异性.本研究制备的BVDV重组E2蛋白兔源多克隆抗体可应用于BVDV的检测,同时为进一步建立检测BVDV E2蛋白的ELISA方法奠定基础.

  8. Analysis of The Causes of Bovine Viral Diarrhea Virus Epidemic Transmission in Beijing Area%北京地区牛病毒性腹泻病毒流行传播的原因分析

    Institute of Scientific and Technical Information of China (English)

    张俊杰; 李栋梁; 马翀; 曹杰

    2013-01-01

    在对北京地区20个奶牛群进行抗体水平检测和全群抗原筛查的基础上,将牛群中牛病毒性腹泻病毒流行状况同牛群在3年内是否引入过动物进行相关性比较.结果显示,20个牛群中,引入过动物的8个牛群中后备牛群血清抗体平均水平为80.0%,抗原筛查的总体阳性率为0.61%;未引入过动物的12个牛群中后备牛血清抗体平均水平为65.5%,抗原筛查的总体阳性率为0.15%,两组数据之间差异极显著,表明牛群在3年内是否引进过动物对牛群持续感染牛的分布影响极显著,动物引进可能是造成BVDV在北京地区流行传播的主要原因之一.%Based on the detection of the antibody level and the whole group antigen screening in 20 dairy herds in Beijing area,the prevalence of bovine viral diarrhea virus in herds was compared with the introduction of animal into herds in 3 years.Within the 20 herds,the 8 herds that had introduced animal from other herds have an average serum antibody level of 80% in replacement group,the positive rate of antigen screening is 0.61%; the average serum antibody level of replacement group in 12 herds that had not introduced animal was 65.5%,the overall positive rate of antigen screening is 0.15%,the difference between two sets of data is extremely significant.The results show,whether introduced animals into a herd within 3 years influence the distribution of PI animals in herds significantly,and the introduction of animal may be one of the main causes of BVDV epidemic transmission in Beijing area

  9. Establishment of Nested-PCR for detecting bovine viral diarrhea viruses from pig%猪源牛病毒性腹泻病毒套式PCR检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    邓宇; 蔺涛; 张荣; 丛雁方; 张建武; 袁世山; 文心田; 郑浩

    2012-01-01

    根据GenBank中BVDV-1、BVDV-2、CSFV及新出现瘟病毒的基因序列,设计2对特异引物,建立了能鉴别诊断BVDV与CSFV的Nested-PCR检测方法;确定其方法的特异性、敏感性、稳定性。建立的Nested-PCR能从BVDV标准毒株、猪源BVDV毒株中扩增198bp特异性片段,对猪瘟病毒、猪繁殖与呼吸综合征病毒检测结果为阴性。结果表明,该方法具有较好特异性;其敏感性可检测到0.195fg RNA模板量;经重复性试验表明该方法具有良好稳定性。初步应用检测表明,猪BVDV阳性率较高,达36.0%;牛血清及其制品、猪瘟活疫苗BVDV污染严重。本试验建立的Nested-PCR具有敏感、特异和稳定等特点,可用于临床诊断和流行病学调查。%The study aimed at establishing Nested-PCR to differentiate pig bovine viral diarrhea viruse(BVDV) and classical swine fever virus(CSFV),for clinical diagnosis,epidemiological survey and quality control of classical swine fever live vaccine.Two pairs of primers were designed according to the genomic sequences of BVDV-1,BVDV-2,CSFV and a new member of the pestivirus genus,and a Nested-PCR were developed for the differentiation of pig BVDVs and CSFV.This PCR assay could respectively amplify a 198 bp fragment from BVDV NADL,pig BVDV,but not from CSFV,porcine reproductive and respiratory syndrome virus (PRRSV).Sensitivity was determined as 0.195 fg RNA.And in the field trail of 100 pig specimen,36% of them were positive.The results showed that the Nested-PCR with high sensitivity and specificity could provide a new and alternative tool for the detection of pig BVDVs and CSFV.

  10. Epidemiology of bovine viral diarrhea virus.

    Science.gov (United States)

    Houe, H

    1995-11-01

    Prevalence studies around the world show that BVDV is widespread in most cattle raising countries. There are significant differences, however, in prevalence between areas, probably the result of differences in cattle population structure and management practice. Direct contact with PI animals is probably the most important method of transmission of infection; however, field studies have shown that some limited spread of infection also occurs in the absence of PI animals. This may be due to contact with acutely infected animals or contact with other species infected with BVDV. Different ways of indirect transmission such as contaminated needles and equipment have been proven experimentally, and indirect transmission is considered to have some importance. If a PI animal is introduced directly into a dairy herd, most animals will be infected within a few months. On many occasions, however, a herd gets infected by other means than direct introduction of PI animals. In these cases, the infection is often spread to only a few animals after which the infections stops. The infection is then reinforced when PI animals are born. Slow and hence prolonged spread of infection in herds without PI animals also has been described, but the mechanism is not fully understood. Family lines of PI animals delivering PI calves are fairly common and can cause the infection to continue for several years. The clinical manifestations, acute BVDV, reproductive disorders, birth of malformed, weak and undersized calves, unthrifty PI animals, and mucosal disease often appear within certain periods. Large variation, however, can occur between herd outbreaks due to variation in virulence of the BVDV strain, housing of the cattle, and variation in transmission patterns. The extensive transmission of infection from PI animals makes different surveillance methods possible. Thus testing of a screening sample of a few young stock of antibodies and determination of antibody titer in bulk milk will often give good indication of presence of PI animals in herds not using BVDV vaccines. In herds using killed vaccine, determination of antibody titers among few young stock can show the presence of PI animals. The high incidence of infection combined with all the different damages that are seen after BVDV infection cause huge economical losses, which on a national level in the UK and Denmark (i.e., areas with widespread occurrence of infection) has been calculated as between 7 and 27 million pounds (between $11 and $42 million) per million calvings. Epidemiologic studies are important as a basis for selection of control strategy. Because of the variation in epidemiology between geographic areas, evaluation of a control strategy in an area preferentially should be based on epidemiologic studies in the same area.

  11. The Situation of Bovine Viral Diarrhea Virus Pollution in Swine Fever Vaccine and Research Progress of its Detection Method%牛病毒性腹泻病毒在猪瘟疫苗中的污染情况及其检测方法的研究进展

    Institute of Scientific and Technical Information of China (English)

    毛晓娜; 缪芬芳; 季伟

    2013-01-01

    牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)和猪瘟病毒(classical swine fever virus,CSFV)同属黄病毒科瘟病毒属,猪瘟疫苗中污染BVDV可引起免疫失败.但由于两者在病毒粒子结构、基因组结构和抗原特性等方面均很接近,在血清学上存在交叉反应,因此难以检测猪瘟疫苗中污染的BVDV.文章对BVDV在猪瘟疫苗中的污染情况和检测方法进行了论述,旨在为猪瘟疫苗污染BVDV的检测提供理论基础.%Bovine viral diarrhea virus(BVDV) and classical swine fever virus(CSFV) both belong to Pestivirus, Flaviviri-dae. It can cause immune failure by BVDV pollution in swine fever vaccine. But because of the similar of their virus structure, genome structure, and antigen characteristics, they have cross reaction. So it is difficult to detect the BVDV pollution in swine fever vaccine. In this paper, we have discussed the situation of BVDV pollution in swine fever vaccine and its detection methods, aiming to provide theoretical basis for detecting BVDV pollution in swine fever vaccines.

  12. RT-PCR em pools de soros sangüíneos para o diagnóstico da infecção aguda e de animais persistentemente infectados pelo vírus da diarréia viral bovina RT-PCR in pools of bovine blood serum to detect acute infection and persistently infected animals with bovine viral diarrhea virus

    Directory of Open Access Journals (Sweden)

    D. Pilz

    2007-02-01

    Full Text Available Utilizou-se a técnica da RT-PCR para a detecção da região 5' UTR do genoma do vírus da diarréia viral bovina (BVDV em pools de soros sangüíneos provenientes de um rebanho, constituído por 226 animais, que apresentava distúrbios da reprodução. A partir das amostras individuais de soro e de acordo com a categoria dos animais e o número de animais por categoria foram formados 10 pools (A a J de soros. A primeira avaliação revelou a amplificação de um produto com 290pb nas reações referentes aos grupos D (35 vacas e H (25 bezerros lactentes que, após o desmembramento em amostras individuais, resultou na identificação de 11 vacas lactantes e 12 bezerros em amamentação positivos. Para a identificação de animais persistentemente infectados (PI entre os 23 positivos na primeira avaliação, realizou-se a segunda colheita de soros sangüíneos, três meses após. A RT-PCR das amostras individuais de soro revelou resultado positivo em cinco bezerros. Em dois, foi possível isolar o BVDV em cultivo de células MDBK. A especificidade das reações da RT-PCR foi confirmada pelo seqüenciamento dos produtos amplificados a partir do soro de uma vaca com infecção aguda, de um bezerro PI e das duas amostras do BVDV isoladas em cultivo celular. A utilização da RT-PCR em pools de soros sangüíneos demonstrou ser uma estratégia rápida de diagnóstico etiológico e de baixo custo tanto para a detecção de infecção aguda quanto de animais PI.The 5' untranslated region of the bovine viral diarrhea virus (BVDV genome was detected by RT-PCR assay in pools of blood sera samples collected from a cattle herd (n=226 animals with reproductive failures. Based on the classes of animal and the number of animals per class, the individual blood serum samples were distributed in 10 sera pools (A to J. During the first evaluation a 290bp amplicon was amplified in reactions from groups D (35 cows and H (25 sucking calves. The individual analysis

  13. Muestreo predial pequeño para predecir una infección activa por virus diarrea viral bovina (VDVB en planteles lecheros de la Xª Región de Chile A small herd sample to predict an active infection with bovine viral diarrhea virus (BVDV in dairy herds of X Region, Chile

    Directory of Open Access Journals (Sweden)

    G. REINHARDT

    2002-01-01

    Full Text Available La diarrea viral bovina está distribuida mundialmente y la mayoría del ganado es seropositivo, aunque la seroprevalencia varía entre predios y grupos de edad. Los animales con infección persistente son los transmisores más eficientes, pasan desapercibidos y son la fuente más importante para la perpetuación de la infección. Este trabajo entrega los resultados del análisis serológico de una muestra predial de 10 animales entre 6 y 12 meses de edad de 44 predios lecheros de la X Región de Chile. Se constató que en 35 planteles (79.5% existiría infección activa con virus diarrea viral bovina, pues al menos 6 de los 10 sueros estudiados presentaron anticuerpos. De esta manera, mediante una muestra pequeña de animales jóvenes es posible predecir, con certeza, la presencia de infección activa en los plantelesBovine viral diarrhea virus (BVDV has a worldwide distribution and most cattle are seropositive, although the prevalence may vary among herds and among different age groups. Persistently infected (PI animals are the most efficient transmitters of infection often remaining unnoticed in the herds thus, becoming the most important source to perpetuate the infection. In each of the 44 dairy herds studied from X Region, Chile, ten young stock aged 6 _ 12 months were tested for antibodies against BVDV. In 35 dairy herds (79.5% BVDV active infection was predicted because at least 6 over ten sera were antibody carriers. Thus, based on few blood samples, herds with PI animals and herds without PI animals could be distinguished with a high degree of accuracy

  14. Molecular detection of bovine coronavirus in a diarrhea outbreak in pasture-feeding Nellore steers in southern Brazil.

    Science.gov (United States)

    Ribeiro, Juliane; Lorenzetti, Elis; Alfieri, Alice Fernandes; Alfieri, Amauri Alcindo

    2016-03-01

    Worldwide diarrhea outbreaks in cattle herds are more frequently detected in calves being that diarrhea outbreaks in adult cattle are not common. Winter dysentery (WD) is a bovine coronavirus (BCoV) enteric infection that is more reported in Northern hemisphere. Seasonal outbreaks of WD in adult cattle occur mainly in dairy cows. WD has not been described in beef cattle herds of tropical countries. This study describes the molecular detection of BCoV in a diarrhea outbreak in beef cattle steers (Nellore) raised on pasture in Parana, southern Brazil. During the outbreak, the farm had about 600 fattening steers. Watery and bloody diarrhea unresponsive to systemic broad-spectrum antibiotic therapy reveals a morbidity rate of approximately 15 %. The BCoV N gene was identified in 42.9 % (6/14) of the diarrheic fecal samples evaluated by semi-nested polymerase chain reaction (SN-PCR) technique. Other enteric microorganisms occasionally identified in adult cattle and evaluated in this study such as bovine groups A, B, and C rotavirus, bovine viral diarrhea virus, bovine torovirus, aichivirus B, and Eimeria sp. were not identified in the fecal samples. To the best knowledge of the authors, this is the first description of the BCoV diagnosis in fecal samples collected in a diarrhea outbreak in adult beef cattle grazing in the grass in a tropical country.

  15. 苦马豆素抗牛病毒性腹泻病毒的研究%Study on Inhibitory Effect of the Swainsonine from Alkaloid of Astragalus strictus Grah.Ex Bend on Bovine Viral Diarrhea Virus

    Institute of Scientific and Technical Information of China (English)

    郝宝成; 武凡琳; 邢小勇; 项海涛; 温峰琴; 王学红; 权晓弟; 胡永浩; 梁剑平

    2014-01-01

    [目的]疯草是中国西部影响草地生态和畜牧业健康发展的毒草之一。主要有豆科棘豆属和黄芪属有毒植物等,在冬季牧草严重缺乏的时节,牛羊等牲畜被迫采食后可引起中毒和生产性能下降,甚至死亡,每年给我国草地生态和畜牧业造成的直接经济损失达几十亿元,是危害最严重的毒草。目前,我国西部天然草地动物因疯草中毒死亡所造成的经济损失仍持续剧增。苦马豆素被认为是引起动物疯草中毒的主要毒性成分。由于疯草分布广泛,资源丰富,如何改善草地生态,合理开发利用疯草成为研究的课题和方向。近几十年来,随着对疯草研究的深入和扩展,人们发现苦马豆素不仅具有良好的抗肿瘤效果,还可作为免疫调节剂、抗HIV及扩散抑制剂、抗病毒和细胞保护剂等药物使用。目前,国内在对苦马豆素抗肿瘤、调节机体免疫方面研究较热,但是,对苦马豆素在抗病毒活性、作用机理方面的研究尚无相关报道。为了探讨茎直黄芪中生物碱苦马豆素(swainsonine,SW)抗牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)的作用机制,明确其体外抗病毒作用效果,为抗BVDV的药物筛选提供参考依据。[方法]利用细胞培养技术,采用CPE观察法和MTT比色法相结合的方法测定不同浓度SW对牛肾原代细胞(madin-darby bovine kidney cells,MDBK)的毒性作用,确定药物的安全浓度和TD 50,并分别采用先加药后感染病毒、先感染病毒后加药、药毒作用2 h后加入、感染病毒同时给药后再加药四种作用方式,检测不同浓度SW对BVDV入侵的阻断作用、复制的抑制作用、直接杀伤作用和综合作用,并计算不同作用方式下的治疗指数。[结果]结果显示:SW浓度小于0.256μg·mL-1范围内对MBDK细胞无毒性作用,在大于0.512μg·mL-1浓度下MBDK细胞表

  16. Vaccination of cattle against bovine viral diarrhoea

    NARCIS (Netherlands)

    Oirschot, van J.T.; Bruschke, C.J.M.; Rijn, van P.A.

    1999-01-01

    This brief review describes types and quality (efficacy and safety) of bovine viral diarrhoea virus (BVDV) vaccines that are in the market or under development. Both conventional live and killed vaccines are available. The primary aim of vaccination is to prevent congenital infection, but the few va

  17. Técnica rápida de neutralização viral para a detecção de anticorpos contra o vírus da Diarréia Viral Bovina (BVDV no leite A rapid virus-neutralization test for detection of antibodies against bovine viral diarrhea virus (BVDV in milk

    Directory of Open Access Journals (Sweden)

    Scherer Charles Fernando Capinos

    2002-04-01

    positivos através do leite enviado rotineiramente para contagem de células somáticas (CCS, reduzindo significativamente os custos com a coleta individual, transporte e teste de amostras.The identification of bovine viral diarrhea virus (BVDV positive herds through detection of antibodies in milk may viabilize large scale control/eradication programs. With this objective, the virus neutralization test (VN was adapted to detect BVDV antibodies in milk. The adaptation consisted of a reduction in the time of incubation followed by detection of viral antigens in the indicator cells by immunofluorescence (IFA and allowed readings at 24 hours. The rapid virus neutralization test (RVN was initially tested in 1,335 serum samples, showing a 93.7% sensitivity and 91.1% agreement with the traditional VN. The RVN was also used to test 423 bovine sera that were toxic for cell culture in the traditional VN test, detecting 316 (74.7% positive samples. Testing of matched serum and milk samples from BVDV seropositive cows showed that the VNR can detect antibodies in the milk of cows with serum neutralizing titers as low as 10. Anti-BVDV neutralizing activity was detected in milk of 97.4% (191/196 of cows with serum titers ³320; in 92.9% (79/85 of cows with titers of 160; in 88% (59/67 of cows with serum titers of 80. The frequency of BVDV antibodies in milk was 76.9% (40/52 for cows with serum titers of 40; 61.3% (19/31 for cows with titers of 20 and 33.3% (10/30 for cows with serum titers of 20. These results demonstrate that the RVN test is adequate for detecting BVDV antibodies in milk, mainly in cows having moderate to high serum titers, and therefore may be used for testing bulk milk samples to identify herds with viral activity. The use of this test may viabilize large scale programs for control/eradication of BVDV infection. It allows to assay a large number of samples and identify positive herds through testing milk routinely submitted for somatic cell counts (SCC, reducing costs

  18. O polietilenoglicol aumenta a penetração do vírus da diarréia viral bovina, do vírus da estomatite vesicular e do vírus sincicial respiratório bovino em células de cultivo Polyethylene glycol increases the penetration of bovine viral diarrhea virus, vesicular stomatitis virus and bovine respiratory syncytial virus in cultured cells

    Directory of Open Access Journals (Sweden)

    Renata Dezengrini

    2009-06-01

    bovine enveloped viruses in culture cells. Penetration efficiency was measured by counting the number of viral plaques produced in bovine kidney cells (MDBK. The addition of 5% PEG (molecular weight 6.000 to the viral inoculum containing 100 TCID50 mL-1 (tissue culture median infectious dosis of each virus, during adsorption for 2h at 37°C, resulted in a significant increase in the number of plaques for bovine viral diarrhea virus (BVDV (increase of 3.4-fold, vesicular stomatitis virus (VSV (2.2-fold and bovine respiratory syncytial virus (BRSV (1.5-fold. The addition of 5% PEG to the inoculum of bovine herpesviruses 1, 2 and 5 (BoHV-1, BoHV-2 and BoHV-5 did not increase the number of viral plaques. On the other hand, PEG produced a reduction in the number of plaques by bovine parainfluenza virus (bPI-3V (1.4-fold. Furthermore, the addition of 5% PEG produced a 10- to 1000-fold increase in the sensitivity of BVDV detection in the serum of three persistently infected calves; and doubled the sensitivity of detection of BRSV in nasal secretions of two experimentally infected sheep. These results demonstrate that PEG enhances the efficiency of infection by BVDV, VSV and BRSV in cultured bovine cells and therefore may be used to increase the sensitivity of virus detection in clinical samples (viral isolation, and/or to increase virus titers in cell cultures.

  19. 牛病毒性腹泻病毒荧光标记单克隆抗体的制备及鉴定%Preparation and identification of fluorescein-labelled monoclonal antibodies against bovine viral diarrhea virus

    Institute of Scientific and Technical Information of China (English)

    徐树兰; 李少英; 赵俊; 武华

    2011-01-01

    To obtain fluorescein-labelled monoclonal antibodies against different genotypes of bovine viral diarrhea virus(BVDV),specific monoclonal antibodies(McAbs) against BVDV type Ⅰ(BVDV-Ⅰ),type Ⅱ(BVDV-Ⅱ) or both were prepared.And in order to obtain large amount of the McAbs for fluorescein-labelling,BALB/c mice were intraperitoneally injected with hybridomas BV1,BV2,or BV12,respectively.Ascites for the three McAbs were purified with caprylic acid-ammonium method,and then identified by immunocompetence,protein concentration,and purity test.The purified McAbs were conjugated with fluorescein isothiocyanate(FITC) by stirring method.The F/P values of the three labeled McAbs were between 2.0 and 4.0,and their DFA titers to the respective BVDV strains were≥1∶200.Result of antigen cross reaction showed fine specificity of labeled-antibodies to their respective types of BVDV.In BVDV titration TCID50 test,fluorescent assay using the labeled McAbs can detect 1 TCID50 unit of BVDV virion.Preliminary diagnostic test to clinical samples showed that the fluorescence antibodies detected ≥95% of the positive samples,with a 100% concordance rate between the self-made and the imported fluorescence antibodies.Therefore,the three fluorescence antibodies established in the present study will be very useful for the identification of BVDV genotype,detection of NCP-BVDV and differential diagnosis of BVDV and CSFV,and worthy to be researched and developed in future.%为制备可以区分牛病毒性腹泻病毒(BVDV)基因型的荧光标记单克隆抗体,分别利用分泌抗牛病毒性腹泻Ⅰ型病毒(BVDV-Ⅰ)、牛病毒性腹泻Ⅱ型病毒(BVDV-Ⅱ)、牛病毒性腹泻病毒(BVDV-Ⅰ和BVDV-Ⅱ)单克隆抗体的杂交瘤细胞株BV1、BV2、BV12,腹腔接种BALB/c小鼠,大量制备单克隆抗体,用辛酸-硫酸铵法提纯,经活性、浓度、纯度检测合格后,利用搅拌法与异硫氰酸荧光素(FITC)偶联,制备成3

  20. Genetic characterization of Brazilian bovine viral diarrhea virus isolates by partial nucleotide sequencing of the 5'-UTR region Caracterização genética de amostras brasileiras do vírus da diarréia viral bovina através do seqüenciamento parcial da Região 5'UTR

    Directory of Open Access Journals (Sweden)

    Adriana Cortez

    2006-12-01

    Full Text Available Nineteen isolates of bovine viral diarrhea virus (BVDV from Brazil were genetically characterized through partial nucleotide sequencing and analysis of the 5'UTR region. The isolates were grouped as BVDV-1 (11/19, BVDV-2 (6/19 or "atypical" pestivirus (2/19. Among the BVDV-1, eight isolates were classified as subgenotype BVDV-1a, whereas most (4 out of 6 BVDV-2 belonged to subgenotype 2b. Two isolates from aborted fetuses were not classified into any genetic group, being considered atypical BVDVs. Genetic diversity among Brazilian BVDV isolates may be responsible for vaccination and diag-nostic failure and therefore may influence the control strategies for BVDV infection in the country.Dezenove amostras do vírus da diarréia viral bovina (BVDV foram caracterizadas geneticamente através do seqüenciamento parcial de nucleotídeos da Região 5'UTR. As amostras foram agrupadas em BVDV-1 (11/19, BVDV-2 (6/19 e num terceiro grupo de amostras denominadas "atípicas" (2/19. Das onze amostras genotipadas como BVDV-1, oito amostras foram sub-genotipadas como BVDV-1a, enquanto que a maioria (4/6 das amostras de BVDV-2 foi agrupada como BVDV-2b. Duas amostras provenientes de fetos bovinos abortados foram classificadas como atípicas, não BVDV-1 e 2. A presença da diversidade genética de BVDV detectada nas amostras estudadas pode ser responsável por falhas vacinais e de diagnóstico e deve influenciar nas estratégias de controle do BVDV aplicadas nas diferentes regiões brasileiras.

  1. Análisis de técnicas de recuperación antigénica para la detección inmunohistoquímica del virus de la diarrea viral bovina Analysis of antigen retrieval techniques for the immunohistochemical detection of the Bovine Viral Diarrhea virus

    Directory of Open Access Journals (Sweden)

    M.R. Marini

    2006-12-01

    Full Text Available En el presente trabajo se evaluaron diferentes métodos de recuperación antigénica para permitir la inmunodetección del virus de la Diarrea Viral Bovina (vDVB en materiales fijados con formol. Se analizó el efecto de la aplicación de digestión proteolítica (ficina, tripsina, pepsina, proteinasa K y su combinación con tratamiento en un horno microondas empleando diferentes concentraciones y tiempos de incubación para el anticuerpo primario. El tratamiento más efectivo fue el realizado con proteinasa K, utilizando diluciones de 1:30 para los anticuerpos primarios, con incubación durante toda la noche a temperatura ambiente. Concluimos que la fijación formólica afecta la antigenicidad del vDVB pero estos efectos pueden ser revertidos por digestión proteolítica específica permitiendo la utilización de la inmunohistoquímica como técnica de rutina para el diagnóstico de la enfermedad.In the present work different methods of antigen retrieval were evaluated to allow the immunodetection of the Bovine Viral Diarrhea virus (BVDv in materials fixed with formol. The effect of the application of proteolytic digestion was analyzed (ficin, trypsin, pepsin, proteinase K and its combination with treatment in a microwave oven using different concentrations and times of incubation for the primary antibody. The most effective treatment was the one carried out with proteinase K, using 1:30 dilution for the primary antibodies, with overnight incubation at room temperature. We conclude that the formol fixation affects the antigenicity of the BVDv but these effects can be reverted by specific proteolytic digestion allowing the use of the immunohistochemical routine technique for the diagnosis of the disease.

  2. Utilización del Método de Elisa en la detección directa de antígeno de virus diarrea viral bovina en muestras de suero sanguíneo de bovinos Use of an ELISA test in the direct diagnosis of viral antigens of Bovine Viral Diarrhea Virus (BVDV in bovine blood serum samples

    Directory of Open Access Journals (Sweden)

    G. REINHARDT

    2003-01-01

    Full Text Available El virus de la Diarrea Viral Bovina (VDVB es un agente infeccioso importante del ganado bovino y está distribuido ampliamente en el mundo, produciendo pérdidas económicas sustanciales en la producción pecuaria. La principal fuente de contagio de los animales susceptibles está en las secreciones y excreciones de los animales infectados persistentes e inmunotolerantes (PI, condición que se produce en la etapa gestacional, específicamente antes de los 120 días de preñez, período en que el sistema inmune del embrión aún no se desarrolla adecuadamente. El propósito de este estudio fue aplicar la utilización de un método inmunoenzimático (ELISA-antígeno para detectar la presencia de animales PI en planteles lecheros de la Xª Región de Chile, a partir de muestras de suero sanguíneo. Para ello se examinaron 335 sueros de bovinos provenientes de 9 predios. Los resultados obtenidos indican que el 33.3% de los planteles analizados presentaron algún animal PI y que a nivel de prevalencia intrapredial, ella varió entre 0.7 y 1.0%. Se concluyó que el método utilizado permite detectar animales PI en forma rápida y sencilla, pudiendo utilizarse en gran cantidad de muestrasBVDV is an important virus of cattle worldwide that induces to substantial economic losses in dairy farms. The major source of infection are secretions and excretions of immunotolerant and persistent infected cattle. That condition is adquired during the early gestational period. The scope of this communication is to inform the use of an ELISA test to detect BVDV persistent infected bovine using blood serum samples in cattle of 9 dairy farms from the Xth. Region of Chile. The results indicated that 0.3% of the serum samples were positive to the ELISA test, and 33.3% of the dairy herds with persistently infected animals. It is concluded that this method diagnose persistently infected cattle, and is very easy to manipulate therefore, is possible to test many animals in

  3. Detection of bovine viral diarrhea virus from suspicious hog cholera cases in Jiangxi%江西部分地区猪瘟疑似病例中牛病毒性腹泻病毒感染情况的初步调查

    Institute of Scientific and Technical Information of China (English)

    戴益民; 张文波; 刘文峰; 闫学艳; 邓舜洲

    2010-01-01

    参照资料选择并验证了一对牛病毒性腹泻病毒(bovine viral diarrhea virus,简称BVDV)特异性引物,摸索了检测BVDV的RT-PCR方法的不同反应条件,选择出了最佳条件,建立了特异性的RT-PCR检测方法.并对已做过CSFV检测的52份病料进行BVDV的凹归性检测,其中BVDV检测结果为阳性的病料7份,表明江两省部分猪群存在BVDV的感染,结合之前做过的猪瘟病毒(CSFV)检测结果,可说明江西省部分猪群存在BVDV的单纯感染和BVDV与CSFV的混合感染.

  4. Development of Antigen Capture ELISA of Detection Bovine Viral Diarrhea Virus%牛病毒性腹泻病毒抗原捕获 ELISA 方法的建立

    Institute of Scientific and Technical Information of China (English)

    范晴; 谢芝勋; 谢志勤; 刘加波; 庞耀珊; 邓显文; 谢丽基; 罗思思

    2015-01-01

    A antigen capture enzyme‐linked immunosorbent assay(ELISA) method was developed to detect antigen of bovine virus diarrhea(BVDV) using mouse monoclonal antibody against NS3 protein of bovine virus diarrhea as capture antibody and polyclonal antiserum (rabbit serum against BVDV ) as coating anti‐body .The optimum conditions were achieved :coating antibody was diluted for 1 :1600 ,the mouse mono‐clonal antibody was diluted for 1∶2 000 and the enzyme‐label antibody was diluted for 1 :2000 .BRV , IBRV and MB were detected by the Ag‐capture ELISA and the result showed that there was no crossing‐reaction with BVDV .The method has a minimum detection concentration is 7 .9 × 103 TCID50 .The result of positive detection by Ag‐capture ELISA were consistent with RT‐PCR .The result showed that the Ag‐capture ELISA was highly rapid ,specific and sensitive ,and it could be the basic for controlling BVDV .%用兔抗牛病毒性腹泻病毒(BVDV )多抗作为包被抗体,BVDV NS3单克隆抗体作为捕获抗体,建立了检测BVDV抗原的捕获ELISA 方法,对各项反应条件进行优化,最终获得最佳工作条件为兔多抗1∶1600稀释包被,NS3单抗1∶2000稀释,酶标抗体工作浓度为1∶4000稀释。特异性和敏感性试验结果表明,该方法对牛轮状病毒、牛传染性鼻气管炎病毒、牛结核杆菌无特异性交叉反应,其最低可检测7.9×103个TCID50的病毒量,与RT‐PCR方法的相比较,符合率为100%。所建立的BVDV抗原捕获ELISA 方法快速、特异、敏感可用于BVDV抗原的检测。

  5. Risk Assessment and Epidemiological Characteristics of Infectious Bovine Rhinotracheitis and Bovine Viral Diarrhea in Dairy Herds in Beijing%北京地区奶牛场牛传染性鼻气管炎、牛病毒性腹泻病风险评估及流行情况分析

    Institute of Scientific and Technical Information of China (English)

    李栋梁; 赵景义; 沈俊乐; 史苍桀; 曹杰

    2013-01-01

    The epidemiology of the infectious bovine rhinotracheitis (IBR) and bovine viral diarrhea (BVD) were assessed in the medium and small dairy herds and the farming community in Beijing.The percent of IBR antibodypositive heifers was 50.52%,and the result was 70.98% for BVDV.At the herds level,the antibody positive rate reached 50% (12/24) for IBR and 87.50% (21/24) for BVD.IBR high-risk herds were 9,BVD high-risk herds were 12,seven herds were combination risk of IBR and BVDV.The serological investigation and analysis showed that IBR prevalence is relatively serious in Beijing,and vaccine should be considered; Most of herds had BVDV history or contact with acute infection,high-risk herds need to start BVDV eradication program.%本研究对北京地区中小奶牛场及养殖小区进行了牛传染性鼻气管炎(IBR)、牛病毒性腹泻病(BVD)的风险评估.结果表明,参试牛场及小区的后备牛IBR血清阳性率达50.52%,BVD血清抗体阳性率达70.98%;从牛场水平看,24个牛场中IBR抗体场间阳性率达到50% (12/24),BVD抗体场间阳性率达到87.50%(21/24),IBR高风险牛场9个,BVD高风险牛场12个,双高风险牛场7个.此次评估结果表明,北京地区IBR流行情况较为严重,应考虑疫苗免疫;参试的大部分牛场有BVDV的急性感染史或接触史,高风险牛场应启动BVDV清除计划.

  6. Seroprevalencia de las infecciones por el virus Diarrea Vírica Bovina en ganado bovino en Andalucía (Seroprevalence of infection with bovine viral diarrhea virus in cattle in Andalucia

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    Gómez-Pacheco, Juan M:

    2009-02-01

    Full Text Available ResumenSe ha realizado un estudio seroepidemiológico frente al virus de la Diarrea Vírica Bovina (vDVB en la cabaña bovina andaluza, tilizando para ello un ELISA indirecto para la detección de anticuerpos frente a una proteína altamente conservada (p80. Después de eliminar los animales vacunados, la encuesta se realizó sobre 4.768 individuos pertenecientes a 227 colectivos no vacunados frente al vDVB, mediante muestreo estadístico para un nivel de confianza del 95 por ciento. La seropositividad obtenida ha sido del 42,3 por ciento de los individuos analizados, mientras que la prevalencia estimada de rebaños seropositivos alcanzó el 70,9 por ciento. La proporción de bovinos persistentemente infectados (IPencontrada en la muestra (0,063 % de los individuos y 1,32 de los colectivos, ha sido más baja de la esperada en función de la alta seroprevalencia detectada, hecho que demuestra que la supervivencia de estos animales lógicamente está condicionada.SummaryAn epidemiological survey of the Bovine Viral Diarrhoea (BVD on bovine farms from Andalusia Region was carried out. An indirect enzyme-linked inmunosorbent assay was used for the detection of p80 specific antibodies. After elimination of vaccinated animals, the study was performed with 4768 serum samples obtained from non vaccinated animals belonged to 227 farm selected by statistical sampling, on the population base of the official registry of bovine herds in this region. The prevalence obtained for the BVD infections in farms were 70.9 percent. This prevalence for individuals resulted in 42.3 percent. A number of Persistent Infected (PI was detected in this study (0.063% animals and 1.325% in farms frequency lower than expected according toseroprevalence obtained.

  7. Atividade antiviral do extrato de própolis contra o calicivírus felino, adenovírus canino 2 e vírus da diarréia viral bovina Antiviral activity of propolis extracts against feline calicivirus, canine adenovirus 2, and bovine viral diarrhea virus

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    Ana Paula Cueto

    2011-10-01

    Full Text Available Dentre as propriedades biológicas da própolis, a atividade antimicrobiana tem merecido destacada atenção. Neste artigo, descreve-se a atividade antiviral de dois extratos etanólicos de própolis (EP1 e EP2 frente aos vírus: calicivírus felino (FCV, adenovírus canino tipo 2 (CAV-2 e vírus da diarréia viral bovina (BVDV. Um dos extratos (EP1 foi obtido por extração etanólica de própolis obtida da região central do Estado do Rio Grande do Sul e o segundo (EP2, obtido comercialmente de uma empresa de Minas Gerais. A análise dos extratos de própolis através da cromatografia líquida de alta eficiência (CLAE identificou a presença de flavonóides como: rutina, quercetina e ácido gálico. A atividade antiviral bem como a citotoxicidade dos extratos aos cultivos celulares foram avaliadas através do MTT [3- (4,5 dimetiltiazol-2yl-2-5-difenil-2H tetrazolato de bromo]. Ambos os extratos evidenciaram atividade antiviral frente ao BVDV e CAV-2 quando acrescidos ao cultivo celular anteriormente à inoculação viral. Os extratos foram menos efetivos contra o FCV em comparação aos resultados obtidos com os outros vírus, e a atividade antiviral neste caso foi observada apenas quando a própolis estava presente após a inoculação viral. O extrato obtido no laboratório (EP1 apresentou valores mais altos de índice de seletividade (IS=CC50/ CE50, quando comparado à outra amostra (EP2. Em resumo, a própolis apresentou atividade antiviral frente a três diferentes vírus, o que a torna alvo para o desenvolvimento de novos compostos naturais com atividade antiviral.Propolis is a resinous substance produced by bees for which several biological activities have been attributed. In this article, the antiviral activity of two propolis extracts was tested against bovine viral diarrhea virus (BVDV, canine adenovirus type 2 (CAV-2, and feline calicivirus (FCV. One of the extracts was obtained by ethanolic extraction of propolis from the Santa

  8. Padronização da técnica de imunoperoxidase para detecção do vírus da diarréia bovina a vírus em cultura de células: Standardization of immunoperoxidase test to detection bovine viral diarrhea virus in cell culture

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    G.I. Andrade

    2002-12-01

    Full Text Available Este estudo teve como objetivo a padronização do ensaio de imunoperoxidase em monocamada de células (IPM para o diagnóstico etiológico da diarréia bovina a vírus (DBV. O teste foi padronizado em monocamada de cultivo primário de pulmão fetal bovino (PFB inoculada com as amostras clássicas, citopatogênica (CP e não citopatogênica (NCP, do vírus da DBV e testado em amostras biológicas suspeitas processadas no teste clássico de isolamento viral (IV. O método de IPM identificou o vírus da DBV, apresentando melhores resultados com a utilização do calor como agente fixador, a soroalbumina bovina a 4% em PBS como bloqueador e a revelação com o cromógeno 3-amino-9-etil-carbazol (AEC. Como anticorpos primários, tanto o anticorpo policlonal como o monoclonal forneceram bons resultados.The aim of this study was to standardize the immunoperoxidase in cell monolayer assay (IPMA for the etiological diagnosis of bovine viral diarrhea (BVD. The method was standardized in monolayer of primary bovine fetal lung culture inoculated with cytophatic and non-cytophatic classical strains of BVD virus and tested using samples that were considered suspected in the classical technique of viral isolation. The IPMA successfully identified BVD virus and presented better results when heat was used for fixation, BSA 4% solution in PBS was used for blocking and AEC chromogen was used for revelation. Both monoclonal and polycloral antibodies gave good results when used as primary antibodies.

  9. Development of Indirect ELISA of Bovine Viral Diarrhea Virus by Using Tandem Epitopes of NS3%牛病毒性腹泻病病毒NS3表位串联蛋白表达及抗体间接ELISA方法的建立

    Institute of Scientific and Technical Information of China (English)

    贾莹; 李岩; 尹鑫; 温凯; 刘华; 张文龙; 王君伟

    2011-01-01

    为建立一种有效的牛病毒性腹泻病病毒(Bovine viral diarrhea virus,BVDV)抗体检测方法,作者将BVDVNS3蛋白2个抗原表位的编码核酸序列进行串联,构建重组表达载体pET-30a-EXnN,并在原核表达系统中表达了重组蛋白.选取包含12个表位肽的重组蛋白( r-BVDV-EX6 N)作为包被抗原建立NS3蛋白抗体间接ELISA方法.该方法的特异性和稳定性良好,与2种商品化试剂盒的符合率分别为96.05%和78.95%.试验结果表明建立的ELISA方法可用于BVDV NS3蛋白抗体的检测.%The present experiment was performed to establish an indirect ELISA for detection of antibodies against Bovine viral diarrhea virus (BVDV). Tandem epitopes of NS3 of BVDV was cloned into pET-30a and expressed in E. coli Rosetta. SDS-PAGE analysis revealed a band of protein correspondent with molecular weight of the target protein, and the result of western blot showed that the recombinant protein was recognized specifically by anti-BVDV positive serum. An indirect ELISA (r-BVDV-EX6N-ELISA) was developed using purified protein which contains 12 epitopes as coating antigen to detect BVDV antibodies in cattle. The assay was highly specific and showed no cross-reaction with positive sera of other bovine diseases. Comparison with two commercial kit showed a coincidence rate of 96. 05% and 78. 95%. The results demonstrated that the indirect ELISA established in this study works well in BVDV detection.

  10. Effects of bovine viral diarrhea viruses in vitro on transcription of interferon-al- pha, beta, gamma mRNA in bovine peripheral blood mononuclear cells%牛病毒性腹泻病毒感染牛外周血单核细胞对IFN-α、β、γmRNA转录的影响

    Institute of Scientific and Technical Information of China (English)

    韩猛立; 黄新; 钟发刚

    2012-01-01

    The study was done to survery the interferon-alpha, beta and gamma mRNA transcription profiles of bovine viral diarrfea viruse(BVDV) infection,and to investigate the host-BVDV interaction. The clinically healthy Holstein cows tested negative for bovine viral diarrhea virus(BVDV) in peripheral blood mononuclear cells(PBMC) were in- fected with noncytopathic(NCP) and cytopathic(NCP) BVDV. The mRNA levels of IFN-α,β and γ genes were ana lyzed using a reaPtime fluorescent quantitative PCR(reaPtime FQ-PCR). The results indicated that the transcription of I type(IFN-α,β) mRNA showed a different increasing levels (P〈0.01) ,after infected CP- and NCP BVDV in PBMC;only IFN-α decreased at 4,12 h(P〈0. 05) after infected CP-BVDV. And IFN-γ was increased throughout the infection process of CP and NCP BVDV in PBMC (P〈0. 05). The transcription levels of IFN mRNA were in- creased when two biotype of BVDV infected in PBMC.%为了解牛病毒性腹泻病毒(BVDV)感染对干扰素(IFN)mRNA转录时相的影响,探讨宿主-病毒之间的相互关系,用非致细胞病变(noncytopathic,NCP)和致细胞病变(cytopathic,CP)型BVDV感染临床健康BVDV检测阴性的荷斯坦奶牛外周血单核细胞(PBMC),利用实时荧光定量PCR技术对感染后IFN-α、β、γmRNA转录水平的变化进行定量分析。结果表明,CP型和NCP型BVDV感染PBMC后,Ⅰ型IFN(IFN-α、β)均呈现出不同程度的转录水平上调,且差异极显著(P〈0.01);只有IFN-α在CP型BVDV感染后4,12h(P〈0.5)出现转录下调。IFN-γ在整个感染过程中均呈现出不同程度的转录水平上调,且差异显著(P〈0.05)。这表明2种生物型BVDV感染可引起PBMC中IFN mRNA转录水平升高。

  11. 牛病毒性腹泻病毒感染牛外周血单核细胞对CD80和CD86 mRNA转录的影响%Effects of bovine viral diarrhea viruses in vitro on transcription of CD80 and CD86 mRNA in bovine peripheral blood mononuclear cells

    Institute of Scientific and Technical Information of China (English)

    韩猛立; 黄新; 钟发刚

    2012-01-01

    The clinically healthy Holstein bovine tested negative for bovine viral diarrhea virus(BVDV) in peripheral blood mononuclear cells(PBMC) were infected with noncytopathic(NCP) and cytopathic(CP) BVDV.The changes levels of mRNA of CD80 and CD86 genes were analyzed using a real-time fluorescent quantitative PCR(real-time FQ-PCR).The results showed that the transcription of CD80 mRNA exhibited two transcription high at 4 h(P0.05) and 12,24 h(P0.01) post-inoculation(PI),and CD86 mRNA reached the highest level at 6 h(P0.05)PI in NCP BVDV-infected PBMC;then the mRNAs transcriptions of CD80 peaked at 24 h(P0.05)and CD86 peaked at 6 h(P0.05) in CP BVDV-infected PBMC with signifficant differences compared to that of the other PI.While the transcription on CD80 mRNAs witnessed more kinetic changes,it indicats NCP and CP BVDV could significantly supress the transcription of CD80-CD86 genes early during the infection,and the situation might weaken the antigen presentation of PBMC in the inoculated bovine.%用非致细胞病变(noncytopathic,NCP)和致细胞病变(cytopathic,CP)型牛病毒性腹泻病毒(BVDV)感染临床健康BVDV检测阴性的荷斯坦奶牛外周血单核细胞(PBMC),利用实时荧光定量PCR技术对感染后共刺激分子CD80和CD86mRNA转录水平的变化进行定量分析。结果表明,在NCP型BVDV感染牛PBMC后CD80在4h(P〈0.05)和12,24h(P〈0.01)出现2次转录高峰,CD86在6h(P〈0.05)出现转录高峰;CP型BVDV感染后,CD80在24h(P〈0.05)出现转录高峰,CD86在6h(P〈0.05)出现转录高峰。尽管CD80在NCP型BVDV感染后呈现较复杂的动态变化,但结果提示NCP型和CP型BVDV感染均可导致牛PBMC的共刺激分子CD80和CD86基因转录在感染早期明显受到抑制,PBMC的抗原呈递能力受到影响。

  12. Neonatal diarrhea by bovine coronavirus (BCoV in beef cattle herds

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    Elis Lorenzetti

    2014-02-01

    Full Text Available Bovine coronavirus (BCoV is the second most important viral agent involved in neonatal diarrhea in calves worldwide. The reports on the frequency of BCoV infection in beef cattle herds under extensive management are uncommon in Brazil. The present study analyzed 93 diarrheic fecal samples of calves up to 60 days of age from 13 commercial beef cattle herds located in the states of Mato Grosso, Mato Grosso do Sul, Minas Gerais, Paraná, and Rondônia. The fecal samples were collected during 2009-2012 and were previously analyzed for the presence of bovine rotavirus group A (BoRVA, with negative results. The presence of BCoV in the fecal samples was evaluated by the partial amplification of the N gene by using the semi-nested PCR technique. The expected products of 251 bp length were amplified 33.3% (31/93 of the analyzed diarrheic fecal samples. The results revealed that coronaviruses has important participation in the neonatal diarrhea complex of beef cattle herds reared extensively from the different geographical regions of Brazil.

