A framework for assessing confidence in freedom from infection in animal disease control programmes.

In the Surveillance Tool for Outcome-based Comparison of FREEdom from infection (STOC free) project (https://www.stocfree.eu), a data collection tool was constructed to facilitate standardised collection of input data, and a model was developed to allow a standardised and harmonised comparison of the outputs of different control programmes (CPs) for cattle diseases. The STOC free model can be used to evaluate the probability of freedom from infection for herds in CPs and to determine whether these CPs comply with the European Union's pre-defined output-based standards. Bovine viral diarrhoea virus (BVDV) was chosen as the case disease for this project because of the diversity in CPs in the six participating countries. Detailed BVDV CP and risk factor information was collected using the data collection tool. For inclusion of the data in the STOC free model, key aspects and default values were quantified. A Bayesian hidden Markov model was deemed appropriate, and a model was developed for BVDV CPs. The model was tested and validated using real BVDV CP data from partner countries, and corresponding computer code was made publicly available. The STOC free model focuses on herd-level data, although that animal-level data can be included after aggregation to herd level. The STOC free model is applicable to diseases that are endemic, given that it needs the presence of some infection to estimate parameters and enable convergence. In countries where infection-free status has been achieved, a scenario tree model could be a better suited tool. Further work is recommended to generalise the STOC free model to other diseases.

Dans le cadre du projet européen STOC free (Surveillance Tool for Outcome-based Comparison of FREEdom from infection, outil de surveillance permettant de comparer les probabilités d'absence d'infection sur la base des résultats, https://www.stocfree.eu), un outil de recueil des données a été construit pour faciliter une collecte normalisée des données d'entrée ; un modèle a également été élaboré pour permettre une comparaison normalisée et harmonisée des données sur les résultats des différents programmes de contrôle des maladies des bovins. Le modèle STOC free peut être utilisé pour évaluer la probabilité d'absence d'infection au sein des troupeaux dans le cadre des programmes de contrôle et déterminer si ces programmes sont conformes aux normes définies par l'Union européenne en termes de résultats attendus. L'infection par le virus de la diarrhée virale bovine a été choisie comme maladie d'étude pour ce projet en raison de la diversité des programmes de contrôle dans les six pays participants. Les informations relatives aux programmes de contrôle et aux facteurs de risque d'infection ont été recueillies à l'aide de l'outil de collecte des données. Les aspects clés et valeurs par défaut ont été quantifiés en vue d'être inclus dans le modèle STOC free. Un modèle de Markov caché dont les paramètres sont estimés par inférence bayésienne a été considéré comme le plus adapté et développé pour une application aux données issues des programmes de contrôle de la diarrhée virale bovine. Ce modèle a été testé et validé en utilisant des données réelles des programmes de contrôle du virus de la diarrhée virale bovine des pays participants ; le code informatique correspondant a été rendu public. Le modèle STOC free utilise des données au niveau des troupeaux, même si des données au niveau des animaux individuels peuvent être incluses une fois agrégées au niveau du troupeau. Le modèle STOC free s'applique aux maladies endémiques, puisqu'un certain niveau de présence de l'infection est nécessaire pour estimer les paramètres et permettre la convergence. Dans les pays ayant obtenu le statut indemne d'infection, un modèle du type arbre de scénario pourrait être un outil plus adapté. Des travaux supplémentaires sont recommandés pour généraliser le modèle STOC free à d'autres maladies.

