A longitudinal study of the dynamics of bovine corona virus and respiratory syncytial virus infections in dairy herds.

The objective of this study was to examine the dynamics of bovine corona virus (BCV) and bovine respiratory syncytial virus (BRSV) infections in dairy herds over a 3-year period. The status of 79 dairy herds located in two Northern and two Southern Regions of Sweden were surveyed by measuring antibody concentrations to BCV and BRSV in pooled milk samples from primiparous cows, and in bulk-tank milk twice annually. In the Southern Regions the percentage of antibody-positive herds remained persistently high (75-100%), whereas in herds based in the Northern Region, the percentage of positive herds for BCV and BRSV was 38-80% and 0-80%, respectively, with antibody levels to BRSV decreasing over time. Pooled milk samples of 'home-bred' primiparous animals were found to be most useful in terms of monitoring herd status but could gradually be replaced by bulk-tank sampling once freedom from infection was established.

Genetic detection and characterization of atypical bovine pestiviruses in foetal bovine sera claimed to be of Australian origin.

Two European laboratories independently detected atypical bovine pestiviral nucleic acids in three commercial batches of foetal bovine serum (FBS) that was claimed by the producers to be of Australian origin. Additional batches of FBS were obtained directly from Australia to exclude possible contamination of the Australian FBS with that of South American origin during manufacturing/packaging in European countries. RT-PCR amplification of partial 5'untranslated region and the complete N(pro) gene yielded a specific band with expected size, which was confirmed by DNA sequencing. Bayesian analysis of sequence data demonstrated a closer phylogenetic relation of the newly detected atypical bovine pestiviruses to those of South American origin, which were related to the recognized bovine pestivirus species, bovine viral diarrhoea virus. Taken together, the results indicated the presence of atypical bovine pestiviruses in the Australian FBS, and most likely in Australian Continent.

Risk factors for seropositivity to bovine coronavirus and bovine respiratory syncytial virus in dairy herds.

A cross-sectional study was conducted to evaluate the association between herd-level characteristics, management routines and farm-level antibody status of bovine coronavirus (BCV) and bovine respiratory syncytial virus (BRSV) in 257 Swedish dairy herds. The possible spatial clustering of positive herds compared with negative herds was also investigated. For each herd, a pooled milk sample from five primiparous cows was analysed for the presence of antibodies to BCV and BRSV. Herd-level information was obtained by a questionnaire. Logistic regression was used to evaluate the association between predictors and antibody status to BCV and BRSV. Large herd size, being located in southern Sweden, and not providing boots for visitors were found to be associated with being antibody-positive to BCV and BRSV. A short distance to the nearest cattle herd was an additional risk factor for BCV. One of the studied areas was suitable for spatial analysis. Positive herds were not spatially autocorrelated when compared with negative herds as estimated by the K-function regarding both BCV and BRSV. This indicates that local factors such as daily visiting milk trucks and wild animals were unlikely to be important sources of infection in this area. Moran's I statistics and semi-variogram showed no evidence of spatial autocorrelation in the residuals, indicating that remaining unidentified factors are not spatially dependent in the areas under study.

Atypical 'HoBi'-like pestiviruses--recent findings and implications thereof.

In 2004, an atypical pestivirus named D32/00_'HoBi', isolated from foetal calf serum (FCS) originating from Brazil, was described (Schirrmeier et al., 2004). A few years later, a closely related virus (Th/04_KhonKaen) was detected in serum from a calf in Thailand, indicating that this group of atypical pestiviruses already is spread in cattle populations in various regions of the world. At the Friedrich-Loeffler-Institute, Insel Riems, Germany, FCS batches are regularly tested for pestivirus contamination, in general with positive PCR results, and in some cases the contaminants have been typed as 'HoBi'-like. At the National Veterinary Institute (SVA) in Uppsala, Sweden, a recent event with contaminated FCS ruined much of the ongoing cell culture work. From the FCS and the contaminated cells we were able to amplify and sequence nucleic acid from three different pestivirus strains, including BVDV-1, -2 and 'HoBi'-like; this in a commercial FCS that had been tested free from pestivirus by the manufacturer. In this short communication we review the current status of atypical 'HoBi'-like pestiviruses, describe recent findings and discuss the implications thereof.

Comparison between dairy cow disease incidence in data registered by farmers and in data from a disease-recording system based on veterinary reporting.

