A Comparison of Dynamics in Two Models for the Spread of a Vector-Borne Disease.

In 2007, bluetongue virus (BTV) was introduced to both Denmark (DK) and the United Kingdom (UK). For this reason, simulation models were built to predict scenarios for future incursions. The DK and UK models have a common description of within-herd dynamics, but differ greatly in their descriptions of between-herd spread, one using an explicit representation of vector dispersal, the other a transmission kernel. Here, we compare model predictions for the dynamics of bluetongue in the UK, based on the 2007 incursion and vaccination rollout in 2008. We demonstrate how an agent-based model shows greater sensitivity to the level of vaccine uptake and has lower variability compared with a kernel-based model. However, a model using a transmission kernel requires less detailed data and is often faster.

Decisions on control of foot-and-mouth disease informed using model predictions.

The decision on whether or not to change the control strategy, such as introducing emergency vaccination, is perhaps one of the most difficult decisions faced by the veterinary authorities during a foot-and-mouth disease (FMD) epidemic. A simple tool that may predict the epidemic outcome and consequences would be useful to assist the veterinary authorities in the decision-making process. A previously proposed simple quantitative tool based on the first 14 days outbreaks (FFO) of FMD was used with results from an FMD simulation exercise. Epidemic outcomes included the number of affected herds, epidemic duration, geographical size and costs. The first 14 days spatial spread (FFS) was also included to further support the prediction. The epidemic data was obtained from a Danish version (DTU-DADS) of a pre-existing FMD simulation model (Davis Animal Disease Spread - DADS) adapted to model the spread of FMD in Denmark. The European Union (EU) and Danish regulations for FMD control were used in the simulation. The correlations between FFO and FFS and the additional number of affected herds after day 14 following detection of the first infected herd were 0.66 and 0.82, respectively. The variation explained by the FFO at day 14 following detection was high (P-value<0.001). This indicates that the FFO may take a part in the decision of whether or not to intensify FMD control, for instance by introducing emergency vaccination and/or pre-emptive depopulation, which might prevent a "catastrophic situation". A significant part of the variation was explained by supplementing the model with the FFS (P-value<0.001). Furthermore, the type of the index-herd was also a significant predictor of the epidemic outcomes (P-value<0.05). The results of the current study suggest that national veterinary authorities should consider to model their national situation and to use FFO and FFS to help planning and updating their contingency plans and FMD emergency control strategies.

Comparing control strategies against foot-and-mouth disease: will vaccination be cost-effective in Denmark?

Recent outbreaks of foot-and-mouth disease (FMD) in Europe have highlighted the need for assessment of control strategies to optimise control of the spread of FMD. Our objectives were to assess the epidemiological and financial impact of simulated FMD outbreaks in Denmark and the effect of using ring depopulation or emergency vaccination to control these outbreaks. Two stochastic simulation models (InterSpreadPlus (ISP) and the modified Davis Animal Disease Simulation model (DTU-DADS)) were used to simulate the spread of FMD in Denmark using different control strategies. Each epidemic was initiated in one herd (index herd), and a total of 5000 index herds were used. Four types of control measures were investigated: (1) a basic scenario including depopulation of detected herds, 3 km protection and 10 km surveillance zones, movement tracing and a three-day national standstill, (2) the basic scenario plus depopulation in ring zones around detected herds (Depop), (3) the basic scenario plus protective vaccination within ring zones around detected herds, and (4) the basic scenario plus protective vaccination within ring zones around detected herds. Disease spread was simulated through direct animal movements, medium-risk contacts (veterinarians, artificial inseminators or milk controllers), low-risk contacts (animal feed and rendering trucks, technicians or visitors), market contacts, abattoir trucks, milk tanks, or local spread. The two simulation models showed different results in terms of the estimated numbers. However, the tendencies in terms of recommendations of strategies were similar for both models. Comparison of the different control strategies showed that, from an epidemiological point of view, protective vaccination would be preferable if the epidemic started in a cattle herd in an area with a high density of cattle, whereas if the epidemic started in an area with a low density of cattle or in other species, protective vaccination or depopulation would have almost the same preventive effect. Implementing additional control measures either 14 days after detection of the first infected herd or when 10 herds have been diagnosed would be more efficient than implementing additional control measures when more herds have been diagnosed. Protective vaccination scenarios would never be cost-effective, whereas depopulation or suppressive vaccination scenarios would most often be recommended. Looking at the median estimates of the cost-benefit analysis, depopulation in zones would most often be recommended, although, in extreme epidemics, suppressive vaccination scenarios could be less expensive. The vast majority of the costs and losses associated with a Danish epidemic could be attributed to export losses.

Experimental airborne transmission of porcine postweaning multisystemic wasting syndrome.