  13. Perfil da infecção pelo vírus da diarreia viral bovina (BVDV em um rebanho bovino leiteiro de alta produção e com programa de vacinação contra o BVDV Bovine viral diarrhea virus (BVDV infection profile in a high production dairy herd with vaccination program against BVDV

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    Stelamaris Dezen

    2013-02-01

    Full Text Available A infecção pelo vírus da diarreia viral bovina (BVDV foi avaliada em um rebanho bovino leiteiro de alta produção com histórico de problemas reprodutivos e de vacinação regular contra o BVDV. A identificação do vírus foi realizada por RT-PCR em soro sanguíneo e o perfil sorológico por vírus-neutralização. Inicialmente, 100% (n=692 dos animais do rebanho foram avaliados com relação à presença de infecção ativa pelo BVDV por meio da RT-PCR. Quatro meses após, todos os animais positivos (n=29 na primeira avaliação foram avaliados novamente pela RT-PCR, assim como todos os animais que nasceram (n=72 e os que apresentaram problemas reprodutivos (n=36 no intervalo entre a primeira e a segunda colheita de sangue. Os resultados finais do estudo possibilitaram identificar 27 animais transitoriamente infectados e três animais persistentemente infectados (PI. A sorologia, realizada apenas nos animais positivos na primeira avaliação pela RT-PCR e nas vacas que apresentaram problemas reprodutivos entre a primeira e a segunda RT-PCR, demonstrou grande flutuação nos títulos de anticorpos neutralizantes, além de soroconversão na maioria dos animais. Foram identificados aumentos nos títulos de anticorpos neutralizantes que variaram entre 3 e 8 log2, indicando infecção ativa no rebanho. A circulação viral no rebanho avaliado foi responsável pela expressão de sinais clínicos da esfera reprodutiva em animais com baixo título de anticorpos e consequente falha na proteção fetal. Os resultados demonstram que o controle da infecção pelo BVDV apenas por meio da vacinação regular em rebanhos com animais PI pode não ser eficaz na profilaxia dessa virose.The profile of bovine viral diarrhea virus (BVDV infection was studies in a high production dairy herd selected based on a history of reproductive failures and regular vaccination against BVDV. Virus identification was performed by RT-PCR and serological profile was

  14. 青海省部分地区3种牛病毒性腹泻病原的感染情况调查%Survey of Three Kinds of Bovine Viral Diarrhea Pathogens Infection in Some Areas of Qinghai Province

    Institute of Scientific and Technical Information of China (English)

    权英存; 刘虎守

    2014-01-01

    为了解青海省部分牛群中牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)、牛轮状病毒(bovine rotavirus,BRV)和牛冠状病毒(bovine coronavirus,BCV)3种牛病毒性腹泻病原的感染现状,本研究采用RT-PCR方法首次对2012~2013年青海省部分地区的32份具有腹泻症状的临床病料及152份健康牛粪便样品进行了BVDV、BRV、BCV的核酸检测与分析.结果显示,32份腹泻牛病料样品中BVDV、BRV、BCV的阳性率分别为65.63%(21/32)、18.75%(6/32)、34.38%(11/32),且存在2种或3种病原的混合感染;152份健康牛粪便样品中BVDV、BRV、BCV的阳性率分别为3.95% (6/152)、1.97%(3/152)、0(0/152).该结果表明青海省部分牛群中普遍存在BVDV、BRV、BCV的感染,且混合感染现象严重,需进一步加强青海省地区牛病毒性腹泻病原的综合防控.

  15. Evidence of mixed persistent infections in calves born to cows challenged with a pool of bovine viral diarrhea virus isolates Evidências de infecção persistente mista em bezerros nascidos de vacas inoculadas com um pool de isolados do vírus da diarréia viral bovina

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    Sandra Arenhart

    2010-12-01

    Full Text Available Pregnant cows infected with noncytopathic (NCP isolates of bovine viral diarrhea virus (BVDV between days 40 and 120 days of gestation frequently deliver immunotolerant, persistently infected (PI calves. We herein report the characterization of PI calves produced experimentally through inoculation of pregnant cows with a pool of Brazilian BVDV-1 (n=2 and BVDV-2 isolates (n=2 between days 60 and 90 of gestation. Two calves were born virus positive, lacked BVDV antibodies, but died 7 and 15 days after birth, respectively. Six other calves were born healthy, seronegative to BVDV, harbored and shed virus in secretions for up to 210 days. Analysis of the antigenic profile of viruses infecting these calves at birth and 30 days later with a panel of monoclonal antibodies indicated two patterns of infection. Whereas three calves apparently harbored only one isolate (either a BVDV-1 or BVDV-2, co-infection by two antigenically distinct challenge viruses was demonstrated in three PI calves. Moreover, testing the viruses obtained from the blood of PI calves by an RT-PCR able to differentiate between BVDV-1 and BVDV-2 confirmed the presence/persistence of two co-infecting viruses of different genotypes (BVDV-1 and BVDV-2 in these animals. These findings indicate that persistent infection of fetuses/calves - a well characterized consequence of fetal infection by BVDV - may be established concomitantly by more than one isolate, upon experimental inoculation. In this sense, mixed persistent infections with antigenically distinct isolates may help in understanding the immunological and molecular basis of BVDV immunotolerance and persistence.Vacas prenhes infectadas com isolados não-citopáticos (NCP do vírus da diarréia viral bovina (BVDV entre os dias 40 e 120 de gestação frequentemente produzem bezerros imunotolerantes, persistentemente infectados (PI. Este artigo relata a caracterização de bezerros PI produzidos experimentalmente, pela inoculação de

  16. Diversidade antigênica de amostras do vírus da diarréia viral bovina isoladas no Brasil: implicações para o diagnóstico e estratégias de imunização Antigenic diversity of Brazilian isolates of bovine viral diarrhea virus (BVDV: implications for diagnosis and immunization strategies

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    E.F. Flores

    2000-02-01

    Full Text Available Seqüenciamento e análise filogenética de 17 amostras do vírus da diarréia viral bovina (BVDV isoladas no Brasil identificaram quatro amostras (23,5% do genótipo 1a (BVDV-1a, nove amostras (52,9% do genótipo 1b (BVDV tipo 1b e quatro amostras (23,5% do genótipo 2 (BVDV tipo 2. As amostras brasileiras de BVDV tipo 2 apresentaram-se genotipicamente distintas dos BVDV tipo 2 até então identificados na América do Norte e Europa, sugerindo pertencerem a um novo subgenótipo. A caracterização antigênica dessas amostras por neutralização cruzada revelou reatividade sorológica muito reduzida com cepas vacinais do BVDV. O anti-soro produzido contra três cepas vacinais do BVDV apresentou atividade neutralizante muito reduzida contra várias amostras brasileiras de BVDV tipos 1 e 2. Diferenças de até 128 vezes nos títulos de anticorpos neutralizantes foram observadas entre cepas vacinais e amostras brasileiras do BVDV. Nos testes de soroneutralização (SN contra o vírus dos tipos 1 e 2, de 1134 amostras testadas, 280 (24,7% possuiam anticorpos neutralizantes anti-BVDV e dessas, 215 (76,8% apresentaram atividade neutralizante contra ambos os vírus, 37 (13,2% reagiram apenas contra o BVDV tipo 2 e 28 amostras (10% foram positivas apenas contra o BVDV tipo 1. Esses resultados demonstram que testes de SN utilizando vírus de apenas um genótipo podem resultar em um número significativo de falsos-negativos e indica a necessidade da formulação de vacinas com amostras locais de BVDV e/ou contendo vírus dos dois genótipos.Nucleotide sequencing and phylogenetic analysis of 17 Brazilian isolates of bovine viral diarrhea virus (BVDV identified four isolates (23.5% belonging to genotype 1a (BVDV-1a, nine isolates (52.9% of the genotype 1b (BVDV-1b and four isolates (23.5% belonging to genotype 2 (BVDV-2. The Brazilian BVDV type 2 isolates were shown to be genotypically different from the BVDV type 2 identified so far in North America and

  17. COMPARACIÓN ENTRE DOS TÉCNICAS DE DIAGNÓSTICO PARA DIARREA VIRAL BOVINA (DVB EN 50 PREDIOS DE LA X REGIÓN, CHILE: SERONEUTRALIZACIÓN Y ENZIMOINMUNOENSAYO INDIRECTO (ELISA-I* Comparison of two diagnostic techniques to bovine viral diarrhea disease (BVD in 50 dairy herds from the Xth Region, Chile: Seroneutralization test and indirect immunosorbent assay (I-ELISA

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    G REINHARDT

    2001-01-01

    Full Text Available Bovine viral diarrhea/Mucosal disease (BVD/MD is a highly spread virosis worldwide and has a great impact in bovine reproduction and production. In Chile, the disease has been reported with over 60% of prevalence and it demands adecuate diagnostic methods. Curently the official serologic diagnostic test in Chile is the serum neutralization test (SNT, this method detects the presence of antibodies against the BVD virus and it is considered to have good specificity and sensitivity, althought, it presents some disadvantages in its interpretation and in its execution. The aim of this investigation was to compare de SNT as gold standard, with a commercial immunosorbent assay (ELISA, in terms of specificity and sensitivity in the detection of antibodies against BVD antigens. A set of 500 bovine sera drawn from 50 milk herds from the Xth Region of Chile were analized. The results showed that the SNT detected 278 serum samples as positives and the ELISA detected 347 serum samples as positives, these represents for ELISA test a relative sensitivity and specificity of 91% and 57%, respectively. Statistically significant differences of the serodiagnosis obtained in both tests were established through the McNemar test (<0.05, and a median concordance between them through the Kappa test. When the SNT titers were related with the optical densities (OD of ELISA, a positive association was detected between this values. It was concluded that ELISA provides good results in comparison with SNT, having the former a higher number of detections because its dignostic higher sensitivity. Therefore, ELISA is an appropiate diagnostic method for large populations of cattle

  18. Viral infections and bovine mastitis: a review

    NARCIS (Netherlands)

    Wellenberg, G.J.; Poel, van der W.H.M.; Oirschot, van J.T.

    2002-01-01

    This review deals with the role of viruses in the aetiology of bovine mastitis. Bovine herpesvirus 1, bovine herpesvirus 4, foot-and-mouth disease virus, and parainfluenza 3 virus have been isolated from milk from cows with clinical mastitis. Intramammary inoculations of bovine herpesvirus 1 or para

  19. Establishment and Application of Real-time Fluorescent Quantitative RT-PCR Method for Detection of Bovine Viral Diarrhea Virus%牛病毒性腹泻病毒实时荧光定量RT-PCR方法的建立及应用

    Institute of Scientific and Technical Information of China (English)

    王荣; 李文文; 王研; 刘亚刚; 余琼; 任永刚

    2014-01-01

    持续性感染和免疫耐受是牛病毒性腹泻病的重要特征,这一特征给该病的诊断、检疫及防控造成很大困难,为此,建立快速实用、高效敏感的牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)检测方法具有重要意义.据GenBank中登录的BVDV 5' UTR保守区域设计并合成1对特异性引物,以SYBR Green Ⅰ染料为扩增指示剂,建立了BVDV实时荧光定量RT-PCR检测方法.结果表明,该方法具有快速、敏感、特异、重复性好等优点.该方法的建立为牛病毒性腹泻病的早期快速诊断和有效检出持续性感染动物提供了手段,是该病检疫和诊断方法的补充和完善.

  20. Expression of P20 and P14 protein genes of bovine viral diarrhea virus and antigenicity analysis of the expressed proteins%牛病毒性腹泻病毒P20及P14蛋白基因的表达及其产物的抗原性分析

    Institute of Scientific and Technical Information of China (English)

    李慧昕; 王君伟

    2006-01-01

    将牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)p20和p14基因分别亚克隆至原核表达载体pGEX-6p-1和pPROEX-HTb,并转化至相应的宿主菌大肠杆菌BL21(DE3)pLysS和DH5α中表达.P20蛋白在2个宿主中都获得了高效表达;P14蛋白在BL21(DE3)pLysS中表达量较低,而在DH5α中未见表达.对P14蛋白诱导表达过程的监测表明,P14蛋白对大肠杆菌是一种毒性蛋白.用Western-blotting分析未能检测到两种蛋白的反应条带,推测这两种蛋白可能存在构象表位.

  1. Isolation and Identification of Bovine Viral Diarrhea Virus JN Strain and Sequence Analysis on E2 Gene%牛病毒性腹泻病毒JN株的分离鉴定及E2基因的序列分析

    Institute of Scientific and Technical Information of China (English)

    汤中和

    2013-01-01

    本研究从疑似牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)感染牛的分泌物与排泄物中分离鉴定1株牛病毒性腹泻病毒,并进行E2基因序列分析.结果表明,分离株病毒命名为JN株;Reed-Muench法测定分离株病毒TCID50为10-7.5/0.1 mL;病毒中和试验结果表明,BVDV JN分离株可被BVDV阳性血清特异性中和,而不能被BVDV阴性血清中和;分离株病毒E2基因序列测序结果表明,该分离毒株属于BVDV Ⅰ a亚型.

  2. 牛病毒性腹泻BVDV2/JZ05-1基础毒株的安全性和免疫原性研究%Safety and Immunogenicity Study of Bovine Viral Diarrhea Virus 2 Strain JZ05-1

    Institute of Scientific and Technical Information of China (English)

    李海涛; 苗利光; 刘艳环; 朱言柱; 王松; 王文昊; 安亚雄

    2012-01-01

    本试验用已分离到的牛病毒性腹泻病毒BVDV2/JZ05-1基础毒株对犊牛免疫的安全性和免疫原性进行了研究.通过对基础毒种单剂量、超剂量、单剂量重复安全试验和免疫原性试验研究证实,BVDV2/JZ05-1对犊牛免疫是安全的,对犊牛感染BVDV有很好的免疫保护效果.%The bovine viral diarrhea virus 2 strain JZ05-1 (BVDV2/JZ05-1) was isolated in this experiment. Single dose, overdose and single dose reduplication safety experiment and immunogenicity experiment were carried out in this experiment. When the calves were immunized with BVDV2/JZ05-1, the safety and immunogenicity of BVDV2/JZ05-1 were examined. The results suggested that the immunization of BVDV2/JZ05 is safe for calves, and it can well protect calves from the BVDV.

  3. Study on Immune Interference of Infectious Bovine Rhinotracheitis Virus and Bovine Viral Diarrhea Virus%牛传染性鼻气管炎弱毒与牛病毒性腹泻弱毒抗原免疫干扰的研究

    Institute of Scientific and Technical Information of China (English)

    郭利; 张淑琴; 王炜; 冷雪; 武华

    2012-01-01

    本试验使用3~6月龄健康易感牛9头(牛传染性鼻气管炎病毒(IBRV)和牛病毒性腹泻病毒(BVDV)抗原、抗体均阴性),共分3组,每组3头犊牛.第1组首免肌肉注射IBRV-LNM弱毒疫苗株种毒,接种1周后,每头牛接种BVDV-SM弱毒疫苗株;第2组只接种BVDV-SM弱毒疫苗株种毒,接种时间同第1组;第3组为对照组,接种MDBK细胞培养液.接种BVDV-SM疫苗毒后每周采血至疫苗毒接种后28 d,测定接种后BVDV抗体效价,并采用BVDV-JL检验用强毒进行攻毒试验.结果表明,第1组与第2组试验动物血清中牛病毒性腹泻病毒抗体水平无明显差异,能够抵抗BVDV-JL强毒攻击达到免疫保护的效果,说明牛传染性鼻气管炎病毒IBRV-LNM弱毒疫苗株接种后在牛体内对牛病毒性腹泻病毒BVDV-SM疫苗毒不产生免疫干扰作用.%This test using totle 9 head of catties, which were 3 to 6 month old healthy and susceptible (IBRVand BVDV antigen negtive,antibody titer is less than or equal to 2) .were divided into 3 groups,each group of 3 cows. The first group was injected with IBRV-LNM attenuated vaccine strain,one weeks later each animal was inoculated with BVDV-SM attenuated vaccine strain. The second group was only inoculated with BVDV-SM attenuated vaccine strain at the same time with the first group. The third group was the control group and inoculated with MDBK cell culture fluid, the determination of BVDV antibody titer post-vaccination every week until 28 days,and then challenged with BVDV-JL virulent strain. The results showed that the serum of bovine viral diarrhea virus antibody levels had no significant differences between the first and the second group, and observation after the challenge showed that the vaccine effectively protected calves from the challenge. This suggested that IBRV attenuated strain was not restrained BVDV attenuated vaccine virus in vivo.

  4. A infecção pelo vírus da diarréia viral bovina (BVDV no Brasil: histórico, situação atual e perspectivas Bovine viral diarrhea virus (BVDV infection in Brazil: history, current situation and perspectives

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    Eduardo F. Flores

    2005-09-01

    BVDV-1 e 2. O conhecimento sobre a infecção pelo BVDV no Brasil tem aumentado consideravelmente nos últimos anos, à medida em que cresce o número de laboratórios envolvidos em diagnóstico e pesquisa sobre esse vírus. Diagnóstico sorológico, virológico ou molecular; estudos sobre epidemiologia sorológica e molecular, patogenia e produção de reagentes para diagnósitco têm contribuído para o aumento no conhecimento sobre a infecção pelo BVDV no país.Bovine viral diarrhea virus (BVDV is one of the most important pathogens of cattle worldwide. BVDV infection and associated diseases have been reported in Brazil since the late 1960ties. Several serological, virological, clinical and pathological reports demonstrate the widespread distribution of BVDV infection among Brazilian cattle. In addition to variable levels of positive serology in beef and dairy cattle, BVDV antibodies have been occasionally detected in swine, wild boars, goats, cervids and water buffaloes. BVDV infection has been diagnosed in aborted fetuses, buffy coats of persistently infected (PI animals, clinical specimens from animals suffering from different clinical syndromes, semen of bulls of artificial insemination (AI centers, in healthy fetuses and in commercial fetal bovine serum and/or cultured cells. About 50 isolates have been genetically and/or antigenically characterized up to date, whilst roughly an equivalent number of isolates awaits characterization. Most of the characterized isolates belong to BVDV-1 genotype, non-cytopathic (NCP biotype, yet some BVDV-2 (and some CP BVDV have been identified as well. Brazilian BVDV isolates display a high antigenic variability and are markedly different from North American vaccine strains. A few inactivated, polyvalent vaccines are currently licensed in the country, yet vaccination is still incipient in many regions: only about 2.5 million doses were marketed in 2003. The low serological cross-reactivity between vaccine strains and field

  5. 一株2型牛病毒性腹泻病毒的分离鉴定及其在MDBK细胞上的克隆纯化%Isolation, Identification and Purification of a Genotype 2 Bovine Viral Diarrhea Virus in MDBK Cells

    Institute of Scientific and Technical Information of China (English)

    聂兆晶; 田夫林; 姜世金

    2012-01-01

    In order to study the contamination status of Bovine viral diarrhea virus(BVDV) on the process of live vaccine production for swine, the calf blood serum which served as primary cell was detected by antigen capture ELISA. The positive serum was inoculated on MDBK cells and reproduced blindly for 3 generations, and then the virus caused cells pathological changes. Through the RT-PCR and the gene sequencing, a BVDV of genotype 2 named JN-2 was obtained. By plaque assay and TCID.o detection, the JN-2 isolate was purified and its viral titer rose significantly in MDBK cells%为了解牛病毒性腹泻病毒(BVDV)在猪用活疫苗生产过程中的污染情况,利用抗原捕获ELISA方法对用于制作原代睾丸细胞的犊牛血清进行检测.将检测结果为阳性的1份犊牛血清,接种于MDBK细胞上,盲传3代,出现明显细胞病变.用BVDV 5′-UTR端特异性引物进行RT-PCR扩增,并对扩增片段进行克隆测序和序列分析.结果表明,分离得到一株2型牛病毒性腹泻病毒,并将其命名为JN-2.将该病毒用蚀斑方法进行克隆纯化,并对纯化前后病毒的TCID50进行测定比较,结果发现纯化后病毒滴度显著提高.

  6. Seroprevalence of some bovine viral respiratory diseases among non vaccinated cattle in Saudi Arabia

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    Mohamed Abd El Fatah Mahmoud

    2013-02-01

    Full Text Available Aim: Four viral pathogens, bovine viral diarrhea virus (BVDV, and bovine herpes virus type 1 (BHV-1, bovine parainfluenza type 3 virus (PI-3V, bovine respiratory syncytial virus (BRSV are mainly associated with bovine respiratory diseases that cause major economic losses in the dairy cattle industry. This study aimed to document exposure of cattle in Saudi Arabia to infectious BVDV, BHV-1, PI-3V and BRSV viruses in non vaccinated cattle in order to obtain epidemiological and immunological information. Materials and Methods: In the present study, 460 random serum samples obtained from non vaccinated cattle in five districts (Riyadh, Eastern Province, Jizan, Najran, Asir of Saudi Arabia between January to March 2011. These samples were tested for presence of antibodies against BVDV, BHV-1, BRSV and PIV-3 by commercial indirect ELISA kits. Results: Our findings displayed that Seropositivity rates were 26 % for BVD, 17.4 % for BHV-1, 69.1 % for PI-3V and 75.6 % for BRSV in the sampled population. In addition, coinfections with more than one virus were considerably common among non-vaccinated dairy cattle. Conclusion: These results indicate that exposure to these agents is common within the study areas. Preventive and control measures against these infectious agents should therefore be adopted. [Vet World 2013; 6(1.000: 1-4

  7. 猪瘟病毒和牛病毒性腹泻病毒双重RT-PCR方法的建立和初步应用%Establishment and Initially Application of Double RT-PCR Detection Method for Both Classical Swine Fever Virus and Bovine Viral Diarrhea Virus

    Institute of Scientific and Technical Information of China (English)

    陈静; 张小飞; 范红结; 黄显明

    2011-01-01

    根据GenBank上已发表的猪瘟病毒(CSFV)和牛病毒性腹泻病毒(BVDV)的全基因序列,进行对比分析,分别设计合成两对能特异性扩增CSFV、BVDV的引物.经过条件优化后,建立了检测(SFV和BVDV的双重RT-PCR方法,扩增两种病毒的片段,大小分别为938、650 bp.应用该方法对11批牛睾丸细胞、7批胎牛血清、60个批次的猪瘟细胞苗、10份全血样及10份组织样进行检测.通过试验证明,所建立的方法具有良好的特异性和敏感性,为防止猪瘟细胞苗的污染及进行CSFV和BVDV鉴别诊断提供了有效方法.%According to the complete genome sequences of classical swine fever virus (CSFV) and bovine viral diarrhea virus (BVDV) in GenBank, two pairs of primers were designed and synthesized. A double RT-PCR was developed based on these two pairs of primers which amplified the CSFV virus-specific segment with 938 bp and the BVDV virus-specific segment with 650 bp in sizes after the conditions of PCR were optimized. Approved this method of 11 batches of bovine testicular cells,7 batches of fetal bovine serum,60 batches of swine fever vaccine, 10 full-blood and 10 tissue samples for testing. The experiment showed that the developed method has good specificity and sensitivity of cell vaccine to prevent the pollution of BVDV and the diagnosis of CSFV and BVDV provides an effective method.

  8. MAGNITUDE, DURAÇÃO E ESPECIFICIDADE DA RESPOSTA SOROLÓGICA EM BOVINOS VACINADOS CONTRA O VÍRUS DA DIARRÉIA VIRAL BOVINA (BVDV MAGNITUDE DURATION AND SPECIFICITY OF THE SEROLOGICAL RESPONSE IN CATTLE VACCINATED AGAINST BOVINE VIRAL DIARRHEA VIRUS (BVDV

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    Fernanda Silveira Flores Vogel

    2002-02-01

    frente à grande diversidade antigênica das amostras de BVDV.The serological response induced by three inactivated commercial vaccines against bovine viral diarrhea virus (BVDV was evaluated in calves immunized three times (days 0, 30 and 180 and examined at different intervals after vaccination. Thirty days after the second vaccination, 74.5% (70/94 of the calves had developed neutralizing antibodies titers against BVDV-1 and 52.1% (49/94 against BVDV-2. The geometric mean titers (GMT and the number of animals with antibodies to BVDV-1 were 109.3(32/36; 54.6(22/28 and 25.5(16/30 for vaccines A, B e C, respectively and 19(27/36, 42.3(12/28 and 18.4(10/30 to BVDV-2. The antibody titers decreased by day 180, when 31.9%(30/94 of the calves showed no neutralizing activity against BVDV-1 and 63.8%(60/94 to BVDV-2. At that time, the GMTs and the number of animals seropositive to BVDV-1 were 28.3 (30/36, 28.3(20/28 and 16.1(14/30; and to BVDV-2 were 16.8(18/36, 21.6(10/28 and 28.3(6/30 for vaccines A, B and C, respectively. After the third dosis (day 180, the antibody titers to BVDV-1 increased significantly in all groups whereas a significant increase in BVDV-2 titers was observed only for vaccine A. At day 210, GMTs and the number of positive animals to BVDV-1 were 104.8(23/24, 50.3(24/26 and 43.7(24/28 and to BVDV-2 were 33.4(23/24, 23.3(22/26 and 15.7(22/28 for vaccines A, B and C. The neutralizing activity against BVDV-1 was higher than to BVDV-2 in all groups at day 210. Sera from five seropositive animals of each vaccine group were tested against four Brazilian BVDV-1 and two BVDV-2 isolates. In addition to the low and moderate titers, the cross-neutralization assays revealed a highly variable neutralizing activity against the Brazilian field isolates. These results demonstrated that the vaccination did not induce an adequate serological response in most animals, mainly due to the high antigenic variability of BVDV.

  9. Proteção fetal frente a desafio com o vírus da Diarréia Viral Bovina (BVDV em ovelhas imunizadas com duas amostras de vírus modificadas experimentalmente Fetal protection against challenge with bovine viral diarrhea virus (BVDV in pregnant ewes immunized with two strains experimentally attenuated

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    Mário C.S. Brum

    2002-04-01

    Full Text Available Duas amostras do vírus da Diarréia Viral Bovina (BVDV submetidas a múltiplas passagens em cultivo celular e exposição à radiação ultravioleta (UV a cada passagem foram avaliadas como candidatos a vírus vacinais. As amostras foram testadas quanto à sua atenuação para bezerros e fetos ovinos, reatividade antigênica contra isolados de campo, e capacidade de induzir proteção fetal em ovelhas prenhes. Inoculação intramuscular (IM dos vírus modificados em quatro bezerros produziu apenas uma elevação discreta e passageira da temperatura corporal, seguida de produção de altos títulos de anticorpos neutralizantes. O vírus não foi detectado em secreções nasais ou sangue nos dias seguintes à inoculação. Porém, a inoculação IM desses vírus em quatro ovelhas prenhes foi seguida de transmissão transplacentária e infecção em todos os fetos. Para os testes de proteção fetal, ovelhas prenhes previamente imunizadas com duas doses vacinais, foram inoculadas por via intranasal com amostras de BVDV-1 (SV-126.8, n=6 ou BVDV-2 (SV-260, n=5. No dia do desafio (134 dias após a segunda dose, todos os animais apresentavam altos títulos de anticorpos neutralizantes (256 a >4096 contra os vírus vacinais; além de títulos variados (8 a >4096 contra várias isolados brasileiros de BVDV-1 e BVDV-2. Quinze dias após o desafio, as ovelhas foram sacrificadas e os tecidos fetais foram examinados para a presença de vírus. Todos os fetos das ovelhas controle não-vacinadas apresentaram-se (n=4 positivos para os vírus utilizados no desafio. Em contraste, nenhum feto das ovelhas imunizadas (n=11 foi positivo para vírus, indicando que a resposta imunológica induzida pela vacinação com os vírus modificados foi capaz de prevenir a infecção fetal. Estes resultados indicam que é possível obter-se forte resposta imunológica e proteção fetal contra o BVDV com o uso de vacinas vivas modificadas.Two isolates of bovine viral diarrhea

  10. Anticorpos neutralizantes contra o vírus da Diarréia Viral Bovina (BVDV: comparação entre um imunógeno experimental atenuado e três vacinas comerciais inativadas Vaccination-induced neutralizing antibodies against bovine viral diarrhea virus (BVDV: comparison between an experimental modified-live vaccine and three comercial inactivated vaccines

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    Marcelo de Lima

    2005-02-01

    Full Text Available Os títulos e duração de anticorpos neutralizantes contra o vírus da Diarréia Viral Bovina (BVDV induzidos por uma vacina experimental atenuada (vacina A: dose única foram comparados com os induzidos por três vacinas comerciais inativadas (B, C e D: duas doses com intervalo de 30 dias. Trinta dias após a vacinação (vacina A ou após a segunda dose (vacinas B, C e D, anticorpos neutralizantes contra o BVDV-1 foram detectados em todos os animais (12/12 do grupo A (título médio geométrico GMT=1612,7; em 32 de 36 animais do grupo B (GMT=14,3; 22 de 28 do grupo C (GMT=25,1; e em 16 de 30 do grupo D (GMT=40,0. Anticorpos frente ao BVDV-2 foram detectados em todos os animais do grupo A (GMT=151,0; em 27 de 36 do grupo B (GMT=10,0; 12 de 28 do grupo C (GMT=11,5 e em 10 de 30 animais do grupo D (GMT=10,0. No dia 180 após a vacinação, o número de animais que ainda apresentava anticorpos contra o BVDV-1 e os GMTs para cada grupo foram: vacina A (12/12, GMT=905,0; vacina B (30/36, GMT=28,3; vacina C (20/28, GMT=28,3; vacina D (14/30, GMT=16,1; e contra o BVDV-2 foram: vacina A (12/12, GMT=56,6; vacina B (18/36, GMT=16,8; vacina C (10/28, GMT=21,6 e vacina D (6/30, GMT=16,1. Os títulos médios (GMTs induzidos pela vacina A foram significativamente superiores aos demais, tanto para o BVDV-1 (PThe titers and duration of neutralizing antibodies against bovine viral diarrhea virus (BVDV induced by an experimental attenuated vaccine (vaccine A: one dose were compared to those induced by three commercial inactivated ones (B, C and D: two doses at a 30 day interval. Thirty days after vaccination (vaccine A or the second dose (vaccines B, C and D, neutralizing antibodies to BVDV-1 were detected in all calves (12/12 from group A (mean geometric titer GMT=1612.7; in 32 out of 36 from group B (GMT=14.3; 22/28 from group C (GMT=25.1; 16/30 from group D (GMT=40.0. Antibodies reacting with BVDV-2 were detected in all animals from group A (GMT=151.0; 27

  11. Preparation of monoclonal antibodies against bovine viral diarrhea virus and the double antibody sandwich ELISA develpoment%抗牛病毒性腹泻病毒单克隆抗体的制备及双抗夹心ELISA检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    蒋颖; 朱国强; 林燕清; 陶洁; 孟祥升; 段小丽; 王娟; 王银; 张信军; 王建业

    2012-01-01

    为建立牛病毒性腹泻病毒(BVDV)双抗夹心ELISA检测方法,本研究将BVDV 890病毒株(BVDV-2)浓缩并纯化后免疫BALB/c小鼠,经常规技术进行细胞融合并筛选得到一株稳定分泌IgG的杂交瘤细胞株,命名为3F9.以BVDV单克隆抗体(MAb) IgM为捕获抗体,生物素标记的3F9为检测抗体,初步建立BVDV特异的双抗体夹心ELISA检测方法(BAS-ELISA).采用建立的BAS-ELISA方法检测35份临床病牛血清样品,检出阳性样本13份;与RT-PCR的符合率达到94.29%;表明本研究所建立的BAS-ELISA方法可用于BVDV感染的临床诊断,为BVDV的免疫学研究奠定了基础.%To establish a double monoclonal antibody (MAb)-mediated sandwich ELISA (DAS-ELISA) for detection of bovine viral diarrhea virus (BVDV), the MAb 3D8 (IgM) and biotin-labelled 3F9 (IgG) were used as capture antibody and detection antibody respectively after obtaining a hybridoma cell line stably secreting antibody (3F9). Thirty-five clinically suspected sera samples were detected by this method and the results of 13 samples were positive. Compared with RT-PCR in the paralled experiment with 11 samples positive, the coincidence rate was 94.25%. It is suggested that the DAS-ELISA was a potentially valuable method with high sensitivity and specificity against BVDV.

  12. 快速检测牛病毒性腹泻病毒实时荧光定量PCR技术建立及应用%The Establishment and Application of Real-time Fluorescent Quantitative PCR Method to Rapidly Detect Bovine Viral Diarrhea Virus

    Institute of Scientific and Technical Information of China (English)

    郭锐; 陈平; 田永祥; 杨克礼; 刘泽文; 段正赢; 梁望旺

    2011-01-01

    以牛病毒性腹泻病毒(BVDV)的保守基因序列为参考.设计、优化出一对特异的PCR引物和一奈TaqMan荧光探针,建立一种快速定量检测牛病毒性腹泻病毒的实时荧光定量PCR技术.该方法检测灵敏度比RT-PCR高100倍,并且避免了常规PCR电泳检测所带来的高污染率.因此.该方法具有快速、灵敏、特异、重复性好和能定量检测等优点,适用于牛场BVDV感染的快速定量检测和肉类食品进出口检疫.%A fluorescent quantitative PCR (FQ-PCR) method based on sequences of the BVDV genome was established to rapidly detect the bovine viral diarrhea virus. It included the disignation and optimization of a pair of specific primers and a fluorescent TaqMan probe for convseved gene. Comparation test showed that the sensitivity of this method was 100 times higher than RT-PCR test; and it could decrease the contamination usually caused by other conventional PCR. In conclusion,the FQ-PCR method was rapid, sensitive, specific and accurate; and thus could be used for rapidly detection of BVDV from cattle's tissues and other meat products.

  13. 牛病毒性腹泻病毒E2蛋白单克隆抗体的制备及初步鉴定%Preparation and Preliminary Characterization of Monoclonal Antibodies against E2 Protein of Bovine Viral Diarrhea Virus

    Institute of Scientific and Technical Information of China (English)

    王姝; 朱远茂; 蔡红; 马磊; 史鸿飞; 吕闯; 董秀梅; 高欲燃; 薛飞

    2013-01-01

    为制备牛病毒性腹泻病毒(BVDV)糖蛋白E2单克隆抗体(MAb),利用原核表达并且纯化的重组糖蛋白E2(rE2)免疫BALB/c小鼠,取免疫后小鼠脾细胞与骨髓瘤细胞SP2/0融合.采用以BVDV为检测抗原的间接ELISA筛选阳性细胞克隆,经3次克隆纯化后获得2株稳定分泌抗E2特异性MAb的杂交瘤细胞株,分别命名为4E3与1G11.用4E3与1G11杂交瘤细胞株接种BALB/c小鼠制备腹水,采用rE2及BVDV包被的ELISA测得的效价分别是6.21×106和6.83×105及6.83×105和7.5×104.间接ELISA、Western blot、IFA试验表明两株杂交瘤细胞所分泌的MAb具有良好的反应性和特异性.经抗体亚类鉴定4E3与1G11均为IgM/K.特异性试验表明4E3与1G11这2株MAb均不与牛传染性鼻气管炎病毒、牛副流感病毒3型、牛腺病毒3型反应;其中4E3不与猪瘟病毒反应,而1G11则可与猪瘟病毒发生交叉反应,这种反应特性可试用于BVDV与猪瘟病毒的鉴别诊断.所制备的4E3与1G11 MAb可以用于BVDV抗原的检测,为建立检测BVDV E2蛋白血清抗体的ELISA奠定了基础.%To prepare monoclonal antibody (MAb) against glycoprotein E2 of bovine viral diarrhea virus ( BVDV) , BALB/c mice were immunized with purified recombinant E2 ( rE2) expressed in E. coli and two hybridomas secreting MAb were screened from fusing the SP2/0 cells with the spleen cells of the immunized BALB/c mice by indirect ELISA coated with BVDV. The titers of MAb 4E3 and 1G11 in ascites were 6. 21 × 106,6. 83 × 105, 6. 83 × 105, and 7. 5 × 104 as detected by rE2 and BVDV , respectively. The MAbs secreted by hybridomas 4E3 and 1G11 had highly reactivity and specificity in indirect ELISA, Western blotting, IFA and identified as IgM with a light chain of κ by indirect ELISA. The specific tests indicated that the MAbs 4E3 and 1G11 had no reaction with bovine infectious rhinotracheitis virus, bovine parainfluenza virus type 3 and bovine adenovirus type 3. Meanwhile the MAb 4E3 had no

  14. Viral Agents of Diarrhea in Young Children in Two Primary Health Centers in Edo State, Nigeria.

    Science.gov (United States)

    Imade, Paul Erhunmwunse; Eghafona, Nosakhare Odeh

    2015-01-01

    Enteric viruses have been shown to be responsible for diarrhea among children during their early childhood. This study was carried out to determine the prevalence of rotavirus, adenovirus, and norovirus infection in young children with diarrhea in two primary health centers in Edo State, Nigeria. A total of 223 stool specimens were collected from children aged 0-36 months with clinical signs of diarrhea and 59 apparently healthy age-matched children as control. These specimens were investigated for three viral agents using immunochromatographic technique (ICT). The overall results showed that at least one viral agent was detected in 95/223 (42.6%) of the children with diarrhea while the control had none. The prevalence of rotavirus was 28.3%, adenovirus 19.3%, and norovirus 3.6%. There was a significant association between age group and infection (P < 0.0001). Seasonal pattern of enteric viruses was not statistically significant (P = 0.17). The overall coinfection rate was 7.6% and rotavirus-adenovirus coinfection had the highest with 5.4%. Rotavirus was the most prevalent viral agent. Coinfections are not uncommon among the population studied. The most commonly associated clinical symptom of viral diarrhea in this study was vomiting. Viral diagnostic tests are advocated for primary health care facilities in this locality.

  15. 9 CFR 113.215 - Bovine Virus Diarrhea Vaccine, Killed Virus.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Bovine Virus Diarrhea Vaccine, Killed Virus. 113.215 Section 113.215 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE VIRUSES, SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS...

  16. Frequência de anticorpos e fatores de risco para a infecção pelo vírus da diarreia viral bovina em fêmeas bovinas leiteiras não vacinadas na região Amazônica Maranhense, Brasil Frequency of antibodies and risk factors of bovine viral diarrhea virus infection in non-vaccinated dairy cows in the Maranhense Amazon region, Brazilfonte 10

    Directory of Open Access Journals (Sweden)

    Nancyleni Pinto Chaves

    2010-06-01

    Full Text Available O presente estudo teve como objetivo determinar a frequência e os fatores de risco para a infecção pelo vírus da diarreia viral bovina (BVDV em fêmeas bovinas leiteiras na região amazônica maranhense. Amostras de soro de animais não vacinados contra o BVDV e provenientes de 40 propriedades foram submetidas à técnica de ELISA indireto. Em cada propriedade avaliada, aplicou-se ainda um questionário epidemiológico para investigar os fatores de risco que poderiam estar associados à infecção pelo vírus. As amostras foram coletadas de animais que apresentavam ou não sinais clínicos sugestivos da infecção pelo BVDV. Das 400 amostras de soro analisadas, 61,5% (n=246 foram reagentes com a detecção de bovinos sorologicamente positivos em 95% (n=38 das propriedades. Dentre os fatores de risco avaliados, produção de leite (1-5L, ausência de assistência veterinária, uso de monta natural e monta natural associada à inseminação artificial apresentaram significância estatística (PThis study was performed in order to determine the frequency of bovine viral diarrhea virus (BVDV, and the main factors related to the frequency of the infection in dairy cows not vaccinated against BVDV from Maranhense Amazon region. Serum samples were submitted to an indirect ELISA test for detection of BVDV antibodies. An epidemiological questionnaire was applied for each herd to investigate variables that could beassociated with this infection. Serum samples were collected from 40 farms, with or without clinical signs of BVDV infection. From 400 serum samples examined, 61.5% (n=246 in 95% (n=38 of the herds were positive for ELISA. Variables identified as risk factors such as milk production (1-5L, absence of veterinary assistance, use of natural breeding or natural breeding associated with artificial insemination. These risk factors presented statistical significance (P<0.05 associated to BVDV infection. These results indicate that BVDV infection is

  17. PREVALÊNCIA DE LEUCOSE ENZOÓTICA BOVINA, DIARRÉIA VIRAL BOVINA RINOTRAQUEÍTE INFECCIOSA BOVINA E NEOSPOROSE BOVINA EM 26 PROPRIEDADES LEITEIRAS DA REGIÃO NORDESTE DO RIO GRANDE DO SUL, BRASIL PREVALENCE OF ENZOOTIC BOVINE LEUKOSIS, BOVINE VIRAL DIARRHEA, INFECTIOUS BOVINE RHINOTRACHEITIS AND BOVINE NEOSPOROSIS IN 26 DAIRY CATTLE FARMS FROM THE NORTHEAST REGION OF RIO GRANDE DO SUL , BRAZIL

    Directory of Open Access Journals (Sweden)

    Cleber Fiori

    2008-12-01

    .936 and sex (p = 0.562. Of the 360 sheep samples, nine (2.5% were reactive. There also was no significant association between the analyzed variables and the seropositiveness for brucellosis: age group (p = 0.522; race (p = 0.576 and sex (p = 0.461. Significant association was observed (p = 0.042 among the studied species and seropositiveness for brucellosis in the investigated animals. The seropositiveness for Brucella abortus in goats and sheep was traced for the first time in the “Sertão” (dry interior region, backlands of Pernambuco, fact that can hinder the success of the National Program of Control and Erradication of Brucellosis, due to the fact that it is common to raise small ruminants with bovines in this area, besides representing risks to Public Health.
     
