Jenny Frössling

Validation of a Neospora caninum iscom ELISA without a gold standard.

Neospora caninum is an intracellular parasite which causes abortion in cattle worldwide. One problem in the validation of the different methods for demonstration of this parasite is the lack of an appropriate gold standard. To validate an immunostimulating complex (iscom) enzyme-linked immunoassay (ELISA) used to detect antibodies to N. caninum, sera from 244 cattle in five Swedish dairy herds infected with N. caninum were analysed. The sera also were analysed by a standard indirect-fluorescent antibody test (IFAT). The results obtained by the two tests were compared using the Gibbs sampler. Gibbs sampling is a latent-class approach based on Bayesian statistics; neither test is assumed to be more correct in stating the true status of infection. The Gibbs sampler was run using both informative and non-informative prior probabilities. We also simulated different cut-offs in the iscom ELISA (providing data to inform selection of optimal cut-off values for different applications). The ELISA produced fewest incorrect test results over all at a cut-off value of 0.200. The sensitivity and specificity at this cut-off were 99 and 96%, respectively. The IFAT had a high specificity (99%) but a lower sensitivity (78%) than expected-confirming that the IFAT cannot be treated as a true gold standard. Sensitivity and specificity of the ELISA were presented in a two-graph receiver operating characteristic (TG-ROC) plot. Any cut-off between 0.150 and 0.300 will have both sensitivity and specificity > or =95%. Optical densities of < or =0.150 and > or =0.550 (or > or =0.350) were suggested as limits to rule out and rule in infection, respectively.

Application of the Neospora caninum IgG avidity ELISA in assessment of chronic reproductive losses after an outbreak of neosporosis in a herd of beef cattle

Point-source infections are most likely the cause for Neospora caninum–induced abortion outbreaks in cattle, whereas an increased annual abortion rate may be a consequence of vertical transmission. The aims of the present study were to examine the reproductive effects of neosporosis in a beef herd for 3 years, after a point-source outbreak and to use IgG avidity serology to examine the chronicity of infections and patterns of transmission. During the study, 76–78% of animals were seropositive for N. caninum. The pregnancy rate varied from 88% to 94%, without any reduction in the pregnancy rate of seropositive cows compared with seronegative cows. The annual abortion rate was 2.5–5.5%, and all but 1 abortion occurred in seropositive dams. The efficiency of vertical transmission was estimated to be 85%. Several calves, born to seronegative dams, were seropositive at 6–13 months of age, indicating a 22% mean annual rate of horizontal transmission. The mean avidity in seropositive cows increased from 30 during the initial outbreak to 74 after 3 years. The mode of IgG avidity was 21–40 during the initial abortion outbreak, 41–60 after 1 year, and 61–80 after 2 and 3 years. The results reveal high annual rates of both vertical and horizontal transmission of N. caninum in a herd of beef cows and provide further validation on the ability of the N. caninum IgG avidity ELISA to accurately assess the chronicity of infection.

Application of network analysis parameters in risk-based surveillance - Examples based on cattle trade data and bovine infections in Sweden

Abstract Financial resources may limit the number of samples that can be collected and analysed in disease surveillance programmes. When the aim of surveillance is disease detection and identification of case herds, a risk-based approach can increase the sensitivity of the surveillance system. In this paper, the association between two network analysis measures, i.e. ‘in-degree’ and ‘ingoing infection chain’, and signs of infection is investigated. It is shown that based on regression analysis of combined data from a recent cross-sectional study for endemic viral infections and network analysis of animal movements, a positive serological result for bovine coronavirus (BCV) and bovine respiratory syncytial virus (BRSV) is significantly associated with the purchase of animals. For BCV, this association was significant also when accounting for herd size and regional cattle density, but not for BRSV. Examples are given for different approaches to include cattle movement data in risk-based surveillance by selecting herds based on network analysis measures. Results show that compared to completely random sampling these approaches increase the number of detected positives, both for BCV and BRSV in our study population. It is concluded that network measures for the relevant time period based on updated databases of animal movements can provide a simple and straight forward tool for risk-based sampling.