Sweden has a national disease-recording system based on veterinary reporting. From this system, all cattle-disease records are transferred to the dairy industry cattle database (DDD) where they are used for several purposes including research and dairy-health statistics. Our objective was to evaluate the completeness of this data source by comparing it with disease data registered by dairy farmers. The proportion of veterinary-treated disease events was estimated, by diagnosis. Disease incidence in the DDD was compared, by diagnosis and age, with disease data registered by the farmers. Comparison was made, by diagnosis, for (i) all disease events and (ii) those reported as veterinary-treated. Disease events, defined as "observed deviations in health, from the normal" were recorded by the farmers during January, April, July and October 2004. For the diagnoses calving problems, peripartum disorders, puerperal paresis and retained placenta, incidence proportions (IP) with 95% confidence intervals (CIs) were estimated. For all other disease problems, incidence rates (IR) were used. In total, 177 farmers reported at least 1 month and 148 reported all 4 months. Fifty-four percent of all disease events in the farmers' data were reported as veterinary-treated. For several of the most common diagnoses, the IRs and IPs for all events were significantly higher in farmers' data than in the DDD. Examples are, in cows: clinical mastitis, cough, gastro-intestinal disorders and lameness in hoof and limb; and in young stock: cough and gastro-intestinal disorders. For veterinary-treated events only, significant differences with higher IR in the farmers' data were found in young stock for sporadic cough and sporadic gastro-intestinal disorders. The diagnosis "other disorders" had significantly more events in the DDD than in farmers' data, i.e. veterinarians tended to choose more unspecific diagnoses than the farmers. This result indicates that the true completeness is likely to be higher than our estimate. We conclude that for the time period studied there was differential under-reporting associated with the diagnosis, the age of the animal and whether the herd was served by a state-employed or private veterinarian.

A six-year study on respiratory viral infections in a bull testing facility.

Viral infection dynamics and bovine respiratory disease (BRD) treatment rates were studied over six years at a Swedish bull testing station with an 'all in, all out' management system. In August of each of the years 1998-2003, between 149 and 185 4-8-month-old calves arrived at the station from 99 to 124 different beef-breeding herds, and remained until March the following year. Only calves that tested free from bovine viral diarrhoea virus (BVDV) were allowed to enter the station and original animal groups were kept isolated from new cattle in their original herds for three weeks before admission. Although neither prophylactic antibiotics, nor BRD vaccines were used, less than 0.7-13.2% (mean 5%) of the calves (n=970) required treatment for BRD during the first five weeks following entry. This was probably due, at least in part, to the season (the summer months) when the animals were commingled. In the six-month period August-February, 38% of the animals were treated one or more times for BRD and mortality was 0.7%. Hereford and Aberdeen Angus calves had significantly higher treatment rates than Charolais, Simmental and Blonde d'Aquitaine. Serological testing on samples obtained in August, November and January indicated that bovine parainfluenza virus 3 (PIV-3) infections occurred each year before November after entry. Bovine coronavirus (BCoV) infections also occurred every year, but in 3/6 years this was not until after November. Bovine respiratory syncytial virus (BRSV) infections occurred only every second year and were associated with a treatment peak and one death on one occasion (December). The herd remained BVDV free during the entire study period. The infection patterns for PIV-3 and BCoV indicated a high level of infectivity amongst bovine calves, whereas the incidence for BRSV was observed at a lower level. Although the rearing of the animals differed from conventional beef production, the study has shown that commingling animals from many sources is not necessarily associated with high morbidity within the first few weeks after arrival. By preventing BRD soon after commingling the prerequisites for protective vaccination at entry might be improved. Applied management routines are discussed.

Dynamics of virus infections involved in the bovine respiratory disease complex in Swedish dairy herds.

The dynamics of bovine respiratory syncytial virus (BRSV), bovine parainfluenza virus 3 (PIV-3), bovine corona virus (BCoV) and bovine viral diarrhoea virus (BVDV) infections were studied in 118 dairy herds in south western Sweden. By using serology on paired samples from three approximately 7 vs. approximately 15-month-old calves per herd, the propagation of infections was investigated over about a 1-year period. The results implied that at least 74% of calves had experienced one or more of the monitored infections at the age of approximately 7 months (Sample 1, Spring); 30%, 48%, 34% and 8% were seropositive to BRSV, PIV-3, BCoV and BVDV, respectively. Seroconversions to BRSV, PIV-3, BCoV and BVDV occurred in 26%, 38%, 50% and 3% of seronegative animals and 63% had antibodies against two or more infections at approximately 15 months (Sample 2). In total, 90-97% of animals that were seropositive in Sample 1 remained positive in Sample 2. A significant association was found between BVDV and BCoV (P = 0.01). Moreover, a significantly higher proportion of herds in which no calves had a recorded history of respiratory disease (n = 15) were classified as negative to all four infections monitored when compared to herds in which disease was observed (P = 0.0002). This study showed a high infection burden in young animals and effective spread of BRSV, PIV-3 and BCoV in one area of Sweden. BVDV infections were restricted to a few herds, reflecting the effect of a voluntary control program against BVDV in Sweden.