    KEY WORDS: Brucellosis, ovines, caprines, small ruminants, serodiagnosis.

  18. 猪源牛病毒性腹泻病毒SH-28分离株的全基因组序列及遗传进化分析%Complete genomic sequencing and analysis of the pig bovine viral diarrhea virus strain SH-28

    Institute of Scientific and Technical Information of China (English)

    陶洁; 王银; 廖金虎; 杨颖; 羊扬; 朱国强

    2013-01-01

    The complete genome of a pig bovine viral diarrhea virus (BVDV) SH-28 was sequenced in order to analyze the genetic difference between pig BVDV and bovine BVDV. Amplified by RT-PCR,18 fragments of SH-28 were cloned into pGEM-T and then sent for sequencing by commerical serves. The retrieved sequences were timed and aligned with CLUSTX(1.8) and Lasergene. Ultimately, we obtained the complete sequence of SH-28. The genome of SH-28 comprises 12 279 nucleotides and contains a large open reading frame (ORF) encoding a polyprotein of 3 895 amino acids. The 5',3'-untranslated region (5'-UTR,3'-UTR) are 385 nt and 206 nt, respectively. The phylogenetic tree of complete sequence showed that SH-28 and HLJ-10 were in the same branch,yet it was most similar to XJ-04 (92. 3%) and divergent with the other pig BVDV-1 (ZM-95,SD0806)(70. 0%,70. 1%). The phylogenetic tree of 5'-UTR showed that SH-28 belonged to BVDV-2a2 while HLJ-10 and XJ-04 were classified into BVDV-2al. Therefore, we speculate that SH-28 is a new BVDV-2 isolate different from the previous BVDV-2 strains and may originate from bovine BVDV-2.%为了解猪源牛病毒性腹泻病毒(BVDV)与牛源BVDV在遗传特性上的差异,本试验对实验室分离保存的1株猪源BVDV(SH-28)进行了全基因组测序.利用反转录-聚合酶链式反应(RT-PCR)方法分段扩增了SH-28分离株的18条cDNA片段,分别克隆于pGEM-T质粒载体并进行测序,用CLUSTX(1.8)和Lasergene软件进行序列的剪切和拼接,最终获得SH-28基因组序列全长为12 279个核苷酸(nt),开放阅读框(ORF)长11 685 nt,编码3 895个氨基酸(aa),5'-UTR和3’-UTR分别长385 nt和206 nt.全基因组进化树结果表明,虽然SH-28与HLJ-10同处于一个分支上,但其与XJ-04的全基因组核苷酸同源性最高(92.3%),而与另外2株猪源BVDV-1(ZM-95、SD0806)的亲缘性较远(70.0%、70.1%).5’-UTR进化树结果显示,SH-28分离株属于BVDV-2a2基因亚型,而HLJ-10和J-04株属于BVDV-2a1.

  19. 牛病毒性腹泻病毒E0和E2蛋白的融合表达及纯化%Fusion expression and purification of fusion protein of bovine viral diarrhea virus E0 and E2

    Institute of Scientific and Technical Information of China (English)

    陈为宏; 周玉龙; 尹辉; 高佳滨; 梁宏儒; 乔波; 陈楠楠; 翟军军; 朱战波

    2014-01-01

    目的 在大肠埃希菌中融合表达牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)E0和E2基因,并进行纯化.方法 采用RT-PCR法分别扩增BVDV BA株的E0和E2基因片段,通过酶切位点将二者串联并克隆至原核表达载体pET-28a上,构建重组表达质粒pET-28a-E0-E2,转化至大肠埃希菌BL21(DE3),IPTG诱导表达.表达的融合蛋白E0-E2经镍离子亲和层析纯化后,进行Western blot分析.结果 重组表达质粒pET-28a-E0-E2经双酶切和测序证明构建正确;表达的融合蛋白E0-E2相对分子质量约为68 500,表达量约占菌体总蛋白的20%,主要以包涵体形式存在;纯化的融合蛋白纯度达95%,可与牛病毒性腹泻病毒阳性血清发生特异性反应.结论 在大肠埃希菌中融合表达了BVDV E0和E2蛋白,纯化后的融合蛋白纯度较高,为BVDV抗体ELISA检测方法的建立及新型疫苗的研制奠定了基础.

  20. 东北地区规模化奶牛场牛病毒性腹泻/黏膜病血清学调查%Serological investigation of bovine viral diarrhea-mucosal disease on scale dairy farms in the Northeast China

    Institute of Scientific and Technical Information of China (English)

    商云鹏; 刘华; 张海丽; 高明春; 张文龙; 王君伟

    2013-01-01

    为了解东北地区牛病毒性腹泻/黏膜病病毒(BVDV)的流行情况,本研究采用间接ELISA试验与套式RT-PCR分型检测方法对东北地区19个规模化奶牛场的1 198份血清进行了BVDV的抗体与核酸检测.结果表明,BVDV在东北地区呈散发性流行,抗体阳性率为23.5%,各奶牛场抗体阳性率在0~40%之间;BVDV核酸阳性的奶牛场均包括在抗体阳性的奶牛场中,并且均为BVDV Ⅰ型.结果提示,东北地区大部分奶牛场中存在BVDV污染,并且可能存在有持续性感染牛,应采取相应的净化措施对该病进行控制.%To investigate the prevalence of bovine viral diarrhea-mucosal disease virus (BVDV) in the Northeast China,a total of 1,198 serum samples,which were collected from nineteen dairy farms in the Northeast China from 2011 to 2012,were detected by an indirect ELISA for antibodies and multiplex nested RT-PCR for the genetype of BVDV.The results showed that BVDV in the Northeast China was sporadic epidemic and the average antibody positive rate was 23.5%,ranging from 0 to 40% in those dairy farms which was identical to the PCR detection.In addition,the BVDVs were all belong to the genotype Ⅰ.The results suggest that the majority of dairy farms in the Northeast China exists BVDV infection and the appropriate measures should be taken to control the disease.

  1. Apoptosis induction by miRNA-125b in bovine viral diarrhea virus infected MDBK cells%miRNA-125b在牛病毒性腹泻病毒感染MDBK细胞中诱导细胞凋亡的研究

    Institute of Scientific and Technical Information of China (English)

    王讲德; 程婷婷; 陈创夫; 任艳; 张辉; 王远志; 李蕊; 李跃峰; 倪伟

    2013-01-01

    To study the apoptosis mechanism of Madin-Darby bovine kidney (MDBK) cells induced by miRNA-125b in the process of the cells infected with cytopathic bovine viral diarrhea virus (cpBVDV),MDBK cells were infected with cpBVDV or transfected with pcDNA-miR-125b as a positive control.The transcrption levels of miRNA-125b and B cell lymphoma/lewkmia-2 (Bcl-2) mRNA were detected by real-time RT-PCR (qRT-PCR),and the cell apoptosis was examined by Flow Cytometry.The results showed the miRNA-125b levels in MDBK cells infected with cpBVDV at 12 hours and 24 hours were 2.01 and 3.85 times higher and the Bcl-2 mRNA levels were also 0.71 and 0.31 times higher than that in normal cells,respectively.In addition,the Bcl-2 mRNA level in MDBK cells transfected with pcDNA-miR-125b at 48 hours was 0.42 times higher compared with that in normal cells.Furthermore,The apoptosis rates of cells infected with cpBVDV at 12 hours and 24 hours were 15.06% and 36.43%,respectively; and apoptosis rate of cells transfected with pcDNA-miR-125b at 48 hours was 25.56%.In conclusion,these data demonstrated that cpBVDV infection triggered the high level transcription of miRNA-125b which induced the cells to undergo apoptosis by inhibiting transcription level of Bcl-2 mRNA.%为研究miRNA-125b在致细胞病变牛病毒性腹泻病毒(cpBVDV)感染牛肾细胞(MDBK细胞)过程中诱导细胞凋亡的机制,本研究将cpBVDV感染MDBK细胞,并将构建的pcDNA-miR-125b转染MDBK细胞作为阳性对照.采用荧光定量RT-PCR (qRT-PCR)检测细胞中miRNA-125b和B细胞淋巴瘤/白血病-2(Bcl-2) mRNA转录水平,通过流式细胞仪检测细胞凋亡率.检测结果显示,cpBVDV感染MDBK细胞12 h和24 h,细胞中miRNA-125b的转录水平分别为正常细胞对照组的2.01倍和3.85倍,Bcl-2 mRNA的转录水平分别为对照组的0.71倍和0.31倍;pcDNA-miR-125b转染MDBK细胞48 h后,Bcl-2 mRNA的转录水平为正常细胞对照组的0.42倍.细胞凋亡检测

  2. Diarrhea

    Science.gov (United States)

    ... with diarrhea should drink water, fruit juices, sports drinks, sodas without caffeine, and salty broths. As your symptoms improve, you can eat soft, bland food. Children with diarrhea should be given oral rehydration solutions ...

  3. Structured literature review of responses of cattle to viral and bacterial pathogens causing bovine respiratory disease complex.

    Science.gov (United States)

    Grissett, G P; White, B J; Larson, R L

    2015-01-01

    Bovine respiratory disease (BRD) is an economically important disease of cattle and continues to be an intensely studied topic. However, literature summarizing the time between pathogen exposure and clinical signs, shedding, and seroconversion is minimal. A structured literature review of the published literature was performed to determine cattle responses (time from pathogen exposure to clinical signs, shedding, and seroconversion) in challenge models using common BRD viral and bacterial pathogens. After review a descriptive analysis of published studies using common BRD pathogen challenge studies was performed. Inclusion criteria were single pathogen challenge studies with no treatment or vaccination evaluating outcomes of interest: clinical signs, shedding, and seroconversion. Pathogens of interest included: bovine viral diarrhea virus (BVDV), bovine herpesvirus type 1 (BHV-1), parainfluenza-3 virus, bovine respiratory syncytial virus, Mannheimia haemolytica, Mycoplasma bovis, Pastuerella multocida, and Histophilus somni. Thirty-five studies and 64 trials were included for analysis. The median days to the resolution of clinical signs after BVDV challenge was 15 and shedding was not detected on day 12 postchallenge. Resolution of BHV-1 shedding resolved on day 12 and clinical signs on day 12 postchallenge. Bovine respiratory syncytial virus ceased shedding on day 9 and median time to resolution of clinical signs was on day 12 postchallenge. M. haemolytica resolved clinical signs 8 days postchallenge. This literature review and descriptive analysis can serve as a resource to assist in designing challenge model studies and potentially aid in estimation of duration of clinical disease and shedding after natural pathogen exposure.

  4. 猪繁殖与呼吸综合征病毒和牛病毒性腹泻病毒双重RT-PCR检测方法的建立%Establishment of duplex RT-PCR assay for detection of bovine viral diarrhea virus and porcine reproductive and respiratory syndrome virus

    Institute of Scientific and Technical Information of China (English)

    王克伟; 李玉峰; 王先炜; 马涛; 张国龙; 姜平

    2011-01-01

    根据牛病毒性腹泻病毒(BVDV)5'端非编码区基因序列,设计合成了1对特异性引物,参考本实验室针对猪繁殖与呼吸综合征病毒(PRRSV)N蛋白设计的引物,经过PCR反应条件的优化,建立了BVDV和PRRSV双重RT-PCR的检测方法.对于PRRSV和BVDV的cDNA最低检测量分别为3.8×10-4 ng和7×1014 ng,对于猪瘟病毒(CFSV)、脑心肌炎病毒(EMCV)和猪圆环病毒2型(PCV-2)的PCR扩增结果均为阴性;用该方法对江苏省不同地区采集的75份仔猪的肺脏、脾脏和淋巴结等病料进行了检测,结果PRRSV有55份阳性,BVDV有14份阳性,PRRSV和BVDV混合感染的有12份,与PRRSV和BVDV单一RT-PCR的检测结果符合率分别为89.3%和92%.证明建立的双重RT-PCR检测方法可用于临床样品中BVDV和PRRSV的检测.%According to the sequence of 5' untranslated region (5'UTR) of the bovine viral diarrhea virus ( BVDV) genome, a pair of primers was designed and synthesized. Combined with the established primers of the N protein of porcine reproductive and respiratory syndrome vims ( PRRSV) , a duplex RT-PCR was established for detection of the two viruses simultaneously based on the optimization of the reaction condition. The specific PCR products were 290 bp for BVDV and 374 bp for PRRSV in length. The detection limit of cDNA template in the RT-PCR was 3. 8×1O-4 ng for PRRSV and 7×10-4 ng for BVDV. The high specificity of the method was demonstrated by detecting classical swine fever virus ( CSFV) , encephalomyocarditis virus ( FMCV) and porcine circovirus 2 ( PCV-2). Among 75 clinical samples,there were 55 samples with PRRSV positive, 14 samples with BVDV positive, and 12 samples with coinfection of BVDV and PRRSV. The coincidence rates were 89. 3% and 92% with the single RT-PCR for PRRSV and BVDV, respectively. It suggested that the duplex RT-PCR assay in this study could be applied for the detection of BVDV and PRRSV in clinical samples.

  5. Development of Real-time Fluorescence Quantitative PCR Assay for Detection of Bovine Viral Diarrhea Virus in Pigs%猪源牛病毒性腹泻病毒实时荧光定量PCR检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    邓宇; 张荣; 丛雁方; 张建武; 袁世山; 文心田; 郑浩

    2011-01-01

    This study was aimed at developing a real-time fluorescence quantitative PCR assay for detecting and discriminating pig bovine viral diarrhea virus-1 (BVDVs) and classical swine fever virus (CSFV). According to the conserved gene sequence of 5 -UTR of BVDVs and CSFV, pairs of primer and specific TaqMan probe were designed. The positive standard plasmids were used as quantitative template to make the standard curves, through the method was optimized. Specificity, sensitivity, and conformity were tested. By sensitivity analysis, the FQ-PCR indicated that a minimum of 10 copies of plasmid DNA was detected. The assay exhibited reproducibility and specificity, and all negative controls such as CSFV and porcine reproductive and respiratory syndrome virus (PRRSV) showed negative detection. As a result of the sensitivity and specificity of the assay with a relatively rapid and simple procedure, the real-time FQ-PCR can be used as a routine assay for the diagnosis of pig BVDV infection and CSFV vaccine polluted by BVDVs.%本研究旨在建立一种荧光定量PCR检测方法,用于猪源牛病毒性腹泻病毒(BVDVs)检测,同时能鉴别诊断猪瘟病毒(CSFV).根据BVDVs与CSFV 5 '-UTR保守序列,设计一套引物和特异TaqMan探针,以本实验室构建的猪源BVDVs 5′-UTR阳性重组质粒为标准品,通过优化反应条件,建立了标准曲线.以构建的标准品为模板进行了特异性、敏感性、重复性试验、并应用于临床检测.结果显示,该方法检测CSFV、猪繁殖与呼吸综合征病毒均为阴性;最低可检测到每个反应相当于10拷贝的标准品DNA;重复性试验的批内变异系数为0.20%~0.95%;应用所建立方法对临床样品进行检测,其结果与普通RT-PCR结果的符合率为86.7%.本研究建立的实时荧光定量PCR具有特异、敏感、快速、重复性好等优点,可用于猪感染BVDVs、猪瘟疫苗污染BVDVs的监测.

  6. Construction and immune effect of DNA vaccine against bovine viral diarrhea-mucosal virus%牛病毒性腹泻/粘膜病病毒核酸疫苗的构建及其免疫效果研究

    Institute of Scientific and Technical Information of China (English)

    宫玉玲; 冉多良; 季新成; 刘建华

    2013-01-01

    为提高牛病毒性腹泻/粘膜病(BVD/MD)核酸疫苗的免疫效力,本实验应用PCR方法扩增BVD病毒(BVDV) E0基因,构建真核表达质粒pVAX1-E0,转染293T细胞,经RT-PCR和western blot分析显示,转染细胞能够瞬时表达E0蛋白.并分别将pVAX1、pVAX1-E0或将pVAX1-E0分别与一种表达细胞因子基因的重组质粒作为佐剂(pVAX1-IL-2、pVAX1-IL-4及pVAX1-IFN-γ)免疫小鼠,采用间接ELISA法检测免疫小鼠BVDV抗体效价,以MTT法检测免疫小鼠脾淋巴细胞的增殖活性.实验结果表明,与pVAX1-E0相比,接种pVAX1-E0/pVAX1-IL-2小鼠血清E0抗体水平及淋巴细胞增殖水平显著提高(p<0.01),表明细胞因子基因佐剂IL-2能够有效提高BVDV E0核酸疫苗免疫效果,可以刺激小鼠产生良好的免疫应答.%To improve the immune efficacy of DNA vaccine against the bovine viral diarrhea virus-mucosal disease virus (BVDV/MDV),BVDV E0 gene was amplified by PCR and inserted into pVAX1 eukaryotic expression vector to construct recombinant plasmid pVAX1-E0.In addition,the BVDV E0 gene transiently expressed in 293T cells transfected with pVAX1-E0 was verified by RT-PCR and western blot.Furthermore,the mice were immunized by co-inoculation of pVAV1-E0 with one of three recombinant plasmids expressing cytokine gene as adjuvant (pVAX1-IL-2,pVAX1-IL-4 and pVAX1-IFN-γ),and single inoculation with pVAX1 and pVAV1-E0,respectively.The results showed that the antibody titers against E0 and the ability to promote lymphocytes proliferation were significantly higher in mice group co-inoculated with pVAX1-E0 and pVAX1-IL-2 than that the mice in control groups (p<0.01),indicating the IL-2 was able to improve the immune effect of DNA vaccine.

  7. Prevalencia y distribución espacial de brucelosis, leucosis bovina, diarrea viral bovina y rinotraqueítis infecciosa bovina a partir del análisis ELISA de estanques prediales en lecherías de la IX Región, Chile Prevalence and space distribution of brucellosis, bovine leukaemia, bovine viral diarrhea and infectious bovine rhinotracheitis by using bulk milk ELISA test in dairy herds of the IX Region, Chile

    Directory of Open Access Journals (Sweden)

    R Felmer

    2009-01-01

    of the OIE List, including foot and mouth disease and classical swine fever. However, several infectious diseases are known to remain among herds, which produce a major effect in production due to losses by abortion, decrease of fertility and what it is more important, some of them represent barriers for export and constitute a risk of zoonosis for the population. In this work, a monitoring system based on the analysis of bulk milk antibodies by means of ELISA test, was implemented to study the epidemiology and distribution of 4 of the main bovine diseases that currently affect the IX Region of Chile (brucellosis, bovine leukaemia, IBR and BVD. The system allowed the surveillance of 279 dairies, which represented 43% of the dairies registered in IX the Region, and included 19,635 milking cows (14%. With this system, a high prevalence for leukaemia (59%, IBR (76% and BVD (96% could be established, whereas it was confirmed that brucellosis is restricted to a few dairies (5%. The surveillance system coupled to the satellite geographic information analysis, allowed to establish the space distribution of these diseases in the different communes of the Region, demonstrating to be an excellent and low cost support tool for the monitoring of the diseases in the herd, which guarantees the possibility of establishing this platform in the Region and its feasibility to project it at national level.

  8. Progress on Yak Viral Diarrhea/Mucosal Disease%牦牛病毒性腹泻/黏膜病研究进展

    Institute of Scientific and Technical Information of China (English)

    李家奎

    2013-01-01

    牛病毒性腹泻/黏膜病(BVD)是由牛病毒性腹泻病毒(BVDV)引起的一种极为复杂,呈多种临床症状类型表现的疾病.目前,该病毒在世界范围内广泛分布,是造成全球乳/肉牛业经济损失的主要病原.本文针对我国牦牛BVD的发病状况进行分析,并对今后牦牛BVD防控进行了展望.%Bovine viral diarrhea (BVD) is a disease caused by bovine viral diarrhea virus (BVDV),this disease is extremely complex with various clinical symptoms. Currently, BVDV is widely distributed in the world, and is a major pathogen causing economic losses of global milk/beef cattle industry. In this paper, the incidence of yak BVD in China is outlined, and how to prevent and control yak BVD is also prospected.

  9. Comparison of different protocols for the bovine viral diarrhea virus detection by RT-PCR in pools of whole blood and blood serum artificially contaminated/ Comparação de diferentes protocolos para a detecção do vírus da diarréia viral bovina por RT-PCR em grupos de sangue total e de soro sangüíneo, artificialmente contaminados

    Directory of Open Access Journals (Sweden)

    Amauri A. Alfieri

    2005-06-01

    Full Text Available The RT-PCR technique was optimized and evaluated to detect the 5’ untranslated region of bovine viral diarrhea virus (BVDV from clinical samples that consisted of blood serum and whole blood artificially contaminated with the NADL strain of BVDV. To optimization of technique, the following conditions were evaluated: i two pairs of primers, 103 / 372 (WEINSTOCK et al., 2001 and 324 / 326 (VILCEK et al., 1994, ii four methods of nucleic acid extraction (phenol/chloroform/isoamyl alcohol; silica/guanidine isothiocyanate; a combination of the two previous methods; and TRIzol™ and iii different concentrations and compositions of reagents and time/temperature of the reactions. Between the alternatives tested that resulted in the amplification of the 290 bp product that was easily visualized in ethidium bromide stained 2% agarose gel was that presented the following conditions: i primers 103 and 372; ii initial volume and clinical sample: 50 mL of blood serum; iii extraction of nucleic acid: silica/guanidine isothiocyanate method; iv reverse transcription: 9 mL extracted nucleic acid, 1xPCR buffer (20 mM Tris-HCl pH 8.4 and 50 mM KCl, 1.5 mM MgCl2; 60 units of reverse transcriptase enzyme M-MLV, RNA denaturation at 97°C / 4 min, and reverse transcription at 42°C / 30 min; v PCR: primers 103 / 372 with anneling temperature at 59°C. The utilization of RT-PCR within these optimized conditions allowed the amplification of the BVDV NADL strain (103,56 TCID50 from pools of artificially contaminated blood serum until the dilution 1:160.A técnica da RT-PCR foi otimizada e avaliada para a detecção da região 5’ terminal não-traduzida do genoma do vírus da diarréia viral bovina (BVDV em amostras clínicas de bovinos, constituídas por soro sangüíneo e sangue total, artificialmente contaminadas com a estirpe NADL do BVDV. Para a otimização da técnica foram avaliados: i dois pares de primers, 103 / 372 (WEINSTOCK et al., 2001 e 324 / 326

  10. [Viral diarrhea].

    Science.gov (United States)

    Stalder, H

    1982-04-12

    Viruses are one of the most frequent causes of acute infectious gastroenteritis. Thus, rotaviruses are responsible for 40-60%--and in winter sometimes for over 90% - of diarrhoea in infants. These viruses may also cause disease in adults, but small viruses of a size of about 27 nm, such as the Norwalk agent, play a much more important role. Several other non-cultivable viruses such as adeno-, calici- and astroviruses have been implicated as a cause of infectious diarrhoea. Knowledge of rotaviruses is so far advanced that routine laboratory diagnosis can easily be performed and active immunization against rotavirus infection envisaged.

  11. Atividade antiviral do extrato de própolis contra o calicivírus felino, adenovírus canino 2 e vírus da diarréia viral bovina Antiviral activity of propolis extracts against feline calicivirus, canine adenovirus 2, and bovine viral diarrhea virus

    OpenAIRE

    Ana Paula Cueto; Sydney Hartz Alves; Marciele Pilau; Rudi Weiblen; Thaís Felli Kubiça; Luciane Teresinha Lovato

    2011-01-01

    Dentre as propriedades biológicas da própolis, a atividade antimicrobiana tem merecido destacada atenção. Neste artigo, descreve-se a atividade antiviral de dois extratos etanólicos de própolis (EP1 e EP2) frente aos vírus: calicivírus felino (FCV), adenovírus canino tipo 2 (CAV-2) e vírus da diarréia viral bovina (BVDV). Um dos extratos (EP1) foi obtido por extração etanólica de própolis obtida da região central do Estado do Rio Grande do Sul e o segundo (EP2), obtido comercialmente de uma e...

  12. Viral metagenomics analysis of picobirnavirus-positive feces from children with sporadic diarrhea in China.

    Science.gov (United States)

    Sun, Guangming; Zang, Qinbo; Gu, Yu; Niu, Guoping; Ding, Chen; Zhang, Peiying

    2016-04-01

    Picobirnaviruses (PBVs) infect humans and a wide range of animals and may cause diarrhea. The aim of this study was to investigate PBV infection and its association with diarrhea. Here, seven PBV RT-PCR-positive fecal samples from diarrheic children and four fecal samples from healthy children as controls were analyzed by viral metagenomics. The results indicated that all the seven diarrheic fecal samples contain high titers of PBV sequences, while three of the controls were negative, and one had low titers of PBV. Three of the diarrheic fecal samples were also positive for other viruses, including anellovirus, human gyrovirus, human parechovirus, and porcine stool-associated circular virus. PBV sequences from the seven patients were assembled, generating seven large contigs with the complete ORF of RNA-dependent RNA polymerase (RdRp). Phylogenetic analysis of the amino acid sequences of RdRp indicated that the seven PBVs in the present study belonged to three different genogroups. Our data suggest that PBV might have been the cause of diarrhea in these seven children.

  13. Detection and characterization of viruses as field and vaccine strains in feedlot cattle with bovine respiratory disease

    Science.gov (United States)

    This study investigated viruses in bovine respiratory disease (BRD) cases in feedlots, including bovine herpesvirus-1 (BoHV-1), bovine viral diarrhea virus (BVDV), bovine respiratory syncytial virus (BRSV), bovine coronaviruses (BoCV) and parainfluenza-3 virus (PI3V). Nasal swabs were collected fro...

  14. 牛病毒性腹泻病毒和牛轮状病毒TaqMan二重实时荧光RT-PCR检测方法的建立%Detection of bovine viral diarrhea virus and bovine rotavirus by TaqMan based real-time RT-PCR

    Institute of Scientific and Technical Information of China (English)

    范晴; 谢芝勋; 刘加波; 庞耀珊; 邓显文; 谢志勤; 谢丽基; 彭宜

    2011-01-01

    根据牛病毒性腹泻病毒(BVDV)5′端非编码区和牛轮状病毒(BRV)VP6基因序列,设计特异性引物和探针。通过对引物和探针浓度、Mg2+浓度、dNTP浓度和Taq酶用量以及反应条件等因素的优化筛选,建立了能同时鉴别BVDV和BRV的二重荧光RT-PCR方法。该方法特异性好,与其他病原如CSFV、MB和IBRV不发生交叉反应;敏感性高,能够检测100个BVDV RNA和100个BRV RNA;稳定性好,批内重复和批间重复变异系数小;干扰性试验表明该方法能同时检测2个模板的不同浓度组合。本研究建立的二重荧光RT-PCR方法可用于BVDV和BRV检测,具有特异、敏感、快速、稳定等优点,是BVDV和BRV基础研究、流行病学调查和临床检测的良好工具。%Two pairs of primers and two TaqMan probes were designed and synthesized according to the conserved gene sequence of BVDV 5′ untrascription region and BRV VP6.The reaction parameters such as the concentration of two pair of primers,two probes and other conditions were optimized to develop a duplex real-time RT-PCR assay for rapid detection of BVDV and BRV.It was found that the specificity of this assay was high,and be able to detected BVDV and BRV without other any cross-reactions to CSFV,MB and IBRV.The detection limit of the real-time RT-PCR assay was 100 copies of BVDV viral RNA and BRV viral RNA,indicating a good sensitivity of the assay.The coefficients of variation were both low for the intra-assay and inter-assay tests respectively,indicating a good reliability.When different concentration of BVDV and BRV was mixed together,the result was without any interference.All the resuls indicate that this duplex real-time PCR assay is a specific,sensitive,rapid and reproducible method for detection of BVDV and BRV,and is could applied in fundamental research,clinical detection and epidemiological investigation of BVDV and BRV.

  15. Viral and Bacterial Pathogens in Bovine Respiratory Disease in Finland

    Directory of Open Access Journals (Sweden)

    Soveri T

    2004-12-01

    Full Text Available Pathogens causing bovine respiratory tract disease in Finland were investigated. Eighteen cattle herds with bovine respiratory disease were included. Five diseased calves from each farm were chosen for closer examination and tracheobronchial lavage. Blood samples were taken from the calves at the time of the investigation and from 86 calves 3–4 weeks later. In addition, 6–10 blood samples from animals of different ages were collected from each herd, resulting in 169 samples. Serum samples were tested for antibodies to bovine parainfluenza virus-3 (PIV-3, bovine respiratory syncytial virus (BRSV, bovine coronavirus (BCV, bovine adenovirus-3 (BAV-3 and bovine adenovirus-7 (BAV-7. About one third of the samples were also tested for antibodies to bovine virus diarrhoea virus (BVDV with negative results. Bacteria were cultured from lavage fluid and in vitro susceptibility to selected antimicrobials was tested. According to serological findings, PIV-3, BAV-7, BAV-3, BCV and BRSV are common pathogens in Finnish cattle with respiratory problems. A titre rise especially for BAV-7 and BAV-3, the dual growth of Mycoplasma dispar and Pasteurella multocida, were typical findings in diseased calves. Pasteurella sp. strains showed no resistance to tested antimicrobials. Mycoplasma bovis and Mannheimia haemolytica were not found.

  16. Diarrhea - overview

    Science.gov (United States)

    ... cause of diarrhea is the stomach flu ( viral gastroenteritis ). This mild viral infection goes away on its own within a few days. Eating or drinking food or water that contains certain types of bacteria or parasites can also lead to diarrhea. This ...

  17. Bovine virus diarrhea virus in free-living deer from Denmark.

    Science.gov (United States)

    Nielsen, S S; Roensholt, L; Bitsch, V

    2000-07-01

    Free-living deer are suggested as a possible source of infection of cattle with bovine virus diarrhea (BVD) virus. To examine this hypothesis blood samples from 476 free-living deer were collected during two different periods and tested for BVD virus and antibody in Denmark. In 1995-96, 207 animals were tested. These included 149 roe deer (Capreolus capreolus), 29 fallow deer (Dama dama), 20 red deer (Cervus elaphus) and one sika deer (Cervus sika). For the remaining eight animals no species information was available. In 1998-99, 269 animals were tested including 212 roe deer and 57 red deer. The animals were selected from areas with a relatively high prevalence of cattle herds with a BVD persistent infection status in 1997 and 1998. All 207 samples from 1995-96 were found antibody-negative except two samples from red deer. Only 158 of the 207 samples were tested for virus and were all found negative. Of the 269 samples from 1998-99 all but one were antibody negative. The positive sample was from a red deer. All samples were virus-negative. It appears that BVD infection does not occur in roe deer in Denmark. The presence of antibody in a few red deer from various districts in Jutland probably results from cattle to deer transmission, rather than spread among deer. Hence, the possibility of free-living deer as a source of infection for cattle in Denmark seems to be remote.

  18. Safety and efficacy of an E2 glycoprotein subunit vaccine produced in mammalian cells to prevent experimental infection with bovine viral diarrhoea virus in cattle.

    Science.gov (United States)

    Pecora, Andrea; Aguirreburualde, María Sol Pérez; Aguirreburualde, Alejandra; Leunda, Maria Rosa; Odeon, Anselmo; Chiavenna, Sebastián; Bochoeyer, Diego; Spitteler, Marcelo; Filippi, Jorge L; Dus Santos, Maria J; Levy, Susana M; Wigdorovitz, Andrés

    2012-09-01

    Bovine viral diarrhea (BVD) infection caused by bovine viral diarrhea virus (BVDV), a Pestivirus of the Flaviviridae family, is an important cause of morbidity, mortality and economical losses in cattle worldwide. E2 protein is the major glycoprotein of BVDV envelope and the main target for neutralising antibodies (NAbs). Different studies on protection against BVDV infection have focused on E2, supporting its putative use in subunit vaccines. A truncated version of type 1a BVDV E2 (tE2) expressed in mammalian cells was used to formulate an experimental oleous monovalent vaccine. Immunogenicity was studied through immunisation of guinea pigs and followed by trials in cattle. Calves of 8-12 months were vaccinated, twice with a 4 week interval, with either a tE2 subunit vaccine (n = 8), a whole virus inactivated vaccine (n = 8) or left untreated as negative control group (n = 8). Four weeks after the last immunisation the animals were experimentally challenged intranasally with a non-cythopathic BVDV strain. Following challenge, BVDV was isolated from all unvaccinated animals, while 6 out of 8 animals vaccinated with tE2 showed complete virological protection indicating that the tE2 vaccine presented a similar performance to a satisfactory whole virus inactivated vaccine.

  19. Research outline of bovine virus diarrhea%牛病毒性腹泻的研究概况

    Institute of Scientific and Technical Information of China (English)

    谢西锋; 崔保安

    2001-01-01

    @@ 牛病毒性腹泻/粘膜病(Bovine viral diarrhea/mucosal disease,BVD/MD),简称牛病毒性腹泻(BVD)或牛粘膜病(BMD),是由牛病毒性腹泻/粘膜病病毒(BVD/MDV)感染牛引起的以发热、粘膜糜烂溃疡、白细胞减少、腹泻、咳嗽及怀孕母牛流产或产出畸形胎儿为主要特征的一种传染病.1946年Olafson等首次报道病毒性腹泻病.1953年Ramsey和Chiver发现粘膜病.1961年Gillespie等研究证明,这两种病毒是有共同抗原性的同种病毒[1],1971年由美国兽医协会将其统一命名为“牛病毒性腹泻/粘膜病”.BVD/MD呈世界性分布,在许多养牛发达国家,如美国、新西兰、加拿大等尤其严重[2].1980年李佑民首先证明我国也有本病存在[3].自80年代以来,我国已有15个省、市、自治区查出该病,感染动物包括牛、羊、猪、骆驼、鹿等[4,5,6],而且其致病机理与临床类型较为复杂,现将本病的病原、流行病学、致病机理、病理变化、临床症状、防治等方面作一综述.

  20. Establishment of a Bovine Herpesvirus 4 based vector expressing a secreted form of the Bovine Viral Diarrhoea Virus structural glycoprotein E2 for immunization purposes

    Directory of Open Access Journals (Sweden)

    Donofrio Gaetano

    2007-10-01

    Full Text Available Abstract Background The biological characteristics of BoHV-4 make it a good candidate as a gene delivery vector for vaccination purposes. These characteristics include little or no pathogenicity, unlikely oncogenicity, the capability to accommodate large amounts of foreign genetic material, the ability to infect several cell types from different animal species, and the ability to maintain transgene expression in both undifferentiated and differentiated cells. Results A recombinant bovine herpesvirus 4 (BoHV-4CMV-IgKE2-14ΔTK expressing an enhanced secreted form of the bovine viral diarrhea virus (BVDV structural glycoprotein E2 (gE2-14, obtained by the removal of the putative transmembrane domain and addition of a 14 amino acids peptide at its carboxyl terminal and an immunoglobulin K signal peptide to the amino terminal, was successfully constructed using a Recombineering (recombination -mediated genetic engineering approach on BoHV-4 cloned as bacterial artificial chromosome. The galactokinase – based recombineering system was modified by the introduction of a kanamycin expression cassette and a kanamycin selection step that allowed a significant reduction of the untargeted background clones. BoHV-4CMV-IgKE2-14ΔTK infected cell lines highly expressed gE2-14, which maintained native antigenic properties in a serum neutralization inhibition test. When rabbits and sheep were immunized with BoHV-4CMV-IgKE2-14ΔTK, high levels of serum neutralized antibodies against BVDV were generated. Conclusion This work highlights the engineerization of BoHV-4 genome as a vector for vaccine purposes and may provide the basis for BVDV vaccination exploiting the BoHV-4- based vector that delivers an improved secreted version of the BVDV structural glycoprotein E2.

  1. Survivability of porcine epidemic diarrhea virus (PEDV) in bovine plasma submitted to spray drying processing and held at different time by temperature storage conditions.

    Science.gov (United States)

    Pujols, Joan; Segalés, Joaquim

    2014-12-05

    Bovine plasma was inoculated with porcine epidemic diarrhea virus (PEDV) at an average final titer of 4.2 log10 TCID50/mL to determine the effect of spray drying on viral inactivation. Using a laboratory scale drier, inoculated plasma was spray dried at 200 °C inlet temperature and either 70 or 80 °C throughout substance. Both liquid and dried samples were subjected to three passages on VERO cell monolayers to determine PEDV infectivity. Results indicated liquid samples contained infective virus, but none of the spray dried samples were infectious. Also, survivability of PEDV inoculated on spray dried bovine plasma (SDBP) and stored at 4, 12 or 22 °C was determined for 7, 14 and 21 days. Commercial SDBP powder was inoculated with PEDV to an average final titer of 2.8 log10 TCID50/g. Five samples per time and temperature conditions were subjected to three passages on VERO cell monolayers to determine PEDV infectivity. The virus was non-infectious for all samples stored at 22 °C at 7, 14 and 21 days. PEDV was infective in 1 out of 5 samples stored at 12 °C at 7 days, but none of the samples stored for 14 and 21 days were infectious in cell culture. For samples stored at 4 °C, 4 out of 5 samples were infectious at 7 days, 1 out of 5 samples were infectious at 14 days, but none were infectious at 21 days. In summary, PEDV was not infectious on cell culture within 7 days when stored at room temperature and within 21 days when stored at refrigerated temperature.

  2. Effect of the bovine viral diarrhoea virus (BVDV) infection on dairy calf rearing.

    Science.gov (United States)

    Diéguez, Francisco J; Yus, Eduardo; Vilar, María J; Sanjuán, María L; Arnaiz, Ignacio

    2009-08-01

    The aim of this study was to compare the cumulative incidence of mortality, clinical diarrhoea and respiratory disease in calves, during their first six months of age, in herds with different bovine viral diarrhoea virus (BVDV) infection status. Calves' health indicators were tested by comparing proportions in 101 farms with dissimilar infection condition. The results indicate that there was a significant relationship between the BVDV status (actively infected herd or not) and the cumulative incidence of mortality and respiratory disorders.

  3. Vertical transmission of bovine viral diarrhoea virus (BVDV) in mousedeer (Tragulus javanicus) and spread to domestic cattle

    DEFF Research Database (Denmark)

    Uttenthal, Åse; Høyer, M.J.; Grøndahl, C.;

    2006-01-01

    This study investigates the transmission of bovine viral diarrhoea virus (BVDV) 1f from a persistently infected (PI) lesser Malayan mousedeer to two bovine calves. Different contact routes to two calves were analysed: 1) aerosol contact between two adjacent pens without physical contact; 2...

  4. 牛病毒性腹泻-粘膜病病毒TaqMan荧光定量RT-PCR检测方法的建立及初步应用%Establishment and Initial Application of Fluorescent Quantitative RT -PCR for Detection of Bovine Viral Diarrhea Virus

    Institute of Scientific and Technical Information of China (English)

    陈其兵; 薛霜; 漆世华; 朱薇; 张萍; 谢红玲; 温文生; 吴玉石

    2012-01-01

    为建立-种牛病毒性腹泻-粘膜病病毒(BVDV)的快速检测方法,根据GenBank中登录的BVDV基因序列,针对5’UTR的保守序列,设计合成了一对特异性引物和一条TaqMan荧光探针。通过对反应条件和反应体系进行优化,建立了一种能快速定量检测BVDV的荧光定量RT—PCR检测方法。通过对20份胎牛血清样品和猪瘟细胞苗半成品进行检测,对该方法的特异性、敏感性和重复性进行试验。结果显示,建立的方法能检测到BVDV,而对CSFV、PRRSV和MDBK细胞的扩增结果均为阴性,具有高度的特异性。在所检测的20份样品中,BVDV阳性6份(30%)。对所构建的标准品进行检测,在10^2-10^8拷贝/μL的范围内可以得到良好的动力学曲线,能检测低至10^3-10^4拷贝/mL的病毒量。表明所建立的检测方法具有快速、特异、灵敏、重复性好等特点,可用于临床及科研中对BVDV的快速定量检测和对牛血清及猪瘟兔化弱毒疫苗等生物制品中是否污染BVDV进行监测。%According to the genomic sequences within 5 ' UTR of BVDV which published in Genbank, a pair of primers and a Taqman probe were designed. Then a rapid fluorescent quantitative RT - PCR method was developed by optimization of the reaction conditions and system. This method was established by specificity, sensibility, reproducibility. Twenty samples of fetal bovine serum and swine fever vaccine were detected by this methed. The experiment showed that six samples were positive and the ratio was 30%, and the detection result of CSFV, PRRSV and MDBK cells were all negative. A standard curve was achieved and the result showed that the sensitivity of this method was 10^3 - 10^4copies/mL and the linear relation was excellent. The results show that this method is fast, specific, sensitive, repeatable and can be used to monitor the pollution of BVDV in biological products and quantitative detection of BVDV.

