Serological evidence of Coxiella burnetii, Leptospira interrogans Hardjo, Neospora caninum and bovine pestivirus infections in a dairy cattle herd from the United Arab Emirates.

The serostatus of five abortigenic agents and the association between abortion history and Coxiella burnetii seropositivity were assessed in 350 dairy cattle from Al Ain, UAE. The bovine sera were ELISA-screened for C. burnetii, Leptospira Hardjo, Neospora caninum, and Brucella abortus antibodies, plus bovine pestivirus (BVDV) antigen. The serology data were collated and the level of significance between the proportions of C. burnetii-seropositive cattle with and without abortion history assessed by the Z test of two proportions. Of the 350 cattle, 41.4%, 1.7%, 1.4%, 0.3%, and 0.0% were seropositive to the above pathogens, respectively. Besides, 61.9%, 2.9%, 1.0%, 0.0%, and 0.0% of the 105 cattle with history of abortion and 32.7%, 1.2%, 1.6%, 0.0% and 0.0% of the 245 seropositive cattle with no history of abortion were also seropositive for the above pathogens respectively. Moreover, the proportion of C. burnetii-seropositive cattle with history of abortion were significantly higher than the C. burnetii-seropositive ones without abortion history (p-value < 0.01). Apparent C. burnetii infections were relatively higher than the other four pathogens suggesting this bacterium contributed to abortion in the herd. Additional research on the public and bovine health implications of C. burnetii and Leptospira in the UAE are urgently needed.

[PESTIVIRUSES IN RUMINANTS].

The genus Pestivirus includes four species: bovine viral diarrhea virus 1, bovine viral diarrhea virus 2, classical swine fever disease virus, and ovine border disease virus. Pestiviruses infect many species of domestic and wild animals. Bovine viral diarrhea virus is a prototypical representative of the pestiviruses of ruminant animals. Recently, new candidates appeared for including in this genus: two viruses of the wild ruminant animals that have not been officially classified and one HoBi-like virus discovered for the first time in the bovine fetal serum. The circulation of the ruminant animal pestiviruses within population of domestic and wild animals, the presence of these viruses in bioproducts stimulates studies of the infection reservoirs and their influence on the effect of the bovine viral diarrhea control programs.

First report of bovine viral diarrhoea virus-2 infection in cattle in Poland.

This report describes the first identification in Poland of bovine viral diarrhoea virus (BVDV)-2 in a dairy herd where severe clinical disease with losses of young animals was observed. The virus was readily cultivated in cell culture and a phylogenetic analysis of the nucleotide sequences and secondary structures of the viral genomic 5' untranslated region confirmed virus identity. The economic impact of the infection was significant compared to the previously prevalent BVDV-1 infections confirming that this genotype of BVDV can cause severe sickness in affected herds. The use of BVDV-1 vaccine did not prevent the infection with the BVDV-2 genotype.

Effect of the vaccination scheme on PregSure® BVD induced alloreactivity and the incidence of Bovine Neonatal Pancytopenia.

Bovine Neonatal Pancytopenia (BNP) is a new neonate-maternal incompatibility phenomenon caused by vaccine-induced, maternal alloantibodies. The syndrome affects newborn calves at the approximate age of ten days and is characterized by spontaneous bleeding, severe anemia with an almost complete destruction of the red bone marrow. During the past two years the causal role of bioprocess impurities in PregSure(®)BVD, a strongly adjuvanted, inactivated vaccine against Bovine Virus Diarrhoea (BVD), in the induction of BNP causing alloantibodies has clearly been established. Despite intensive research efforts that have elucidated the basic principles of the BNP immunopathology still a number of questions remain open. In the current manuscript we address the puzzling observation that BNP incidences vary widely between different regions: as an example we compare the BNP incidences in the German Federal States of Bavaria and Lower Saxony. In Bavaria the BNP-incidence reaches 100 cases per 100,000 doses PregSure(®)BVD, while in Lower Saxony the incidence is as low as 6 cases per 100,000 doses. In Bavaria the vaccine has always been used according to the instructions for use. By contrast, in Lower Saxony BVD-immunization was performed according to a two-step vaccination protocol including a first immunization with an inactivated BVD-vaccine followed by booster immunizations with a live-attenuated BVD-vaccine. As a consequence, those cattle that received PregSure(®)BVD received in general more than two doses in Bavaria, while in Lower Saxony cows received at maximum one dose. By experimental immunization we can show that the two-step regimen including PregSure(®)BVD as a priming vaccine results in significantly lower alloantibody titers as compared to repetitive immunizations with the inactivated vaccine. The lower alloantibody titer after two-step vaccination explains the lower BNP-incidence in Lower Saxony and - generally speaking - indicates that variations in the vaccination regimen have a great influence on the induction of adverse reactions through bioprocess impurities.

Bovine respiratory syncytial virus seroprevalence and risk factors in endemic dairy cattle herds.