Indirect transmission of bovine viral diarrhoea virus at calving and during the postparturient period.

Two trials were designed to investigate whether bovine viral diarrhoea virus (BVDV) could be transmitted after the birth of persistently infected calves, even if they were removed immediately after birth. In trial 1, 11 calves were actively exposed to fetal fluids and uterine lochia collected from cows that had delivered calves persistently infected with type 1 BVDV. One calf that was exposed to a sample taken on the day of calving seroconverted. In trial 2, six calves were housed in stables where persistently infected calves were being born and then removed immediately from their dams and from the stable unit within two to three hours. One of four calves in close contact with the cows after delivery seroconverted and one of two calves housed within the same stable unit became infected.

BVDV and BHV-1 infections in dairy herds in northern and northeastern Thailand.

Bulk milk samples from 220 dairy herds were collected at 9 public milk collection centres in the northeastern and northern Thailand, and a subset of 11 herds was selected for individual testing. The samples were tested for presence of antibodies to BVDV and BHV-1 using an indirect ELISA. The results from the bulk milk testing demonstrated a moderate level of exposure to BVDV and BHV-1 (73% and 67%, respectively). However, the low proportion of herds with high BVDV antibody-levels (13%) and the low within-herd seroprevalence of BVDV and BHV-1 in the 11 herds (24% and 5%, respectively), particularly among the young stock (15% and 0%, respectively), demonstrated a low prevalence of active BVDV infection and a low rate of reactivation of latent BHV-1. The presence of a self-clearance process was also indicated by the results from the individual testing. Moreover, a surprisingly low prevalence of BVDV and BHV-1 antibody-positive herds at one of the milk centres was found. This centre was established 5-10 years before the others. Our impression is that this reflects the self-clearance process, where consecutive replacement of imported infected animals without further spread has resulted in a nearly total elimination of the infections. Based on our experiences and on these results we are convinced that this process can continue if there is awareness of herd biosecurity. This is especially important in the context of a future intensification of the dairy production.

No effect of a homeopathic preparation on neonatal calf diarrhoea in a randomised double-blind, placebo-controlled clinical trial.

A double-blind, placebo-controlled clinical trial of a homeopathic treatment of neonatal calf diarrhoea was performed using 44 calves in 12 dairy herds. Calves with spontaneously derived diarrhoea were treated with either the homeopathic remedy Podophyllum (D30) (n = 24) or a placebo (n = 20). No clinically or statistically significant difference between the 2 groups was demonstrated. Calves treated with Podophyllum had an average of 3.1 days of diarrhoea compared with 2.9 days for the placebo group. Depression, inappetence and fever were presented equally in the 2 groups. These results support the widely held opinion that scientific proof for the efficacy of veterinary homeopathy is lacking. In the European Union this implies a considerable risk for animal welfare, since in some countries priority is given to homeopathic treatments in organic farming.

Experimental infection of cows with bovine viral diarrhoea virus in early pregnancy - findings in serum and foetal fluids.

Nineteen pregnant cows were experimentally infected with bovine viral diarrhoea virus (BVDV) between day 74 and 81 of pregnancy. All cows became infected and developed serum antibodies. Sixteen of the cows delivered persistently infected (PI) offspring, whereas the remaining three gave birth to calves with detectable serum antibodies and free from BVDV. The 16 cows with PI foetuses developed higher levels of antibodies in serum during pregnancy than did their three peers carrying non-PI calves. Multivariate analysis showed that the antibody levels in these two groups of cows were significantly different from day 135 of pregnancy. Foetal fluid was successfully collected from 18 of the 19 infected cows and from five uninfected control cows between 10 and 24 days before delivery by use of a percutaneous, blind puncture technique. No negative effects were observed in the cows or their offspring. BVDV was isolated and detected with an immunoperoxidase test in foetal fluid from 13 of the 16 cows carrying PI foetuses, and from 15 of the cows when a quantitative fluorescent polymerase chain reaction (PCR) technique was used. The negative sample in the PCR assay was positive for BVDV antibodies. The number of viral copies per microlitre in foetal fluids varied between 103 and 1080 in the positive samples. All samples taken from the cows carrying non-PI foetuses were negative for BVDV in both assays. In this experiment, examination of either serum or foetal fluids could identify the cows carrying a PI foetus. Examination of serum for BVDV antibodies was a reliable indicator of a PI foetus if the serum was collected during the last 2 months of pregnancy. For examination of foetal fluids, both viral and serological analyses should be performed. For viral analysis, PCR should be the test of choice. High levels of BVDV antibodies in conjunction with a negative result in the PCR may be indicative of a false-negative virus result. Further experience with the method of collection of foetal fluids is necessary for evaluation of its safety. Investigation of pregnant cows in order to discover a PI offspring before it is born could be a useful tool in control and eradication of BVDV.