The objective of these studies was to investigate if porcine postweaning multisystemic wasting syndrome (PMWS) could be induced in healthy pigs following contact with air from pigs with clinical signs of PMWS. The pigs were housed in different units. Either 31 (study I) or 25 (study II) pigs with clinical symptoms of PMWS from a PMWS-affected herd and 25 healthy pigs from a PMWS-free, but PCV2-positive, herd were housed in unit A. Fifty pigs from a PMWS-free herd were housed in unit B, which were connected by pipes to unit A. In unit C, 30 pigs from a PMWS-free herd were housed as controls. In study II, the pigs in units A and B from the PMWS-free herd developed clinical signs of PMWS 2-3 weeks after arrival. PMWS was confirmed at necropsy and the diseased pigs had increased PCV2 load and increased antibody titers against PCV2 in serum that coincided with the development of clinical signs typical of PMWS. Sequence analysis revealed that the PCV2 isolate belonged to genotype 2b. In conclusion, the present study showed that PMWS can be induced in pigs from a PMWS-free herd by airborne contact with pigs from a PMWS-affected herd.

Meta-analysis on the efficacy of foot-and-mouth disease emergency vaccination.

The objectives of this study were to provide a summary quantification of the efficacy of FMD emergency vaccination based on a systematic review and a meta-analysis of available literature, and to further discuss the suitability of this review and meta-analysis to summarize and further interpret the results. Peer-reviewed, symposium, and unpublished studies were considered in the analysis. Clinical protection and virological protection against FMD were used as parameters to assess the efficacy of emergency vaccination. The clinical protection was estimated based on the appearance of clinical signs including FMD lesions and fever, while the virological protection parameter was estimated based on the outcome of laboratory tests that were used to diagnose FMD infection. A meta-analysis relative risk was calculated per protection parameter. Results of the meta-analyses were examined using publication bias tests. In total, 31 studies were included in the analyses, of which 26 were peer-reviewed studies, 1 was a symposium study and 4 were unpublished studies. Cattle, swine and sheep were well protected against clinical disease and FMD infection following the use of emergency vaccine. Fortunately, no significant bias that would alter the conclusions was encountered in the analysis. Meta-analysis can be a useful tool to summarize literature results from a systematic review of the efficacy of FMD emergency vaccination.

Inter-laboratory and inter-assay comparison on two real-time PCR techniques for quantification of PCV2 nucleic acid extracted from field samples.

Several real-time PCR assays for quantification of PCV2 DNA (qPCR) have been described in the literature, and different in-house assays are being used by laboratories around the world. A general threshold of 10(7) copies of PCV2 per millilitre serum for postweaning multisystemic wasting syndrome (PMWS) diagnosis has been suggested. However, neither inter-laboratory nor inter-assay comparisons have been published so far. In the present study, two different qPCR probe assays used routinely in two laboratories were compared on DNA extracted from serum, nasal and rectal swabs. Results showed a significant linear association between the assays (p<0.0001), and a systematic difference of 1.4 log10 copies of PCV2 per millilitre of sample (p<0.0001). This difference indicated that the assay from laboratory 1 yielded a higher output than the one from laboratory 2. Results also showed that there was no linear association between the amount of PCV2 DNA and the amount of total DNA, neither in nasal (p=0.86) nor in rectal (p=0.78) swabs, suggesting that normalizing of PCV2 DNA load in swab samples to total DNA concentration is not suitable. The present exploratory study highlights the need for the performance of ring trials on qPCV2 protocols between laboratories. Meanwhile, the proposed thresholds for PMWS diagnosis should only be considered reliable for each particular laboratory and each particular assay.

Postweaning multisystemic wasting syndrome in Danish pig herds: productivity, clinical signs and pathology.

A case-control study of 74 herds with postweaning multisystemic wasting syndrome (pmws) and 74 matched control herds was carried out. In the case herds the mortality rates of weaner and finisher pigs were 11.2 and 5.2 per cent respectively, compared with 3.1 and 3.2 per cent in the control herds. In most case herds, pmws developed within the first four weeks after weaning. Wasting, diarrhoea and respiratory signs were observed in 10 per cent of the weaner pigs (7 to 30 kg) in the case herds compared with 7 per cent in the control herds. The average daily gains of the weaner pigs and finisher pigs were 36 g and 52 g less in the case herds than in the control herds. By examining three weaner pigs from each herd the pmws diagnosis was confirmed by histopathology and immunohistochemistry in 78 per cent of the case herds, but at least one pmws-positive weaner pig was found in 19 of the control herds. The prevalence of pmws-positive pigs among illthriven weaner pigs was 45 per cent (101/222) in the case herds, and 12 per cent (27/222) in the control herds. Specific gross pathological findings were associated with a positive pmws diagnosis; pigs with heavy, rubber-like lungs, atonic intestines, and enlarged bronchial and inguinal lymph nodes, had a 0.7 probability of a positive pmws diagnosis by laboratory examinations. However, for illthriven pigs, this probability of having pmws was equal in the case herds and the control herds.