  5. A Multicenter, Prospective, Randomized Controlled Trial to Evaluate the Additional Benefit of a Multistrain Synbiotic (Prodefen®) in the Clinical Management of Acute Viral Diarrhea in Children

    Science.gov (United States)

    García-Menor, Emilia; García-Marín, Fátima; Vecino-López, Raquel; Horcajo-Martínez, Gloria; de Ibarrondo Guerrica-Echevarría, María-José; Gómez-González, Pedro; Velasco-Ortega, Syra; Suárez-Almarza, Javier; Nieto-Magro, Concepción

    2016-01-01

    This randomized, open-label study evaluated the additional benefits of the synbiotic Prodefen® in the clinical management of acute diarrhea of suspected viral origin in children between 6 months and 12 years of age. Study outcomes included the duration of diarrhea, the recovery from diarrhea, and the tolerability and acceptance of the treatment. The proportion of patients without diarrhea over the study period was greater in the synbiotic group than in the control group at all study time points, showing a statistically significant difference on the fifth day (95% vs 79%, p diarrhea (median and interquartile range) was reduced by 1 day in the synbiotic-treated patients (3 [2-5] vs 4 [3-5], p = 0.377). The tolerability of the treatment regimen, as evaluated by the parents, was significantly better in those receiving the synbiotic than in the control group. Overall, 96% of the parents of children receiving the synbiotic reported being satisfied to very satisfied with the treatment regimen. The results of this study indicate that the addition of the synbiotic Prodefen® is a well-tolerated and well-accepted approach that provides an additional benefit to the standard supportive therapy in the management of acute viral diarrhea in children. PMID:28229091

  6. A Multicenter, Prospective, Randomized Controlled Trial to Evaluate the Additional Benefit of a Multistrain Synbiotic (Prodefen® in the Clinical Management of Acute Viral Diarrhea in Children

    Directory of Open Access Journals (Sweden)

    Emilia García-Menor MD

    2016-11-01

    Full Text Available This randomized, open-label study evaluated the additional benefits of the synbiotic Prodefen® in the clinical management of acute diarrhea of suspected viral origin in children between 6 months and 12 years of age. Study outcomes included the duration of diarrhea, the recovery from diarrhea, and the tolerability and acceptance of the treatment. The proportion of patients without diarrhea over the study period was greater in the synbiotic group than in the control group at all study time points, showing a statistically significant difference on the fifth day (95% vs 79%, p < 0.001. The duration of diarrhea (median and interquartile range was reduced by 1 day in the synbiotic-treated patients (3 [2-5] vs 4 [3-5], p = 0.377. The tolerability of the treatment regimen, as evaluated by the parents, was significantly better in those receiving the synbiotic than in the control group. Overall, 96% of the parents of children receiving the synbiotic reported being satisfied to very satisfied with the treatment regimen. The results of this study indicate that the addition of the synbiotic Prodefen® is a well-tolerated and well-accepted approach that provides an additional benefit to the standard supportive therapy in the management of acute viral diarrhea in children.

  7. 川西北牦牛3种病毒性腹泻病血清学调查%Serological Survey on Three Viral Diarrhea Diseases of Yaks in Northwest Sichuan Province

    Institute of Scientific and Technical Information of China (English)

    何美琳; 张焕容; 王永; 王言轩; 王远微; 汤承

    2014-01-01

    本研究旨在了解川西北牦牛病毒性腹泻病的流行情况,为防制这类疫病提供一定科学依据.采用酶联免疫吸附试验(ELISA)对来源于川西北阿坝州8县的1070份牦牛血清中牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)、牛冠状病毒(bovine coronavirus,BCV)和牛轮状病毒(bovine rotavirus,BRV)进行了抗体检测.结果显示,BVDV、BCV和BRV抗体平均阳性率分别为44.3%、84.1%和94.4%.结果表明,BVDV、BCV和BRV感染在川西北牦牛中普遍存在,需进一步加强该地区牦牛病毒性腹泻疾病的综合防制.

  8. Seroprevalence of bovine viral diarrhoea virus in Hungary - situation before launching an eradication campaign.

    Science.gov (United States)

    Kővágó, Csaba; Forgách, Petra; Szabára, Ágnes; Mándoki, Míra; Hornyák, Ákos; Duignan, Conor; Pásztiné Gere, Erzsébet; Rusvai, Miklós

    2015-06-01

    Bovine viral diarrhoea (BVD) is a viral disease appearing in various forms and causing high economic losses in the cattle stocks of Hungary. The aim of the present study was to determine the prevalence of bovine viral diarrhoea virus (BVDV) in Hungary through a monitoring survey carried out on samples collected in cattle-keeping units throughout the country. Since no such survey had been carried out in Hungary during the last thirty years, our study may serve as a basis for later monitoring investigations aimed at following the progress of an expected eradication campaign of BVD. The tests were carried out using an ELISA method, on a total of 1200 blood samples submitted from 54 cattle herds. The herds had not been vaccinated against BVDV before the sampling. Out of the 1200 samples, 521 proved to be positive (43.4%), 40 gave doubtful result (3.3%) and 639 were negative (53.3%). In some stocks the samples were collected from cows having completed several lactation periods, and therefore the seronegativity indicates the BVDV-free status of the given stock. Moreover, among the positive herds we found a few where the seropositivity rate was rather low (campaign launched in the near future, or carried out parallel to the IBR eradication programme, are better than previously expected.

  9. A Two-Tube Multiplex Reverse Transcription PCR Assay for Simultaneous Detection of Viral and Bacterial Pathogens of Infectious Diarrhea

    Directory of Open Access Journals (Sweden)

    Ji Wang

    2014-01-01

    Full Text Available Diarrhea caused by viral and bacterial infections is a major health problem in developing countries. The purpose of this study is to develop a two-tube multiplex PCR assay using automatic electrophoresis for simultaneous detection of 13 diarrhea-causative viruses or bacteria, with an intended application in provincial Centers for Diseases Control and Prevention, China. The assay was designed to detect rotavirus A, norovirus genogroups GI and GII, human astrovirus, enteric adenoviruses, and human bocavirus (tube 1, and Salmonella, Vibrio parahaemolyticus, diarrheagenic Escherichia coli, Campylobacter jejuni, Shigella, Yersinia, and Vibrio cholera (tube 2. The analytical specificity was examined with positive controls for each pathogen. The analytical sensitivity was evaluated by performing the assay on serial tenfold dilutions of in vitro transcribed RNA, recombinant plasmids, or bacterial culture. A total of 122 stool samples were tested by this two-tube assay and the results were compared with those obtained from reference methods. The two-tube assay achieved a sensitivity of 20–200 copies for a single virus and 102-103 CFU/mL for bacteria. The clinical performance demonstrated that the two-tube assay had comparable sensitivity and specificity to those of reference methods. In conclusion, the two-tube assay is a rapid, cost-effective, sensitive, specific, and high throughput method for the simultaneous detection of enteric bacteria and virus.

  10. Genetic diversity of bovine viral diarrhoea viruses (BVDV) in Denmark during a 10-year eradication period

    DEFF Research Database (Denmark)

    Uttenthal, Åse; Stadejek, T.; Nylin, B.

    2005-01-01

    A 243 base-pair fragment of the 5'- untranslated region (5'-UTR) of bovine viral diarrhoea virus (BVDV) was RT-PCR amplified from tissue samples (after one passage) or from plasma collected from Danish cattle in 1962 (1), 1993 (7), or in 2002-03 (28) when BVD was almost extinct as a result of a 6...... subtype, the samples collected in 2002-2003 belonged to Id (22 samples), 1b (5 samples) and le (I sample) subtypes. In five herds, materials from two animals were obtained for PCR analysis. In four of five herds the sequences of the two viruses were identical, but in one herd the obtained sequences...

  11. Molecular epidemiology and phylogenetic analysis of diverse bovine astroviruses associated with diarrhea in cattle and water buffalo calves in China.

    Science.gov (United States)

    Alfred, Niyokwishimira; Liu, Huan; Li, Mu Lan; Hong, Shao Feng; Tang, Hai Bo; Wei, Zu Zhang; Chen, Ying; Li, Fa Kai; Zhong, Yi Zhi; Huang, Wei Jian

    2015-06-01

    Astroviruses are the principal causative agents of gastroenteritis in humans and have been associated with diarrhea in other mammals as well as birds. However, astroviral infection of animals had been poorly studied. In the present study, 211 rectal swabs collected from cattle and water buffalo calves with mild to severe diarrhea were tested for bovine astrovirus (BAstV) by RT-PCR. Results: 92/211 (43.6%) samples were positive for BAstV, at a rate of 46.10% (71/154) in cattle and 36.84% (21/57) in water buffalo. Phylogenetic analysis based on the partial and full-length of 25 ORF2 amino acid sequences obtained in this study classified the Guangxi BAstVs isolates into five subgroups under the genus of Mamastrovirus, genotype MAstV33, which suggested that the water buffalo was a new host of this genogroup that previously included only cattle and roe deer. Despite the origin of the host, the Guangxi BAstV isolates were closely related to the BAstV Hong Kong isolates (B18/HK and B76-2/HK), but highly divergent from the BAstV NeuroS1 isolate previously associated with neurologic disease in cattle in the U.S.A. Nucleotide sequence-based characterization of the ORF1b/ORF2 junction and corresponding overlapping regions showed distinctive properties, which may be common to BAstVs. Our results suggested that cattle and water buffalo are prone to infection of closely related astroviruses, which probably evolved from the same ancestor. The current study described astroviruses in water buffalo for the first time and is thus far among the largest epidemiological investigations of BAstV infection in cattle conducted in China.

  12. Steps can be taken to keep bovine viral diarrhea (BVD) out of your herd

    Science.gov (United States)

    This is a review, written for a lay publication whose core audience in dairy producers. Control of BVD in any dairy operation must rely on the implementation of an organized strategy combining biosecurity, surveillance and increased herd resistance. This article discusses the design and implementati...

  13. Innate and adaptive immune responses to in utero infection with bovine viral diarrhea virus

    Science.gov (United States)

    Infection of pregnant cows with noncytopathic (ncp) BVDV induces rapid innate and adaptive immune responses resulting in clearance of the virus in less than 3 weeks. Seven to 14 days after inoculation of the cow, ncpBVDV crosses the placenta and induces a fetal viremia. Establishment of persistent ...

  14. Absolute quantification of Bovine Viral Diarrhea Virus (BVDV) RNA by the digital PCR technique

    Science.gov (United States)

    Flatschart, R. B.; Almeida, D. O.; Heinemann, M. B.; Medeiros, M. N.; Granjeiro, J. M.; Folgueras-Flatschart, A. V.

    2015-01-01

    The quality control of cell lines used in research and industry is critical to ensure confidence in experimental results and to guarantee the safety of biopharmaceuticals to consumers. The BVDV is a common adventitious agent in many cell lines. We preliminarly evaluate the use of Digital Droplet PCR (ddPCR) for the detection and enumeration of genome copies of BVDV in cell culture and on FBS. The application of a commercial Real-Time PCR kit with the ddPCR technique was successful on different matrices. The technique allowed the absolute quantification of the genome without the use of calibration standards, suggesting its promising application on the development of reference materials for quantification of nucleic acids.

  15. Optimization of surveillance opf Bovine Viral Diarrhea in Danish dairy herds

    DEFF Research Database (Denmark)

    Foddai, Alessandro

    ELISA (Rønsholt et al., 1997; Bitsch et al., 1997) and the SVANOVIR ELISA (Juntti at al., 1987; Niskanen, 1993) were compared on milk and serum samples. The prevalence of antibody positive milking cows, which can be detected by each of those tests, was estimated by diluting positive individual milk...... dairy herds (e.g. after import of infected cattle) could be more difficult to detect compared to the past, due to the lower prevalence of antibody positive milking cows and the (expected) higher dilution of antibodies in bigger milk containers. Therefore, an evaluation and an eventual optimization...... of the BVD surveillance system in Danish dairy herds were considered necessary by the Danish Cattle Federation. In study I, we verified how the BVD herd prevalence, the herd size and the dilution of individual milk within the bulk tank milk (BTM) changed, between 2003 and 2010. Moreover, the Danish blocking...

  16. Quantitative assessment of the risk of introduction of bovine viral diarrhea virus in Danish dairy herds

    DEFF Research Database (Denmark)

    Foddai, Alessandro; Boklund, Anette; Stockmarr, Anders;

    2014-01-01

    of live cattle, aswell as use of vaccines were analyzed. Information regarding the application of biosecuritymeasures, by veterinarians and hoof trimmers practicing in Denmark and in other countries,was obtained by contacting several stakeholders, public institutions and experts. Stochas-tic scenario...

  17. Evaluation of commercial ELISA kits for detection of antibodies against bovine atypical pestivirus

    DEFF Research Database (Denmark)

    Larska, Magdalena; Polak, Mirosław P.; Uttenthal, Åse;

    A group of emerging bovine pestiviruses becomes a possible threat to Bovine Viral diarrhea virus (BVDV) control and eradication programs in the countries of their origin and in the new continents due to the lack of validated detection methods. The use of ELISA kits may be acheaper, time saving...

  18. HoBi-like viruses – the typical 'atypical bovine pestivirus'

    Science.gov (United States)

    HoBi-like viruses, also referred to as bovine viral diarrhea virus 3 (BVDV-3) and atypical pestivirus, have been proposed as a new putative bovine pestivirus species. These viruses were first identified in the last decade and are currently distributed in at least three continents. Published findings...

  19. Physiology of acid-base balance in bovines with diarrhea backgrounds from Monteria, Colombia

    Directory of Open Access Journals (Sweden)

    César Betancur H

    2015-05-01

    Full Text Available ABSTRACT Objective. Evaluate the acid-base balance (ABB in bovines with diarrheic backgrounds in four areas of Montería, Colombia. Materials and methods. From a total of 300 pregnant cows, 60 were selected with their newborns. A direct inspection was performed of vital signs on the calves and the ABB indicators were determined using a gasometric method. Data were processed by means of descriptive statistics and the Duncan test was used to differentiate between the averages. The degree of association was established between the ABB indicators in cows and calves by using the Pearson correlation and a comparison of proportions was performed on the indexes of the newborns. Results. Regarding the cows, the ABB indicators were found within the reference values; however, in the calves the pH, pCO2, HCO3 - , the anion gap (AG and the bases excess (BE varied. A correlation was found between AG, BE and metabolic hydrogen ions (M*H. The AG in cows and calves showed notable differences (p<0.05 among the farms in the study. According to the numeric classification system, the suction reflect indicated a greater percentage of calves in group one. Conclusions. The ABB analyte measurement in cows was similar to the consulted reference; however, in calves some analytes did not coincide. This suggests metabolic acidosis in newborn calves due to the increase of AG and the decrease of BE. Additionally, its correlation with M*H opens the possibility of new proposals to determine ABB in bovines.

  20. Characterisation of bovine viral diarrhoea virus (BVDV) isolates from an outbreak with haemorrhagic enteritis and severe pneumonia.

    Science.gov (United States)

    Yeşilbağ, Kadir; Förster, Christine; Ozyiğit, M Ozgür; Alpay, Gizem; Tuncer, Pelin; Thiel, Heinz-Jürgen; König, Matthias

    2014-02-21

    During 2007 a disease outbreak occurred in cattle in the Marmara region of western Turkey characterised by severe pneumonia and haemorrhagic enteritis in calves. Cases from three farms at different locations were examined and bovine viral diarrhoea virus (BVDV) isolated in all cases. Phylogenetic characterisation of the virus isolates allocated them in a new cluster tentatively named as BVDV-1r.

  1. Human-, Ovine-, and Bovine-Specific Viral Source Tracking Tools to Discriminate Between the Major Fecal Sources in Agricultural Waters.

    Science.gov (United States)

    Rusiñol, Marta; Moriarty, Elaine; Lin, Susan; Bofill-Mas, Sílvia; Gilpin, Brent

    2016-03-01

    This study evaluated the sources of fecal contamination in different river catchments, using a combination of microbial source tracking tools, for human, ruminant, ovine and bovine livestock, in order to define appropriate water management strategies. Every source of waterway pollution was evaluated in river water samples from one urban river catchment and two important farming regions in New Zealand. Fecal pollution was initially measured by testing Escherichia coli and evaluating the presence of human- and ruminant-associated DNA markers of Bacteroidales (BiAdo, BacHum-UCD, BacH, and BacR) and human and ruminant fecal sterols/stanols ratios. Then specific fecal pollution sources were assessed with previously reported quantitative PCR assays targeting human-, bovine-, and ovine-specific viruses: human adenoviruses (HAdV), human JC polyomaviruses, bovine polyomaviruses (BPyV), and ovine polyomaviruses (OPyV). High level of ruminant fecal contamination was detected all over the farming areas, whereas no ruminant sources were identified in the urban river sampling sites. BacR was the most frequently observed ruminant marker and OPyV and BPyV allowed the identification of ovine and bovine fecal sources. The human fecal viral marker (HAdV) was the most frequently observed human marker, highly abundant in the urban sites, and also present in farming areas. This is the first study using simultaneously the ovine and the bovine viral markers to identify and quantify both bovine and ovine fecal pollution.

  2. Bovine viral diarrhoea, bovine herpesvirus and parainfluenza-3 virus infection in three cattle herds in Egypt in 2000.

    Science.gov (United States)

    Aly, N M; Shehab, G G; Abd el-Rahim, I H A

    2003-12-01

    This study reported field outbreaks of bovine viral diarrhoea virus (BVDV) infection, either alone or mixed with bovine herpesvirus-1 (BHV-1) and/or parainfluenza-3 virus (PI-3V) in Egypt during 2000. In Lower Egypt, young calves in three cattle herds in El-Minufiya Province, El-Fayoum Province and in governmental quarantine in El-Behira Province, showed symptoms of enteritis, either alone or accompanied by respiratory manifestations. The affected herds were visited and the diseased animals were clinically examined. Many epidemiological aspects, such as morbidities, mortalities and case fatalities, as well as the abortive rate, were calculated. Ethylenediamine tetra-acetic acid-blood samples, sterile nasal swabs and serum samples were obtained for virological and serological diagnosis. The laboratory investigations revealed that the main cause of calf mortalities in the three herds was infection with BVDV, either alone, as on the El-Minufiya farm, or mixed with PI-3V, as on the El-Fayoum farm, or mixed with both BHV-1 and PI-3V, as in the herd in governmental quarantine in El-Behira Province. A total of nine dead calves from the three herds were submitted for thorough post-mortem examination. Tissue samples from recently dead calves were obtained for immunohistochemical and histopathological studies. The most prominent histopathological findings were massive degeneration, necrosis and erosions of the lining epithelium of the alimentary tract. Most of the lymphoreticular organs were depleted of lymphocytes. In pneumonic cases, bronchopneumonia and atypical interstitial pneumonia were evident. The present study suggested that the immunosuppressive effect of BVDV had predisposed the animals to secondary infection with BHV-1 and PI-3V. This study concluded that concurrent infection with BVDV, BHV-1 and PI-3V should be considered as one of the infectious causes of pneumoenteritis and, subsequently, the high morbidities and mortalities among young calves in Egypt

  3. Analysis of Pan-European attitudes to the eradication and control of bovine viral diarrhoea.

    Science.gov (United States)

    Heffernan, C; Misturelli, F; Nielsen, L; Gunn, G J; Yu, J

    2009-02-07

    At present, national-level policies concerning the eradication and control of bovine viral diarrhoea (BVD) differ widely across Europe. Some Scandinavian countries have enacted strong regulatory frameworks to eradicate the disease, whereas other countries have few formal policies. To examine these differences, the attitudes of stakeholders and policy makers in 17 European countries were investigated. A web-based questionnaire was sent to policy makers, government and private sector veterinarians, and representatives of farmers' organisations. In total, 131 individuals responded to the questionnaire and their responses were analysed by applying a method used in sociolinguistics: frame analysis. The results showed that the different attitudes of countries that applied compulsory or voluntary frameworks were associated with different views about the attribution or blame for BVD and the roles ascribed to farmers and other stakeholders in its eradication and control.

  4. Prevalence of bovine viral diarrhoea virus in cattle farms in Hungary.

    Science.gov (United States)

    Szabára, Ágnes; Lang, Zsolt; Földi, József; Hornyák, Ákos; Abonyi, Tamás; Ózsvári, László

    2016-06-01

    A study was performed to survey the virological prevalence of bovine viral diarrhoea (BVD) virus (BVDV) in cattle herds in Hungary between 2008 and 2012. A total of 40,413 samples for BVDV detection and 24,547 samples for antibody testing were collected from 3,247 herds (570,524 animals), thus representing approximately 75% of the cattle population in Hungary. Retrospective Bayesian analysis demonstrated that (1) the herd-level true virus prevalence was 12.4%, (2) the mean individual (within-herd) true virus prevalence was 7.2% in the herds having at least one virus-positive animal and 0.89% for all investigated herds with a mean apparent prevalence of 1.15% for the same population. This is the first study about BVDV prevalence in Hungary.

  5. The role of viral population diversity in adaptation of bovine coronavirus to new host environments.

    Directory of Open Access Journals (Sweden)

    Monica K Borucki

    Full Text Available The high mutation rate of RNA viruses enables a diverse genetic population of viral genotypes to exist within a single infected host. In-host genetic diversity could better position the virus population to respond and adapt to a diverse array of selective pressures such as host-switching events. Multiple new coronaviruses, including SARS, have been identified in human samples just within the last ten years, demonstrating the potential of coronaviruses as emergent human pathogens. Deep sequencing was used to characterize genomic changes in coronavirus quasispecies during simulated host-switching. Three bovine nasal samples infected with bovine coronavirus were used to infect human and bovine macrophage and lung cell lines. The virus reproduced relatively well in macrophages, but the lung cell lines were not infected efficiently enough to allow passage of non lab-adapted samples. Approximately 12 kb of the genome was amplified before and after passage and sequenced at average coverages of nearly 950×(454 sequencing and 38,000×(Illumina. The consensus sequence of many of the passaged samples had a 12 nucleotide insert in the consensus sequence of the spike gene, and multiple point mutations were associated with the presence of the insert. Deep sequencing revealed that the insert was present but very rare in the unpassaged samples and could quickly shift to dominate the population when placed in a different environment. The insert coded for three arginine residues, occurred in a region associated with fusion entry into host cells, and may allow infection of new cell types via heparin sulfate binding. Analysis of the deep sequencing data indicated that two distinct genotypes circulated at different frequency levels in each sample, and support the hypothesis that the mutations present in passaged strains were "selected" from a pre-existing pool rather than through de novo mutation and subsequent population fixation.

  6. Viral and Bacterial Etiology of Acute Diarrhea among Children under 5 Years of Age in Wuhan, China

    OpenAIRE

    Xu-Hui Zhu; Lei Tian; Zhong-Ju Cheng; Wei-Yong Liu; Song Li; Wei-Ting Yu; Wen-Qian Zhang; Xu Xiang; Zi-Yong Sun

    2016-01-01

    Background: Acute diarrhea remains the serious problem in developing countries, especially among children under 5 years of age. Currently, only two or three common diarrhea pathogens were screened at most hospitals in China. The aim of this study was to provide a wide variety of diarrhea pathogens and their antimicrobial resistance patterns in children under 5 years of age. Methods: Totally 381 stool samples collected from Tongji Hospital between July 1, 2014 and June 30, 2015 were tested by ...

  7. Phylogenetic study on the 5'-untranslated region of bovine viral diarrhoea virus isolates from Iran

    Directory of Open Access Journals (Sweden)

    Majid Esmaelizad

    2014-09-01

    Full Text Available Bovine viral diarrhoea virus is a pathogen of bovids associated with reproduction system, causing in infected animals a range of ailments, from abortion to congenital defects. In this article, the nucleotide structure of the 5'-untranslated region (5-UTR from 7 Iranian bovine diarrhoea virus (BVDV isolates was characterized and subjected to comparative analysis against a panel of BVDV isolates from different sources. To this end, a 288 bp-long stretch of the internal ribosome entry site was amplified by RT-PCR. The PCR products subsequently cloned into PTZ57T vector and sequenced using T7 promoter primers. This resulted in detection of 3 new point mutations G→A and G→T in 2 isolates. When these findings were phylogenetically assessed, all the examined Iranian isolates were deemed to belong to the type1 of BVDV. Besides, 2 subtypes were identified among these isolates. In group A, a high level of similarity (99.2% between Iranian isolates with a cytopathic Australian strain of BVDV-1c was detected; while in group B, the 4 Iranian isolates proved to be very similar to NADL-like BVDV-1a strains. We believe that the surprisingly high level of similarity between group A Iranian isolates and their corresponding Australian strain is likely to be an indication of a shared common ancestor. If correct, the most likely explanation of this observation is the introduction of such strains from Australia to Iran, possibly through exportation of infected live animals or animal productions (e.g. semen and meat at some points in the past. Nevertheless, this hypothesis remains to be proved as further epidemiological work at genomic level is required to understand population of BVDV in Iran.

  8. Bovine Mx1 enables resistance against foot-and-mouth disease virus in naturally susceptible cells by inhibiting the replication of viral RNA.

    Science.gov (United States)

    Wang, H-M; Xia, X-Z; Hu, G-X; Yu, L; He, H-B

    2016-03-01

    Innate immunity, especially the anti-viral genes, exerts an important barrier function in preventing viral infections. Myxovirus-resistant (Mx) gene take an anti-viral role, whereas its effects on foot-and-mouth disease virus (FMDV) in naturally susceptible cells are still unclear. The bovine primary fetal tracheal epithelial cell line BPTE-siMx1, in which bovine Mx1 gene was silenced, was established and treated with IFN alpha for 6 hr before FMDV infection. The copy numbers of the negative and positive strand viral RNA were determined by strand-specific real-time fluorescence quantitative RT-PCR. The TCID50 of BPTE-siMx1 cells increased at least 17-fold as compared to control cells BPTE-LacZ at 8 hr post infection, thus silencing of bovine Mx1 could promote the replication of FMDV. The amount of both the negative and positive strand viral RNA in BPTE-siMx1 cells significantly increased as compared to BPTE-LacZ cells, indicating that the replication levels of viral RNA were promoted by silencing bovine Mx1. The bovine Mx1 gene could provide resistance against FMDV in the bovine primary fetal tracheal epithelial cells via suppressing the replication of viral RNA.

  9. Bovine Viral Diarrhoea Virus (BVDV) in Dairy Cattle: A Matched Case-Control Study.

    Science.gov (United States)

    Machado, G; Egocheaga, R M F; Hein, H E; Miranda, I C S; Neto, W S; Almeida, L L; Canal, C W; Stein, M C; Corbellini, L G

    2016-02-01

    Bovine viral diarrhoea virus (BVDV) causes one of the most important diseases of cattle in terms of economic costs and welfare. The aims were to estimate herd prevalence and to investigate the factors associated with antibodies in bulk tank milk (BTM) in dairy herds through a matched case-control study. To estimate herd prevalence, BTM samples were randomly selected (n = 314) from a population (N = 1604). The true prevalence of BVDV was 24.3% (CI 95% = 20.1-29.3%). For the case-control study, BVDV antibody-positive herds (high antibody titres) were classified as cases (n = 21) and matched (n = 63) by milk production with herds presenting low antibody titres (ratio of 1 : 3). Three multivariable models were built: 1) full model, holding all 21 variables, and two models divided according to empirical knowledge and similarity among variables; 2) animal factor model; and 3) biosecurity model. The full model (model 1) identified: age as a culling criteria (OR = 0.10; CI 95% = 0.02-0.39; P cattle of neighbouring farms (OR = 5.78; CI 95% = 1.41-23.67; P = 0.04). We recommend the application of grouping predictors as a good choice for model building because it could lead to a better understanding of disease-exposure associations.

  10. Establishing a pilot bovine viral diarrhoea virus eradication scheme in Somerset.

    Science.gov (United States)

    Booth, R E; Brownlie, J

    2012-01-21

    Beginning in April 2006, 41 farms were recruited onto a pilot Bovine viral diarrhoea virus (BVDV) eradication programme across the south of England with the majority of study herds concentrated in Somerset. Each herd was assessed and where relevant cleared of persistently infected (PI) animals. Seven farms dropped out before whole herd screening could be performed. Of the remaining 34 farms, 20 (59 per cent) were classified as infected although two of these were initially misclassified as BVDV-free. Over the course of three years, 61 PIs were identified across 16 of the 20 infected farms. 72 per cent of PIs indentified on the first herd test were below two years of age. PI prevalence ranged from 0.2 to 3.1 per cent of infected herds and was highest in herds that did not vaccinate. By the end of 2009, 24/34 (71 per cent) of study farms were BVDV-free while 10 (29 per cent) remained infected.

  11. Molecular analysis of bovine viral diarrhoea virus isolates from South Africa

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    N. Kabongo

    2003-11-01

    Full Text Available The presence of bovine viral diarrhoea virus in South Africa has been confirmed by several serological surveys. However, little is known about its biological properties. Twenty five isolates obtained by isolation in tissue culture and detected by means of the antigen capture ELISA from clinically sick cattle and from foetal calf serum in South Africa were characterized on the basis of analysis of the 5' non-translated (NTR region of the genome. A reverse-transcription polymerase chain reaction (RT-PCR was used to amplify specific sequences from the 5'NTR of the genome. The oligonucleotide primers corresponding to positions 105-125 and 399-378, respectively, in the sequence of BVDV strain NADL were used to generate the PCR products. Both strands were sequenced directly with these primers and fluorescence-labelled dideoxynucleotides in an automated nucleic acid sequencer. Reference strains of pestiviruses [(BVDV type I, BVDV type II, border disease virus (BDV and hog cholera virus (HCV] and isolates from a previous investigation on BVDV in southern Africa were included for comparative purposes. All the BVDV strains obtained during this study belong to subgroups of BVDV genotype I. No association could be demonstrated between the geographic origin of the isolates. A number of isolates formed another branch separate from the existing branches Ia, Ib and Ic. These findings suggest that extensive genetic diversity can be found within BVDV type I isolates from southern Africa. Isolates that group with the classical BVDV type I strains, particularly of American origin, coexist with variants that appear to represent a local genetic pool and or variants evolving from the classical strains.

  12. Evaluation of long-term antibody responses to two inactivated bovine viral diarrhoea virus (BVDV) vaccines.

    Science.gov (United States)

    González, Ana M; Arnaiz, Ignacio; Yus, Eduardo; Eiras, Carmen; Sanjuán, María; Diéguez, Francisco J

    2014-03-01

    The aim of the present study was to determine the serological response of heifers after vaccination with two inactivated bovine viral diarrhoea virus (BVDV) vaccines by means of various ELISA tests. Three dairy farms were selected from the Galicia region of Spain. In each herd, a batch of heifers to be vaccinated for the first time was selected and followed for 15 months. Heifers from farm 1 (n=25) were vaccinated with Vaccine A, whereas heifers from farm 2 (n=16) were vaccinated with Vaccine B. Heifers from farm 3 (n=17), where no BVDV vaccines were used, acted as controls. Blood samples were analyzed periodically for BVDV antibodies, using five commercial ELISAs, based on BVDV p80 antigen or whole virus. At the end of the study, none of the animals vaccinated with Vaccine A seroconverted according to p80 antibody status, whereas up to 80% tested positive by ELISA against whole virus antigen. For the animals vaccinated with Vaccine B, 2/16 animals seroconverted according to p80 antibody ELISAs, whereas all had seroconverted according to the ELISA against whole virus antigen. In most cases, based on the use of ELISAs to detect specific antibodies against the p80 protein, at 15 months post-vaccination with inactivated BVDV vaccines the responses did not seem to interfere with detection of antibody to BVDV infection. However, the finding of a small proportion of vaccinated animals seropositive against BVDV p80 antigen suggests that antibodies that interfere with diagnosis of BVDV infection within the herd could exist, even when using p80 ELISAs.

  13. Repertoire of bovine miRNA and miRNA-like small regulatory RNAs expressed upon viral infection.

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    Evgeny A Glazov

    Full Text Available MicroRNA (miRNA and other types of small regulatory RNAs play a crucial role in the regulation of gene expression in eukaryotes. Several distinct classes of small regulatory RNAs have been discovered in recent years. To extend the repertoire of small RNAs characterized in mammals and to examine relationship between host miRNA expression and viral infection we used Illumina's ultrahigh throughput sequencing approach. We sequenced three small RNA libraries prepared from cell line derived from the adult bovine kidney under normal conditions and upon infection of the cell line with Bovine herpesvirus 1. We used a bioinformatics approach to distinguish authentic mature miRNA sequences from other classes of small RNAs and short RNA fragments represented in the sequencing data. Using this approach we detected 219 out of 356 known bovine miRNAs and 115 respective miRNA* sequences. In addition we identified five new bovine orthologs of known mammalian miRNAs and discovered 268 new cow miRNAs many of which are not identifiable in other mammalian genomes and thus might be specific to the ruminant lineage. In addition we found seven new bovine mirtron candidates. We also discovered 10 small nucleolar RNA (snoRNA loci that give rise to small RNA with possible miRNA-like function. Results presented in this study extend our knowledge of the biology and evolution of small regulatory RNAs in mammals and illuminate mechanisms of small RNA biogenesis and function. New miRNA sequences and the original sequencing data have been submitted to miRNA repository (miRBase and NCBI GEO archive respectively. We envisage that these resources will facilitate functional annotation of the bovine genome and promote further functional and comparative genomics studies of small regulatory RNA in mammals.

  14. Morphological changes in cultured bovine lymphoid cell lines associated with bovine viral diarrhea virus (BVDV) single and dual infections with bovine leukemia virus (BLV)

    Science.gov (United States)

    Currently, American Type Culture Collection (ATCC) makes available two cell lines derived from the same lymphoblast-like suspension cell that have been confirmed by next-generation sequencing and RT-PCR to have either a single contaminate of BVDV2a (CRL-8037) or dual contaminates of both BVDV and BL...

  15. In vivo delivery of bovine viral diahorrea virus, E2 protein using hollow mesoporous silica nanoparticles

    Science.gov (United States)

    Mahony, D.; Cavallaro, A. S.; Mody, K. T.; Xiong, L.; Mahony, T. J.; Qiao, S. Z.; Mitter, N.

    2014-05-01

    Our work focuses on the application of mesoporous silica nanoparticles as a combined delivery vehicle and adjuvant for vaccine applications. Here we present results using the viral protein, E2, from bovine viral diarrhoea virus (BVDV). BVDV infection occurs in the target species of cattle and sheep herds worldwide and is therefore of economic importance. E2 is a major immunogenic determinant of BVDV and is an ideal candidate for the development of a subunit based nanovaccine using mesoporous silica nanoparticles. Hollow type mesoporous silica nanoparticles with surface amino functionalisation (termed HMSA) were characterised and assessed for adsorption and desorption of E2. A codon-optimised version of the E2 protein (termed Opti-E2) was produced in Escherichia coli. HMSA (120 nm) had an adsorption capacity of 80 μg Opti-E2 per mg HMSA and once bound E2 did not dissociate from the HMSA. Immunisation studies in mice with a 20 μg dose of E2 adsorbed to 250 μg HMSA was compared to immunisation with Opti-E2 (50 μg) together with the traditional adjuvant Quillaja saponaria Molina tree saponins (QuilA, 10 μg). The humoral responses with the Opti-E2/HMSA nanovaccine although slightly lower than those obtained for the Opti-E2 + QuilA group demonstrated that HMSA particles are an effective adjuvant that stimulated E2-specific antibody responses. Importantly the cell-mediated immune responses were consistently high in all mice immunised with Opti-E2/HMSA nanovaccine formulation. Therefore we have shown the Opti-E2/HMSA nanoformulation acts as an excellent adjuvant that gives both T-helper 1 and T-helper 2 mediated responses in a small animal model. This study has provided proof-of-concept towards the development of an E2 subunit nanoparticle based vaccine.Our work focuses on the application of mesoporous silica nanoparticles as a combined delivery vehicle and adjuvant for vaccine applications. Here we present results using the viral protein, E2, from bovine viral

  16. Serological Study on Bovine Viral Diarrhoea Virus Infection in Pig Population in Poland Between 2008 and 2011

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    Lipowski Andrzej

    2014-10-01

    Full Text Available In total, 14 608 pig sera, collected between 2008 and 2011, were tested with ELISA using antibodies specific for bovine viral diarrhoea virus (BVDV. All doubtful and positive samples were retested by virus neutralisation test (neutralising peroxidase-linked assay. The BVDV seroreagents were detected in 11 (68.75% out of 16 provinces, the seroprevalence varied from 0.1% to 1.04% (average 0.31%. The obtained results indicate that the prevalence of BVDV infection in pig population in Poland is low.

  17. Comparative Suitability of Ear Notch Biopsy and Serum Pairs for Detecting Nature of Bovine Viral Diarrhoea Virus Infection in Dairy Herds

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    Arfan Ahmad, Masood Rabbani, Muhammad Zubair Shabbir, Khushi Muhammad1, Tahir Yaqub, Muhammad Kashif Saleemi2, Rana Khurram Khalid, Muhammad Abu Bakr Shabbir3 and Shumaila Yousaf Alvi4

    2012-06-01

    Full Text Available Suitability of ear notch biopsy (EN and serum pairs (n= 307 collected from 10 Holstein dairy herds located in Charlottetown, Canada was evaluated for simultaneous detection, nature of bovine viral diarrhea virus (BVDV infection and genotype of the prevailing BVDV through Real time RT-PCR. Depending upon vaccination status and age, the sampled animals were categorized into two groups, A (n=123, ≤ 6 month of age and B (n=184, ≥ 6 months of age originating from 4 vaccinated (n=108 and 3 non-vaccinated (n=76 animal herds. On first round of testing a discrepancy between ear notch biopsies and sera pairs (3.25 and 6.50%; P<0.05 of groups A was observed, however, a complete harmony (50% for EN and sera each, P<0.01 was found on second round of testing that confirmed the presence of 4 persistent infection (PI animals harboring genotype 1 of BVDV. Complete concordance between EN and sera pairs (P<0.01 on first and follow up testing in group B was observed (2.77%, each, depicting 3 PI animals with the same genotype as in group A. In the study, ear notch biopsies did not detect any transient infection (TI but sera samples detected 3.25% transiently infected animals in group A that was 1.30 % among all the test samples (n=307 while no TI animal was found in group B. It may be concluded that both the serum and ear notch biopsy can be used to detect PI animals and that, serum samples are more sensitive than ear notch (P < 0.05 for detection of TI using real time RT-PCR.

  18. Expression and In Silico Analysis of the Recombinant Bovine Papillomavirus E6 Protein as a Model for Viral Oncoproteins Studies

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    J. Mazzuchelli-de-Souza

    2013-01-01

    Full Text Available Bovine papillomaviruses (BPVs are recognized as the causal agents of economical relevant diseases in cattle, associated with the development of tumors in skin and mucosa. The oncogenesis process is mainly associated with different viral oncoprotein expressions, which are involved in cell transformation. The expression and characterization of recombinant viral oncoproteins represent an attractive strategy to obtain biotechnological products as antibodies and potential vaccines, Thus, the aim of this work was to clone and express the BPV-1 and BPV-2 E6 recombinant proteins and perform in silico analysis in order to develop a strategy for the systematic study of other papillomaviruses oncoproteins. The results demonstrated that BPV-1 and BPV-2 E6 recombinant proteins were expressed and purified from bacterial system as well as its in silico analysis was performed in order to explore and predict biological characteristics of these proteins.

  19. The viral polymerase inhibitor 2'-C-methylcytidine inhibits Norwalk virus replication and protects against norovirus-induced diarrhea and mortality in a mouse model.

    Science.gov (United States)

    Rocha-Pereira, Joana; Jochmans, Dirk; Debing, Yannick; Verbeken, Erik; Nascimento, Maria S J; Neyts, Johan

    2013-11-01

    Human noroviruses are a major cause of food-borne illness, accountable for 50% of all-etiologies outbreaks of acute gastroenteritis (in both developing and developed countries). There is no vaccine or antiviral drug for the prophylaxis or treatment of norovirus-induced gastroenteritis. We recently reported the inhibitory effect of 2'-C-methylcytidine (2CMC), a hepatitis C virus polymerase inhibitor, on the in vitro replication of murine norovirus (MNV). Here we evaluated the inhibitory effect of 2CMC on in vitro human norovirus replication through a Norwalk virus replicon model and in a mouse model by using AG129 mice orally infected with MNV. Survival, weight, and fecal consistency were monitored, and viral loads in stool samples and organs were quantified. Intestines were examined histologically. 2CMC reduced Norwalk virus replicon replication in a dose-dependent manner and was able to clear cells of the replicon. Treatment of MNV-infected AG129 mice with 2CMC (i) prevented norovirus-induced diarrhea; (ii) markedly delayed the appearance of viral RNA and reduced viral RNA titers in the intestine, mesenteric lymph nodes, spleen, lungs, and stool; (iii) completely prevented virus-induced mortality; and (iv) resulted in protective immunity against a rechallenge. We demonstrate for the first time that a small-molecule inhibitor of norovirus replication protects from virus-induced disease and mortality in a relevant animal model. These findings pave the way for the development of potent and safe antivirals as prophylaxis and therapy of norovirus infection.

  20. Milk supplemented with immune colostrum: protection against rotavirus diarrhea and modulatory effect on the systemic and mucosal antibody responses in calves experimentally challenged with bovine rotavirus.

    Science.gov (United States)

    Parreño, V; Marcoppido, G; Vega, C; Garaicoechea, L; Rodriguez, D; Saif, L; Fernández, F

    2010-07-01

    Group A bovine rotavirus (BRV) is the major cause of neonatal calf diarrhea worldwide. As a preventive strategy, we evaluated the protection and immunomodulation in two groups of BRV-inoculated calves. All calves received control colostrum (CC; VN=65,536; IgG(1)=16,384) prior to gut closure followed by the milk supplemented with immune colostrum (VN=1,048,576; IgG(1)=262,144), twice a day, for 14 days. Calves received milk supplemented with 0.8% immune colostrum [(Gp 1) VN=16,384; IgG(1)=4096] or milk supplemented with 0.4% immune colostrum [(Gp 2) VN=1024; IgG(1)=1024]. Calves receiving CC or colostrum deprived calves (CD) fed antibody (Ab) free milk served as controls (Gp 3 and 4). Calves were inoculated with virulent BRV IND at 2 days of age. Group 1 calves (milk IgG(1) 4096) showed 80% protection against BRV diarrhea and significantly reduced virus shedding. At 21 post-inoculation days (PID), the antibody secreting cell (ASC) responses of Gp 1 calves were limited mainly to duodenal and jejunal lamina propria (LP) with limited or no responses in systemic sites (spleen and PBL) and mesenteric lymph nodes. The profile of serum and fecal Ab responses as well as the ASC responses was also modulated by the presence of passive IgG(1) Abs and probably other colostrum components, toward higher titers of IgA Ab in serum and feces and a greater number of IgA ASC in the proximal intestine, reflecting positive modulation by colostrum toward this isotype associated with optimal protection of the intestinal mucosa. After challenge, at PID 21, all calves in Gp 1 and 2 were fully protected against diarrhea and only 1 of 5 calves in Gp 1 shed virus asymptomatically, indicating that the passive Ab treatment for 14 days was effective in protecting most of the animals after a first and a second virus exposure. The final outcome was a positive modulation of the mucosal immune responses and a high protection rate against diarrhea and virus shedding during the period of peak

  1. Establishment of Universal RT-PCR for Bovine Virus Diarrhea%牛病毒性腹泻病毒通用型RT-PCR检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    郭春娟; 吴星星; 王璐; 季新成; 冉多良; 买买提·黑牙斯丁; 王克栋; 熊浩

    2012-01-01

    为建立一种快速、特异、简便的检测牛病毒粘膜腹泻病病毒Ⅰ、Ⅱ型抗原的通用RT-PCR方法.根据GenBank上已发表的牛病毒性粘膜腹泻病毒(BVDV)Ⅰ、Ⅱ型5’端非编码区保守序列设计合成了两条引物,其中上游引物为兼并引物.应用一步法RT-PCR技术分别对Ⅰ(Oregon C24V)、Ⅱ型(279)标准株进行扩增,将PCR产物胶回收后连与pMD18-T载体,经PCR、酶切鉴定条带均正确(288 bp),测序结果与预期一致.同时设牛疱疹病毒Ⅰ型、牛传染性鼻气管炎、猪瘟病毒、蓝舌病病毒、口蹄疫病毒、MDBK细胞作对照,结果均为阴性;并对新疆各地州的样品进行检测.优化反应条件后,检测其灵敏度为101 TCID50/mL.经对400份临床样品检测,阳性检出率为10.75%,明显高于ELISA检测.试验表明,该方法具有特异、灵敏、高效、快速、重复性好等特点,可用于牛病毒粘膜腹泻病的临床检测及流行病学检测.%To establish a rapid,specific,simple common RT-PCR method to detect bovine viral diarrhea/ mocosal disease virus type Ⅰ,Ⅱ antigen, the two synthesized primers were designed by Bovine Viral Diar-rhea/Mucosa Birus (BVDV) Ⅰ,Ⅱ type 5'Non-coding Reigion of Conserved Sequence published on Gen-Bank;one of which was an upstream and degenerate primer. I (Oregon C24V), II type (279) standard strainswere were amplified respectively by one-step RT-PCR, the PCR products gel were recovered even with the pMD18-T by PCR,restriction enzyme digestion bands were correct as 288 bp,sequencing results were in accordance with expectations. Also located bovine herpesvirus type I .Infectious Bovine Rhinotra-cheitis,classical swine fever virus,Blue tongue virus,Foot and mouth disease virus,MDBK cells were used as control,the results were negative; and also the samples from different parts of Xinjiang were tested. When reaction conditions were optimized,the sensitivity was detected as 10'TCID 50/mL. In a test of 400

  2. Genetic diversity of Brazilian bovine pestiviruses detected between 1995 and 2014

    Science.gov (United States)

    Pestivirus infections in ruminants result in significant economic losses worldwide. The etiological agents are three species from the genus Pestivirus, family Flaviviridae, including Bovine Viral Diarrhea Virus type 1 (BVDV-1), BVDV-2, Border Disease Virus (BDV), and an atypical pestivirus named HoB...