Probability of freedom from disease after the first detection and eradication of PRRS in Sweden: Scenario-tree modelling of the surveillance system

In July 2007, PRRS was detected for the first time in Sweden. A total of eight positive herds were identified and various measures were taken to eradicate the disease, including restrictions and slaughter of infected herds. Subsequently, both active and passive surveillance activities were undertaken. This study describes stochastic scenario-tree modelling of all the various surveillance system components, to estimate the current probability that Sweden is free from PRRS. The model includes all actions taken after the first positive herd was detected. The surveillance system components included in the model were as follows: investigations undertaken in association with the outbreak, a serological study based on samples collected at slaughter, samples collected in the national PRRS surveillance programme and passive clinical surveillance. The probability of freedom was estimated in time steps of 1 month, from July to December 2007. After each time step, the calculated posterior probability of freedom from the previous month, combined with the probability of introduction, was used as a prior probability for the next month. The result from the model showed a 99.8% probability that Sweden was free from PRRS at the end of December 2007. The estimated total sensitivity of the surveillance system varied between 81.2% and 94.3% and was highest during the first months after the outbreak. For sensitivity analysis purposes, the model was also applied using higher risks of introduction. However, this did not make considerable difference to the final estimates.

The Swedish breeding cat: population description, infectious diseases and reproductive performance evaluated by a questionnaire.

The questionnaire based study gives a combined description of management, infectious diseases and reproductive performance in breeding catteries during 1 year. The mean number of cats per cattery was 6.1, and 25% of the breeders let some of their cats have free access to outdoors. Breeders reported that infection with feline panleukopenia virus, feline immunodeficiency virus or feline leukaemia virus was uncommon, but 8% of the breeders had sold or had themselves owned a cat that died of feline infectious peritonitis. Presence of conjunctivitis was reported by 33.3% of the breeders. Mean litter size was 3.7±1.5, with 9.7% stillbirths and 8.3% kitten mortality week 1–12. The percentage of stillborn kittens increased with the age of the queen and litter size, and also differed among breeds. Kitten mortality differed among breeds, but did not increase with age of the queen. Seven percent of the litters were delivered by caesarean section, significantly more during winter and positively associated with presence of stillborn kittens.

Q fever infection in dairy cattle herds: increased risk with high wind speed and low precipitation

SUMMARY Ruminants are considered the main reservoir for transmission of Coxiella burnetii (Cb) to humans. The implementation of effective control measures against Cb in ruminants requires knowledge about potential risk factors. The objectives of this study were (i) to describe the spatial distribution of Q fever-infected dairy cattle herds in Sweden, (ii) to quantify the respective contributions of wind and animal movements on the risk of infection, while accounting for other sources of variation, and (iii) to investigate the possible protective effect of precipitation. A total of 1537 bulk milk samples were collected and tested for presence of Cb antibodies. The prevalence of test-positive herds was higher in the south of Sweden. For herds located in areas with high wind speed, open landscape, high animal densities and high temperature, the risk of being infected reached very high values. Because these factors are difficult to control, vaccination could be an appropriate control measure in these areas. Finally, the cumulated precipitation over 1 year was identified as a protective factor.

Surveillance system sensitivities and probability of freedom from Mycobacterium avium subsp. paratuberculosis infection in Swedish cattle.

Previous investigations suggest that the prevalence of Mycobacterium avium subsp. paratuberculosis (MAP) in Swedish cattle is low and all recent cases have been linked to imported animals. The aim of this study was to evaluate the surveillance system for MAP infection in Swedish cattle and to estimate the probability that the Swedish cattle population is free from this infection. Calculations of surveillance sensitivities and probability of freedom were made using stochastic scenario-tree modelling, which allows inclusion of information from several different sources, of complex surveillance data including results from non-representative sampling, as well as of documentations of differences in risk of being infected. The surveillance components included in the model were: (1) clinical surveillance, (2) fallen stock investigations, (3) the national surveillance programme (mainly beef herds), (4) a survey involving dairy herds and (5) a risk-based survey targeting herds with imported cattle. Previous or current presence of imported animals and participation in the on-going control programme was specified for each tested herd, in order to adjust for differences in risk. Calculations were made for each year from the start of 2005 to the end of 2008, and this formed the basis for a final estimate covering the whole study period and predictions of future probabilities of freedom from MAP. Results show that when applying a design prevalence of one animal in 0.1% of the herds, the probability of freedom at the end of 2008 was 0.63. At the design prevalence of one animal in 0.5% of herds, the estimated probability is >95% and it is demonstrated that the prevalence of MAP in Swedish cattle is below this level or absent. In order to increase the annual surveillance sensitivity in the future and thereby improve the probability of freedom, new surveillance activities or an intensification of current ones are needed.