Population dynamics and intra-litter transmission patterns of Isospora suis in suckling piglets under on-farm conditions.

The aim of this study was to investigate the intra-litter infection dynamics of Isospora suis under natural conditions, and to study any association between parasite transmission and the contamination level of the farrowing pen by applying different interventions in order to reduce the transmission of I. suis infection within the litter. The study was divided in 2 trials including in total 22 litters (254 piglets). The first trial included 4 litters (where standard procedures practiced routinely on the farm piglets were applied) and the piglets were followed coprologically from farrowing until 2 weeks after weaning. The sows of those litters were also examined at various intervals before and after farrowing. The second trial included the application of 3 different management procedures: (A) standard farm hygiene and management procedures, (B) standard farm hygiene and management procedures+the first piglets found to excrete I. suis oocysts in each pen were removed from the pen, and (C) reduced cleaning. Each procedure was studied in 2 litters. This was replicated 3 times to yield a total of 18 litters. The results suggested that (i) the sow does not play an important role in transmission of I. suis in the farrowing pen; (ii) in natural infections, both the age of the piglet age at onset of oocyst excretion and the oocyst excretion patterns may vary considerably; (iii) the course of oocyst excretion or development of diarrhoea is related to the time of initial infection and (iii) piglets, which are heavy at birth, are more prone to acquire I. suis infection. Moreover, it was demonstrated that cleaning could be an effective means of restricting the spread of the parasite within the litter and thus the development of diarrhoea.

Bayesian estimation of sensitivity and specificity of serum ELISA and faecal culture for diagnosis of paratuberculosis in Greek dairy sheep and goats.

Latent class models were used to estimate the sensitivity (Se) and the specificity (Sp) of a serum ELISA and a faecal culture (FC) method for the diagnosis of paratuberculosis separately, in sheep and goats. The estimates were obtained by a Bayesian method. Possible dependence of diagnostic errors was investigated by comparing models where independence was assumed to models allowing for conditional dependence given the true disease status. ROC analysis for the serum ELISA was also performed and optimized cut-off values based on the misclassification cost term were determined. No evidence of conditional dependence was found. Assuming independence, posterior medians and 95% credible intervals for the Se(ELISA), Sp(ELISA), Se(FC) and Sp(FC), were 63% (42, 93%), 95% (90, 98%), 8% (2, 17%) and 98% (95, 100%) in goats and 37% (10, 80%), 97% (93, 99%), 16% (2, 48%) and 97% (95, 99%) in sheep. AUC was calculated 0.702 for sheep and 0.847 for goats. For the serum ELISA, there is need of species- and purpose-specific cut-off selection. For instance, with 20% prevalence situation and assuming equal and five-fold cost of a false negative to a false positive test result, the optimal cut-off is 0.3 and 0.05 in sheep, respectively, while it is 0.6 and 0.1 in goats, respectively. Serum ELISA performed better in goats than in sheep. Lowering the cut-off, in relation to the one recommended by the manufacturer, improved Se(ELISA) without seriously compromising Sp(ELISA), in either species.

Prevalence and diversity of Campylobacter jejuni in pig herds on farms with and without cattle or poultry.

The prevalence and diversity of Campylobacter jejuni was investigated in pig herds on farms with and without cattle or poultry production. A bacteriological screening of pig cecal samples from 247 finisher herds was carried out at the slaughter-house. Subsequently, a follow-up study was conducted in 24 herds (either with or without prior C. jejuni isolation from pigs) in which fecal samples were collected from pigs and, if present, cattle and poultry. Samples were analyzed for presence of Campylobacter, and subsequent analysis included species identification, serotyping, and, for selected strains, pulsed-field gel electrophoresis typing. In the slaughterhouse screening, C. jejuni was isolated from pigs in 21 (8.5%) herds, but no significant difference in prevalence was found between herd types (pigs, pigs and cattle, pigs and poultry). At the slaughterhouse, C. jejuni and Campylobacter coli prevalence in pigs was 2.3 and 90.1%, respectively. In the follow-up study, herd prevalence of C. jejuni was 8.3%, whereas C. jejuni and C. coli were isolated from 0.8 and 92.0% of pigs, respectively. In mixed production herds, C. jejuni predominated in cattle (42.7%) and poultry (31.6%), whereas C. jejuni was only isolated from 1.3 to 2.5% of pigs in these herds. There were no significant differences in C. jejuni or C. coli prevalence in pigs, cattle, and poultry between herds with and without prior C. jejuni isolation at the slaughterhouse. Pulsed-field gel electrophoresis typing did not yield evidence of C. jejuni transmission between cattle or poultry and pigs in mixed production herds. In contrast, pulsed-field gel electrophoresis analysis showed indistinguishable serotypes of C. coli in pigs and cattle in two herds. Verification of C. jejuni-positive pig samples showed that individual pigs can excrete high levels of C. jejuni and that mixed infection with C. jejuni and C. coli was common in C. jejuni-positive pigs. The results of our study suggest that transmission of C. jejuni between pigs and cattle or poultry in mixed production herds occurs infrequently. Detection of indistinguishable C. coli isolates in two herds, however, might indicate the existence of low-level transmission between pigs and cattle in herds of mixed production.