  3. Direct production losses and treatment costs from bovine viral diarrhoea virus, bovine leukosis virus, Mycobacterium avium subspecies paratuberculosis, and Neospora caninum.

    Science.gov (United States)

    Chi, Junwook; VanLeeuwen, John A; Weersink, Alfons; Keefe, Gregory P

    2002-09-30

    Our purpose was to determine direct production losses (milk loss, premature voluntary culling and reduced slaughter value, mortaliy loss, and abortion and reproductive loss) and treatmetn costs (veterinary services, medication cost, and extra farm labour cost) due to four infectious diseases in the maritime provinces of Canada: bovine viral diarrhoea (BVD), enzootic bovine leukosis (EBL), Johne's Disease (JD), and neosporosis. We used a partial-budget model, and incorporated risk and sensitivity analyses to identify the effects of uncertainty on costs. Total annual costs for an average, infected, 50 cow herd were: JD$ 2472; BVD$ 2421; neosporosis $ 2304; EBL$ 806. The stochastic nature of the proportion of infected herds and prevalence of infection within a herd were used to estimate probability distributions for these ex post costs. For all diseases, these distributions were right skewed. A sensitivity analysis showed the largest effect on costs was due to milk yield effects. For example, changing milk production loss from 0 to 5% for BVD increased the costs for the disease by 266%.

  4. Immunisation of Sheep with Bovine Viral Diarrhoea Virus, E2 Protein Using a Freeze-Dried Hollow Silica Mesoporous Nanoparticle Formulation

    OpenAIRE

    Donna Mahony; Mody, Karishma T.; Antonino S Cavallaro; Qiuhong Hu; Mahony, Timothy J.; Shizhang Qiao; Neena Mitter

    2015-01-01

    Bovine viral diarrhoea virus 1 (BVDV-1) is arguably the most important viral disease of cattle. It is associated with reproductive, respiratory and chronic diseases in cattle across the world. In this study we have investigated the capacity of the major immunological determinant of BVDV-1, the E2 protein combined with hollow type mesoporous silica nanoparticles with surface amino functionalisation (HMSA), to stimulate immune responses in sheep. The current work also investigated the immunogen...

  5. In vitro inhibition of the bovine viral diarrhoea virus by the essential oil of Ocimum basilicum (basil) and monoterpenes.

    Science.gov (United States)

    Kubiça, Thaís F; Alves, Sydney H; Weiblen, Rudi; Lovato, Luciane T

    2014-01-01

    The bovine viral diarrhoea virus (BVDV) is suggested as a model for antiviral studies of the hepatitis C virus (HCV). The antiviral activity of the essential oil of Ocimum basilicum and the monoterpenes camphor, thymol and 1,8-cineole against BVDV was investigated. The cytotoxicities of the compounds were measured by the MTT (3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide) test, and the antiviral activities were tested by the plaque reduction assay. The oil or compounds were added to the assay in three different time points: a) pre-treatment of the virus (virucidal assay); b) pre-treatment of the cells; or c) post-treatment of the cells (after virus inoculation). The percentage of plaques inhibition for each compound was determined based on the number of plaques in the viral control. The results were expressed by CC50 (50% cytotoxic concentration), IC50 (inhibitory concentration for 50% of plaques) and SI (selectivity index = CC50/IC50). Camphor (CC50 = 4420.12 μg mL(-1)) and 1,8-cineole (CC50 = 2996.10 μg mL(-1)) showed the lowest cytotoxicities and the best antiviral activities (camphor SI = 13.88 and 1,8-cineol SI = 9.05) in the virucidal assay. The higher activities achieved by the monoterpenes in the virucidal assay suggest that these compounds act directly on the viral particle.

  6. Herd-level prevalence and associated risk factors for Toxoplasma gondii, Neospora caninum, Chlamydia abortus and bovine viral diarrhoea virus in commercial dairy and beef cattle in eastern, northern and northeastern China.

    Science.gov (United States)

    Sun, Wu-Wen; Meng, Qing-Feng; Cong, Wei; Shan, Xiao-Feng; Wang, Chun-Feng; Qian, Ai-Dong

    2015-11-01

    Although the seroprevalence of Toxoplasma gondii, Neospora caninum, Chlamydia abortus and bovine viral diarrhea virus infection in cattle have been reported in some areas in China, most of them were conducted with small number of cattle samples and very limited districts and neglected the assessment of herd management factors associated with herd-level prevalence of these pathogen infections. Thus, from September 2013 to December 2014, a large-scale seroprevalence study was conducted to determine the animal-level and herd-level seroprevalence and identify herd-level risk factors associated with these pathogen infections in 4487 cattle from 134 herds in five provinces (Heilongjiang, Jilin, Liaoning, Shandong, Hebei) and Inner Mongolia Autonomous Region of China. At animal level, the true prevalence of antibodies against T. gondii, N. caninum, C. abortus and bovine viral diarrhoea virus (BVDV) was 10.48, 17.14, 11.92 and 50.10%, respectively. At herd level, the true prevalence of antibodies against T. gondii, N. caninum, C. abortus and BVDV was 27.16, 29.10, 37.31 and 40.30%, respectively. Multivariate analysis of these characteristics showed that source of water and presence of felids were significantly associated with T. gondii infection in the studied cattle herds. Source of water was significantly associated with N. caninum infection in the studied cattle herds. While herd size and management system were significantly associated with BVDV infection in the studied cattle herds, this is the first report of herd-level prevalence and associated risk factors of T. gondii, N. caninum, C. abortus and BVDV infection in cattle in China.

  7. Advanced Research of Bovine Viral Diarrhea%牛病毒胜腹泻病的研究进展

    Institute of Scientific and Technical Information of China (English)

    范俊; 姜露

    2009-01-01

    牛病毒性腹泻.粘膜病是由牛病毒性腹泻病毒引起的一种呈多种临床症状的疾病,给世界养牛业造成了巨大的经济损失.本文从BVD的病原学、发病机理、诊断、治疗和预防等方面进行了详细阐述.

  8. Comparison of temperature fluctuations at multiple anatomical locations in cattle during exposure to bovine viral diarrhea virus

    Science.gov (United States)

    Rectal temperature is generally considered the “gold standard” for monitoring temperature changes associated with environmental, immunological or endocrine stimuli in cattle. With the development of new telemetry systems, other anatomical locations and methods can be utilized to help continuously m...

  9. Puesta en evidencia del virus diarrea viral bovina en bovinos clínicamente afectados Isolation of the bovine viral diarrhoea virus from tissue of clinically affected cattle

    Directory of Open Access Journals (Sweden)

    M.O CELEDON

    1997-01-01

    Full Text Available Para conocer la presencia del virus diarrea viral bovina (VDVB en animales sospechosos de estar cursando un cuadro clínico provocado por este virus, se trabajó con un total de 33 animales, correspondiendo a 23 fetos abortados, 2 mortinatos, un nonato, 3 vacas: una madre de mortinato, una madre de aborto y una muerta, 2 novillos muertos y 2 terneros muertos. Muestras de órganos se inocularon en cultivos primarios de pulmón fetal bovino (PFB y en la línea MDBK. Después del primer pasaje en células de PFB, se detectó la presencia de antígenos del VDVB por la prueba de inmunoperoxidasa indirecta (IPI. Todas las muestras con reacción positiva a IPI se inocularon por segunda y tercera vez en células de PFB, aplicándose la prueba de IPI en el tercer pasaje. Sobre un cuarto pasaje se aplicó la prueba de inmunofluorescencia direccta (IFD. Todas las muestras, positivas y negativas a IPI, se inocularon en 3 pasajes seriados en las células MDBK. En 23 de los 33 animales se aisló VDVB cepas no citopatogénicas (NCP, correspondiendo a 14 fetos abortados, un nonato, un mortinato, 3 vacas, 2 novillos y 2 terneros. En 6 fetos abortados, independiente de los infectados con el VDVB, se aisló el virus de la rinotraqueítis infecciosa bovina (RIB. Se concluye que la presencia del VDVB es de alta frecuencia en muestras clínicas de ganado bovino con patologías asociables al VDVB, desconociéndose el rol patógeno del virus en estos aislados.Cattle infected with the bovine viral diarrhoea (BVD virus can present a variety of clinical signs. This research studied the presence of BVD virus in cattle by virus isolation in primary cell cultures of bovine embryo lungs. Virus identification was done using the immunoperoxidase staining assay and the direct fluorescent antibody staining. As a result, 23 out of 33 animals were identified as positive to BVD virus: 14 foetal abortions, 2 stillbirths, 3 dams, 2 steers and 2 calves. No cytopathogenic isolates were

  10. The papain-like protease of porcine epidemic diarrhea virus negatively regulates type I interferon pathway by acting as a viral deubiquitinase.

    Science.gov (United States)

    Xing, Yaling; Chen, Jianfei; Tu, Jian; Zhang, Bailing; Chen, Xiaojuan; Shi, Hongyan; Baker, Susan C; Feng, Li; Chen, Zhongbin

    2013-07-01

    Porcine epidemic diarrhea virus (PEDV) is the cause of an economically important swine disease. Previous studies suggested that PEDV does not elicit a robust IFN response, but the mechanism(s) used to evade or block this innate immune response was not known. In this study, we found that PEDV infection blocked synthetic dsRNA-induced IFN-β production by interfering with the activation of interferon regulatory factor 3 (IRF3). We identified PEDV replicase encoded papain-like protease 2 (PLP2) as an IFN antagonist that depends on catalytic activity for its function. We show that levels of ubiquitinated proteins are reduced during PEDV infection and that PEDV PLP2 has deubiquitinase (DUB) activity that recognizes and processes both K-48 and K-63 linked polyubiquitin chains. Furthermore, we found that PEDV PLP2 strongly inhibits RIG-I- and STING-activated IFN expression and that PEDV PLP2 can be co-immunoprecipitated with and deubiquitinates RIG-I and STING, the key components of the signalling pathway for IFN expression. These results show that PEDV infection suppresses production of IFN-β and provides evidence indicating that the PEDV papain-like protease 2 acts as a viral DUB to interfere with the RIG-I- and STING-mediated signalling pathway.

  11. The prevalence of bovine viral diarrhoea antibodies in selected South African dairy herds, and control of the disease

    Directory of Open Access Journals (Sweden)

    G.M. Ferreira

    2000-07-01

    Full Text Available The prevalence of bovine viral diarrhoea (BVD serologically positive animals in 18 dairy herds with clinical and pathological lesions suggestive of BVD infection, the post-vaccinal seroconversion rates in negative animals vaccinated twice with an inactivated BVD vaccine, and the control measures taken, are described. The pathological and histopathological findings in 6 necropsies performed on animals that died in 5 separate herds closely resembled published descriptions. Positive immunohistochemistry results in 3 cases confirmed the diagnosis in those animals. In 1 herd the prevalence of prevaccinal BVDantibodies was only 36.8 %, while the prevalence varied from 79.85 to 100 % in the remainder. Control measures taken included immunoprophylaxis with an inactivated vaccine, culling animals that were serologically negative after vaccination that were regarded as probably persistently infected (PI and the implementation of additional biosecurity measures. The prevalence of serologically negative PI animals in 10 herds varied from 0.38 to 4.04 %, with 8 herds less than 1 %and 2 herds at 2.79 %and 4.04 %, respectively. Methods based on vaccinating the herd, followed by serological testing and culling cattle that did not develop an antibody titre, are not reliable. The identification of PI animals should be confirmed by isolation of the virus or identification of the antigen.

  12. Serological survey of bovine viral diarrhoea virus in Namibian and South African kudu (Tragelaphus strepsiceros and eland (Taurotragus oryx

    Directory of Open Access Journals (Sweden)

    Terence P. Scott

    2013-02-01

    Full Text Available Bovine viral diarrhoea virus (BVDV is a pestivirus that affects members of the order Artiodactyla, including members of the subfamily Bovinae. Little is known about the seroprevalence of BVDV in southern Africa, especially the prevalence in wild ruminant populations such as kudu (Tragelaphus strepsiceros. A handful of random surveys suggested that seroprevalence ranged between 6% and 70% in southern African wild ruminants. The present study aimed to determine the seroprevalence of BVDV amongst kudu and eland (Taurotragus oryx from Namibia and South Africa. A BVDV-specific enzyme-linked immunosorbent assay was performed on 50 serum samples from kudu and eland from South Africa and Namibia. The seroprevalence of BVDV in South African kudu was 71%, identical to that in Namibian kudu. The seroprevalence in Namibian eland was 40%. The kudu and cattle farming (free ranging regions in Namibia predominantly overlap in the central regions, ensuring ample opportunity for cross-species transmission of BVDV. It is therefore important to determine the true prevalence of BVDV in southern Africa in both domesticated and wild animals. In addition, a potential link between BVDV incidence and a devastating rabies epidemic in Namibian kudu was proposed and such a notion could be supported or discredited by comparative prevalence data.

  13. Respiratory disease associated with bovine coronavirus infection in cattle herds in Southern Italy.

    Science.gov (United States)

    Decaro, Nicola; Campolo, Marco; Desario, Costantina; Cirone, Francesco; D'Abramo, Maria; Lorusso, Eleonora; Greco, Grazia; Mari, Viviana; Colaianni, Maria Loredana; Elia, Gabriella; Martella, Vito; Buonavoglia, Canio

    2008-01-01

    Four outbreaks of bovine respiratory disease (BRD) associated with bovine coronavirus (BCoV) infection in Italian cattle herds were reported. In 3 outbreaks, BRD was observed only in 2-3-month-old feedlot calves, whereas in the remaining outbreak, lactating cows, heifers, and calves were simultaneously affected. By using reverse transcription polymerase chain reaction (RT-PCR), BCoV RNA was detected in all outbreaks without evidence of concurrent viral pathogens (i.e., bovine respiratory syncytial virus, bovine herpesvirus type 1, bovine viral diarrhea virus, bovine parainfluenza virus). Common bacteria of cattle were recovered only from 2 outbreaks of BRD: Staphylococcus spp. and Proteus mirabilis (outbreak 1) and Mannheimia haemolytica (outbreak 4). A recently established real-time RT-PCR assay showed that viral RNA loads in nasal secretions ranged between 3.10 x 10(2) and 7.50 x 10(7) RNA copies/microl of template. Bovine coronavirus was isolated from respiratory specimens from all outbreaks except outbreak 1, in which real-time RT-PCR found very low viral titers in nasal swabs.

  14. 天津市病毒性腹泻的病原学分析%Pathogenic Analysis on Viral Diarrhea in Tianjin

    Institute of Scientific and Technical Information of China (English)

    田宏; 雷玥; 刘杨; 张颖; 李佳萌

    2011-01-01

    目的 了解天津市病毒性腹泻的病原学分布情况.方法 于2009年3-10月从天津市儿童医院和宝坻医院收集感染性腹泻患者的粪便标本,采用酶联免疫技术(ELISA)检测轮状病毒,采用逆转录-聚合酶链反应(RT-PCR)方法检测杯状病毒、星状病毒,并对轮状病毒ELISA检测阳性标本进行分型.采用聚合酶链反应(PCR)检测腺病毒.结果 从天津市儿童医院收集儿童粪便标本124份,四种病毒检测阳性率顺位依次为杯状病毒(17.7%)>腺病毒(16.9%)>轮状病毒(12.1%)>星状病毒(1.6%),总检出率为48.4%.从宝坻医院收集成人粪便标本102份,四种病毒检测阳性率顺位依次为杯状病毒(8.8%)>腺病毒(3.9%)>轮状病毒(1.0%)>星状病毒(1.0%),总检出率为14.7%.16株轮状病毒G血清型阳性8株,G1为5株(31.2%),G3为3株(18.8%),8株未能分型(50%);P基因型分型结果阳性7株,P[4]为2株(12.5%),P[6]为1株(6.2%),P[8]为4株(25%),9株未能分型(56.3%).G型与P型组合P[4]G1为1株(6.2%),P[8]G3为1株(6.2%),P[8]G1为3株(18.8%).31株诺如病毒全部为GⅡ型.腺病毒主要是Ad40和Ad41型.星状病毒是1型.结论 本次调查的该市儿童病毒性腹泻发病率明显高于成年人,病毒分布呈现出多样性.%Objective To investigate the main pathogens of viral diarrhea in Tianjin. Methods The feces specimens of infectious diarrhea patients were collected from Tianjin Children's Hospital and Baodi Hospital. Rotavirus was detected by ELISA,calicivirus and astrovirus were detected by RT-PCR. Further strain characterization of rotavirus was carried out with RT-PCR and adenovirus were detected by PCR. Results Tested feces specimens with amount of 124 were collected from Tianjin Children's Hospital. Calicivirus was detected in 17.7% of the cases, adenoviruses in 16.9% , rotavirus in 12.1% and astrovirus in 1.6 %.The total positive rate was 48.4%. Tested feces specimens with amount of 102 were collected from Baodi

  15. Management factors related to seroprevalences to bovine viral-diarrhoea virus, bovine-leukosis virus, Mycobacterium avium subspecies paratuberculosis, and Neospora caninum in dairy herds in the Canadian Maritimes.

    Science.gov (United States)

    Chi, Junwook; VanLeeuwen, John A; Weersink, Alfons; Keefe, Gregory P

    2002-09-10

    Bovine viral-diarrhoea (BVD), enzootic bovine leukosis (EBL), Johne's disease (JD), and neosporosis lower on-farm productivity, reduce export competitiveness, and increase consumer concerns regarding safety. Our purpose was to examine the relationship between 27 control practices and the estimated true seroprevalences for these four diseases for 2604 cattle in 90 dairy herds in the Maritimes provinces of Canada. Overall, 37.8, 20.4, 3.4, and 19.2% of all sampled cattle were truly exposed to the agents of BVD, EBL, JD, and neosporosis, respectively. The median within-herd true prevalences were 0, 9.3, 0, and 12.3%, respectively. Factor analysis reduced the 27 control practices to two highly correlated factors. Tobit-regression analyses determined that vaccination practices were associated with reduced prevalence of exposure for Bovine viral-diarrhoea and EBL. Also, farms that tended to purchase their dairy animals were associated with higher seroprevalence for Johnes' disease. Neither of these two factors was associated with the seroprevalence of Neospora caninum infection. The few routine biosecurity measures that were investigated in this study were generally not related to the seroprevalences of these farms.

  16. Engineered Lactobacillus rhamnosus GG expressing IgG-binding domains of protein G: Capture of hyperimmune bovine colostrum antibodies and protection against diarrhea in a mouse pup rotavirus infection model.

    Science.gov (United States)

    Günaydın, Gökçe; Zhang, Ran; Hammarström, Lennart; Marcotte, Harold

    2014-01-16

    Rotavirus-induced diarrhea causes more than 500,000 deaths annually in the world, and although vaccines are being made available, new effective treatment strategies should still be considered. Purified antibodies derived from hyperimmune bovine colostrum (HBC), from cows immunized with rotavirus, were previously used for treatment of rotavirus diarrhea in children. A combination of HBC antibodies and a probiotic strain of Lactobacillus (L. rhamnosus GG) was also found to be more effective than HBC alone in reducing diarrhea in a mouse model of rotavirus infection. In order to further improve this form of treatment, L. rhamnosus GG was engineered to display surface expressed IgG-binding domains of protein G (GB1, GB2, and GB3) which capture HBC-derived IgG antibodies (HBC-IgG) and thus target rotavirus. The expression of IgG-binding domains on the surface of the bacteria as well as their binding to HBC-IgG and to rotavirus (simian strain RRV) was demonstrated by Western blot, flow cytometry, and electron microscopy. The prophylactic effect of engineered L. rhamnosus GG and anti-rotaviral activity of HBC antibodies was evaluated in a mouse pup model of RRV infection. The combination therapy with engineered L. rhamnosus GG (PG3) and HBC was significantly more effective in reducing the prevalence, severity, and duration of diarrhea in comparison to HBC alone or a combination of wild-type L. rhamnosus GG and HBC. The new therapy reduces the effective dose of HBC between 10 to 100-fold and may thus decrease treatment costs. This antibody capturing platform, tested here for the first time in vivo, could potentially be used to target additional gastrointestinal pathogens.

  17. Single Pathogen Challenge with Agents of the Bovine Respiratory Disease Complex

    Science.gov (United States)

    Gershwin, Laurel J.; Van Eenennaam, Alison L.; Anderson, Mark L.; McEligot, Heather A.; Toaff-Rosenstein, Rachel; Taylor, Jeremy F.; Neibergs, Holly L.; Womack, James

    2015-01-01

    Bovine respiratory disease complex (BRDC) is an important cause of mortality and morbidity in cattle; costing the dairy and beef industries millions of dollars annually, despite the use of vaccines and antibiotics. BRDC is caused by one or more of several viruses (bovine respiratory syncytial virus, bovine herpes type 1 also known as infectious bovine rhinotracheitis, and bovine viral diarrhea virus), which predispose animals to infection with one or more bacteria. These include: Pasteurella multocida, Mannheimia haemolytica, Mycoplasma bovis, and Histophilus somni. Some cattle appear to be more resistant to BRDC than others. We hypothesize that appropriate immune responses to these pathogens are subject to genetic control. To determine which genes are involved in the immune response to each of these pathogens it was first necessary to experimentally induce infection separately with each pathogen to document clinical and pathological responses in animals from which tissues were harvested for subsequent RNA sequencing. Herein these infections and animal responses are described. PMID:26571015

  18. Single Pathogen Challenge with Agents of the Bovine Respiratory Disease Complex.

    Directory of Open Access Journals (Sweden)

    Laurel J Gershwin

    Full Text Available Bovine respiratory disease complex (BRDC is an important cause of mortality and morbidity in cattle; costing the dairy and beef industries millions of dollars annually, despite the use of vaccines and antibiotics. BRDC is caused by one or more of several viruses (bovine respiratory syncytial virus, bovine herpes type 1 also known as infectious bovine rhinotracheitis, and bovine viral diarrhea virus, which predispose animals to infection with one or more bacteria. These include: Pasteurella multocida, Mannheimia haemolytica, Mycoplasma bovis, and Histophilus somni. Some cattle appear to be more resistant to BRDC than others. We hypothesize that appropriate immune responses to these pathogens are subject to genetic control. To determine which genes are involved in the immune response to each of these pathogens it was first necessary to experimentally induce infection separately with each pathogen to document clinical and pathological responses in animals from which tissues were harvested for subsequent RNA sequencing. Herein these infections and animal responses are described.

  19. Simultaneous Concentration of Bovine Viruses and Agricultural Zoonotic Bacteria from Water Using Sodocalcic Glass Wool Filters.

    Science.gov (United States)

    Abd-Elmaksoud, Sherif; Spencer, Susan K; Gerba, Charles P; Tamimi, Akrum H; Jokela, William E; Borchardt, Mark A

    2014-12-01

    Infiltration and runoff from manured agricultural fields can result in livestock pathogens reaching groundwater and surface waters. Here, we measured the effectiveness of glass wool filters to simultaneously concentrate enteric viruses and bacteria of bovine origin from water. The recovery efficiencies were determined for bovine viral diarrhea virus types 1 and 2, bovine rotavirus group A, bovine coronavirus, poliovirus Sabin III, toxigenic Escherichia coli ,and Campylobacter jejuni seeded into water with three different turbidity levels (0.5, 215, and 447 NTU). Twenty liters of dechlorinated tap water (pH 7) were seeded with the test organisms, and then passed through a glass wool filter using a peristaltic pump (flow rate = 1 liter min(-1)). Retained organisms were eluted from the filters by passing beef extract-glycine buffer (pH 9.5) in the direction opposite of sample flow. Recovered organisms were enumerated by qPCR except for C. jejuni, which was quantified by culture. Mean recovery efficiencies ranged from 55 to 33% for the bacteria and 58 to 16% for the viruses. Using bootstrapping techniques combined with Analysis of Variance, recovery efficiencies were found to differ among the pathogen types tested at the two lowest turbidity levels; however, for a given pathogen type turbidity did not affect recovery except for C. jejuni. Glass wool filtration is a cost-effective method for concentrating several waterborne pathogens of bovine origin simultaneously, although recovery may be low for some specific taxa such as bovine viral diarrhea virus 1.

  20. AIDS-associated diarrhea and wasting in northeast Brazil is associated with subtherapeutic plasma levels of antiretroviral medications and with both bovine and human subtypes of Cryptosporidium parvum

    Directory of Open Access Journals (Sweden)

    Richard K. Brantley

    2003-02-01

    Full Text Available Advanced HIV infection is frequently complicated by diarrhea, disruption of bowel structure and function, and malnutrition. Resulting malabsorption of or pharmacokinetic changes in antiretroviral agents might lead to subtherapeutic drug dosing and treatment failure in individual patients, and could require dose adjustment and/or dietary supplements during periods of diarrheal illness. We determined the plasma levels of antiretroviral medications in patients that had already been started on medication by their physicians in an urban infectious diseases hospital in northeast Brazil. We also obtained blood samples from patients hospitalized for diarrhea or AIDS-associated wasting, and we found reduced stavudine and didanosine levels in comparison with outpatients without diarrhea or wasting who had been treated at the same hospital clinic. There was a predominance of the protozoal pathogens Cryptosporidium and Isospora belli, typical opportunistic pathogens of AIDS-infected humans, in the stool samples of inpatients with diarrhea. We conclude that severe diarrhea and wasting in this population is associated with both protozoal pathogens and subtherapeutic levels of antiretroviral medications.

  1. Whole Genomic Analysis of an Unusual Human G6P[14] Rotavirus Strain Isolated from a Child with Diarrhea in Thailand: Evidence for Bovine-To-Human Interspecies Transmission and Reassortment Events.

    Science.gov (United States)

    Tacharoenmuang, Ratana; Komoto, Satoshi; Guntapong, Ratigorn; Ide, Tomihiko; Haga, Kei; Katayama, Kazuhiko; Kato, Takema; Ouchi, Yuya; Kurahashi, Hiroki; Tsuji, Takao; Sangkitporn, Somchai; Taniguchi, Koki

    2015-01-01

    An unusual rotavirus strain, SKT-27, with the G6P[14] genotypes (RVA/Human-wt/THA/SKT-27/2012/G6P[14]), was identified in a stool specimen from a hospitalized child aged eight months with severe diarrhea. In this study, we sequenced and characterized the complete genome of strain SKT-27. On whole genomic analysis, strain SKT-27 was found to have a unique genotype constellation: G6-P[14]-I2-R2-C2-M2-A3-N2-T6-E2-H3. The non-G/P genotype constellation of this strain (I2-R2-C2-M2-A3-N2-T6-E2-H3) is commonly shared with rotavirus strains from artiodactyls such as cattle. Phylogenetic analysis indicated that nine of the 11 genes of strain SKT-27 (VP7, VP4, VP6, VP2-3, NSP1, NSP3-5) appeared to be of artiodactyl (likely bovine) origin, while the remaining VP1 and NSP2 genes were assumed to be of human origin. Thus, strain SKT-27 was found to have a bovine rotavirus genetic backbone, and thus is likely to be of bovine origin. Furthermore, strain SKT-27 appeared to be derived through interspecies transmission and reassortment events involving bovine and human rotavirus strains. Of note is that the VP7 gene of strain SKT-27 was located in G6 lineage-5 together with those of bovine rotavirus strains, away from the clusters comprising other G6P[14] strains in G6 lineages-2/6, suggesting the occurrence of independent bovine-to-human interspecies transmission events. To our knowledge, this is the first report on full genome-based characterization of human G6P[14] strains that have emerged in Southeast Asia. Our observations will provide important insights into the origin of G6P[14] strains, and into dynamic interactions between human and bovine rotavirus strains.

  2. Both viral E2 protein and the cellular factor PEBP2 regulate transcription via E2 consensus sites within the bovine papillomavirus type 4 long control region.

    OpenAIRE

    Jackson, M E; Campo, M S

    1995-01-01

    The bovine papillomavirus type 4 (BPV4) long control region (LCR) contains three consensus binding sites, E2(1), E2(2), and E2(3) (ACCN6GGT), for the viral E2 transcription factor and a fourth degenerate site, dE2 (ATCN6GGT), which lies 3 bp upstream of E2(3). The E2(2) site was found to bind the cellular transcription factor PEBP2, and mutations at this site reduced basal promoter activity by as much as 60%, indicating an important role for PEBP2 in LCR function. Mutation of the E2(3) or dE2...

  3. Endotoxin-free purification for the isolation of Bovine Viral Diarrhoea Virus E2 protein from insoluble inclusion body aggregates

    Directory of Open Access Journals (Sweden)

    Mahony Timothy J

    2011-07-01

    Full Text Available Abstract Background Protein expression in Escherichia coli may result in the recombinant protein being expressed as insoluble inclusion bodies. In addition, proteins purified from E. coli contain endotoxins which need to be removed for in vivo applications. The structural protein, E2, from Bovine Viral Diarrhoea Virus (BVDV is a major immunogenic determinant, and is an ideal candidate as a subunit vaccine. The E2 protein contains 17 cysteine residues creating difficulties in E. coli expression. In this report we outline a procedure for successfully producing soluble and endotoxin-free BVDV E2 protein from inclusion bodies (IB. Results The expression of a truncated form of BVDV-E2 protein (E2-T1 in E. coli resulted in predominantly aggregated insoluble IB. Solubilisation of E2-T1 with high purity and stability from IB aggregates was achieved using a strong reducing buffer containing 100 mM Dithiothreitol. Refolding by dialysis into 50 mM Tris (pH 7.0 containing 0.2% Igepal CA630 resulted in a soluble but aggregated protein solution. The novel application of a two-phase extraction of inclusion body preparations with Triton X-114 reduced endotoxin in solubilised E2-T1 to levels suitable for in vivo use without affecting protein yields. Dynamic light scattering analyses showed 37.5% of the protein was monomeric, the remaining comprised of soluble aggregates. Mice immunised with E2-T1 developed a high titre antibody response by ELISA. Western hybridisation analysis showed E2-T1 was recognised by sera from immunised mice and also by several BVDV-E2 polyclonal and monoclonal antibodies. Conclusion We have developed a procedure using E. coli to produce soluble E2-T1 protein from IB, and due to their insoluble nature we utilised a novel approach using Triton X-114 to efficiently remove endotoxin. The resultant protein is immunogenic and detectable by BVDV-E2 specific antibodies indicating its usefulness for diagnostic applications and as a subunit

  4. Silica Vesicle Nanovaccine Formulations Stimulate Long-Term Immune Responses to the Bovine Viral Diarrhoea Virus E2 Protein.

    Directory of Open Access Journals (Sweden)

    Karishma T Mody

    Full Text Available Bovine Viral Diarrhoea Virus (BVDV is one of the most serious pathogen, which causes tremendous economic loss to the cattle industry worldwide, meriting the development of improved subunit vaccines. Structural glycoprotein E2 is reported to be a major immunogenic determinant of BVDV virion. We have developed a novel hollow silica vesicles (SV based platform to administer BVDV-1 Escherichia coli-expressed optimised E2 (oE2 antigen as a nanovaccine formulation. The SV-140 vesicles (diameter 50 nm, wall thickness 6 nm, perforated by pores of entrance size 16 nm and total pore volume of 0.934 cm3 g(-1 have proven to be ideal candidates to load oE2 antigen and generate immune response. The current study for the first time demonstrates the ability of freeze-dried (FD as well as non-FD oE2/SV140 nanovaccine formulation to induce long-term balanced antibody and cell mediated memory responses for at least 6 months with a shortened dosing regimen of two doses in small animal model. The in vivo ability of oE2 (100 μg/SV-140 (500 μg and FD oE2 (100 μg/SV-140 (500 μg to induce long-term immunity was compared to immunisation with oE2 (100 μg together with the conventional adjuvant Quil-A from the Quillaja saponira (10 μg in mice. The oE2/SV-140 as well as the FD oE2/SV-140 nanovaccine generated oE2-specific antibody and cell mediated responses for up to six months post the final second immunisation. Significantly, the cell-mediated responses were consistently high in mice immunised with oE2/SV-140 (1,500 SFU/million cells at the six-month time point. Histopathology studies showed no morphological changes at the site of injection or in the different organs harvested from the mice immunised with 500 μg SV-140 nanovaccine compared to the unimmunised control. The platform has the potential for developing single dose vaccines without the requirement of cold chain storage for veterinary and human applications.

  5. Toddler's Diarrhea

    Science.gov (United States)

    ... days when stools are more formed. Despite the diarrhea, the child continues to grow and gain weight appropriately as ... could suggest other causes such as infection. Toddler’s diarrhea is not considered a disease, and children with this condition will get better on their ...

  6. Effect of cantharidin, cephalotaxine and homoharringtonine on "in vitro" models of hepatitis B virus (HBV) and bovine viral diarrhoea virus (BVDV) replication.

    Science.gov (United States)

    Romero, Marta R; Serrano, Maria A; Efferth, Thomas; Alvarez, Marcelino; Marin, Jose J

    2007-06-01

    The effect as antiviral agents versus viral hepatitis B and C of three compounds purified from natural products commonly used as remedies in traditional Chinese medicine, cantharidin, cephalotaxine and homoharingtonine, was investigated. To assess the activity of these compounds against flavivirus, we used bovine viral diarrhoea virus (BVDV) as a surrogate for hepatitis C virus (HCV). Anti-BVDV activity was determined by reduction in BVDV-RNA production and protection of infected embryonic bovine trachea (EBTr) cells against the cytopathic effect of BVDV. The effect versus hepatitis B virus (HBV) was investigated by measuring HBsAg and HBV-DNA release from hepatoblastoma HepG2 2.2.15 cells infected with HBV. As positive control we used the standard anti-HBV and anti-HCV drugs, lamivudine and ribavirin, respectively. Up to 100 microM lamivudine and ribavirin did not induce cell toxicity, whereas they induced dose-dependent anti-HBV and anti-BVDV effects, respectively. In the same range, cantharidin, cephalotaxine and homoharringtonine induced toxicity in EBTr cells and had no protective effect against BVDV. In contrast, they were able to inhibit HBV production at concentrations 10- to 100-fold lower than those inducing cell toxicity, which suggests that they are useless for the treatment of infection by flaviviruses, but potentially useful in combined therapy against hepatitis B.

  7. Retrospective epidemiological evaluation of molecular and animal husbandry data within the bovine viral diarrhoea virus (BVDV) control programme in Western Austria during 2009-2014.

    Science.gov (United States)

    Schoepf, Karl; Revilla-Fernández, Sandra; Steinrigl, Adolf; Fuchs, Reinhard; Sailer, Andreas; Weikel, Joachim; Schmoll, Friedrich

    2016-01-01

    A retrospective epidemiological investigation of molecular and animal husbandry data collected over an observation period of five years (2009-2014) within the compulsory bovine viral diarrhoea virus (BVDV) control programme in Western Austria, covering the federal provinces of Tyrol and Vorarlberg is presented in this study. Samples collected from 232 infected calves were phylogenetically classified based on the 5' untranslated region (5'UTR). All but 13 samples, which were typed as border disease virus subtype 3 (BDV-3), belonged to the bovine viral diarrhoea virus genotype 1 (BVDV-1) and clustered within six different subtypes (1b, 1e, 1f, 1h, 1d and 1k). Movement data and survival times from infected individual animals were analysed because of their potential of passing on infection to naive herds. From the moment of submission of the laboratory results, 180 animals were culled within the first month, 13 lived longer than two but not longer than six months and seven infected animals lived longer than one year. 13 of the infected animals were born on alpine pastures and eleven infected animals were grazed on mountain pastures during summer. The movement of infected animals and the role of trade in alpine areas are a possible source for spreading the infection, thus hampering the progress of eradication.

  8. Antibody response against three widespread bovine viruses is not impaired in Holstein cattle carrying bovine leukocyte antigen DRB3.2 alleles associated with bovine leukemia virus resistance.

    Science.gov (United States)

    Juliarena, M A; Poli, M; Ceriani, C; Sala, L; Rodríguez, E; Gutierrez, S; Dolcini, G; Odeon, A; Esteban, E N

    2009-01-01

    Due to the wide dissemination of bovine leukemia virus (BLV) infection among dairy cattle, control and eradication programs based on serological detection of infected cattle and subsequent culling face a major economic task. In Argentina, genetic selection of cattle carrying alleles of the bovine leukocyte antigen (BoLA) DRB3.2 gene associated with BLV-infection resistance, like *0902, emerges as the best additional tool toward controlling virus spread. A potential risk in expanding or segregating BoLA selected populations of cattle is that it might increase susceptibility to other common viruses. Special concern raises the strong association found between low proviral load and low antibody titer against major BLV structural proteins. This phenomenon might depend on host genetic factors influencing other viruses requiring, unlike BLV, strong and long-lasting humoral immune response to prevent infection. In this study, we demonstrate that there is no association among neutralizing antibody titers against foot and mouth disease virus, bovine viral diarrhea virus, or bovine herpesvirus type 1 and polymorphism of the BoLA DRB3.2 gene. Conversely, there is strong association between BoLA DRB3.2*0902 and low antibody titers against 2 BLV structural proteins--env gp51 and gag p24--to date, the best BLV resistance marker. There is also significant association between low antibody titers against gp51 and p24 and BoLA DRB3.2*1701 and low antibody titers against p24 and BoLA DRB3.2*1101 or 02. Our data suggest that increasing BoLA-selected BLV-resistant cattle or segregating BoLA-associated alleles to BLV susceptibility would not affect the resistance or the predisposition to bovine viral diarrhea virus, bovine herpesvirus type 1, or foot and mouth disease virus infection.

  9. [Persistent diarrhea

    Science.gov (United States)

    Andrade, J A; Moreira, C; Fagundes Neto, U

    2000-07-01

    INTRODUCTION: Persistent diarrhea has high impact on infantile morbidity and mortality rates in developing countries. Several studies have shown that 3 to 20% of acute diarrheal episodes in children under 5 years of age become persistent. DEFINITION: Persistent diarrhea is defined as an episode that lasts more than 14 days. ETIOLOGY: The most important agents isolated in persistent diarrhea are: Enteropathogenic E. coli (EPEC), Salmonella, Enteroaggregative E. coli (EAEC), Klebisiella and Cryptosporidium. CLINICAL ASPECTS: In general, the clinical characteristics of patients with persistent diarrhea do not change with the pathogenic agent. Persistent diarrhea seems to represent the final result of a several insults a infant suffers that predisposes to a more severe episode of diarrhea due to a combination of host factors and high rates of enviromental contamination. Therefore, efforts should be made to promptly treat all episodes of diarrhea with apropriate follow-up. THERAPY: The aim of the treatment is to restore hydroelectrolytic deficits and to replace losses until the diarrheal ceases. It is possible in the majority of the cases, using oral rehydration therapy and erly an appropriate type of diet. PREVENTION: It is imperative that management strategies also focus on preventive aspects. The most effective diarrheal prevention strategy in young infants worldwide is promotion of exclusive breast feeding.

  10. Diarrhea among children in developing countries.

    Science.gov (United States)

    Nataro, James P

    2013-01-01

    Diarrhea continues to stand among the most important causes of global morbidity and mortality in children under 5 years of age. Although the introduction of oral rehydration and other case-management strategies have reduced acute diarrhea fatalities, many of the survivors develop persistent diarrhea and/or deficiencies of growth and cognition. Thus understanding the true global burden of diarrhea requires attention to acute diarrhea as well is its sequelae. To understand the etiology of moderate to severe diarrhea among children in high mortality areas of sub-Saharan Africa and south Asia we performed a comprehensive case-control study of children under 5 years of age at seven sites. Each site employed an identical case-control study design and each utilized a uniform comprehensive set of microbiological assays to identify the likely bacterial, viral and protozoal etiologies. Results of the studies will inform diarrhea prevention and management efforts worldwide.

  11. Travelers' Diarrhea

    Science.gov (United States)

    ... Visiting Friends and Family in Areas with Chikungunya, Dengue, or Zika Visiting Friends or Family in an ... East, Africa, Mexico, and Central and South America. Prevention In otherwise healthy adults, diarrhea is rarely serious ...

  12. THE CLINICAL STUDY OF XIYANPING COMBINED WITH BIFIDOBACTERIUM IN TREATMENT WITH INFANTILE VIRAL DIARRHEA%喜炎平联合双歧杆菌治疗小儿病毒腹泻的临床研究

    Institute of Scientific and Technical Information of China (English)

    张雪松

    2011-01-01

    [Objective] To the clinical effect of xiyanping combined with bifidobacterium in treatment with infantile viral diarrhea. [Methods] From February 2009 to February 2010, 112 cases with infantile viral diarrhea were randomly classified into two groups, 56 cases in control group was treated with ribavirin and 56 cases in observation group was treated with xiyanping combined with bifidobacterium. The curative effect, antidiarrheal time, fever clearance time, and total treatment time between the two groups were compared. [Results] The total effective rate in observation group were significantly higher than that in control group (P < 0.05). The antidiarrheal time, fever clearance time, and total treatment time in observation group were significantly shorter than those in control group (P < 0.05). [Conclusion] Xiyanping combined with bifidobacterium in treatment with infantile viral diarrhea can improve clinical curative effect, improve clinical symptom, which can be applied in clinic.%[目的]观察喜炎平联合双歧杆菌治疗小儿病毒腹泻的临床疗效.[方法]2010年2月~2011年2月,112例病毒腹泻患儿随机分为两组,对照组56例采用病毒唑治疗,观察组56例采用喜炎平联合双歧杆菌治疗,比较观察两组的临床疗效、止泻、退热时间和住院时间.[结果]观察组的总有效率明显高于对照组(P<0.05).观察组的平均止泻时间、平均退热时间和住院时间均明显短于对照组(P<0.05).[结论]喜炎平联合双歧杆菌治疗小儿病毒腹泻进行治疗,能够明显提高疗效,改善患儿临床症状,值得临床推广应用.