The herd seroprevalence of bovine respiratory syncytial virus (BRSV) was studied in 59 dairy cattle herds using serology on random selected animals stratified by two age classes (heifers, cows). Risk factors for primary infections in heifers were investigated using a questionnaire on management conditions and data on bovine viral diarrhoea (BVD) status. At least one seropositive cow was present in all the herds. In 25% of the herds all individual were seropositive and 22% of herds had all heifers seronegative. Analysis of the influence of risk factors retained summer pasture and BVD status. In particular, absence of summer pasture and the BVD positive status of heifers were associated with an increased risk of BRSV infection in heifers group.

Epidemiology and control of BVD in the U.S.

The apparent prevalence of bovine viral diarrhea virus (BVDV) persistently infected cattle has been found to be low in U.S. dairies, beef herds and feedlots. Current management practices within U.S. cattle industries that impact the epidemiology of BVDV infections include purchasing untested cattle, lack of biosecurity procedures, large herd sizes, mixing cattle from multiple sources, high cattle densities in dairy and feedlot operations, synchronous breeding of beef herds, communal grazing and widespread vaccination. Evidence for BVDV infection has been found in farmed and free-ranging wildlife in North America; however the risk of BVDV transmission from wildlife to cattle is not known. The perception of a low prevalence of BVDV herd infections, the unrestricted sale of PI cattle, lack of economic data, intensive marketing of vaccines, reluctance to accept federal regulations, and a "gambler's" attitude among producers are impediments to implementation of a national systematic BVD control program. Since 2004, voluntary BVDV control programs have been organized in nine states reflecting the recognition of BVD as an important and preventable problem in the U.S.

In vitro amplification of BVDV field strains isolated in Argentina: effect of cell line and culture conditions.

The aim of this work was to study the in vitro amplification of BVDV (Pestivirus, Flaviridae) field isolates from Argentina in MDBK, BoTur and BHK-21 continuous cell lines. Field isolates 99/134 (mucosal disease), 00/693 (mucosal disease), 04P7016 (respiratory disease) and 04/89 (mucosal disease), genotype 1b, were used and compared with the Singer and NADL reference strains, genotype 1a. Additionally, cell lines derived from explants of bovine testis (RD-420), bovine uterus (NCL-1) and porcine kidney (PKZ) were tested as alternative substrates for BVDV propagation in vitro. The effect of cell line, harvest time and infection protocol was evaluated. The viral titers observed depended on the virus and harvest time but not on the infection protocol. We found that MDBK and BoTur cell lines were susceptible to the infection whereas BHK-21 and PKZ were not. NADL viral titers, 00/693 and 04/89, increased from 24 to 48 h p.i. in BoTur cells and then reached a plateau, whereas those of 99/134 and 04P7016 remained constant between 24 and 72 h p.i. BVDV Singer, on the other hand, presented a maximum titer at 24 h p.i. and then decreased. BVDV-NADL titers increased in MDBK and NCL-1 but not in RD-420 between 24 and 48 h p.i., and then decreased at 72 h p.i. These facts lead us to conclude that neither the subgenotypes (1a, 1b) nor the clinical symptoms of the animal from the virus had been isolated seem to affect the virus cell line kinetics of viral replication in vitro. On the other hand, the most homogenous behavior, the most similar replication curves, and highest titers observed in MDBK and NCL-1 seem to indicate that these lines are generally more susceptible to BVDV replication.

In vitro amplification of BVDV field strains isolated in Argentina: effect of cell line and culture conditions / Amplificación in vitro de cepas de campo de virus de la diarrea viral bovina (VDVB) aisladas en Argentina: efecto de la línea celular y las condicioes de cultivo

The aim of this work was to study the in vitro amplification of BVDV (Pestivirus, Flaviridae) field isolates from Argentina in MDBK, BoTur and BHK-21 continuous cell lines. Field isolates 99/134 (mucosal disease), 00/693 (mucosal disease), 04P7016 (respiratory disease) and 04/89 (mucosal disease), genotype 1b, were used and compared with the Singer and NADL reference strains, genotype 1a. Additionally, cell lines derived from explants of bovine testis (RD- 420), bovine uterus (NCL-1) and porcine kidney (PKZ) were tested as alternative substrates for BVDV propagation in vitro. The effect of cell line, harvest time and infection protocol was evaluated. The viral titers observed depended on the virus and harvest time but not on the infection protocol. We found that MDBK and BoTur cell lines were susceptible to the infection whereas BHK-21 and PKZ were not. NADL viral titers, 00/693 and 04/89, increased from 24 to 48 h p.i. in BoTur cells and then reached a plateau, whereas those of 99/134 and 04P7016 remained constant between 24 and 72 h p.i. BVDV Singer, on the other hand, presented a maximum titer at 24 h p.i. and then decreased. BVDV-NADL titers increased in MDBK and NCL-1 but not in RD-420 between 24 and 48 h p.i., and then decreased at 72 h p.i. These facts lead us to conclude that neither the subgenotypes (1a, 1b) nor the clinical symptoms of the animal from the virus had been isolated seem to affect the virus cell line kinetics of viral replication in vitro. On the other hand, the most homogenous behavior, the most similar replication curves, and highest titers observed in MDBK and NCL-1 seem to indicate that these lines are generally more susceptible to BVDV replication.