A survey of visitors on Swedish livestock farms with reference to the spread of animal diseases

BackgroundIn addition to livestock movements, other between-farm contacts such as visitors may contribute to the spread of contagious animal diseases. Knowledge about such contacts is essential for contingency planning. Preventive measures, risk-based surveillance and contact tracing may be facilitated if the frequency and type of between-farm contacts can be assessed for different types of farms. The aim of this study was to investigate the frequency and types of visitors on farms with cloven-hoofed animals in Sweden and to analyse whether there were differences in the number of visitors attributable to region, season, and type of herd. Data were collected from Swedish farmers through contact-logs covering two-week periods during four different seasons.ResultsIn total, 482 (32%) farmers filled in the contact log for at least one period and the data represent 18,416 days. The average number of professional and non-professional visitors per day was 0.3 and 0.8, respectively. Whereas the number of professional visitors seemed to increase with increasing herd size, this relation was not seen for non-professional visits. The mean numbers of visitors per day were highest in the summer and in the farm category ‘small mixed farm’. Reports of the visitors’ degree of contact with the animals showed that veterinarians, AI-technicians, animal transporters and neighbours were often in direct contact with the animals or entered the stables and 8.8% of the repairmen were also in direct contact with animals, which was unexpected. In a multivariable analysis, species, herd size and season were significantly associated with the number of professional visitors as well as the number of visitors in direct contact with the animals.ConclusionIn conclusion there was a large variation between farms in the number and type of contacts. The number of visitors that may be more likely to spread diseases between farms was associated with animal species and herd size.

Spatial patterns of Bovine Corona Virus and Bovine Respiratory Syncytial Virus in the Swedish beef cattle population

BackgroundBoth bovine coronavirus (BCV) and bovine respiratory syncytial virus (BRSV) infections are currently wide-spread in the Swedish dairy cattle population. Surveys of antibody levels in bulk tank milk have shown very high nationwide prevalences of both BCV and BRSV, with large variations between regions. In the Swedish beef cattle population however, no investigations have yet been performed regarding the prevalence and geographical distribution of BCV and BRSV. A cross-sectional serological survey for BCV and BRSV was carried out in Swedish beef cattle to explore any geographical patterns of these infections.MethodsBlood samples were collected from 2,763 animals located in 2,137 herds and analyzed for presence of antibodies to BCV and BRSV. Morans I was calculated to assess spatial autocorrelation, and identification of geographical cluster was performed using spatial scan statistics.ResultsAnimals detected positive to BCV or BRSV were predominately located in the central-western and some southern parts of Sweden. Morans I indicated global spatial autocorrelation. BCV and BRSV appeared to be spatially related: two areas in southern Sweden (Skaraborg and Skåne) had a significantly higher prevalence of BCV (72.5 and 65.5% respectively); almost the same two areas were identified as being high-prevalence clusters for BRSV (69.2 and 66.8% respectively). An area in south-east Sweden (Kronoberg-Blekinge) had lower prevalences for both infections than expected (23.8 and 20.7% for BCV and BRSV respectively). Another area in middle-west Sweden (Värmland-Dalarna) had also a lower prevalence for BRSV (7.9%). Areas with beef herd density > 10 per 100 km2 were found to be at significantly higher risk of being part of high-prevalence clusters.ConclusionThese results form a basis for further investigations of between-herds dynamics and risk factors for these infections in order to design effective control strategies.

Toxoplasma gondii seroprevalence in wild boars ( Sus scrofa ) in Sweden and evaluation of ELISA test performance

SUMMARY Toxoplasma gondii is a zoonotic protozoan parasite, infecting a wide range of warm-blooded animals. The Swedish wild boar population is expanding and increased hunting provides its meat to a growing group of consumers. We performed a spatio-temporal investigation of T. gondii seroprevalence in Swedish wild boars. An ELISA was set up and evaluated against a commercial direct agglutination test, using Bayesian latent class analysis. The ELISA sensitivity and specificity were estimated to 79% and 85%, respectively. Of 1327 serum samples, 50% were positive. Thirty-four per cent of young wild boars and 55% of adults were positive (P < 0·001). The total seroprevalence ranged from 72% in 2005 to 38% in 2011 (P < 0·001), suggesting a declining trend. The highest seroprevalence, 65%, was recorded in South Sweden. In other regions it varied from 29% in Stockholm to 46% in East Middle Sweden.