  13. Influenza Viral Manipulation of Sphingolipid Metabolism and Signaling to Modulate Host Defense System

    Directory of Open Access Journals (Sweden)

    Madhuvanthi Vijayan

    2014-01-01

    Full Text Available Viruses attempt to create a distinctive cellular environment to favor viral replication and spread. Recent studies uncovered new functions of the sphingolipid signaling/metabolism during pathogenic virus infections. While sphingolipids such as sphingomyelin and ceramide were reported to influence the entry step of several viruses, sphingolipid-metabolizing enzymes could directly alter viral replication processes. Influenza virus was shown to increase the level of sphingosine kinase (SK 1 to promote virus propagation. The mechanism involves regulation of intracellular signaling pathways, leading to the amplification of influenza viral RNA synthesis and nuclear export of viral ribonucleoprotein (RNP complex. However, bovine viral diarrhea virus inhibits SK1 to enhance the efficacy of virus replication, demonstrating the presence of virus-specific strategies for modulation of the sphingolipid system. Therefore, investigating the sphingolipid metabolism and signaling in the context of virus replication could help us design innovative therapeutic approaches to improve human health.

  14. Dexamethasone-induced reactivation of bovine herpesvirus type 5 latent infection in experimentally infected rabbits results in a broader distribution of latent viral DNA in the brain

    Directory of Open Access Journals (Sweden)

    S.V. Mayer

    2006-03-01

    Full Text Available Bovine herpesvirus type 5 (BHV-5 is a major agent of meningoencephalitis in cattle and establishes latent infections mainly in sensory nerve ganglia. The distribution of latent BHV-5 DNA in the brain of rabbits prior to and after virus reactivation was studied using a nested PCR. Fifteen rabbits inoculated intranasally with BHV-5 were euthanized 60 days post-inoculation (group A, N = 8 or submitted to dexamethasone treatment (2.6 mg kg-1 day-1, im, for 5 days and euthanized 60 days later (group B, N = 7 for tissue examination. Two groups of BHV-1-infected rabbits (C, N = 3 and D, N = 3 submitted to each treatment were used as controls. Viral DNA of group A rabbits was consistently detected in trigeminal ganglia (8/8, frequently in cerebellum (5/8, anterior cerebral cortex and pons-medulla (3/8 and occasionally in dorsolateral (2/8, ventrolateral and posterior cerebral cortices, midbrain and thalamus (1/8. Viral DNA of group B rabbits showed a broader distribution, being detected at higher frequency in ventrolateral (6/7 and posterior cerebral cortices (5/7, pons-medulla (6/7, thalamus (4/7, and midbrain (3/7. In contrast, rabbits inoculated with BHV-1 harbored viral DNA almost completely restricted to trigeminal ganglia and the distribution did not change post-reactivation. These results demonstrate that latency by BHV-5 is established in several areas of the rabbit's brain and that virus reactivation leads to a broader distribution of latent viral DNA. Spread of virus from trigeminal ganglia and other areas of the brain likely contributes to this dissemination and may contribute to the recrudescence of neurological disease frequently observed upon BHV-5 reactivation.

  15. Genetic Typing of Bovine Viral Diarrhoea Virus (BVDV by Restriction Fragment Length Polymorphism (RFLP and Identification of a New Subtype in Poland

    Directory of Open Access Journals (Sweden)

    Kuta Aleksandra

    2015-04-01

    Full Text Available Restriction fragment length polymorphism (RFLP analysis was developed for genetic typing of Polish strains of bovine viral diarrhoea virus (BVDV. The method was applied using 60 BVDV isolates, which included BVDV genotype 1, subtypes a, b, d, e, f, and g, and genotype 2a. RT-PCR products of the 5’untranslated region (5’UTR were digested using three enzymes. Restriction patterns classified the strains into seven groups, each with a specific and different pattern from other subtypes. These findings were confirmed by nucleotide sequencing and phylogenetic analysis. The results suggest that RFLP analysis is a simple, reliable, and fast genotyping method for BVDV strains in comparison with sequencing. This method can distinguish six subtypes of BVDV-1 including a new subtype 1e, identified exclusively by this method, and it allows differentiation of BVDV-1 from BVDV-2 genotype.

  16. Antiviral effect of artemisinin from Artemisia annua against a model member of the Flaviviridae family, the bovine viral diarrhoea virus (BVDV).

    Science.gov (United States)

    Romero, Marta R; Serrano, Maria A; Vallejo, Marta; Efferth, Thomas; Alvarez, Marcelino; Marin, Jose J G

    2006-10-01

    The antiviral activity versus flaviviruses of artemisinin, a safe drug obtained from Artemisia annua and commonly used to treat malaria, has been investigated using as an IN VITRO model bovine epithelial cells from embryonic trachea (EBTr) infected with the cytopathic strain Oregon C24V, of bovine viral diarrhoea virus (BVDV), which is a member of the Flaviviridae family. Antiviral activity was estimated by the degree of protection against the cytopathic effect of BVDV on host cells and by the reduction in BVDV-RNA release to the culture medium. To induce an intermediate cytopathic effect in non-treated cells, EBTr cells were first exposed to BVDV for 48 h and then incubated with virus-free medium for 72 h. Ribavirin and artemisinin (up to 100 microM) induced no toxicity in host cells, whereas a slight degree of toxicity was observed for IFN-alpha at concentrations above 10 U/mL up to 100 U/mL. Treatment of infected cells with IFN-alpha, ribavirin and artemisinin markedly reduced BVDV-induced cell death. A combination of these drugs resulted in an additive protective effect. These drugs induced a significant reduction in the production/release of BVDV virions by infected EBTr cells; there was also an additive effect when combinations of them were assayed. These results suggest a potential usefulness of artemisinin in combination with current pharmacological therapy for the treatment of human and veterinary infections by flaviviruses.

  17. Acute diarrhea.

    Science.gov (United States)

    Tobillo, E T; Schwartz, S M

    1998-10-01

    Diarrhea can result from damage to the intestinal lining caused by viruses or bacteria, malabsorption, inflammatory processes, bile salt and pancreatic enzyme deficiency, abnormal motility, or the presence of osmotically active solutes in the gut. While it is important to elicit information to determine the possible cause of diarrhea, be sure to check circulatory status first. Some patients may need rehydration therapy more urgently than they need a diagnosis. The main goals of treatment are to prevent dehydration and correct electrolyte imbalance, to provide supportive and symptomatic therapy, and to treat underlying disease. In most cases, a specific diagnosis is not necessary to guide initial treatment.

  18. Complete suppression of viral gene expression is associated with the onset and progression of lymphoid malignancy: observations in Bovine Leukemia Virus-infected sheep

    Directory of Open Access Journals (Sweden)

    Burny Arsène

    2007-07-01

    Full Text Available Abstract Background During malignant progression, tumor cells need to acquire novel characteristics that lead to uncontrolled growth and reduced immunogenicity. In the Bovine Leukemia Virus-induced ovine leukemia model, silencing of viral gene expression has been proposed as a mechanism leading to immune evasion. However, whether proviral expression in tumors is completely suppressed in vivo was not conclusively demonstrated. Therefore, we studied viral expression in two selected experimentally-infected sheep, the virus or the disease of which had features that made it possible to distinguish tumor cells from their nontransformed counterparts. Results In the first animal, we observed the emergence of a genetically modified provirus simultaneously with leukemia onset. We found a Tax-mutated (TaxK303 replication-deficient provirus in the malignant B-cell clone while functional provirus (TaxE303 had been consistently monitored over the 17-month aleukemic period. In the second case, both non-transformed and transformed BLV-infected cells were present at the same time, but at distinct sites. While there was potentially-active provirus in the non-leukemic blood B-cell population, as demonstrated by ex-vivo culture and injection into naïve sheep, virus expression was completely suppressed in the malignant B-cells isolated from the lymphoid tumors despite the absence of genetic alterations in the proviral genome. These observations suggest that silencing of viral genes, including the oncoprotein Tax, is associated with tumor onset. Conclusion Our findings suggest that silencing is critical for tumor progression and identify two distinct mechanisms-genetic and epigenetic-involved in the complete suppression of virus and Tax expression. We demonstrate that, in contrast to systems that require sustained oncogene expression, the major viral transforming protein Tax can be turned-off without reversing the transformed phenotype. We propose that suppression

  19. 牛病毒性腹泻-黏膜病的诊断与防治%Diagnosis and Cure of Bovine Viral Diarrhea-mucosal Disease

    Institute of Scientific and Technical Information of China (English)

    张宁; 奏建华

    2008-01-01

    牛病毒性腹泻-黏膜病是由牛病毒性腹泻病毒引起的一种复杂、呈多种临床类型的疾病.临床上以发热、黏膜溃疡糜烂、白细胞减少、腹泻、怀孕母牛流产或产畸型胎儿为主要特征.根据发病情况、临床特征、剖检变化、实验室诊断等情况,对牛病毒性腹泻病毒感染的病例进行了诊断,采取了相应的防治措施,取得了较好的效果.

  20. Evaluation of response to bovine viral diarrhea virus type 2 vaccination and timing of weaning on yearling ultrasound body composition, performance, and carcass quality traits in Angus calves

    Science.gov (United States)

    There are concerns about antagonisms between immunity and animal productivity in livestock production. The objective of this study was to evaluate the effect of antibody levels through a response to vaccination protocol, weaning timing, and their interaction on performance and carcass quality traits...

  1. 牛病毒性腹泻检测技术研究进展%Research progress on detection technique of bovine viral diarrhea

    Institute of Scientific and Technical Information of China (English)

    吴明福; 王金良; 孙进华

    2006-01-01

    近年来牛病毒性腹泻病的诊断技术,特别是在分子生物学诊断技术上取得了显著的成就.本文从BVD的病毒分离鉴定、血清学诊断方法、免疫学诊断方法以及分子生物学诊断技术等几个方面的进展进行了综述.

  2. 牛病毒性腹泻病毒RT-PCR检测方法研究%Detection of Bovine Viral Diarrhea Virus by RT-PCR Technique

    Institute of Scientific and Technical Information of China (English)

    高存福; 秦建华; 赵月兰; 左玉柱; 包永占; 李艳琴

    2007-01-01

    [目的]建立快速、简便、特异的检测牛病毒性腹泻病毒抗原的RT-PCR方法;[方法]根据已发表的牛病毒性腹泻病毒株(BVDV)5'端非编码区保守序列设计合成了两条引物,应用RT-PCR技术对两株BVDV标准株(OregonC24V and NADL)进行基因扩增,对扩增产物进行酶切鉴定.同时设猪瘟兔化弱毒疫苗株、轮状病毒株、MDBK细胞作对照;[结果]两株BVDV标准株均扩增出了长度为325bp的片段,扩增产物经酶切后形成长度为111bp和214bp两条片段,而对猪瘟兔化弱毒疫苗株、轮状病毒株、MDBK正常细胞扩增均为阴性,与预期结果一致.[结论]PCR检测方法可以快速准确地对牛病毒性腹泻-黏膜病作出诊断.该方法的敏感性可高达10-1TCID50.

  3. Serum Antibody Detection of Bovine Viral Diarrhea Virus%牛病毒性腹泻病毒血清抗体检测

    Institute of Scientific and Technical Information of China (English)

    王文; 张娟; 员丽娟; 季新成

    2011-01-01

    采用Svanova公司牛病毒性腹泻病毒血清抗体ELISA抗体检测试剂盒对采自新疆10个不同地区的875份牛血清进行了牛病毒性腹泻病毒血清抗体检测,共检出抗体阳性血清759份,最高阳性率为100%,最低为55.0%,平均为86.7%.检测结果表明该病已在新疆普遍存在,感染没有明确的地域分布规律.根据ELISA检测结果,从中选择15份血清用病毒中和试验进行检测,结果显示,所采用的ELISA检测方法具有较好的敏感性.

  4. Immunization against mucosal disease--bovine viral Diarrhea (MD-BVD) in calves applying lapinized vaccine against swine fever and a modified live virus of MD-BVD.

    Science.gov (United States)

    Haralambiev, H; Tsvetkov, P; Nikolov, A; Gaytandgieva, R

    1975-01-01

    Calves vaccinated with the modified strain C24V react by creating neutralizing antibodies in high titres and are resistent to infection while non-immunized calves manifest the clinical symptoms of MD-BVD. Calves, vaccinated with the lapinized strain "K" of the SF virus react by creating neutralizing antibodies in low titres against the vius of MD-BVD and manifest no apparent clinical symptoms after challenge with the virulent strain MD-BVD.

  5. Molecular epidemiology of viral diarrhea in Chengdu infants and young children%成都地区婴幼儿病毒性腹泻分子流行病学研究

    Institute of Scientific and Technical Information of China (English)

    谢晓丽; 张伟; 廖雪春; 邓孝智; 刘丽荣; 张琪琛; 商丽红; 温燕; 任敏

    2012-01-01

    目的 通过分子流行病学研究成都地区婴幼儿病毒性腹泻的病原学特点,掌握本地区病原分布特征,为疫苗研制和疫情控制提供科学依据.方法 采用酶联免疫吸附试验( ELISA)及逆转录聚合酶链反应(RT-PCR)对成都地区2006 -2008年度376例婴幼儿腹泻粪便标本进行轮状病毒(RV)、杯状病毒(HuCV)、星状病毒(AstV)及肠道腺病毒(Adv)检测.结果 RV的检出率为37.76%(142/376),其中42例进行G分型,45例进行P分型,G分型以G3型为主21例(50%),其次为G2和G1型,P分型以P[8]型为主21例,其次是P[4]型19例.RV感染主要为6~23月的婴幼儿,发病高峰在10 ~12月份(75.8%).RT-PCR法检出HuCV、AstV及Adv的检出率分别为15.85%、1.64%及2.04%.结论 RV是成都地区婴幼儿病毒性腹泻的主要病原,其流行的主要血清型为G3型、P[8]、P[4]型.除RV外,HuCV也是重要的病原.%Objective To investigated the molecular epidemiologic features of viral diarrhea in Chengdu infants and young children,and to establish baseline patterns of etiology,provides the scientific basis for the vaccine development and the epidemic situation control.Methods From March,2006 to December,2008,a total of 376 infants and young children from Chengdu area hospitalized for diarrhea in Chengdu Children's Hospital were enrolled in this study.The stool specimen collected from each patient was tested for rotavirus ( RV),Calicivirus ( CV),astrovirus ( AstV ) and adenovirus (Adv) by using enzyme linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR) examination.Results Among those 376 cases,there were 142 cases (37.76%) of RV infections,which scattered predominantly in October to December.Among 234 cases RV negativity,there were 29 cases HuCV infections ( 15.85% ),5 cases AstV infections ( 1.64% ),and 8 cases Adv infections (2.04%).Conclusion RV appeared to be the main etiological agent of viral diarrhea in Chengdu

  6. 2008-2009年天津市病毒性腹泻病例疾病负担分析%The analysis of disease burdern of viral diarrhea cases in Tianjin during 2008-2009

    Institute of Scientific and Technical Information of China (English)

    陈静; 李佳葫; 张颖; 刘辉; 高璐; 徐文体; 苏旭; 田宏; 张之伦

    2012-01-01

    目的 评价天津市病毒性腹泻病例疾病负担,为制定防控政策提供科学依据.方法 采用面对面现场询问的方法,参考家庭负担会谈量表自制问卷,调查被调查对象一般情况和相关费用及家庭负担.采集病例粪便标本进行实验室检测.结果 天津市2008 -2009年951例病毒性腹泻病例人均总费用为178.8元,5岁以下的门诊病例(3 640.5元)和住院病例(4 434.5元)的人均费用分别是5岁及以上病例的3.23倍和6.24倍,5岁以下组病例感染诺如病毒直接医疗费用要高于轮状病毒(P<0.05),感染轮状病毒比诺如病毒对家庭成员收入和家庭娱乐活动影响要大(P<0.05);而5岁及以上组病例,感染轮状病毒的直接费用要高于诺如病毒(P<0.05),而诺如病毒造成的家庭负担最重(P<0.05).结论 病毒性腹泻病例中5岁以下儿童经济负担重,并且感染诺如病毒比轮状病毒的疾病负担高,建议将有效的肠道病毒疫苗列为计划免疫范畴.%Objective To evaluate the disease burden of viral diarrhea cases in Tianjin so as to provide the scientific basis for decision making. Methods To investigate the basic situation of the patients' families, the treatment-related cost and family burden by the face-to-face interview with the self-designed questionnaire of diseases burden scale. Stool specimens were taken from cases for laboratory test. Results Nine hundred and fifty one diarrhea cases in Tianjin during 2008-2009 were investigated and the average total cost was 178.8¥. The average cost (3 640.5¥) of outpatient cases under 5 year-old group was 3.23 times compared with that of cases over 5 year-old group,and the average cost (4 434.5¥) of inpa-tient cases under 5 years was 6.24 times compared with that of cases over 5 year-old. For cases under 5 year-old group, the direct medical cost of norovirus infectious cases was higher than that of rotavirus infectious cases (I^O.05), but ro-tavirus infectious

  7. Antibiotic-Associated Diarrhea

    Science.gov (United States)

    Antibiotic-associated diarrhea Overview By Mayo Clinic Staff Antibiotic-associated diarrhea refers to passing loose, watery stools ... after taking medications used to treat bacterial infections (antibiotics). Most often, antibiotic-associated diarrhea is mild and ...

  8. The effect of bovine viral diarrhoea virus on fertility in dairy cows: two case-control studies in the province of Styria, Austria.

    Science.gov (United States)

    Burgstaller, Johann; Obritzhauser, Walter; Kuchling, Sabrina; Kopacka, Ian; Pinior, Beate; Köfer, Josef

    2016-01-01

    Bovine viral diarrhoea (BVD) leads to substantial economic losses in beef and dairy herds worldwide. Two case-control studies were carried out using production data from 1996 to 2012 to analyse the impact of BVD virus (BVDV) on fertility in dairy herds in the province of Styria during an eradication programme. In study 1, herds in which at least one persistently BVDV-infected (PI) animal was detected (case herds) were compared to a group of control herds proven free from BVDV infection (contro herds). In study 2, within BVD infected herds the period during which P animals were present (exposed period) was compared to the period after successful BVD eradication (unexposed period). Calving interval (CAl) and the probability of a first service conception (FSC) were used as indicators in a mixed regression model to investigate the impact of BVD on reproductive performance. The model results indicated that BVD had a significant influence on CAl and FSC. Cows from control herds were 1.1 times more likely to conceive at first service compared to cows from case herds and cows served during the BVDV unexposed period were 1.3 times more likely to conceive at first service than those inseminated during the exposed period. In BVD-infected herds the CAI averaged seven days shorter in unexposed periods than in exposed periods. Besides BVD the animal breed and the parity substantially impact the analysed fertility indicators.

  9. Evidence of a humoral immune response against the prokaryotic expressed N-terminal autoprotease (Npro) protein of bovine viral diarrhoea virus

    Indian Academy of Sciences (India)

    Niranjan Mishra; Katherukamem Rajukumar; Shruti Shrikant Pitale; Anil Prakash; Ram Kumar Nema; Sthita Pragnya Behera; Shiv Chandra Dubey

    2010-03-01

    Bovine viral diarrhoea virus (BVDV) is an economically important pathogen of cattle and sheep belonging to the genus Pestivirus of the family Flaviviridae. Although the BVDV non-structural N-terminal protease (Npro) acts as an interferon antagonist and subverts the host innate immunity, little is known about its immunogenicity. Hence, we expressed a recombinant BVDV Npro–His fusion protein (28 kDa) in E. coli and determined the humoral immune response generated by it in rabbits. The antigenicity of the Npro protein was confirmed by western blot using anti-BVDV hyperimmune cattle, sheep and goat serum, and anti-Npro rabbit serum. When rabbits were immunized with the Npro protein, a humoral immune response was evident by 4 weeks and persisted till 10 weeks post immunization as detected by ELISA and western blot. Despite Npro-specific antibodies remaining undetectable in 80 serum samples from BVDV-infected sheep and goats, BVDV hyperimmune sera along with some of the field cattle, sheep and goat sera with high BVDV neutralizing antibody titres were found positive for Npro antibodies. Our results provide evidence that despite the low immunogenicity of the BVDV Npro protein, a humoral immune response is induced in cattle, sheep and goats only with repeated BVDV exposure.

  10. New parvovirus in child with unexplained diarrhea, Tunisia.

    Science.gov (United States)

    Phan, Tung G; Sdiri-Loulizi, Khira; Aouni, Mahjoub; Ambert-Balay, Katia; Pothier, Pierre; Deng, Xutao; Delwart, Eric

    2014-11-01

    A divergent parvovirus genome was the only eukaryotic viral sequence detected in feces of a Tunisian child with unexplained diarrhea. Tusavirus 1 shared 44% and 39% identity with the nonstructural protein 1 and viral protein 1, respectively, of the closest genome, Kilham rat parvovirus, indicating presence of a new human viral species in the Protoparvovirus genus.

  11. Acute diarrhea in children

    Directory of Open Access Journals (Sweden)

    Radlović Nedeljko

    2015-01-01

    Full Text Available Acute diarrhea (AD is the most frequent gastroenterological disorder, and the main cause of dehydration in childhood. It is manifested by a sudden occurrence of three or more watery or loose stools per day lasting for seven to 10 days, 14 days at most. It mainly occurs in children until five years of age and particularly in neonates in the second half-year and children until the age of three years. Its primary causes are gastrointestinal infections, viral and bacterial, and more rarely alimentary intoxications and other factors. As dehydration and negative nutritive balance are the main complications of AD, it is clear that the compensation of lost body fluids and adequate diet form the basis of the child’s treatment. Other therapeutic measures, except antipyretics in high febrility, antiparasitic drugs for intestinal lambliasis, anti-amebiasis and probiotics are rarely necessary. This primarily regards uncritical use of antibiotics and intestinal antiseptics in the therapy of bacterial diarrhea. The use of antiemetics, antidiarrhetics and spasmolytics is unnecessary and potentially risky, so that it is not recommended for children with AD.

  12. Distribution pattern of bovine viral diarrhoea virus type 1 genome in lymphoid tissues of experimentally infected sheep.

    Science.gov (United States)

    Karikalan, M; Rajukumar, K; Mishra, N; Kumar, M; Kalaiyarasu, S; Rajesh, K; Gavade, V; Behera, S P; Dubey, S C

    2016-06-01

    In this study, cellular localization and the distribution pattern of BVDV genome in lymphoid tissues during the course of experimental acute BVDV-1 infection of sheep was investigated. Tonsils, mesenteric lymph nodes (MLN) and spleen were collected on 3, 6, 9, 12 and 15 days post infection (dpi) from twenty 4-month-old lambs, experimentally inoculated intra-nasally with 5 × 10(5) TCID50 of a non-cytopathic (ncp) BVDV-1 isolate, Ind-17555. Tissues collected from ten mock-infected lambs served as controls. In situ hybridization (ISH) was carried out in paraformaldehyde fixed paraffin embedded tissue sections using digoxigenin labelled riboprobe targeting 5'-UTR of BVDV-1. BVDV genome was detected at all the intervals from 3 dpi to 15 dpi in the lymphoid tissues with variations between the intervals and also amongst the infected sheep. During the early phase of acute infection, presence of viral genome was more in tonsils than MLN and spleen, whereas the distribution was higher in MLN during later stages. BVDV-1 genome positive cells included lymphocytes, macrophages, plasma cells, reticular cells and sometimes crypt epithelial cells. Genome distribution was frequently observed in the lymphoid follicles of tonsils, MLN and spleen, besides the crypt epithelium in tonsils, paracortex and medullary sinus and cords of MLN. Most abundant and widespread distribution of BVDV-1 genome was observed on 6 dpi while there was a reduction in number and intensity of positive signals by 15 dpi in most of the infected animals. This is the first attempt made to study the localisation of BVDV-1 in lymphoid tissues of acutely infected sheep by in situ hybridization. The results show that the kinetics of BVDV-1 distribution in lymphoid tissues of experimentally infected non-pregnant sheep follows almost a similar pattern to that demonstrated in BVDV infected cattle.

  13. Immunisation of Sheep with Bovine Viral Diarrhoea Virus, E2 Protein Using a Freeze-Dried Hollow Silica Mesoporous Nanoparticle Formulation.

    Directory of Open Access Journals (Sweden)

    Donna Mahony

    Full Text Available Bovine viral diarrhoea virus 1 (BVDV-1 is arguably the most important viral disease of cattle. It is associated with reproductive, respiratory and chronic diseases in cattle across the world. In this study we have investigated the capacity of the major immunological determinant of BVDV-1, the E2 protein combined with hollow type mesoporous silica nanoparticles with surface amino functionalisation (HMSA, to stimulate immune responses in sheep. The current work also investigated the immunogenicity of the E2 nanoformulation before and after freeze-drying processes. The optimal excipient formulation for freeze-drying of the E2 nanoformulation was determined to be 5% trehalose and 1% glycine. This excipient formulation preserved both the E2 protein integrity and HMSA particle structure. Sheep were immunised three times at three week intervals by subcutaneous injection with 500 μg E2 adsorbed to 6.2 mg HMSA as either a non-freeze-dried or freeze-dried nanoformulation. The capacity of both nanovaccine formulations to generate humoral (antibody and cell-mediated responses in sheep were compared to the responses in sheep immunisation with Opti-E2 (500 μg together with the conventional adjuvant Quil-A (1 mg, a saponin from the Molina tree (Quillaja saponira. The level of the antibody responses detected to both the non-freeze-dried and freeze-dried Opti-E2/HMSA nanoformulations were similar to those obtained for Opti-E2 plus Quil-A, demonstrating the E2 nanoformulations were immunogenic in a large animal, and freeze-drying did not affect the immunogenicity of the E2 antigen. Importantly, it was demonstrated that the long term cell-mediated immune responses were detectable up to four months after immunisation. The cell-mediated immune responses were consistently high in all sheep immunised with the freeze-dried Opti-E2/HMSA nanovaccine formulation (>2,290 SFU/million cells compared to the non-freeze-dried nanovaccine formulation (213-500 SFU/million cells

  14. Genomic characterization of a rotavirus G8P[1] detected in a child with diarrhea reveal direct animal-to-human transmission.

    Science.gov (United States)

    Martinez, Magaly; Phan, Tung Gia; Galeano, Maria Eugenia; Russomando, Graciela; Parreno, Viviana; Delwart, Eric; Parra, Gabriel I

    2014-10-01

    Group A rotavirus is a major cause of severe gastroenteritis in children and young animals. During a retrospective analysis of samples collected from Paraguayan children under 5 years old with diarrhea, and previously negative for rotavirus and norovirus, we detected the presence of bovine rotavirus sequences by viral metagenomics. Nucleic acid was extracted direct from stool sample and determined to be G8P[1]. The genomic analyzes revealed that the strain presents an Artiodactyl-like genome (G8-P[1]-I2-R2-C2-M1-Ax-N2-T6-E12-H3) suggesting a direct animal-to-human transmission.

  15. Relative associations of cattle movements, local spread, and biosecurity with bovine viral diarrhoea virus (BVDV) seropositivity in beef and dairy herds.

    Science.gov (United States)

    Gates, M C; Woolhouse, M E J; Gunn, G J; Humphry, R W

    2013-11-01

    The success of bovine viral diarrhoea virus (BVDV) eradication campaigns can be undermined by spread through local transmission pathways and poor farmer compliance with biosecurity recommendations. This work combines recent survey data with cattle movement data to explore the issues likely to impact on the success of BVDV control in Scotland. In this analysis, data from 249 beef suckler herds and 185 dairy herds in Scotland were studied retrospectively to determine the relative influence of cattle movements, local spread, and biosecurity on BVDV seropositivity. Multivariable logistic regression models revealed that cattle movement risk factors had approximately 3 times greater explanatory power than risk factors for local spread amongst beef suckler herds, but approximately the same explanatory power as risk factors for local spread amongst dairy herds. These findings are most likely related to differences in cattle husbandry practices and suggest that where financial prioritization is required, focusing on reducing movement-based risk is likely to be of greatest benefit when applied to beef suckler herds. The reported use of biosecurity measures such as purchasing cattle from BVDV accredited herds only, performing diagnostic screening at the time of sale, implementing isolation periods for purchased cattle, and installing double fencing on shared field boundaries had minimal impact on the risk of beef or dairy herds being seropositive for BVDV. Only 28% of beef farmers and 24% of dairy farmers with seropositive herds recognized that their cattle were affected by BVDV and those that did perceive a problem were no less likely to sell animals as replacement breeding stock and no more likely to implement biosecurity measures against local spread than farmers with no perceived problems. In relation to the current legislative framework for BVDV control in Scotland, these findings emphasize the importance of requiring infected herds take appropriate biosecurity measures

  16. Validation of a test for dams carrying foetuses persistently infected with bovine viral-diarrhoea virus based on determination of antibody levels in late pregnancy.

    Science.gov (United States)

    Lindberg, A; Groenendaal, H; Alenius, S; Emanuelson, U

    2001-10-11

    Our objective was to estimate, using a generalised linear mixed-model approach, the sensitivity and specificity of an indirect ELISA when used to identify dams pregnant with persistently bovine viral-diarrhoea virus (BVDV)-infected foetuses. Cows that had been tested for antibodies to BVDV with a positive result during their pregnancy and where the offspring had been tested for both antibody and virus were identified by accessing the Swedish BVD database and the official pedigree records. The resulting data set consisted of 2162 cow-calf pairs in 126 herds, of which 281 included virus-positive calves. The sensitivities and specificities at 12 different decision thresholds (corresponding to optical densities (ODs) between 0.5 and 1.6) were estimated using generalised linear mixed models (binomial error, logit link), in which the gold standard (the BVDV status of the calf) was included as a covariate. In each model, the dependent variable was the dichotomous test result at the decision threshold in question. There was a significant positive interaction between the calf's status and gestational stage in all 12 models--indicating that the sensitivity and specificity at any given decision threshold was improved when the the test was performed later in pregnancy. The test should be applied only when samples have been taken in late gestation--not before the seventh month in pregnancy. If applied during the last months of pregnancy, the point estimate of the sensitivity ranges between 0.94 and 1.0 as the decision threshold is moved from 1.0 and downwards to 0.7. Similarly, the specificity ranges between 0.39 and 0.67 as the decision threshold is moved from 0.8 and upwards to 1.1.

  17. A case control study on the risk factors of viral diarrhea in children below 5 years old%5岁以下婴幼儿病毒性腹泻的危险因素配对病例对照研究

    Institute of Scientific and Technical Information of China (English)

    许可; 霍翔; 祖荣强; 李亮; 汤奋扬; 朱凤才; 羊海涛; 汪华

    2011-01-01

    Objective: To assess the risk factors of viral diarrhea among children below 5 years old. Methods:A case-control study was conducted on two groupe of children matched by sex, age with or without viral diarrhea from hospitals in Xuzhou and Zhenjiang area. The pathogen of diarrhea was confirmed by ELISA and RT-PCR methods. Interview was carried out with uniform designed questionnaires. Cox's proportional hazards regression analysis was then performed. Results:Factors that were independently associated with the development of viral diarrhea, would include children's address, weight, parents' Schooling and profession, washing-up sink in kitchen, cutting cooked and uncooked foods by different kitchen knife and target, raising pets, cooker washing hands before cook, the children contacting animals and taking antibiotics one week ago before diarrhea. Anamnesis and rotavims vaccine inoculation were not found associating with viral diarrhea. Multivariate logistic regression model was then fitted with three variables, including washing-up sink in kitchen, cutting cooked and uncooked foods by different kitchen knife and target, taking antibiotics one week ago before diarrhea. The 95% CI of OR was 0.340~0.852, 0.295~0.705 and 2.153~6.227 separately. Conclusion:The risk of viral diarrhea increased with worse hygienic habits and antibiotics taken.%目的:探讨5岁以下婴幼儿病毒性腹泻的危险因素.方法:采用性别、年龄匹配的1:2配对病例对照研究,对江苏省徐州、镇江市县级医疗机构2007年10月至2008年10月收治的5岁以下腹泻儿童500例和对照儿童1 000例进行问卷调查.腹泻的病原体采用酶联免疫吸附剂测定(enzyme linked immunosorbent assay,ELISA)或反转录PCR(reverse transcription-PCR,RT-PCR)进行检测确认.数据采用Cox比例风险模型拟合Logistic回归.结果:单因素分析显示病毒性腹泻患儿的城乡差异、父母受教育程度、职业、家庭饮食卫生(包括厨

  18. Detection of viral nucleic acid of vesicular stomatitis virus and bovine viral diarrhea virus simultaneously by Bi-PCR%二重RT-PCR同时检测VSV与BVDV核酸

    Institute of Scientific and Technical Information of China (English)

    杨桂梅; 徐自忠; 高洪; 花群义; 周晓黎; 杨昌焰

    2003-01-01

    水泡性口炎病毒(VSV)与牛病毒性腹泻病毒(BVDV)具有相近的传播途径与类似的检测方法,本文参照文献报道的基因序列,设计合成了两对能分别扩增vSV(202 bp)、BVDV(341 bp)基因片段的引物,并对PCR扩增条件进行优化,建立了二重RT-PCR方法,可同时检测VSV与BVDV病毒核酸.VSV产物经测序显示与报道的核酸序列同源性为88.6%.二重RT-PCR同时检测VSV与BVDV经济、快速、敏感、特异,可用于实验研究和流行病学调查.

  19. Increase of cells expressing PD-L1 in bovine leukemia virus infection and enhancement of anti-viral immune responses in vitro via PD-L1 blockade

    Directory of Open Access Journals (Sweden)

    Ikebuchi Ryoyo

    2011-09-01

    Full Text Available Abstract The inhibitory receptor programmed death-1 (PD-1 and its ligand, programmed death-ligand 1 (PD-L1 are involved in immune evasion mechanisms for several pathogens causing chronic infections. Blockade of the PD-1/PD-L1 pathway restores anti-virus immune responses, with concomitant reduction in viral load. In a previous report, we showed that, in bovine leukemia virus (BLV infection, the expression of bovine PD-1 is closely associated with disease progression. However, the functions of bovine PD-L1 are still unknown. To investigate the role of PD-L1 in BLV infection, we identified the bovine PD-L1 gene, and examined PD-L1 expression in BLV-infected cattle in comparison with uninfected cattle. The deduced amino acid sequence of bovine PD-L1 shows high homology to the human and mouse PD-L1. The proportion of PD-L1 positive cells, especially among B cells, was upregulated in cattle with the late stage of the disease compared to cattle at the aleukemic infection stage or uninfected cattle. The proportion of PD-L1 positive cells correlated positively with prediction markers for the progression of the disease such as leukocyte number, virus load and virus titer whilst on the contrary, it inversely correlated with the degree of interferon-gamma expression. Blockade of the PD-1/PD-L1 pathway in vitro by PD-L1-specific antibody upregulated the production of interleukin-2 and interferon-gamma, and correspondingly, downregulated the BLV provirus load and the proportion of BLV-gp51 expressing cells. These data suggest that PD-L1 induces immunoinhibition in disease progressed cattle during chronic BLV infection. Therefore, PD-L1 would be a potential target for developing immunotherapies against BLV infection.

  20. Diarrhea (For Parents)

    Science.gov (United States)

    ... hospitalization. Another group of viruses that can cause diarrhea in children, especially during the summer months, are enteroviruses, particularly ... water, this parasite often is the culprit behind diarrhea epidemics in child-care centers and other public places. Cryptosporidiosis often ...

  1. Diarrhea in infants

    Science.gov (United States)

    ... baby food that makes diarrhea worse, such as: Apple juice Milk Fried foods Full-strength fruit juice ... Diarrhea Browse the Encyclopedia A.D.A.M., Inc. is accredited by URAC, also known as the ...

  2. Human and bovine viruses in the Milwaukee River Watershed: hydrologically relevant representation and relations with environmental variables

    Science.gov (United States)

    Corsi, Steven R.; Borchardt, M. A.; Spencer, S. K.; Hughes, Peter E.; Baldwin, Austin K.

    2014-01-01

    To examine the occurrence, hydrologic variability, and seasonal variability of human and bovine viruses in surface water, three stream locations were monitored in the Milwaukee River watershed in Wisconsin, USA, from February 2007 through June 2008. Monitoring sites included an urban subwatershed, a rural subwatershed, and the Milwaukee River at the mouth. To collect samples that characterize variability throughout changing hydrologic periods, a process control system was developed for unattended, large-volume (56–2800 L) filtration over extended durations. This system provided flow-weighted mean concentrations during runoff and extended (24-h) low-flow periods. Human viruses and bovine viruses were detected by real-time qPCR in 49% and 41% of samples (n = 63), respectively. All human viruses analyzed were detected at least once including adenovirus (40% of samples), GI norovirus (10%), enterovirus (8%), rotavirus (6%), GII norovirus (1.6%) and hepatitis A virus (1.6%). Three of seven bovine viruses analyzed were detected including bovine polyomavirus (32%), bovine rotavirus (19%), and bovine viral diarrhea virus type 1 (5%). Human viruses were present in 63% of runoff samples resulting from precipitation and snowmelt, and 20% of low-flow samples. Maximum human virus concentrations exceeded 300 genomic copies/L. Bovine viruses were present in 46% of runoff samples resulting from precipitation and snowmelt and 14% of low-flow samples. The maximum bovine virus concentration was 11 genomic copies/L. Statistical modeling indicated that stream flow, precipitation, and season explained the variability of human viruses in the watershed, and hydrologic condition (runoff event or low-flow) and season explained the variability of the sum of human and bovine viruses; however, no model was identified that could explain the variability of bovine viruses alone. Understanding the factors that affect virus fate and transport in rivers will aid watershed management for minimizing

  3. Some viral and bacterial respiratory tract infections of dairy cattle during the summer season

    Directory of Open Access Journals (Sweden)

    Kale M.

    2013-01-01

    Full Text Available In this research, dairy cattle with respiratory system problems that were brought to a private slaughterhouse in Burdur province were investigated for viral and bacterial infections present in the summer season. The blood samples were collected from 56 animals. The samples were tested for antibodies against bovine herpesvirus 1 (BoHV-1, bovine viral diarrhea virus (BVDV, bovine respiratory syncytial virus (BRSV, bovine parainfluenza virus 3 (BPIV-3 and bovine adenovirus 3 (BAV-3 by ELISA. Bacteriological cultivation was carried out from lung samples taken after cutting the same animals. The seropositivity rates which were determined for 5 viruses in cattle (BoHV- 1, BVDV, BRSV, BPIV-3 and BAV-3 were 7.14%, 50%, 94.64%, 94.64% and 82.14% respectively. The presence of antibodies against the viruses was as follows; 5.36% of cattle had antibodies against only one virus, 14.29% against two, 30.36% against three, 44.64% against four and 5.36% against five viruses. A total of 36 bacterial agents were isolated from 30 out of 56 lung samples. From the lung samples, only one bacterium was isolated from 39.3% (22/56 samples, and more than one bacterium from 14.3% (8/56. Escherichia coli, Staphylococcus aureus and Streptococcus spp. were detected as the most often isolated agents. Compared to bacteria, the rates of viral infections associated with Escherichia coli (BRSV+BPIV-3+BAV- 3+Escherichia coli; 8.92% and BRSV+BPIV-3+Escherichia coli; 5.35% were higher. As a consequence, it was thought that primary agents which were the viruses and bacteria may have attended as secondary factors in respiratory tract infections of dairy cattle.

  4. Enzootic bovine leukosis and Bovine leukemia virus

    OpenAIRE

    Amauri Alcindo Alfieri; Alice Fernandes Alfieri; Luis Álvaro Leuzzi Junior

    2004-01-01

    All over de World the Enzootic Bovine Leukosis is a important viral infection in cattle herds. This revision points out topics relative to the etiological agent, clinical signals, diagnosis methods, control and prophylaxis of the infection.A Leucose Enzoótica Bovina é uma infecção viral amplamente disseminada em rebanhos bovinos de todo o mundo. Esta revisão tem por objetivo apresentar tópicos relacionados ao agente etiológico, à doença clínica e aos métodos de diagnóstico, controle e profila...

  5. Virus survival in slurry: Analysis of the stability of foot-and-mouth disease, classical swine fever, bovine viral diarrhoea and swine influenza viruses

    DEFF Research Database (Denmark)

    Bøtner, Anette; Belsham, Graham

    2012-01-01

    Farm slurry can be highly contaminated with viral pathogens. The survival of these pathogens within slurry is important since this material is often distributed onto farm land either directly or after heat treatment. There is clearly some risk of spreading pathogens in the early stages of an outb...... viruses under all conditions tested. The implications for disease spread are discussed....

  6. Surveillance of rotavirus diarrhea

    Directory of Open Access Journals (Sweden)

    Titis Widowati

    2012-01-01

    Full Text Available Background Rotavirus is a major cause of severe diarrhea and dehydration in children worldwide. Data on the burden of disease in Indonesia is limited. Objective To provide an epidemiological profile of rotavirus infection among children hospitalized for diarrhea in Mohammad Hoesin Hospital, Palembang. Methods In January - December 2006, a prospective, hospital-based surveillance was carried out in children aged less than five years, presenting with diarrhea. Stool samples were examined for rotavirus using enzyme immunoassay (EIA. G- and P-typing were performed on specimens confirmed to be positive by EIA. Results A total of 513 fecal specimens from 534 children were tested for rotavirus. Rotavirus was detected in 64% of the specimens, mostly of the G9 type (62.5%. Incidence of rotavirus diarrhea was highest in the 6 month to 2 years age group (60.4%. Children with rotavirus diarrhea were more likely to present with dehydration, compared to those with non-rotavirus diarrhea (94% vs 70%, respectively, P=0.03. Conclusion Rotavirus was the most common pathogen found in children with diarrhea. Rotavirus was detected in 64% of pediatric diarrheal specimens tested in our study. This finding warrants the use of a large-scale program to prevent disease, such as vaccination against rotavirus. [Paediatr Indones. 2012;52:22-7].