Implementation and evaluation of an ordered subsets reconstruction algorithm for transmission PET studies using median root prior and inter-update median filtering.

An ordered subsets (OS) reconstruction algorithm based on the median root prior (MRP) and inter-update median filtering was implemented for the reconstruction of low count statistics transmission (TR) scans. The OS-MRP-TR algorithm was evaluated using an experimental phantom, simulating positron emission tomography (PET) whole-body (WB) studies, as well as patient data. Various experimental conditions, in terms of TR scan time (from 1 h to 1 min), covering a wide range of TR count statistics were evaluated. The performance of the algorithm was assessed by comparing the mean value of the attenuation coefficient (MVAC) of known tissue types and the coefficient of variation (CV) for low-count TR images, reconstructed with the OS-MRP-TR algorithm, with reference values obtained from high-count TR images reconstructed with a filtered back-projection (FBP) algorithm. The reconstructed OS-MRP-TR images were then used for attenuation correction of the corresponding emission (EM) data. EM images reconstructed with attenuation correction generated by OS-MRP-TR images, of low count statistics, were compared with the EM images corrected for attenuation using reference (high statistics) TR data. In all the experimental situations considered, the OS-MRP-TR algorithm showed: (1) a tendency towards a stable solution in terms of MVAC; (2) a difference in the MVAC of within 5% for a TR scan of 1 min reconstructed with the OS-MRP-TR and a TR scan of 1 h reconstructed with the FBP algorithm; (3) effectiveness in noise reduction, particularly for low count statistics data [using a specific parameter configuration the TR images reconstructed with OS-MRP-TR(1 min) had a lower CV than the corresponding TR images of a 1-h scan reconstructed with the FBP algorithm]; (4) a difference of within 3% between the mean counts in the EM images attenuation corrected using the OS-MRP-TR images of 1 min and the mean counts in the EM images attenuation corrected using the OS-MRP-TR images of 1 h; (5) preservation of "good" image quality for both TR and EM reconstructed images. In conclusion, the OS-MRP-TR algorithm is particularly suitable for WB PET studies, allowing: (1) the acquisition of a very short TR scan (within 1 min), (2) the reconstruction of such TR data in low-noise TR images and (3) the use of the reconstructed OS-MRP-TR images for attenuation correction of corresponding EM data.

Insemination of susceptible heifers with semen from a non-viraemic bull with persistent bovine virus diarrhoea virus infection localized in the testes.

Bulls shedding bovine viral diarrhoea virus (BVDV) in semen and simultaneously having a high concentration of circulating antibodies may cause reproductive problems and spread the viral infection within cattle populations. To investigate this in detail, three heifers were inseminated with BVDV-infected semen from a non-viraemic, seropositive Holstein-Friesian bull, named 'Cumulus'. One control heifer was inseminated with semen from a healthy bull that was free of BVDV. All four heifers remained clinically healthy throughout the experiment. The conception succeeded in the control animal and in two of the three heifers inseminated with semen containing BVDV. The heifer with the failed conception was the only one that became systemically infected with BVDV. This animal was deemed non-pregnant by ultrasonic examination on day 34 after insemination and showed no signs of subsequent oestrus during the entire experimental period. At slaughter, 42 days after insemination, there were no histopathological changes in the ovaries and virus was not detected in ovarian tissue. The fact that seronegative dams served with semen from persistently infected bulls have occasionally produced persistently infected calves together with the present findings and the fact that non-viraemic, seropositive bulls can constantly shed BVDV, suggest that the use of semen from such bulls in BVDV-free herds could have far-reaching consequences, especially if it led to the birth of persistently infected (P1) calves.