Estimation of sensitivity, specificity and predictive values of two serologic tests for the detection of antibodies against Actinobacillus pleuropneumoniae serotype 2 in the absence of a reference test (gold standard).

Latent-class models were used to determine the sensitivity, specificity and predictive values of a polyclonal blocking enzyme-linked immunosorbent assay (ELISA) and a modified complement-fixation test (CFT) when there was no reference test. The tests were used for detection of antibodies against Actinobacillus pleuropneumoniae serotype 2 in a survey of respiratory diseases in Danish finishing pigs. The estimates were obtained by maximum-likelihood and also by a Bayesian method (implemented with Gibbs sampling). Possible dependence of diagnostic errors was investigated by comparing models where independence was assumed to models allowing for conditional dependence, given the true disease status. No strong evidence of conditional dependence in either test sensitivity or specificity was found. Assuming independence, maximum-likelihood estimates and 95% confidence intervals of the sensitivity and specificity of the ELISA were 100% and 92.8% (90.1-95.5%) and the corresponding values of the CFT were 90.6% (85.8-95.4%) and 98.6% (98.0-99.3%), respectively. Bayesian estimates and posterior 95% credible intervals of the sensitivity and specificity of the ELISA were 99.7% (98.7-100%) and 92.7% (89.9-95.3%) and of the CFT were 90.6% (86.0-95.3%) and 98.7% (98.0-99.3%). The sensitivity and specificity of a combined test, where the CFT is subsequently applied to the pig sera that test positive in the ELISA, were estimated at 90.2% (85.6-95.0%) and 99.9% (99.8-100%), respectively. The cost of the combined test was less than the cost of the use of the CFT alone, at prevalences <54%. Prevalences and predictive values and their 95% limits were estimated in six sub-samples of data. The estimates of sensitivity and specificity obtained in the present investigation generally validate those reported from other sources.

Estimation of sensitivity and specificity of diagnostic tests and disease prevalence when the true disease state is unknown.

The performance of a new diagnostic test is frequently evaluated by comparison to a perfect reference test (i.e. a gold standard). In many instances, however, a reference test is less than perfect. In this paper, we review methods for estimation of the accuracy of a diagnostic test when an imperfect reference test with known classification errors is available. Furthermore, we focus our presentation on available methods of estimation of test characteristics when the sensitivity and specificity of both tests are unknown. We present some of the available statistical methods for estimation of the accuracy of diagnostic tests when a reference test does not exist (including maximum likelihood estimation and Bayesian inference). We illustrate the application of the described methods using data from an evaluation of a nested polymerase chain reaction and microscopic examination of kidney imprints for detection of Nucleospora salmonis in rainbow trout.

Prevalence of subclinical Salmonella enterica infection in Danish finishing pig herds.

Our aim was to determine the Salmonella enterica prevalence in 96 randomly selected Danish pig herds, based on serological examination of blood samples and bacteriological examination of faecal samples (collected simultaneously from the same pens). For comparison, 39 high-seroprevalence herds were included in the study. The representativeness of the selected herds was assessed, based on descriptive statistics of herd size and type. Totals of 1330 pen samples and 6814 blood samples were examined.The results from the meat-juice screening in the Danish S. enterica Control Programme were available for 3372 meat-juice samples from 91 of the 96 randomly selected herds and 1195 meat-juice samples from 37 of the 39 high-seroprevalence herds. Of the 96 randomly selected herds, 23 herds had no positive pen samples (out of 10), no positive blood samples (out of 50) and no positive meat-juice samples (out of approximately 30-40 samples in 6 months). Ten herds had one or more positive meat-juice samples but were otherwise negative. S. Typhimurium was isolated from 30 of the 39 high-seroprevalence herds. Our conclusions were: (1) The within-herd seroprevalence among the 96 randomly selected Danish pig herds was low (average within-herd seroprevalence=2%, maximum=32%). (2) Among the 39 high-seroprevalence herds (recently assigned level 3 in the S. enterica Control Programme), S. enterica was isolated from 77% of the herds when 10 pen samples were examined bacteriologically. (3) Seropositivity tended to be related to the presence of S. Typhimurium.