  7. Diarrhea in the immunocompromised patient.

    Science.gov (United States)

    Krones, Elisabeth; Högenauer, Christoph

    2012-09-01

    Diarrhea is a common problem in patients with immunocompromising conditions. The etiologic spectrum differs from patients with diarrhea who have a normal immune system. This article reviews the most important causes of diarrhea in immunocompromised patients, ranging from infectious causes to noninfectious causes of diarrhea in the setting of HIV infection as a model for other conditions of immunosuppression. It also deals with diarrhea in specific situations, eg, after hematopoietic stem cell or solid organ transplantation, diarrhea induced by immunosuppressive drugs, and diarrhea in congenital immunodeficiency syndromes.

  8. Carbamazepine-Induced Diarrhea

    OpenAIRE

    J Gordon Millichap

    1992-01-01

    Intractable diarrhea induced by carbamazepine (CBZ) in 3 patients and necessitating discontinuation of the drug is reported from the Departments of Neurology and Medicine, University of Louisville School of Medicine, Kentucky.

  9. Molecular and antigenic characterization of bovine Coronavirus circulating in Argentinean cattle during 1994-2010.

    Science.gov (United States)

    Bok, M; Miño, S; Rodriguez, D; Badaracco, A; Nuñes, I; Souza, S P; Bilbao, G; Louge Uriarte, E; Galarza, R; Vega, C; Odeon, A; Saif, L J; Parreño, V

    2015-12-31

    Bovine coronavirus (BCoV) is an important viral pathogen associated with neonatal calf diarrhea. Our aim was to investigate the incidence of BCoV in diarrhea outbreaks in beef and dairy herds from Argentina during 1994-2010. A total of 5.365 fecal samples from diarrheic calves were screened for BCoV diagnosis by ELISA. The virus was detected in 1.71% (92/5365) of the samples corresponding to 5.95% (63/1058) of the diarrhea cases in 239 beef and 324 dairy farms. The detection rate of BCoV was significantly higher in dairy than in beef herds: 12.13% (29/239) vs. 4.32% (14/324) respectively. Phylogenetic analysis of the hypervariable S1 region of seven representative samples (from different husbandry systems, farm locations and years of sampling) indicated that BCoV strains circulating in Argentinean beef and dairy herds formed a cluster distinct from other geographical regions. Interestingly, Argentinean strains are distantly related (at both the nucleotide and amino acid levels) with the Mebus historic reference BCoV strain included in the vaccines currently available in Argentina. However, Mebus-induced antibodies were capable of neutralizing the BCoV Arg95, a field strain adapted to grow in vitro, and vice versa, indicating that both strains belong to the same CoV serotype reported in cattle. This work represents the first large survey describing BCoV circulation in Argentinean cattle.

  10. Molecular epidemiological survey on children with viral diarrhea in Tangshan%唐山地区儿童病毒性腹泻分子流行病学调查

    Institute of Scientific and Technical Information of China (English)

    余亮科; 赵文喆; 高捷; 张双; 张志坤

    2011-01-01

    Objective: To explore the current situation of diarrhea in children in Tangshan region from January 2007 to December 2010, and a epidemiological investigation and analysis on the viruses of diarrhea was conducted. Methods: 1 100 children with diarrhea were selected randomly from Tangshan region from January 2007 to December 2010, all the children received stool examination, a variety of methods were used to analyze the common pathogens and peak time of onset Results: Among 1 100 children, 582 children were found with rolavirus, accounting for 52. 91 % ; 258 children were found with norovirus, accounting for 23. 45% ; 89 children were found with sapovirus, accounting for 8. 09% ; 52 children were found with astrovirus, accounting for 4. 73 % ; 119 children were found with adenovirus, accounting for 10. 82%. The peak time of onset of rotavirus was October - December and January, while the peak time of onset of norovirus was July ~ September, the other viruses could be detected in each time period Conclusion: The peak age of onset among the children with diarrhea is below 5 years. The major viruses of diarrhea are rotavirus and norovirus; astrovirus, sapovirus and adenovirus are also common viruses. Different viruses have different peak times of onset, for rotavirus, the peak times of onset are autumn and winter; while for norovirus, the peak times of onset are summer and autumn; the other viruses have no obvious seasonal distribution.%目的:探讨唐山地区2007年1月~2010年12月儿童腹泻发病现状,并对腹泻病毒进行流行病学调查分析.方法:随机抽取唐山地区2007年1月~2010年12月腹泻儿童1 100例资料.所有患儿均经过粪便检查,采用多种方法分析常见病原菌及好发时间.结果:1 100例儿童中,轮状病毒582例,占52.91%;诺如病毒258例,占23.45%;札如病毒89例,占8.09%;星状病毒52例,占4.73%;腺病毒119例,占10.82%.其中轮状病毒在10~12月及1月高发,诺如病毒为7~9月高

  11. Bovine herpesvirus 1 infection and infectious bovine rhinotracheitis

    OpenAIRE

    2007-01-01

    International audience; Bovine herpesvirus 1 (BoHV-1), classified as an alphaherpesvirus, is a major pathogen of cattle. Primary infection is accompanied by various clinical manifestations such as infectious bovine rhinotracheitis, abortion, infectious pustular vulvovaginitis, and systemic infection in neonates. When animals survive, a life-long latent infection is established in nervous sensory ganglia. Several reactivation stimuli can lead to viral re-excretion, which is responsible for the...

  12. Establishment of a new bovine leukosis virus producing cell line.

    Science.gov (United States)

    Beier, D; Riebe, R; Blankenstein, P; Starick, E; Bondzio, A; Marquardt, O

    2004-11-01

    Due to the prevalence of different bovine leukosis virus (BLV) species in the cattle population in Europe, problems may arise in the serological diagnosis of BLV infections. In addition, earlier investigations demonstrated that contamination of the BLV antigen-producing cell culture systems by bovine viral diarrhea virus (BVDV) may give rise to misinterpretation of serological test results after BVDV vaccination of cattle. By co-cultivation of peripheral leukocytes of a BLV-infected cow with a permanent sheep kidney cell line, a new BLV-producing cell line named PO714 was established. This line carries a BLV provirus of the Belgian species and has been tested to be free of a variety of possibly contaminating viruses and mycoplasms. Investigations of a panel of well-characterised sera by agar gel immunodiffusion (AGID) and capture ELISA (cELISA) tests using antigen prepared from this new cell line in comparison with antigen of the well-known cell line FLK/BLV yielded comparable results. False positive results caused by BVDV cross-reactions could be eliminated when tests were carried out with antigen derived from the new cell line.

  13. [Rotavirus and other viruses of diarrhea].

    Science.gov (United States)

    Bajolet, O; Chippaux-Hyppolite, C

    1998-01-01

    Rotaviruses represent 80% of recognized viral etiologies and 140 million cases of diarrhea per year. They strike young children with similar frequency throughout the world, but the mortality rate is high in developing countries only, with some 870,000 deaths per year (WHO, 1997). Rotaviruses belong to the family of Reoviridae; they are segmented bicatenary RNA viruses, which explains their genetic variability, the presence of mixed infections, the establishment for some time already of a molecular epidemiology by electrophore types. The viruses are "naked" and thus resistant to the outside environment; their massive elimination, 10(8) to 10(10)viral particles per gram of faeces, begins with the first day of diarrhea. They are found in used water and can also be concentrated by shellfish; the environment thus constitutes a notable reservoir for the virus. Oral-faecal transmission is facilitated by deficient sanitary conditions. The 11 fragments of the genome each codify for 1 viral protein; 2 surface proteins, VP4 and VP7, bring about the formation of neutralizing antibodies, which are important for the protection and determination of different serotypes. A non structural protein--NSP4--would seem to intervene in the cytopathogenic effect and may act as a veritable viral enterotoxine. Numerous animal species are infected by rotaviruses which are district from the human ones. The pathology as it affects animals is of economic importance and of interest as an experimental and vaccinal model. Between human and animal rotaviruses there can be genetic rematchings and the VP6 protein is an antigen common to the group. The description of the other viruses responsible for diarrhea has benefited from widespread use of electronic microscopes from the very first years of study of rotaviruses. These other viruses belong to 6 different types: adenovirus, calcivirus, astrovirus, Norwalk agent and related viruses, coronavirus, enterovirus. They therefore have a structural and

  14. Detección de bovinos portadores e inmunotolerantes al virus de la diarrea viral bovina en predios lecheros de la Región Metropolitana de Chile Identification of bovine viral diarrhoea virus in persistently infected immunotolerant cows in herds of the Region Metropolitana, Chile

    Directory of Open Access Journals (Sweden)

    M. CELEDON

    1998-01-01

    Full Text Available Se implementó una técnica para detectar la presencia de bovinos portadores e inmunotolerantes (PIT al virus de la diarrea viral bovina (VDVB, a través de la pesquisa de viremia persistente y bajos títulos de anticuerpos, para el VDVB, en dos muestras de sangre tomadas, desde un mismo animal, con tres meses de separación. La presencia de virus en la sangre se evidenció a través del aislamiento viral (AV -desde muestras de plasma y linfocitos- en cultivos primarios de testículo bovino y la identificación del supuesto aislado se efectuó por prueba de inmunoperoxidasa indirecta (IPI en el segundo pasaje del inóculo sobre las células. La detección de anticuerpos séricos se realizó por prueba de seroneutralización (SN, empleando la cepa citopatogénica Singer del VDVB. Se consideró animal PIT al que evidenció presencia de virus en las 2 muestras de sangre y mantuvo títulos de anticuerpos menores o iguales a 8. De 238 bovinos muestreados en 42 se detectó viremia persistente en ausencia o bajos títulos de anticuerpos, considerándose una prevalencia de 18% de bovinos PIT, y de los 34 predios muestreados en 22 se detectó la presencia de animales PIT, considerándose una prevalencia predial de 65%. Hubo diferencias significativas (p The presence of immunotolerant, persistently infected (IPI animals with bovine viral diarrhoea virus (BVDV was detected in several milking herds with abortions, repeat breeders, still births and poor growth rates, by the virus isolation test and measurement of serum neutralising antibodies, in 2 blood samples from each animal. Samples were taken 3 month apart. Virus isolation was done by inoculation of the plasma and lymphocytes from peripheral blood in primary bovine testicle cell culture. The isolates were identified, on the second serial passage, by the indirect immunoperoxidase test. Cows were considered IPI when BVDV was isolated in the two blood samples and serum title neutralising antibodies were

  15. Development and Characterization of A Multiplexed RT-PCR Species Specific Assay for Bovine and one for Porcine Foot-and-Mouth Disease Virus Rule-Out

    Energy Technology Data Exchange (ETDEWEB)

    Smith, S M; Danganan, L; Tammero, L; Vitalis, B; Lenhoff, R; Naraghi-arani, P; Hindson, B

    2007-08-06

    Lawrence Livermore National Laboratory (LLNL), in collaboration with the Department of Homeland Security (DHS) and the United States Department of Agriculture (USDA), Animal and Plant Health Inspection Services (APHIS) has developed candidate multiplexed assays that may potentially be used within the National Animal Health Laboratory Network (NAHLN), the National Veterinary Services Laboratory (Ames, Iowa) and the Plum Island Animal Disease Center (PIADC). This effort has the ability to improve our nation's capability to discriminate between foreign animal diseases and those that are endemic using a single assay, thereby increasing our ability to protect food and agricultural resources with a diagnostic test which could enhance the nation's capabilities for early detection of a foreign animal disease. In FY2005 with funding from the DHS, LLNL developed the first version (Version 1.0) of a multiplexed (MUX) nucleic-acid-based RT-PCR assay that included signatures for foot-and-mouth disease virus (FMDV) detection with rule-out tests for two other foreign animal diseases (FADs) of swine, Vesicular Exanthema of Swine (VESV) and Swine Vesicular Disease Virus (SVDV), and four other domestic viral diseases Bovine Viral Diarrhea Virus (BVDV), Bovine Herpes Virus 1 (BHV-1), Bluetongue virus (BTV) and Parapox virus complex (which includes Bovine Papular Stomatitis Virus [BPSV], Orf of sheep, and Pseudocowpox). In FY06, LLNL has developed Bovine and Porcine species-specific panel which included existing signatures from Version 1.0 panel as well as new signatures. The MUX RT-PCR porcine assay for detection of FMDV includes the FADs, VESV and SVD in addition to vesicular stomatitis virus (VSV) and porcine reproductive and respiratory syndrome (PRRS). LLNL has also developed a MUX RT-PCR bovine assay for detection of FMDV with rule out tests for the two bovine FADs malignant catarrhal fever (MCF), rinderpest virus (RPV) and the domestic diseases vesicular stomatitis

  16. [Parasitic diarrhea in children].

    Science.gov (United States)

    Gendrel, D

    2003-12-01

    In many areas, Entamoeba histolytica and Entamoeba dispar are found together and their microscopic appearance is identical. Biochemical tests which can show cell wall differences are often falsely negative and the only possible way is to treat with metronidazole when amoebiasis is suspected. In case of clinical failure of metronidazole, a bacterial diarrhea is frequently found. Giardia is an other protozoa frequently found in stools of children in endemic areas. Diarrheas due to Giardia are possible in normal children and frequent in malnourished. They can determine severe atrophy of jejunal mucosa and must be treated. Cryptoridiosis is frequently asymptomatic but induces diarrhea in malnourished children. Diarrhea due to helminths is rare and only Strongyloides stercoralis induces severe diarrhea in malnourished child and must be treated in emergency with Ivermectin to avoid dissemination. In immune deficiency induced by corticosteroid treatment or cancer chemotherapy, a prophylactic treatment with Ivermectin against Strongyloides stercoralis must be given in endemic areas or after return, and probably also with metronidazole against Giardia.

  17. Management of chronic diarrhea in HIV-infected patients: current treatment options, challenges and future directions

    Directory of Open Access Journals (Sweden)

    Lidia Elfstrand

    2010-11-01

    Full Text Available Lidia Elfstrand, Claes-Henrik FlorénDepartment of Medicine, Division of Clinical Sciences, Skåne University Hospital, Lund University, Lund, SwedenAbstract: Diarrhea is a common clinical manifestation of HIV infection regardless of whether the patients have AIDS. HIV and malnutrition tend to occur in the same populations, the underprivileged and resource-poor. Malnutrition increases severity and mortality of infection. Occurrence of chronic diarrhea in HIV-infected patients, gut status and pathogenic agents, nutritional status and the crucial role of nutrition are reviewed. Bovine colostrum-based food can be useful for managing chronic diarrhea in HIV-infected patients, enhancing both nutritional and immunological status.Keywords: HIV, diarrhea, nutrition, bovine colostrum, CD4+ 

  18. Traveler’s diarrhea diet

    Science.gov (United States)

    ... risk for getting traveler's diarrhea by avoiding water, ice, and food that may be contaminated. The goal of the traveler's diarrhea diet is to make your symptoms better and prevent you from getting ...

  19. Human and bovine viruses in the Milwaukee River watershed: Hydrologically relevant representation and relations with environmental variables

    Energy Technology Data Exchange (ETDEWEB)

    Corsi, S.R., E-mail: srcorsi@usgs.gov [U.S. Geological Survey, Wisconsin Water Science Center, Middleton, WI 53562 (United States); Borchardt, M.A.; Spencer, S.K. [U.S. Department of Agriculture, Agricultural Research Service, 2615 Yellowstone Dr., Marshfield, WI 54449 (United States); Hughes, P.E.; Baldwin, A.K. [U.S. Geological Survey, Wisconsin Water Science Center, Middleton, WI 53562 (United States)

    2014-08-15

    To examine the occurrence, hydrologic variability, and seasonal variability of human and bovine viruses in surface water, three stream locations were monitored in the Milwaukee River watershed in Wisconsin, USA, from February 2007 through June 2008. Monitoring sites included an urban subwatershed, a rural subwatershed, and the Milwaukee River at the mouth. To collect samples that characterize variability throughout changing hydrologic periods, a process control system was developed for unattended, large-volume (56–2800 L) filtration over extended durations. This system provided flow-weighted mean concentrations during runoff and extended (24-h) low-flow periods. Human viruses and bovine viruses were detected by real-time qPCR in 49% and 41% of samples (n = 63), respectively. All human viruses analyzed were detected at least once including adenovirus (40% of samples), GI norovirus (10%), enterovirus (8%), rotavirus (6%), GII norovirus (1.6%) and hepatitis A virus (1.6%). Three of seven bovine viruses analyzed were detected including bovine polyomavirus (32%), bovine rotavirus (19%), and bovine viral diarrhea virus type 1 (5%). Human viruses were present in 63% of runoff samples resulting from precipitation and snowmelt, and 20% of low-flow samples. Maximum human virus concentrations exceeded 300 genomic copies/L. Bovine viruses were present in 46% of runoff samples resulting from precipitation and snowmelt and 14% of low-flow samples. The maximum bovine virus concentration was 11 genomic copies/L. Statistical modeling indicated that stream flow, precipitation, and season explained the variability of human viruses in the watershed, and hydrologic condition (runoff event or low-flow) and season explained the variability of the sum of human and bovine viruses; however, no model was identified that could explain the variability of bovine viruses alone. Understanding the factors that affect virus fate and transport in rivers will aid watershed management for minimizing

  20. Viral Hepatitis

    Science.gov (United States)

    ... Public Home » For Veterans and the Public Viral Hepatitis Menu Menu Viral Hepatitis Viral Hepatitis Home For ... the Public Veterans and Public Home How is Hepatitis C Treated? Find the facts about the newest ...

  1. Chronic Diarrhea in Children

    Science.gov (United States)

    ... All Content Irritable Bowel Syndrome (IBS) in Children Lactose Intolerance Ménétrier’s Disease Microscopic Colitis Ostomy Surgery of the ... and Diseases Diarrhea Celiac Disease IBS in Children Lactose Intolerance Related Diagnostic Tests Colonoscopy Flexible Sigmoidoscopy Upper GI ...

  2. Diarrhea and Swimming

    Science.gov (United States)

    ... if they swallow the water. If you have diarrhea, the most important thing you can do is to drink plenty of fluids to prevent dehydration. This is especially important for young children, pregnant women, and persons with weakened immune systems (such as those living with HIV/AIDS, ...

  3. Alimentary and respiratory tract lesions in eight medically fragile Holstein cattle with bovine leukocyte adhesion deficiency (BLAD).

    Science.gov (United States)

    Ackermann, M R; Kehrli, M E; Laufer, J A; Nusz, L T

    1996-05-01

    Lesions in the alimentary tract were studied in eight medically fragile Holstein cattle homozygous for the bovine leukocyte adhesion deficiency (BLAD) allele as determined by polymerase chain reaction and restriction endonuclease analysis. These cattle received institutional medical care but died or were euthanatized because of chronic debilitation associated with diarrhea (6/8) and pneumonia (4/8). The six cattle with diarrhea had acute (n = 3) or chronic (n = 3) intestinal ulcers, but the other two remained relatively healthy for 3 years and did not develop intestinal tract ulcers. Ulcerated areas were present in the small intestine in six animals, and two of these also had ulcers in the large intestine. Ulcers were covered by thick exudates that, in chronic lesions, partially occluded the intestinal lumen. Intramural and serosal fibrosis also contributed to lumen constriction. Pseudomonas aeruginosa was isolated from the intestine of four cattle. Bovine viral disease virus and Salmonella were not isolated from the five cattle that were tested. Respiratory tract lesions consisted of dense infiltrates of neutrophils in bronchi, bronchioles, and alveoli. This study suggests that intestinal lesions are integral to the demise of BLAD cattle that receive intensive medical care and that neutrophils do infiltrate the lung and enter airway lumina, despite the adhesion deficiency.

  4. 梅花鹿牛病毒性腹泻病毒的分离及其RT-PCR鉴定%Isolation and RT-PCR Identification of Bovine Viral Diarrhea Virus from China Sike Deer

    Institute of Scientific and Technical Information of China (English)

    郜玉钢; 郑全; 杜锐; 王全凯; 王树志

    2004-01-01

    对梅花鹿的牛病毒性腹泻病毒(BVDV)分离培养及RT-PCR进行了研究.从腹泻鹿粪样中分离到了与BVDV的C24V标准株致细胞病变相同规律的BVDV病毒株,经RT-PCR扩增进一步证实其为牛病毒性腹泻病毒.

  5. Serological Survey of Bovine Viral Diarrhea/Mucosal Disease in the Partial Regions of Chongqing%重庆市部分地区牛病毒性腹泻/粘膜病血清流行病学调查

    Institute of Scientific and Technical Information of China (English)

    杨泽林; 冉智光; 曾政; 黄诚; 熊仲良; 苏承忠; 郭利敏; 米自由

    2009-01-01

    为了解牛病毒性腹泻/粘膜病(BVD/MD)的流行情况,从重庆市辖区内11个区县(自治县)采集奶牛、黄牛、水牛血清共369头份,用酶联免疫吸附试验(ELISA)检测BVD/MD抗体.结果从9个区县(自治县)检测出阳性样品,阳性率介于8.33%~50.00%之间,总阳性率为22.49%;规模奶牛场、散养奶牛、散养黄牛、散养水牛的阳性率分别为42.16%、9.62%、21.31%、9.76%.提示该病在我市各种牛群中存在,污染较广,应引起重视.

  6. A cross-sectional study to estimate the frequency of anti-bovine viral diarrhea virus-1 antibodies in domestic pigs of Mossoró region in the state of Rio Grande do Norte, Brazil

    Directory of Open Access Journals (Sweden)

    Igor Renan Honorato Gatto

    2016-01-01

    Full Text Available ABSTRACT: This study investigated the occurrence of antibodies for BVDV-1 in swine herds located in the region of Mossoró city of the state of Rio Grande do Norte, Brazil. A sample size of 412 animals was estimated assuming unknown prevalence (set at 50%. Virus neutralization assay was used to the detect the presence of antibodies for BVDV-1 and the results found were analysed using multivariable logistic regression model. The obtained prevalence was 4% at animal level and 45% at the animal and herd level. The titers were highly variable between animals and within farms. The multivariable logistic regression analysis showed an association between being housed outside and exposure to BVDV-1 (OR=0.24, 95% CI:0.06, 0.96, P=0.04. Highly correlated data and low prevalence of antibodies at the animal level resulted in insufficient power to detect significant differences with other selected risk factors. In conclusion, the prevalence is within the range reported for other countries.

  7. Bovine viral diarrhea virus infection on the immune effect of classical swine fever%牛病毒性腹泻病毒感染对猪瘟免疫的影响

    Institute of Scientific and Technical Information of China (English)

    张慧英

    2010-01-01

    猪瘟病毒(CSFV)与同属的牛病毒性腹泻病痛毒(BVDV)同源性较高,抗原性上有交叉.本次调查对368份猪瘟免疫猪血清样本进行BVDV抗原检测,其中7份呈阳性,阳性率1.90%.对7份BVDV阳性血清采用ELISA和IHA两种方法检测猪瘟(CSFV)抗体水平,抗体合格率偏低,两者的结果符合率为71%.研究表明:BVDV在一定程度上干扰了猪瘟疫苗的免疫效果,影响抗体水平.

  8. Research Progress of Bovine Viral Diarrhea Virus Antigen and Antibody Detection Methods%牛病毒性腹泻病毒抗原抗体检测方法研究进展

    Institute of Scientific and Technical Information of China (English)

    赵霞; 岳海宁; 马占绔; 李福虹

    2014-01-01

    随着现代分子生物学与免疫学技术的快速发展与应用,牛病毒性腹泻病毒新型抗原抗体检测方法不断发展和完善对近年来对牛病毒性腹泻病毒的RT-PCR、荧光定量RT-PCR、RT-LAMP、ELISA和胶体金技术等抗原抗体检测方法进行综述,以期为不同实验室寻找适合的诊断方法以及发展新型诊断方法提供参考,为我国牛病毒性腹泻病的综合防控提供合理的科学依据.

  9. 牛病毒性腹泻病毒单克隆抗体捕获ELISA方法的建立%Establishment of antigen-captured ELISA using monoclonal antibodies to detect bovine viral diarrhea virus

    Institute of Scientific and Technical Information of China (English)

    钟发刚; 程安春; 王新华; 邓宇; 郭燕

    2008-01-01

    以纯化的牛病毒性腹泻病毒(BVDV)作为抗原免疫BALB/C系小鼠,通过细胞融合和克隆筛选出稳定分泌BVDV抗体的杂交瘤细胞株,将杂交瘤细胞接种于BALB/C系小鼠的腹腔内诱生腹水,腹水单抗与BVDV均呈阳性反应.将5A9、2G3和5C2(分别结合不同的抗原结合位点)诱生的BVDV单抗纯化后等量混合包被酶标板,与鸡抗BVDV IgY(检测抗体)联合应用,建立BVDV抗原捕捉ELISA(AC-ELISA)方法.采用方阵滴定法确定单抗、鸡抗BVDV IgY的最适工作浓度,用阴性组织样品作为判定检测结果的D490临界值,最后以建立的AC-ELISA检测临床粪样棉拭子27份,结果表明:该方法敏感性、特异性高,与全血病毒分离结果的符合率迭86.36%;与全血和粪样RT-PCR检测结果的符合率分别为89.47%和94.74%.阴性对照RT-PCR、病毒分离和AC-ELISA检测均为阴性.

  10. 牛病毒性腹泻病毒不同检测方法的比较研究%Comparative Study of Different Assays for Detection of Bovine Viral Diarrhea Virus

    Institute of Scientific and Technical Information of China (English)

    季新成; 曾新强; 员丽娟; 牛国辉; 于学辉; 段晓东; 王大孝

    2009-01-01

    采用Idexx公司抗原捕获ELISA试剂盒对新疆部分地区160份牛血清进行了牛病毒性腹泻病毒抗原检测,结果有6份样品为阳性,对该6份血清用BVDV阴性血清进行了系列稀释后,分别用抗原捕获ELISA、RT-PCR和荧光RT-PCR进行检测,发现RT-PCR方法比ELISA检测灵敏度高10-100倍,荧光RT-PCR比RT-PCR灵敏度高约10-100倍.

  11. 应用酶联免疫吸附试验检测牛病毒性腹泻-黏膜病病毒%Application of ELISA for the detection of bovine viral diarrhea (mucosal disease) virus

    Institute of Scientific and Technical Information of China (English)

    罗长保; 林志雄; 鱼海琼; 刘琳琳; 陈茹

    2002-01-01

    @@ 牛病毒性腹泻--黏膜病(BVD-MD)是由牛病毒性腹泻病毒(BVDV)引起牛的以黏膜发炎、糜烂、坏死和腹泻为特征的疾病.本病呈全球性分布,各养牛业发达国家均有流行.在自然条件下,可感染家养和野生的反刍兽,主要侵害6~18月龄的幼牛,患牛表现为发病急,体温突然升高至40~42℃,食欲废绝,消化道黏膜损伤严重,最初常见水样腹泻,后期便中带血和黏膜,病牛的死亡率可高达95%.怀孕母牛感染后可造成流产、早产或死胎.由于本病的发病特点及其危害,我国从国外引进牛、羊等反刍动物都必须对该病进行检疫.

  12. Development of RT-PCR assay for rapidly detecting of bovine viral diarrhea virus%RT-PCR快速检测牛病毒性腹泻病毒方法的建立与应用

    Institute of Scientific and Technical Information of China (English)

    张俭伟; 赵宏坤; 杨少华; 高运东; 王长法; 仲跻峰

    2008-01-01

    根据已发表的牛病毒性腹泻病毒(BVDV)5'-UTR序列,设计1对特异性引物,建立了快速检测BVDV的RT-PCR方法.利用该方法能从BVDV中扩增出196 bp特异性目的片段.应用此方法检测了26份疑似病例临床样品,14份为阳性.试验表明,该方法特异性强、敏感性好,能检测到1 pg的病毒RNA,是一种快速准确的检测方法.

  13. Development of an antigen-capture ELISA for detecting bovine viral diarrhea virus%牛病毒性腹泻病毒(BVDV)抗原捕获ELISA检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    张丽颖; 刘畅; 鲍永华; 赵志辉

    2009-01-01

    用牛病毒性腹泻病毒(BVDV)单克隆抗体包被酶标板,以兔多克隆抗体作为夹心抗体,建立BVDV抗原捕获ELISA(AC-ELISA)检测方法,优化反应条件并对该方法的稳定性等指标进行了测试和评价.结果表明,单抗最佳包被质量浓度为5μg/mL,兔多抗血清最佳质量浓度为10μg/mL;单抗在4℃包被12~24 h,多抗在37℃作用1 h为双抗体的最佳反应条件;酶标抗体最适稀释度1∶10000,最适作用时间为1 h;采用1%BSA和1%明胶分别在抗体包被后和加入待检抗原反应后进行两次封闭效果好.用AC-ELISA方法检测临床采集的11份牛腹泻病料和12份健康牛组织样品,同时以病毒分离和RT-PCR检测方法做对比,3种方法符合率很高.研究表明AC-ELISA方法稳定性好,可用于BVDV的临床快速检测.

  14. Establishment of double RT-PCR methodfor both bovine viral diarrhea virusand classical swine fever virus detection%牛病毒性腹泻病毒和猪瘟病毒双重RT-PCR方法的建立

    Institute of Scientific and Technical Information of China (English)

    魏春华; 戴爱玲; 刘建奎; 李晓华; 杨小燕

    2013-01-01

    参照GenBank上已发表的牛病毒性腹泻病毒(BVDV)和猪瘟病毒(CSFV)的全基因序列,针对瘟病毒高度保守的5'-UTR设计3条引物,特异性扩增BVDV、CSFV.经过条件优化后,建立了快速鉴别BVDV和CSFV的双重RT-PCR诊断方法,扩增两种病毒的片段,大小分别为260、200 bp.试验证明,所建立的方法具有良好的特异性和敏感性,利用双重RT-PCR对临床上60份疑似病料进行检测,双重PCR检测的结果与单重PCR检测结果总体符合率为100%,可用于临床病料检测,为防止猪瘟细胞苗的污染及进行CSFV和BVDV的鉴别诊断提供了有效方法.

  15. Establishment of Real-time Flourescent Quantitative PCR for Detection of Bovine Viral Diarrhea Virus%牛病毒性腹泻病毒实时荧光定量RT-PCR检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    范晴; 谢芝勋; 刘加波; 庞耀珊; 邓显文; 谢志勤; 谢丽基; 彭宜

    2010-01-01

    根据GenBank公布的牛病毒性腹泻病毒(BVDV)5′端非编码区核苷酸序列,设计引物和TaqMan荧光探针,建立了BVDV的实时荧光定量PCR检测方法.建立的方法只能检测到BVDV,而与CSFV、BRV、MT及IBRV没有交叉反应,具有高度的特异性,在1010~102模板范围内具有良好的线性关系,所制作的标准曲线相关系数为 0.991,最低可检测到100个拷贝的阳性质粒.所建立的BVDV实时荧光定量PCR检测方法具有快速、特异、灵敏等特点.

  16. 牛病毒性腹泻病毒抗原捕获ELISA检测方法的标准化研究%Development of Ag-capture ELISA for detecting bovine viral diarrhea virus

    Institute of Scientific and Technical Information of China (English)

    邓宇; 王新华; 郭燕; 钟发刚; 沈敏

    2007-01-01

    用纯化牛病毒性腹泻病毒免疫蛋鸡制备出的卵黄抗体作为包被抗体,采用自制的单抗为一抗,建立牛病毒性腹泻病毒抗原捕获ELISA方法.通过试验确定,抗牛病毒性腹泻病毒卵黄抗体最佳包被浓度为1:50;McAb最适稀释浓度为1:10,HRP-羊抗鼠IgG工作浓度为1:800.通过引入牛病毒性腹泻病毒质控血清进行质控检验.该方法所得检测结果均在质量控制范围内,达到预定标准化要求.标准化的抗原捕获ELISA方法具有特异、灵敏、可靠、方便、快捷等特点,可广泛应用推广,为我国牛病毒性腹泻病毒监测提供了行之有效的技术手段.

  17. 牛病毒性腹泻病毒双抗体夹心ELISA检测方法的建立%Detection of bovine viral diarrhea virus by sandwich ELISA

    Institute of Scientific and Technical Information of China (English)

    高存福; 秦建华; 赵月兰; 包永占; 张宁

    2005-01-01

    将牛病毒性腹泻病毒超免疫血清以常规方法提取IgG,采用过碘酸钠法标记辣根过氧化物酶(HRP),建立了从粪样中检测牛病毒性腹泻病毒抗原的双抗体夹心ELISA.结果,抗体的最佳包被量为150μg/mL,酶标抗体最适工作浓度为1∶200;封闭液为50mL/L的兔血清;待检粪样及酶标抗体的感作时间为37℃ 120min;底物显色时间为室温15min.应用建立的检测方法对河北省8个大中型奶牛场298份乳牛腹泻粪样进行了检测,结果,阳性检出率为42.6%.

  18. Establishment of Loop-Mediated Isothermal Amplification for Detection of Bovine Viral Diarrhea Virus%牛病毒性腹泻病毒RT-LAMP检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    范晴; 谢芝勋; 刘加波; 庞耀珊; 邓显文; 谢志勤; 谢丽基; 彭宜

    2010-01-01

    目的:建立牛病毒性腹泻病毒(BVDv)的环介导体外等温扩增(LAMP)快速检测方法.方法:根据BVDv的5'端非编码区序列,在保守区的8个位点设计LAMP特异性引物(2对特异性引物和1对环引物),对反应条件和试剂浓度进行优化,建立恒温(63.5℃)、快速(65 min)的检测方法.结果:建立的方法特异性好,检测其他对照病毒均为阴性;灵敏度高,最低可检测到1个拷贝的阳性质粒,可通过观察浑浊度或加入染料后直接判定扩增结果.结论:建立了用于检测BVDv的LAMP方法,该方法简便、快速、特异性好、灵敏度高,适合基层和现场检测.

  19. Eukaryotic expression of Erns gene of bovine viral diarrhea virus and antigenic detection of the expressed protein%牛病毒性腹泻病毒Erns基因的真核表达及抗原性检测

    Institute of Scientific and Technical Information of China (English)

    吴立君; 师东方; 王丹娜; 何海娟; 吴明福; 王君伟

    2006-01-01

    为研究牛病毒性腹泻病毒(BVDV)Erns基因的生物学功能,将含有牛病毒性腹泻病毒Erns基因的质粒pMD18-T-Erns经BamH Ⅰ/HindⅢ双酶切,获得了Erns片段,再与杆状病毒转移载体pBlueBacHis2A连接,构建成重组质粒.将重组质粒pBlueBacHis2A-Erns与Bac-N-BlueTMDNA共转染至sf9昆虫细胞中,获得了重组病毒,经噬斑筛选纯化,感染sf9昆虫细胞进行表达.SDS-PAGE分析结果表明,表达的目的蛋白大小约30 ku;Western-blotting检测表明,该蛋白具有良好的抗原性.

  20. 青海牦牛BVDV E0基因的表达与抗原性检测%Prokaryotic expression and detection of antigencity of E0 protein of bovine viral diarrhea virus from yak in China

    Institute of Scientific and Technical Information of China (English)

    王光华; 叶成玉; 蔡其刚; 王戈平; 陆艳; 张芳芳; 马利青; 周继章

    2013-01-01

    将青海牦牛病毒性腹泻病毒(BVDV) QHZK株的E0基因亚克隆人原核表达载体pET-32(α),构建了重组表达载体pET-32 (α)-E0,然后用重组质粒转化Rosetta(DE3)感受态细胞,并利用IPTG诱导蛋白表达.表达的蛋白用His-Band镍柱进行亲合层析纯化,Western-blot鉴定表达蛋白.结果显示,E0基因可在大肠杆菌中获得表达,表达产物的分子质量约为44ku,与预计的蛋白分子质量大小一致.Western-blot分析表明,该蛋白可以与BVDV标准阳性血清产生特异性结合反应,具有良好的抗原性,为BVDV亚单位疫苗的研究奠定了基础.

  1. 新疆地区牛病毒性腹泻病毒血清抗体检测%Detection of Serum Antibody of Bovine Viral Diarrhea in Xinjiang

    Institute of Scientific and Technical Information of China (English)

    刘晓娜; 刘晓妮

    2013-01-01

    本次研究主要是采用酶联免疫吸附试验(ELISA)检测试剂盒对采自新疆不同地区的800份血清进行了牛病毒性腹泻病毒(BVDV)血清抗体检测,共检出抗体阳性血清694份,最高阳性率为99%,最低为55%,平均为86.6%,通过数据来看,近几年有过流产史的牛血清BVDV抗体阳性率普遍偏高.感染牛没有明显的年龄差别.该病已在新疆地区普遍存在,且又不断增强的趋势.

  2. Establishment and Application of Nested PCR Assay for Bovine Viral Diarrhea Virus Detection%牛病毒性腹泻病毒巢式 RT-PCR 检测方法的建立及应用

    Institute of Scientific and Technical Information of China (English)

    韩猛立; 李娜; 黄新; 薄新文; 王新华; 赵玉龙; 钟发刚

    2010-01-01

    参照多株不同基因型和基因亚型牛病毒性腹泻病毒(BVDV)5'UTR 相对保守的基因序列,设计 3 对引物,其中1对能够扩增不同基因型 BVDV 5'UTR 片段(288 bp),另2对分别能够扩增 BVDV-1 型和 BVDV-2 型 5'UTR 片段(195 bp和116 bp).扩增片段大小与预计片段一致.对 PCR 反应条件进行优化,建立了一种特异、敏感的 RT-nPCR 检测方法.敏感性试验表明,敏感性为10-2TCID50/mL .特异性试验表明,对猪瘟兔化弱毒疫苗株、牛轮状病毒、牛传染性鼻气管炎病毒的检测结果均为阴性.通过对新疆部分地区无BVD 临床症状的牛场 208 份奶牛血样检测,阳性率为 50.43% ,表明该方法具有快速、简便、特异性强和灵敏度高等特点,可以作为 BVDV 隐性感染临床诊断的有效方法.

  3. Prokaryotic expression of E1 genes of bovine viral diarrhea virus and detection of the expressed protein%牦牛病毒性腹泻病毒E1基因的原核表达

    Institute of Scientific and Technical Information of China (English)

    王盼盼; 刘亚刚; 孙凯; 王文伯; 胡炳峰; 于吉锋; 冯英阳

    2010-01-01

    以牦牛BVDV基因组DNA为模板, 运用聚合酶链反应成功扩增出牦牛BVDV E1基因, 将其插入到pET-28a及pET-32a原核表达载体, 构建重组表达质粒pET-28a-E1和pET-32a-E1并分别转化至Rosetta(DE3)和BL21(DE3)宿主菌中, IPTG诱导表达, 经SDS-PAGE检测, pET-28a-E1和pET-32a-E1在宿主菌Rosetta(DE3)和BL21(DE3)中均表达出约40KU的融合蛋白, 结果分析表明E1基因在两个宿主中都获得了高效表达, 表达出的蛋白大小与预期结果相符.

  4. Detection Methods of Bovine Rotavirus and Viral Diarrhea Virus by Duplex RT-PCR%牛轮状病毒与病毒性腹泻病毒的双重RT-PCR检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    张俭伟; 赵宏坤; 杨少华; 高运东; 王长法; 仲跻峰

    2008-01-01

    建立了能够同时检测牛轮状病毒(BRV)与牛病毒性腹泻病毒(BVDV)的双重RT-PCR方法.应用两对特异性引物进行了双重RT-PCR扩增,这两对引物分别对应于BRV的VP7基因和BVDV的5-UTR中的部分编码序列,其扩增产物分别为342bp和196bp.说明该方法的特异性强、敏感性高,可检测到1pg的病毒RNA,可应用于临床诊断和流行病学研究.

  5. Establishment and application of RT-PCR for detection of bovine viral diarrhea virus%牛病毒性腹泻病毒RT-PCR检测方法的建立及应用

    Institute of Scientific and Technical Information of China (English)

    孟雨; 吴发兴; 朱紫祥; 张志; 张燕霞; 刘爽; 李晓成; 王晶钰

    2010-01-01

    根据GenBank中登录的牛病毒性腹泻病毒(BVDV)5'UTR基因序列,设计合成了1对特异性引物,建立了检测BVDV 218 bp片段的RT-PCR方法.通过对该方法的特异性、敏感性和重复性进行试验,结果显示,该方法从BVDV标准毒株Oregon C_(24)V中扩增出了218 bp的特异性片段,而对猪瘟病毒、蓝舌病病毒、牛传染性鼻气管炎病毒、猪繁殖与呼吸综合征病毒、猪伪狂犬病病毒、MDBK正常细胞的扩增结果均为阴性.经对标准毒株的细胞毒进行检测,其敏感度达10~(-1) TCID_(50).不同人员用该方法进行检测,结果均一致,表明其重复性好.应用该方法对70份临床疑似发病猪样品进行检测,结果检出11份阳性,阳性检出率约为15.7%.在对5份细胞培养用犊牛血清的检测中,亦检出2份阳性.表明,建立的RT-PCR方法具有特异、灵敏、高效、快速的特点,可用于BVDV的临床检测及流行病学监测.

  6. RT-PCR amplification of bovine viral diarrhea virus E0 gene from sika deer%鹿源牛病毒性腹泻病毒E0基因的RT-PCR扩增

    Institute of Scientific and Technical Information of China (English)

    郜玉钢; 李璠瑛; 刘佳佳; 杜锐; 臧埔

    2009-01-01

    用RT-PCR方法,成功扩增了鹿源牛病毒性腹泻病毒保护性抗原E0基因.该RT-PCR最佳反应条件分别为退火温度为55℃、25 mmol/μL[番茄花园1] MgCl2(4 μL)、10 pmol/μL上下游引物(2 μL)、5 U/μL Taq酶(1μL).

  7. Viral marketing

    OpenAIRE

    Král, Jiří

    2015-01-01

    Bachelor's Thesis deals with effective promotional tools called viral marketing. The main contribution of the thesis is the definition and history of viral marketing, making analysis of process of viral marketing, progresses definition and rules for creating a viral campaign. And also aspects are necessary for a successful viral spread. There are analysis of the characteristics of social media which are dividing according to the orientation and marketing tactics. Thesis is especially about so...

  8. Epidemiology of traveler's diarrhea.

    Science.gov (United States)

    Castelli, F; Carosi, G

    1995-01-01

    Annually, over 75 million international passengers travel to tropical areas, more than 20 million of whom come from industrialized countries. They experience a high rate of traveler's diarrhea (TD), varying from 20 to 56%, which may result in serious limitations to their activities. The cause of TD is considered to be infectious in the overwhelming majority of cases and, apart from differences in relative importance, the list of responsible microbial agents is fairly constant regardless of geographic origin. The ingestion of contaminated food or water is considered to be the principal mode of transmission of the enteric pathogens of TD. Several factors have been proposed as playing a role in the etiogenesis of diarrhea in travelers, including personal (age, socioeconomic status, body weight, preexisting gastrointestinal illnesses), behavioral (mode of travel, standard of accommodation, eating in public places, dietary errors) and travel-related (destination, duration of stay, country of origin, season) factors, which are reviewed in detail.