Implementation and evaluation of a 3D one-step late reconstruction algorithm for 3D positron emission tomography brain studies using median root prior.

A fully three-dimensional (3D) one-step late (OSL), maximum a posteriori (MAP) reconstruction algorithm based on the median root prior (MRP) was implemented and evaluated for the reconstruction of 3D positron emission tomography (PET) studies. The algorithm uses the ordered subsets (OS) scheme for convergence acceleration and data update during iterations. The algorithm was implemented using the software package developed within the EU project PARAPET (www.brunel.ac.uk/~masrppet). The MRP algorithm was evaluated using experimental phantom and real 3D PET brain studies. Various experimental set-ups in terms of activity distribution and counting statistics were considered. The performance of the algorithm was assessed by calculating figures of merit such as: contrast, coefficient of variation, activity ratio between two regions and full width at half of maximum for resolution measurements. The performance of MRP was compared with that of 3D ordered subsets-expectation maximisation (OSEM) and 3D re-projection (3DRP) algorithms. In all the experimental situations considered, MRP showed: (1) convergence to a stable solution, (2) effectiveness in noise reduction, particularly for low statistics data, (3) good preservation of spatial details. Compared with the OSEM and 3DRP algorithms, MRP provides comparable or better results depending on the parameters used for the reconstruction of the images.

Validation of a test for dams carrying foetuses persistently infected with bovine viral-diarrhoea virus based on determination of antibody levels in late pregnancy.

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.

Surveillance of ruminant diseases in the Nordic countries.

All the Nordic countries have a basis for their surveillance and disease control in ruminants in national legislation and regulations listing notifiable diseases of concern to the countries. The Nordic countries are a disease-free zone comparing to other parts of the world and the aim of the surveillance is to keep that status and be able to document it. Following is a short summary from each country.

Improvement of brain perfusion SPET using iterative reconstruction with scatter and non-uniform attenuation correction.

Filtered back-projection (FBP) is generally used as the reconstruction method for single-photon emission tomography although it produces noisy images with apparent streak artefacts. It is possible to improve the image quality by using an algorithm with iterative correction steps. The iterative reconstruction technique also has an additional benefit in that computation of attenuation correction can be included in the process. A commonly used iterative method, maximum-likelihood expectation maximisation (ML-EM), can be accelerated using ordered subsets (OS-EM). We have applied to the OS-EM algorithm a Bayesian one-step late correction method utilising median root prior (MRP). Methodological comparison was performed by means of measurements obtained with a brain perfusion phantom and using patient data. The aim of this work was to quantitate the accuracy of iterative reconstruction with scatter and non-uniform attenuation corrections and post-filtering in SPET brain perfusion imaging. SPET imaging was performed using a triple-head gamma camera with fan-beam collimators. Transmission and emission scans were acquired simultaneously. The brain phantom used was a high-resolution three-dimensional anthropomorphic JB003 phantom. Patient studies were performed in ten chronic pain syndrome patients. The images were reconstructed using conventional FBP and iterative OS-EM and MRP techniques including scatter and nonuniform attenuation corrections. Iterative reconstructions were individually post-filtered. The quantitative results obtained with the brain perfusion phantom were compared with the known actual contrast ratios. The calculated difference from the true values was largest with the FBP method; iteratively reconstructed images proved closer to the reality. Similar findings were obtained in the patient studies. The plain OS-EM method improved the contrast whereas in the case of the MRP technique the improvement in contrast was not so evident with post-filtering.

Lack of virus transmission from bovine viral diarrhoea virus infected calves to susceptible peers.

None of 14 calves not previously exposed to BVDV became infected after being forced to have nose-to-nose contact with a group of 5 calves primarily infected with BVDV. These were 5 male calves primarily infected with a type I BVDV strain, after nose-to-nose contact with a persistently viraemic calf. All 5 became infected and were clinically affected. They were slightly depressed and pyretic at 8-9 days post-infection, with a body temperature of up to 41.6 degrees C, but no medical treatment was required. Seroconversions to BVDV were detected in these calves at 14 to 21 days post-infection. The 14 healthy calves, proved to be free from BVD virus--as well as antibodies, were introduced 2 by 2 into the group of 5 primarily infected calves on days 4, 7, 14, 21, 28, 35 and 42 after the 5 calves had been in contact with the persistently BVDV-infected calf. Each pair of calves stayed within the primarily infected group for 2 days. None of these 14 calves seroconverted to BVDV.