Se estudió la interacción de aislamientos de campo de Argentina del VDVB (Pestivirus, Flaviridae) en las líneas celulares continuas MDBK, BoTur y BHK-21. Se utilizaron los virus de campo genotipo 1b, 99/134, 00/693 (casos compatibles con enfermedad de las mucosas) y 04P7016 (cuadro respiratorio) y las cepas de referencia genotipo 1a Singer y NADL. Además se evaluó la interacción de VDVB-NADL con las líneas celulares experimentales de bovino RD-420 y NCL-1 y de riñón porcino (PKZ). Se usaron 2 protocolos de infección. Los títulos virales observados dependieron del virus y del tiempo de infección y no así del modo de infección. Mientras que MDBK y BoTur resultaron susceptibles a la infección, BHK-21 y PKZ no lo fueron. Los virus NADL, 00/693 y 04/89 incrementaron su título entre las 24 y las 48 h p.i. en BoTur para mantenerlo posteriormente; los virus 99/134 y 04P7016 no presentaron variaciones y la cepa Singer presentó título máximo a las 24 h p.i para luego descender. La cinética del virus NADL en las células MDBK, RD-420 y NCL-1 tuvo un incremento de título para MDBK y NCL-1 entre las 24 y 48 h p.i que descendió a las 72 h p.i. La interacción virus-línea celular no estaría relacionada con el sub-genotipo del virus (1a o 1b), ni con el cuadro clínico; las células MDBK y NCL-1 serían más susceptibles a la replicación del VDVB.

Variation in ptaquiloside content In bracken (Pteridium esculentum (Forst. f) Cockayne ) in New Zealand.

AIM: To examine stands of bracken fern (Pteridium esculentum) from throughout New Zealand for the presence and concentration of ptaquiloside (Pta), and to compare the presence and/or concentrations of Pta in areas where bovine enzootic haematuria (BEH) and/or acute haemorrhagic syndrome (AHS) has been known to occur with those where BEH/AHS has not been recorded. METHODS: Stands of bracken fern were sampled from 275 sites throughout New Zealand. Sixty-two stands were from a regional survey predominantly from the Waikato and Coromandel regions, 27 were from a farm in the King Country where BEH/AHS had been investigated previously, and 186 were from a national survey of the North and South Islands. Sampling sites were from a mixture of grazed paddocks, roadsides, and forest and bush areas. Samples comprised whole young fronds, the tops of unfurling young fronds, or, for the Regional Survey, mature green fronds from the previous season. Pta was extracted from the samples, and measured using high-performance liquid chromatography. Information on the occurrence of BEH/AHS at specific locations was obtained from published information and records from animal health laboratories in New Zealand. RESULTS: The 275 samples contained widely varying concentrations of Pta. In the Farm Survey, concentrations ranged from 280-13,300 (mean 3,800) microg/g (on a dry-weight basis) in the 63% of samples that contained Pta. A high proportion of samples from the Regional and National Surveys covering large areas of the country contained no detectable levels of Pta. The majority (61%) of samples from these two surveys which contained Pta were from areas where BEH/AHS was reported to occur. Combining data from all surveys, in areas with reported BEH/AHS, 42% of samples collected contained Pta, compared with 6% where BEH/AHS was not known to occur. CONCLUSIONS: Concentrations of Pta in bracken in New Zealand vary greatly, and in a high proportion of stands Pta is not found. A higher incidence of Pta, and some very high concentrations, are found in areas where BEH/AHS was known to occur.

Genetic analysis of a bovine viral diarrhea virus 2 isolate from Slovakia.

The identification and genetic characterization of bovine viral diarrhea virus (BVDV) isolate 17237 detected in western Slovakia is described. The analysis of 5'-untranslated region (5'-UTR), autoprotease (Npro) gene, and structural genes (C, Erns, E1, E2) was carried out. The percentage of nucleotide and deduced amino acid identity in analyzed genes implied that the isolate was closely related to the bovine viral diarrhea virus 2 (BVDV-2). Furthermore, the phylogenetic analysis revealed that this isolate fall into BVDV-2b subtype that is sporadic in Europe. The cleavage sites between viral proteins were similar to the ones of a reference strain of BVDV-2.

The evolution of bovine viral diarrhea: a review.

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 understanding of pathogenic mechanisms are now being reevaluated because of these "new" virus strains. This shift in virulence has confounded both nomenclature and the significance of current bovine viral diarrhea virus categorization. The purpose of this review is to summarize our current understanding of bovine viral diarrhea virus with a chronological review of prevailing scientific tenets and practices as described in clinical and scientific North American veterinary journals and textbooks. The first part of this review describes how we have arrived at our current understanding of the viruses, the diseases, and their nomenclature. The second part of the review deals with current concepts in virology and how these concepts may both explain and predict bovine viral diarrhea virus pathogenesis. By reviewing how knowledge of bovine viral diarrhea has evolved and the theories of how the virus itself is able to evolve, the interpretation of diagnostic tests are more effectively utilized in the control and treatment of bovine viral diarrhea virus associated disease.