  9. Risk factors against bovine respiratory diseade in suckling calves from Argentina

    OpenAIRE

    García-Bocanegra, Ignacio; Arenas-Montes, Antonio José; Perea-Remujo, J.A.; Arenas-Casas, Antonio; Torralbo, A.; Borge-Rodríguez, Carmen; Carbonero-Martínez, Alfonso; Maldonado García, Alfonso

    2011-01-01

    An observacional cross-sectional study was performed to determine the risk factors associated to the main viral agents of the bovine respiratory disease: bovine herpesvirus type 1 (HVB1), bovine viral diarrhoea virus (VDVB), bovine respiratory syncytial virus (VRSB) and parainfluenza 3 virus (VPI3). Blood samples from dairy calves in the provinces of Cordova and Santa Fe (Argentina) were obtained, and an epidemiological ques-tionnaire was filled. Antibodies against studied viruses were detect...

  10. Nosocomial diarrhea in children:is astrovirus the leading pathogen?

    Institute of Scientific and Technical Information of China (English)

    Anita Chakravarti; Manisha Jain; Mayank Singh Chauhan; Anju Sharma; Sayani Tewari

    2010-01-01

    Objective:To screen for the presence of mixed infection with rotavirus. Methods:The present study included 140 children aged less than 2 years with acute diarrhea. Fecal samples of all these patients were analyzed for the presence of astroviral antigen by enzyme immunoassay. Also 40 rotavirus positive fecal samples were screened for the presence of astrovirus. Results:In case of acute diarrhea in children the prevalence of astrovirus was around 34%(48/140). It was seen that even in rotavirus positive cases astrovirus co infection was 25%. Conclusions:Astrovirus is a growing problem which is often underrecognised. With the rotavirus vaccine licensure being imminent astrovirus will emerge out as the single most important cause of viral diarrhea.

  11. Viral encephalitis

    Directory of Open Access Journals (Sweden)

    Marcus Tulius T Silva

    2013-09-01

    Full Text Available While systemic viral infections are exceptionally common, symptomatic viral infections of the brain parenchyma itself are very rare, but a serious neurologic condition. It is estimated that viral encephalitis occurs at a rate of 1.4 cases per 100.000 inhabitants. Geography is a major determinant of encephalitis caused by vector-borne pathogens. A diagnosis of viral encephalitis could be a challenge to the clinician, since almost 70% of viral encephalitis cases are left without an etiologic agent identified. In this review, the most common viral encephalitis will be discussed, with focus on ecology, diagnosis, and clinical management.

  12. Immunological Response to Single Pathogen Challenge with Agents of the Bovine Respiratory Disease Complex: An RNA-Sequence Analysis of the Bronchial Lymph Node Transcriptome.

    Directory of Open Access Journals (Sweden)

    Polyana C Tizioto

    Full Text Available Susceptibility to bovine respiratory disease (BRD is multi-factorial and is influenced by stress in conjunction with infection by both bacterial and viral pathogens. While vaccination is broadly used in an effort to prevent BRD, it is far from being fully protective and cases diagnosed from a combination of observed clinical signs without any attempt at identifying the causal pathogens are usually treated with antibiotics. Dairy and beef cattle losses from BRD are profound worldwide and genetic studies have now been initiated to elucidate host loci which underlie susceptibility with the objective of enabling molecular breeding to reduce disease prevalence. In this study, we employed RNA sequencing to examine the bronchial lymph node transcriptomes of controls and beef cattle which had individually been experimentally challenged with bovine respiratory syncytial virus, infectious bovine rhinotracheitis, bovine viral diarrhea virus, Pasteurella multocida, Mannheimia haemolytica or Mycoplasma bovis to identify the genes that are involved in the bovine immune response to infection. We found that 142 differentially expressed genes were located in previously described quantitative trait locus regions associated with risk of BRD. Mutations affecting the expression or amino acid composition of these genes may affect disease susceptibility and could be incorporated into molecular breeding programs. Genes involved in innate immunity were generally found to be differentially expressed between the control and pathogen-challenged animals suggesting that variation in these genes may lead to a heritability of susceptibility that is pathogen independent. However, we also found pathogen-specific expression profiles which suggest that host genetic variation for BRD susceptibility is pathogen dependent.

  13. Nonreplicative RNA Recombination of an Animal Plus-Strand RNA Virus in the Absence of Efficient Translation of Viral Proteins

    Science.gov (United States)

    Kleine Büning, Maximiliane; Meyer, Denise; Austermann-Busch, Sophia; Roman-Sosa, Gleyder; Rümenapf, Tillmann

    2017-01-01

    RNA recombination is a major driving force for the evolution of RNA viruses and is significantly implicated in the adaptation of viruses to new hosts, changes of virulence, as well as in the emergence of new viruses including drug-resistant and escape mutants. However, the molecular details of recombination in animal RNA viruses are only poorly understood. In order to determine whether viral RNA recombination depends on translation of viral proteins, a nonreplicative recombination system was established which is based on cotransfection of cells with synthetic bovine viral diarrhea virus (family Flaviviridae) RNA genome fragments either lacking the internal ribosome entry site required for cap-independent translation or lacking almost the complete polyprotein coding region. The emergence of a number of recombinant viruses demonstrated that IRES-mediated translation of viral proteins is dispensable for efficient recombination and suggests that RNA recombination can occur in the absence of viral proteins. Analyses of 58 independently emerged viruses led to the detection of recombinant genomes with duplications, deletions and insertions in the 5′ terminal region of the open reading frame, leading to enlarged core fusion proteins detectable by Western blot analysis. This demonstrates a remarkable flexibility of the pestivirus core protein. Further experiments with capped and uncapped genome fragments containing a luciferase gene for monitoring the level of protein translation revealed that even a ∼1,000-fold enhancement of translation of viral proteins did not increase the frequency of RNA recombination. Taken together, this study highlights that nonreplicative RNA recombination does not require translation of viral proteins. PMID:28338950

  14. Development and Characterization of a Multiplexed RT-PCR Species Specific Assay for Bovine and one for Porcine Foot-and-Mouth Disease Virus Rule-Out Supplemental Materials

    Energy Technology Data Exchange (ETDEWEB)

    Smith, S; Danganan, L; Tammero, L; Lenhoff, R; Naraghi-arani, P; Hindson, B

    2007-08-06

    Lawrence Livermore National Laboratory (LLNL), in collaboration with the Department of Homeland Security (DHS) and the United States Department of Agriculture (USDA), Animal and Plant Health Inspection Services (APHIS) has developed advanced rapid diagnostics that may be used within the National Animal Health Laboratory Network (NAHLN), the National Veterinary Services Laboratory (Ames, Iowa) and the Plum Island Animal Disease Center (PIADC). This effort has the potential to improve our nation's ability to discriminate between foreign animal diseases and those that are endemic using a single assay, thereby increasing our ability to protect animal populations of high economic importance in the United States. Under 2005 DHS funding we have developed multiplexed (MUX) nucleic-acid-based PCR assays that combine foot-and-mouth disease virus (FMDV) detection with rule-out tests for two other foreign animal diseases Vesicular Exanthema of Swine (VESV) and Swine Vesicular Disease (SVD) and four other domestic viral diseases Bovine Viral Diarrhea Virus (BVDV), Bovine Herpes Virus 1 (BHV-1 or Infectious Bovine Rhinotracheitus IBR), Bluetongue virus (BTV) and Parapox virus complex (which includes Bovine Papular Stomatitis Virus BPSV, Orf of sheep, and Pseudocowpox). Under 2006 funding we have developed a Multiplexed PCR [MUX] porcine assay for detection of FMDV with rule out tests for VESV and SVD foreign animal diseases in addition to one other domestic vesicular animal disease vesicular stomatitis virus (VSV) and one domestic animal disease of swine porcine reproductive and respiratory syndrome (PRRS). We have also developed a MUX bovine assay for detection of FMDV with rule out tests for the two bovine foreign animal diseases malignant catarrhal fever (MCF), rinderpest virus (RPV) and the domestic diseases vesicular stomatitis virus (VSV), bovine viral diarrhea virus (BVDV), infectious bovine rhinotracheitus virus (BHV-1), bluetongue virus (BTV), and the Parapox

  15. Bovine coronavirus hemagglutinin protein.

    Science.gov (United States)

    King, B; Potts, B J; Brian, D A

    1985-02-01

    Treatment of purified bovine coronavirus (Mebus strain) with pronase destroyed the integrity of virion surface glycoproteins gp140, gp120, gp100, reduced the amount of gp26 and destroyed the hemagglutinating activity of the virus. Bromelain, on the other hand, destroyed the integrity of gp120, gp100 and gp26 but failed to remove gp140 and failed to destroy viral hemagglutinating activity. These experiments suggest that gp140 is the virion hemagglutinin. Immunoblotting studies using monospecific antiserum demonstrate that gp140 is a disulfide-linked dimeric structure reducible to monomers of 65 kDa.

  16. Viral marketing

    OpenAIRE

    BLÁHOVÁ, Adéla

    2012-01-01

    The aim of my thesis is to provide a comprehensive overview of the viral marketing and to analyze selected viral campaigns. There is a description of advantages and disadvantages of this marketing tool. In the end I suggest for which companies viral marketing is an appropriate form of the promotion.

  17. Diagnóstico virológico de Herpesvirus bovino tipo-1 (Virological diagnostic of Bovine herpesvirus type-1

    Directory of Open Access Journals (Sweden)

    Avila Sánchez, Mislay;

    2008-03-01

    such as: Infectious bovine rhinotracheitis/infectious pustular vulvovaginitis (IBR/IPV, complex bovine viral diarrhea/disease of the mucous and mamillitis, due to its similarity as for the clinical signs. The development reached by the cattle raising in the last years has stimulated the search of new diagnostic methods which guarantee a higher speed, specificity and sensitivity in the detection of the diseases affecting cattle. Among these diseases, those of viral origin are dangerous, since their rapid spread causes great economic damages. Bovine herpesvirus type-1 is IBR/IPV causal agent. The diagnostic of laboratory of this disease is carried out by the classical methods of the infectious agent detection such as the viral isolation and later identification by viral neutralization or immunohistochemistry. However, viral isolation requires cell culture and a lot of time to obtain the results. The Polymerase Chain Reaction (PCR is a method based on the amplification in vitro of the nucleic acid. It is more sensitive, rapid, specific, less laborious and more economic than nucleic acid hybridization and than the traditional detection methods of this infectious agent. PCR allows detecting live and inactivated viruses, carrier and latently infected animals and it facilitates a later molecular characterization of isolates from the nucleic acid amplified.

  18. Investigation of Diarrhea in AIDS

    Directory of Open Access Journals (Sweden)

    Klaus E Mönkemüller

    2000-01-01

    Full Text Available Chronic diarrhea is a common problem in patients with acquired immune deficiency syndrome (AIDS, resulting in significant morbidity and potential mortality. In the early stages of immunodeficiency, human immunodeficiency virus (HIV-infected patients are susceptible to infection with the same enteric pathogens that cause diarrhea in immunocompetent hosts, but with progressive immunodeficiency, these patients become susceptible to numerous opportunistic disorders. The main factor to consider when tailoring the work-up of diarrhea in the HIV-infected patient is the immune status, which is reflected by the total CD4 lymphocyte cell count. A CD4 count of less than 100 cells/µL is significantly correlated with opportunistic disorders. For the HIV-infected patient with diarrhea, repeated stool studies to investigate for bacteria, ova and parasites should be the first step. When either upper or lower gastrointestinal tract symptoms are present and stool studies are negative, endoscopy directed to the probable organ of involvement is appropriate. If localizing symptoms are absent, the most appropriate next test is sigmoidoscopy with biopsies. Not infrequently, despite extensive evaluation, the cause of diarrhea in patients with AIDS remains unexplained. Recently, the widespread use of highly active antiretroviral therapy, including protease inhibitors, has led to a change in the epidemiology of diarrhea in AIDS patients. As their immune status improves, HIV-infected patients treated with combination therapy become less prone to opportunistic disorders. However, diarrhea appears to be frequent because several antiretroviral agents can themselves cause diarrhea.

  19. Pharmacologic Agents for Chronic Diarrhea

    OpenAIRE

    Lee, Kwang Jae

    2015-01-01

    Chronic diarrhea is usually associated with a number of non-infectious causes. When definitive treatment is unavailable, symptomatic drug therapy is indicated. Pharmacologic agents for chronic diarrhea include loperamide, 5-hydroxytryptamine type 3 (5-HT3) receptor antagonists, diosmectite, cholestyramine, probiotics, antispasmodics, rifaximin, and anti-inflammatory agents. Loperamide, a synthetic opiate agonist, decreases peristaltic activity and inhibits secretion, resulting in the reductio...

  20. Estimation of nasal shedding and seroprevalence of organisms known to be associated with bovine respiratory disease in Australian live export cattle.

    Science.gov (United States)

    Moore, S Jo; O'Dea, Mark A; Perkins, Nigel; O'Hara, Amanda J

    2015-01-01

    The prevalence of organisms known to be associated with bovine respiratory disease (BRD) was investigated in cattle prior to export. A quantitative reverse transcription polymerase chain reaction assay was used to detect nucleic acids from the following viruses and bacteria in nasal swab samples: Bovine coronavirus (BoCV; Betacoronavirus 1), Bovine herpesvirus 1 (BoHV-1), Bovine viral diarrhea virus 1 (BVDV-1), Bovine respiratory syncytial virus (BRSV), Bovine parainfluenza virus 3 (BPIV-3), Histophilus somni, Mycoplasma bovis, Mannheimia haemolytica, and Pasteurella multocida. Between 2010 and 2012, nasal swabs were collected from 1,484 apparently healthy cattle destined for export to the Middle East and Russian Federation. In addition, whole blood samples from 334 animals were tested for antibodies to BoHV-1, BRSV, BVDV-1, and BPIV-3 using enzyme-linked immunosorbent assay. The nasal prevalence of BoCV at the individual animal level was 40.1%. The nasal and seroprevalence of BoHV-1, BRSV, BVDV-1, and BPIV-3 was 1.0% and 39%, 1.2% and 46%, 3.0% and 56%, and 1.4% and 87%, respectively. The nasal prevalence of H. somni, M. bovis, M. haemolytica, and P. multocida was 42%, 4.8%, 13.4%, and 26%, respectively. Significant differences in nasal and seroprevalence were detected between groups of animals from different geographical locations. The results of the current study provide baseline data on the prevalence of organisms associated with BRD in Australian live export cattle in the preassembly period. This data could be used to develop strategies for BRD prevention and control prior to loading.

  1. Full genome analysis of bovine astrovirus from fecal samples of cattle in Japan: identification of possible interspecies transmission of bovine astrovirus.

    Science.gov (United States)

    Nagai, Makoto; Omatsu, Tsutomu; Aoki, Hiroshi; Otomaru, Konosuke; Uto, Takehiko; Koizumi, Motoya; Minami-Fukuda, Fujiko; Takai, Hikaru; Murakami, Toshiaki; Masuda, Tsuneyuki; Yamasato, Hiroshi; Shiokawa, Mai; Tsuchiaka, Shinobu; Naoi, Yuki; Sano, Kaori; Okazaki, Sachiko; Katayama, Yukie; Oba, Mami; Furuya, Tetsuya; Shirai, Junsuke; Mizutani, Tetsuya

    2015-10-01

    A viral metagenomics approach was used to investigate fecal samples of Japanese calves with and without diarrhea. Of the different viral pathogens detected, read counts gave nearly complete astrovirus-related RNA sequences in 15 of the 146 fecal samples collected in three distinct areas (Hokkaido, Ishikawa, and Kagoshima Prefectures) between 2009 and 2015. Due to the lack of genetic information about bovine astroviruses (BoAstVs) in Japan, these sequences were analyzed in this study. Nine of the 15 Japanese BoAstVs were closely related to Chinese BoAstVs and clustered into a lineage (tentatively named lineage 1) in all phylogenetic trees. Three of 15 strains were phylogenetically separate from lineage 1, showing low sequence identities, and clustered instead with an American strain isolated from cattle with respiratory disease (tentatively named lineage 2). Interestingly, two of 15 strains clustered with lineage 1 in the open reading frame (ORF)1a and ORF1b regions, while they clustered with lineage 2 in the ORF2 region. Remarkably, one of 15 strains exhibited low amino acid sequence similarity to other BoAstVs and was clustered separately with porcine astrovirus type 5 in all trees, and ovine astrovirus in the ORF2 region, suggesting past interspecies transmission.

  2. How to Do in Persistent Diarrhea of Children?: Concepts and Treatments of Chronic Diarrhea

    OpenAIRE

    Lee, Kun Song; Kang, Dong Soo; Yu, JeeSuk; Chang, Young Pyo; Park, Woo Sung

    2012-01-01

    Chronic diarrhea is defined as passing watery stools that lasts for more than 2 weeks. Persistent diarrhea belongs to chronic diarrhea and is a chronic episode of diarrhea of infectious etiology. The etiology of chronic diarrhea is varied. It is important to consider the child's age and clinical manifestations with alarm signals for an application of proper treatments to children with chronic diarrhea. Vicious cycle is present in chronic diarrhea and nutritional rehabilitation can break the v...

  3. Management of the returning traveler with diarrhea.

    Science.gov (United States)

    de Saussure, Philippe P H

    2009-11-01

    Traveler's diarrhea (TD) strikes 20-60% of travelers visiting developing countries. It occurs shortly after the return and can be distinguished into two categories: acute and persistent TD. Acute TD, mostly caused by bacterial and viral pathogens, is usually mild and self-limited, and deserves empirical symptomatic and/or antibiotic therapy in selected cases. Fluoroquinolones are progressively superseded in this indication by azithromycin, a well tolerated macrolide active against most bacteria responsible for TD, including the quinolone-resistant species of Campylobacter jejuni that are now pervasive, especially in Southeast Asia and India. Persistent TD in the returning traveler is much rarer than its acute counterpart and may be associated with three types of causes. Persistent infections, among which Giardia and possibly Entamoeba predominate, account for a significant proportion of cases. Postinfectious processes represent a second cause and comprise temporary lactose malabsorption and postinfectious irritable bowel syndrome, now considered a major cause of persistent TD. Finally, apparently unrelated chronic diseases causing diarrhea are occasionally unmasked by TD and represent a third type of persistent TD, among which the well established case of incident inflammatory bowel disease poses intriguing pathogenesis questions. This review discusses recent advances in the field and provides practical recommendations for the management of TD in adult, immunocompetent returning travelers.

  4. Factors that explain excretion of enteric pathogens by persons without diarrhea.

    Science.gov (United States)

    Levine, Myron M; Robins-Browne, Roy M

    2012-12-01

    Excretion of enteropathogens by subjects without diarrhea influences our appreciation of the role of these pathogens as etiologic agents. Characteristics of the pathogens and host and environmental factors help explain asymptomatic excretion of diarrheal pathogens by persons without diarrhea. After causing acute diarrhea followed by clinical recovery, some enteropathogens are excreted asymptomatically for many weeks. Thus, in a prevalence survey of persons without diarrhea, some may be excreting pathogens from diarrheal episodes experienced many weeks earlier. Volunteer challenges with Vibrio cholerae O1, enterotoxigenic Escherichia coli (ETEC), enteropathogenic E. coli, Campylobacter jejuni, and Giardia lamblia document heterogeneity among enteropathogen strains, with some inexplicably not eliciting diarrhea. The immune host may not manifest diarrhea following ingestion of a pathogen but may nevertheless asymptomatically excrete. Some human genotypes render them less susceptible to symptomatic or severe diarrheal infection with certain pathogens such as Vibrio cholerae O1 and norovirus. Pathogens in stools of individuals without diarrhea may reflect recent ingestion of inocula too small to cause disease in otherwise susceptible hosts or of animal pathogens (eg, bovine or porcine ETEC) that do not cause human illness.

  5. Prediction of respiratory disease and diarrhea in veal calves based on immunoglobulin levels and the serostatus for respiratory pathogens measured at arrival.

    Science.gov (United States)

    Pardon, Bart; Alliët, Jeroen; Boone, Randy; Roelandt, Sophie; Valgaeren, Bonnie; Deprez, Piet

    2015-06-15

    Failure of passive transfer is a common problem in calves destined for veal production. At present it is unknown whether the risk for respiratory disease (BRD) or neonatal calf diarrhea (NCD) in the veal herd is associated with total immunoglobulin (Ig) and/or on the serostatus for respiratory pathogens measured at arrival. Therefore, the first objective of this prospective longitudinal cohort study was to determine associations between serum protein fractions as determined by routine electrophoresis (total protein, albumin, alpha-1 and -2 globulins, beta-globulins and Ig's) at arrival and BRD and NCD in the first 3 weeks of the production cycle. The second objective was to determine whether the serostatus (seropositive/seronegative) of seven respiratory pathogens (bovine respiratory syncytial virus (BRSV), parainfluenzavirus-3, bovine coronavirus (BCV), bovine herpesvirus-1, bovine viral diarrhea virus, Mannheimia haemolytica and Mycoplasma bovis) of these arrival serum samples could be associated with the risk of having BRD. The third objective was to determine which of the electrophoresis proteins and respiratory serostatuses were associated with average daily gain (ADG) in the study period. The study population consisted of 150 rosé veal calves housed in a single air-space. The study period ended at day 18 post arrival, when BRD incidence was judged to be too high to further postpone a group treatment. A Cox regression model was used to determine the effect of the studied protein fractions and antibodies on the time to BRD and NCD occurrence. The effect of the studied predictors on ADG was determined by linear regression. Calves with Ig levels under 7.5g/L had an increased BRD hazard (hazard ratio (HR)=1.9 (95% confidence interval (CI)=1.2-3.0)). NCD was only positively associated with the alpha-2 globulin concentration. Calves with a negative serostatus for BCV (HR=1.7 (95% CI=1.0-2.8)) or BRSV (HR=2.0 (95% CI=1.0-3.9)) had an increased BRD hazard. Average

  6. Bovine coronavirus antibody titers at weaning negatively correlate with incidence of bovine respiratory disease in the feed yard

    Science.gov (United States)

    Bovine respiratory disease complex (BRDC) is a multifactorial disease caused by complex interactions among viral and bacterial pathogens, stressful management practices and host genetic variability. Although vaccines and antibiotic treatments are readily available to prevent and treat infection caus...

  7. Full-length genomic analysis of porcine rotavirus strains isolated from pigs with diarrhea in Northern Italy.

    Science.gov (United States)

    Monini, Marina; Zaccaria, Guendalina; Ianiro, Giovanni; Lavazza, Antonio; Vaccari, Gabriele; Ruggeri, Franco M

    2014-07-01

    Group A rotaviruses (RVA) cause acute dehydrating diarrhea in young of man and many animal species, including pigs. Swine RVA has an important economic impact on the farming industry, and pigs represent a potential reservoir for zoonotic transmission of RVA to humans. To investigate the genetic diversity of porcine RVA strains in Italy and identify their possible zoonotic characteristics, 25 RVA-positive feces were collected from diarrheic pigs in Northern Italy, in 2009-2010; all viral strains were characterized by G and P genotyping RT-PCR. Three samples were selected for full genome sequencing. Sequencing of the NSP3 genes of all samples was also performed. Rotavirus diagnosis was carried out by ELISA and electron microscopy. RT-PCR and Sanger sequencing were performed in a one-tube format, using primer sets specific for each of the 11 genome segments. Analysis of the G (VP7) and P (VP4) genotypes showed that all strains identified were typical porcine RVAs (G4, G5, G9; P[6], P[13], P[23]). Full-length genome sequencing was performed on selected G9 isolates. Most segments belonged to the genotype constellation 1 (Wa-like), which is shared by most human RVA strains, but gene types such as I5 (VP6) and A8 (NSP1), which are typical of porcine and rare among human RVAs, were also detected. We identified RVA strains showing the T7 genotype, an NSP3 gene type that was previously reported in unusual strains of possible porcine or bovine origin from children with diarrhea. Recent reports suggested that G9 RVA may have been introduced from swine to human populations involving gene reassortment events. The observation that some of the RVA genotypes from swine in Italy were similar to viruses characterized in children underlines the importance of animal RVA surveillance, to clarify and monitor the role of animals as genetic reservoirs of emerging RVA strains pathogenic for humans.

  8. When your child has diarrhea

    Science.gov (United States)

    ... foods, pastries, donuts, and sausage. Avoid giving children apple juice and full-strength fruit juices, as they ... Diarrhea Browse the Encyclopedia A.D.A.M., Inc. is accredited by URAC, also known as the ...

  9. Investigation of Diarrhea in AIDS

    OpenAIRE

    Mönkemüller, Klaus E; C Mel Wilcox

    2000-01-01

    Chronic diarrhea is a common problem in patients with acquired immune deficiency syndrome (AIDS), resulting in significant morbidity and potential mortality. In the early stages of immunodeficiency, human immunodeficiency virus (HIV)-infected patients are susceptible to infection with the same enteric pathogens that cause diarrhea in immunocompetent hosts, but with progressive immunodeficiency, these patients become susceptible to numerous opportunistic disorders. The main factor to consider ...

  10. Viral information.

    Science.gov (United States)

    Rohwer, Forest; Barott, Katie

    2013-03-01

    Viruses are major drivers of global biogeochemistry and the etiological agents of many diseases. They are also the winners in the game of life: there are more viruses on the planet than cellular organisms and they encode most of the genetic diversity on the planet. In fact, it is reasonable to view life as a viral incubator. Nevertheless, most ecological and evolutionary theories were developed, and continue to be developed, without considering the virosphere. This means these theories need to be to reinterpreted in light of viral knowledge or we need to develop new theory from the viral point-of-view. Here we briefly introduce our viral planet and then address a major outstanding question in biology: why is most of life viral? A key insight is that during an infection cycle the original virus is completely broken down and only the associated information is passed on to the next generation. This is different for cellular organisms, which must pass on some physical part of themselves from generation to generation. Based on this premise, it is proposed that the thermodynamic consequences of physical information (e.g., Landauer's principle) are observed in natural viral populations. This link between physical and genetic information is then used to develop the Viral Information Hypothesis, which states that genetic information replicates itself to the detriment of system energy efficiency (i.e., is viral in nature). Finally, we show how viral information can be tested, and illustrate how this novel view can explain existing ecological and evolutionary theories from more fundamental principles.

  11. VIRAL MARKETING

    OpenAIRE

    OLENTSOVA Y.A.

    2016-01-01

    Abstract This project seeks to investigate how the company Gitz can create awareness towards their brand by using viral marketing. To do this we analyze which elements of viral marketing the company can use, to reach their goal. In order to utilize the selected tools of viral marketing best possible, we need to figure out the company’s customer segment and figure out how to reach that segment. This has been done with the use of Henrik Dahl’s Minerva-model that divides the population into f...

  12. Viroses confundíveis com febre aftosa Viral diseases to be differentiated from foot-and-mouth disease

    Directory of Open Access Journals (Sweden)

    Franklin Riet-Correa

    1996-08-01

    Full Text Available Revisam-se as doenças que devem ser consideradas no diagnóstico diferencial de febre aftosa. Dentre as doenças vesiculares ou erosivas, descrevem-se os principais aspectos relacionados ao diagnóstico da estomatite vesicular, diarréia viral bovina, febre catarral maligna, infecções por herpesvírus bovino 1 e 5, e uma estomatite ulcerativa associada a parvovírus bovino, que ocorreu no Rio Grande do Sul; língua azul, para a qual tem sido detectados anticorpos em bovinos e ovinos do Rio Grande do Sul; mamilite herpética que ocorre em outros Estados do País;peste bovina, que foi diagnosticada e erradicada no Estado de São Paulo em 1921; estomatite popular; e duas doenças exóticas:exantema vesicular e doença vesicular do suíno.Diseases to be considered in the differential diagnosis of foot-and-mouth disease are reviewed. The main aspects relating to the diagnosis of vesicular stomatitis, bovine virus diarrhea, malignant catarrhal fever, bovine herpesvirus 1 and 5, andem ulcerative stomatitis associated with bovine parvovirus are described. Bluetongue, that probably occurs in Rio Grande do Sul because antibodies to the virus have been detected in cattle and sheep; is refered. Bovine ulcerative mammilitis, reported in other Brazilian States, rinderpest, reported and eradicated in the State of São Paulo in 1921, and popular stomatitis are also cited, and so are two exotic diseases: vesicular exanthema and swine vesicular disease.

  13. Molecular diagnosis of infectious diarrhea: focus on enteric protozoa.

    Science.gov (United States)

    Verkerke, Hans P; Sobuz, Shihab U; Petri, William A

    2014-11-01

    Robust detection of enteric protozoa is a critical step toward determining the etiology of diarrhea. Widespread use of conventional microscopy, culturing and antigen detection in both industrial and developing countries is limited by relatively low sensitivity and specificity. Refinements of these conventional approaches that reduce turnaround time and instrumentation have yielded strong alternatives for clinical and research use. However, advances in molecular diagnostics for protozoal, bacterial, viral and helminth infections offer significant advantages in studies seeking to understand pathogenesis, transmission and long-term consequences of infectious diarrhea. Quantitation of enteropathogen burden and highly multiplexed platforms for molecular detection dramatically improve predictive power in emerging models of diarrheal etiology, while eliminating the expense of multiple tests.

  14. Viral arthritis

    Science.gov (United States)

    Infectious arthritis - viral ... Arthritis may be a symptom of many virus-related illnesses. It usually disappears on its own without ... the rubella vaccine, only a few people develop arthritis. No risk factors are known.

  15. Symposium on diarrhea. 1. Definition and mechanisms of diarrhea.

    Science.gov (United States)

    Jeejeebhoy, K N

    1977-04-01

    Diarrhea, an increase in frequency of evacuation and in water content of the stool, is the result of three categories of mechanism--solute malabsorption, secretion of fluid and motility disturbance. Before diarrhea is considered an abnormal condition, any alteration in stool frequency and content must be related to an individual person's normal bowel habit and to norms for the population, but more than three bowel movements or the passage of liquid stools exceeding 300 g daily should, in general, be considered abnormal. A useful way of understanding the mechanism of diarrhea is to become familiar with the normal functions of the bowel in regard to water and electrolyte absorption and motility, and then to relate these functions to solute malabsorption, fluid secretion and motility disturbance.

  16. Role of probiotics in antibiotic-associated diarrhea, Clostridium difficile-associated diarrhea, and recurrent Clostridium difficile-associated diarrhea.

    Science.gov (United States)

    Surawicz, Christina M

    2008-07-01

    The role of probiotics in the prevention and treatment of antibiotic-associated diarrhea, Clostridium difficile diarrhea, and recurrent C. difficile diarrhea is reviewed. Various probiotics have variable efficacy. More studies are needed to define further their efficacies, roles, and indications.

  17. Diarrhea in neonatal intensive care unit

    Institute of Scientific and Technical Information of China (English)

    Annalisa; Passariello; Gianluca; Terrin; Maria; Elisabetta; Baldassarre; Mario; De; Curtis; Roberto; Paludetto; Roberto; Berni; Canani

    2010-01-01

    AIM:To investigate the frequency,etiology,and current management strategies for diarrhea in newborn.METHODS:Retrospective,nationwide study involving 5801 subjects observed in neonatal intensive care units during 3 years.The main anamnesis and demographic characteristics,etiology and characteristics of diarrhea,nutritional and therapeutic management,clinical outcomes were evaluated.RESULTS:Thirty-nine cases of diarrhea(36 acute,3 chronic) were identified.The occurrence rate of diarrhea was 6.72 per 1000 hosp...

  18. Zinc therapy for different causes of diarrhea

    OpenAIRE

    Hafaz Zakky Abdillah; Supriatmo Supriatmo; Melda Deliana; Selvi Nafianti; Atan Baas Sinuhaji

    2013-01-01

    Background The incidence of diarrhea in Indonesia has declined in the past five years. In spite of the increasing number of studies on the treatment for acute diarrhea, especially the use of zinc, it is not known if bacterial vs. non-bacterial etiology makes a difference in the reduction of severity of acute diarrhea in children on zinc therapy. Objective To assess the effect of zinc therapy in reducing the severity of acute bacterial and non-bacterial diarrhea. Method...

  19. Calcium ameliorates diarrhea in immune compromised children

    OpenAIRE

    Cheng, Sam X.; Bai, Harrison X.; Gonzalez-Peralta, Regino; Mistry, Pramod K.; Gorelick, Fred S.

    2013-01-01

    Treatment of infectious diarrheas remains a challenge, particularly in immunocompromised patients in whom infections usually persist and resultant diarrhea is often severe and protracted. Children with infectious diarrhea who become dehydrated are normally treated with oral or intravenous rehydration therapy. Although rehydration therapy can replace the loss of fluid, it does not ameliorate diarrhea. Thus, over the past decades, there has been continuous effort to search for ways to safely st...

  20. Viral quasispecies.

    Science.gov (United States)

    Andino, Raul; Domingo, Esteban

    2015-05-01

    New generation sequencing is greatly expanding the capacity to examine the composition of mutant spectra of viral quasispecies in infected cells and host organisms. Here we review recent progress in the understanding of quasispecies dynamics, notably the occurrence of intra-mutant spectrum interactions, and implications of fitness landscapes for virus adaptation and de-adaptation. Complementation or interference can be established among components of the same mutant spectrum, dependent on the mutational status of the ensemble. Replicative fitness relates to an optimal mutant spectrum that provides the molecular basis for phenotypic flexibility, with implications for antiviral therapy. The biological impact of viral fitness renders particularly relevant the capacity of new generation sequencing to establish viral fitness landscapes. Progress with experimental model systems is becoming an important asset to understand virus behavior in the more complex environments faced during natural infections.

  1. Evaluation of viral clearance in the production of HPV-16 L1 virus-like particles purified from insect cell cultures.

    Science.gov (United States)

    Jeong, Hye-Sung; Shin, Jin-Ho; Choi, Jung-Yun; Kim, Young-Lim; Bae, Jei-Jun; Kim, Byoung-Guk; Ryu, Seung-Rel; Kim, Soon-Nam; Min, Hong-Ki; Kim, Hong-Jin; Park, Sue-Nie

    2006-12-01

    Biopharmaceutical products produced from cell cultures have a potential for viral contamination from cell sources or from adventitious introduction during production. The objective of this study was to assess viral clearance in the production of insect cell-derived recombinant human papillomavirus (HPV)-16 type L1 virus-like particles (VLPs). We selected Japanese encephalitis virus (JEV), bovine viral diarrhea virus (BVDV), and minute virus of mice (MVM) as relevant viruses to achieve the aim of this study. A downstream process for the production of purified HPV-16 L1 VLPs consisted of detergent lysis of harvested cells, sonication, sucrose cushion centrifugation, and cesium chloride (CsCl) equilibrium density centrifugation. The capacity of each purification/treatment step to clear viruses was expressed as reduction factor by measuring the difference in log virus infectivity of sample pools before and after each process. As a result, detergent treatment (0.5% v/v, Nonidet P-40/phosphate-buffered saline) was effective for inactivating enveloped viruses such as JEV and BVDV, but no significant reduction ( or = 4.40 log(10) reductions). Given the study conditions we used, overall cumulative reduction factors for clearance of JEV, BVDV, and MVM were > or = 10.50, > or = 9.20, and > or = 6.40 log(10) in 150 ml of starting cell cultures, respectively.

  2. Enteropathogens Associated with Acute Diarrhea in Children from Households with High Socioeconomic Level in Uruguay

    Directory of Open Access Journals (Sweden)

    Gustavo Varela

    2015-01-01

    Full Text Available Infectious diarrhea, a common disease of children, deserves permanent monitoring in all social groups. To know the etiology and clinical manifestations of acute diarrhea in children up to 5 years of age from high socioeconomic level households, we conducted a descriptive, microbiological, and clinical study. Stools from 59 children with acute community-acquired diarrhea were examined, and their parents were interviewed concerning symptoms and signs. Rotavirus, adenovirus, and norovirus were detected by commercially available qualitative immunochromatographic lateral flow rapid tests. Salmonella, Campylobacter, Yersinia, and Shigella were investigated by standard bacteriological methods and diarrheagenic E. coli by PCR assays. We identified a potential enteric pathogen in 30 children. The most frequent causes of diarrhea were enteropathogenic E. coli (EPEC, viruses, Campylobacter, Salmonella, and Shiga-toxin-producing E. coli (STEC. Only 2 patients showed mixed infections. Our data suggest that children with viral or Campylobacter diarrhea were taken to the hospital earlier than those infected with EPEC. One child infected with STEC O26 developed “complete” HUS. The microbiological results highlight the importance of zoonotic bacteria such as atypical EPEC, Campylobacter, STEC, and Salmonella as pathogens associated with acute diarrhea in these children. The findings also reinforce our previous communications about the regional importance of non-O157 STEC strains in severe infant food-borne diseases.

  3. 北京地区规模化奶牛场三种病毒性腹泻病的血清学调查%Serological Survey of Three Viral Diarrheal Diseases in Beijing Large-scale Dairy Cow Farms

    Institute of Scientific and Technical Information of China (English)

    傅彩霞; 杜鹃; 靳兴军; 郑瑞峰; 郭峰; 韩磊; 李佳; 毛惠明; 王玉田; 张跃

    2012-01-01

    为了解近年北京地区奶牛腹泻性疾病的流行情况,采用酶联免疫吸附试验(ELISA)对北京地区密云、怀柔和昌平3个区县的未免疫接种牛病毒性腹泻病毒(Bovine viral diarrhea virus,BVDV)、牛冠状病毒(Bovine coronavirus,BCV)和牛轮状病毒(Bovine rotavirus,BRV)疫苗的31个规模化奶牛场的1 650份血清样品进行了BVDV、BCV、BRV感染抗体检测.结果显示,BVDV抗体平均阳性率为48.2%,BCV抗体平均阳性率为57.2%,BRV抗体平均阳性率为52.2%,BVDV、BCV及BRV感染在密云、怀柔和昌平3个区县的牛群中普遍存在,需进一步加强奶牛腹泻性疾病的综合防控.

  4. Molecular analysis of the bovine coronavirus S1 gene by direct sequencing of diarrheic fecal specimens

    Directory of Open Access Journals (Sweden)

    E. Takiuchi

    2008-04-01

    Full Text Available Bovine coronavirus (BCoV causes severe diarrhea in newborn calves, is associated with winter dysentery in adult cattle and respiratory infections in calves and feedlot cattle. The BCoV S protein plays a fundamental role in viral attachment and entry into the host cell, and is cleaved into two subunits termed S1 (amino terminal and S2 (carboxy terminal. The present study describes a strategy for the sequencing of the BCoV S1 gene directly from fecal diarrheic specimens that were previously identified as BCoV positive by RT-PCR assay for N gene detection. A consensus sequence of 2681 nucleotides was obtained through direct sequencing of seven overlapping PCR fragments of the S gene. The samples did not undergo cell culture passage prior to PCR amplification and sequencing. The structural analysis was based on the genomic differences between Brazilian strains and other known BCoV from different geographical regions. The phylogenetic analysis of the entire S1 gene showed that the BCoV Brazilian strains were more distant from the Mebus strain (97.8% identity for nucleotides and 96.8% identity for amino acids and more similar to the BCoV-ENT strain (98.7% for nucleotides and 98.7% for amino acids. Based on the phylogenetic analysis of the hypervariable region of the S1 subunit, these strains clustered with the American (BCoV-ENT, 182NS and Canadian (BCQ20, BCQ2070, BCQ9, BCQ571, BCQ1523 calf diarrhea and the Canadian winter dysentery (BCQ7373, BCQ2590 strains, but clustered on a separate branch of the Korean and respiratory BCoV strains. The BCoV strains of the present study were not clustered in the same branch of previously published Brazilian strains (AY606193, AY606194. These data agree with the genealogical construction and suggest that at least two different BCoV strains are circulating in Brazil.

  5. EFFECTS OF SUPPLEMENTAL LYSINE ON PERFORMANCE, ANTIBODY TITER AND RECTAL TEMPERATURE IN RESPONSE TO A MODIFIED-LIVE VIRAL VACCINE IN NEONATAL CALVES

    Directory of Open Access Journals (Sweden)

    Kate Sharon

    2014-01-01

    Full Text Available Infectious Bovine Rhinotracheitis (IBR, caused by bovine herpesvirus-1, contributes significantly to economic losses in the dairy and beef cattle industry. Lysine has been shown reduce virulence of herpesviruses in felids and humans. Our objective was to evaluate the effects of supplemental lysine on serum IBR antibody titer and rectal temperature in response to a modified-live Intranasal (IN or Intramuscular (IM respiratory-virus vaccination. Sixty-four neonatal Holstein bull calves (7±2 d of age; BW = 37±4.2 kg were used in a completely randomized design. Calves were fed milk replacer supplemented with either 17 g/d L-lysine monohydrochloride (LYS; 28 calves or an equivalent amount of casein (CAS; 28 calves for 42 d. Calves were then vaccinated with either an IN IBR-Parainfluenza virus-3 (PI3 or an IM (IBR-PI3-bovine viral diarrhea type I and II, bovine respiratory syncytial virus modified-live vaccine on d 36. A control group (8 calves received no supplement or vaccination. All calves were housed in individual calf pens (1.2×2.1 m. Daily feed intakes were monitored and BW measured weekly. Calves were bled on d 0, 35, 36, 37 and 42. Temperature data loggers were attached to rectal probes and temperatures were recorded every 5 min from d 28 to d 42. No significant differences were determined for average performance, rectal temperature, or IBR antibody titers with either IN or IM vaccinations between LYS and CAS treated calves (p>0.10. However, serum urea nitrogen and the ratio of serum lysine: Arginine increased (p<0.05 for LYS compared to CAS calves. These results suggest that supplementing lysine impacts nitrogen metabolism but does not alter the response to IBR vaccination or animal performance in neonatal Holstein calves.

  6. VIRAL ETIOLOGY ACUTE INTESTINAL INFECTIONS MOLECULAR MONITORING IN CHILDREN’S HOSPITAL

    OpenAIRE

    A. V. Sergeeva; L. Y. Poslova; O. V. Kovalishena; A. S. Blagonravova; N. V. Epifanova; T. A. Sashina; Morozova, O.V.; N. A. Novikova

    2015-01-01

    On the territory of the Russian Federation in the overall structure of acute intestinal infections the proportion of viral diarrhea among children varies from 24 to 78% of cases depending on the season. The acute viral intestinal infections etiological confirmation is performed mainly among patients of infectious hospitals. The prevalence of viral acute intestinal infections in non-infectious hospitals, including infections associated with medical care, remains unclear. Currently estimation o...

  7. 77 FR 29914 - Bovine Spongiform Encephalopathy; Importation of Bovines and Bovine Products

    Science.gov (United States)

    2012-05-21

    ... RIN 0579-AC68 Bovine Spongiform Encephalopathy; Importation of Bovines and Bovine Products AGENCY... live bovines and products derived from bovines with regard to bovine spongiform encephalopathy. This... with regard to bovine spongiform encephalopathy. Comments on the proposed rule were required to......

  8. Expression of Bovine Leukemia Virus Genome is Blocked by a Nonimmunoglobulin Protein in