Como parte del proyecto europeo STOC free (Surveillance Tool for Outcome-based Comparison of FREEdom from infection, herramienta de vigilancia para comparaciones por resultados respecto a la ausencia de infecciones, https://www.stocfree.eu), se confeccionó una herramienta de obtención de datos para facilitar la recogida normalizada de datos entrantes y se elaboró un modelo que posibilitara una comparación normalizada y armonizada de los resultados (datos salientes) de distintos programas de control de enfermedades bovinas. El modelo STOC free puede servir para calcular la probabilidad de ausencia de infección en los rebaños como parte de los programas de control y para determinar si estos programas se ajustan a las normas predefinidas de resultados de la Unión Europea. Como ejemplo de estudio para el proyecto se eligió el virus de la diarrea viral bovina (virus DVB) por la diversidad que presentaban los correspondientes programas de control de los seis países participantes. Empleando la herramienta de obtención de datos, se reunió información pormenorizada de los programas de control del virus DVB y los factores de riesgo. Para incluir los datos en el modelo STOC free, se cifraron unos aspectos clave y valores predeterminados Juzgando conveniente el empleo de un modelo oculto de Markov cuyos parámetros se estiman por inferencia bayesiana, se elaboró un modelo de esta índole aplicable a los programas de control del virus DVB. Para ensayar y validar el modelo se utilizaron datos reales de los programas de control del virus DVB de los países participantes, tras lo cual se hizo público el correspondiente código informático. El modelo STOC free trabaja con los datos por rebaño, aunque tras la agregación por rebaños pueden incluirse también datos por individuo. Para que este modelo sea aplicable a una enfermedad es preciso que esta sea endémica, pues el modelo requiere la presencia de cierto nivel de infección para calcular los parámetros y determinar convergencias. En aquellos países donde ya esté reconocida la ausencia de infección, sería más apropiado utilizar como herramienta un modelo de árbol de hipótesis. Los autores recomiendan ahondar en esta línea de trabajo para poder extender a otras enfermedades el uso del modelo STOC free.

Output-based assessment of herd-level freedom from infection in endemic situations: Application of a Bayesian Hidden Markov model.

Countries have implemented control programmes (CPs) for cattle diseases such as bovine viral diarrhoea virus (BVDV) that are tailored to each country-specific situation. Practical methods are needed to assess the output of these CPs in terms of the confidence of freedom from infection that is achieved. As part of the STOC free project, a Bayesian Hidden Markov model was developed, called STOC free model, to estimate the probability of infection at herd-level. In the current study, the STOC free model was applied to BVDV field data in four study regions, from CPs based on ear notch samples. The aim of this study was to estimate the probability of herd-level freedom from BVDV in regions that are not (yet) free. We additionally evaluated the sensitivity of the parameter estimates and predicted probabilities of freedom to the prior distributions for the different model parameters. First, default priors were used in the model to enable comparison of model outputs between study regions. Thereafter, country-specific priors based on expert opinion or historical data were used in the model, to study the influence of the priors on the results and to obtain country-specific estimates. The STOC free model calculates a posterior value for the model parameters (e.g. herd-level test sensitivity and specificity, probability of introduction of infection) and a predicted probability of infection. The probability of freedom from infection was computed as one minus the probability of infection. For dairy herds that were considered free from infection within their own CP, the predicted probabilities of freedom were very high for all study regions ranging from 0.98 to 1.00, regardless of the use of default or country-specific priors. The priors did have more influence on two of the model parameters, herd-level sensitivity and the probability of remaining infected, due to the low prevalence and incidence of BVDV in the study regions. The advantage of STOC free model compared to scenario tree modelling, the reference method, is that actual data from the CP can be used and estimates are easily updated when new data becomes available.

Evaluation of herd-level sampling strategies for control of Salmonella in Swedish cattle.

Based on Swedish legislation, all herds where Salmonella of any serotype is detected are put under restrictions, and measures aiming at eradication are required. Costs for sampling and control have increased in recent years and the aim of this study was to investigate the efficiency of different sampling strategies. We also compiled test results from recent surveillance activities and used these to complement and compare with calculated results. Sensitivities and specificities at group and herd level were calculated for different test strategies. A scenario-tree modeling approach was used to account for the hierarchy of animals within herds, and different relative risk of salmonella in different age groups. Negative and positive predictive values (NPV and PPV), and probability of freedom from Salmonella were calculated to compare the added value of different sampling strategies. Results showed that more fecal samples than serological samples per group are needed to reach a group sensitivity >0.50. This also means that serological testing leads to a higher NPV. For example, with 10 negative test-results from a group of 25 animals in a herd with a suspicion of Salmonella, the NPV based on serology was 0.75 and based on culture was 0.56. For the PPV, testing based on culture from fecal sampling was superior, as specificity of such testing was close to perfect. By changing the threshold for considering a group positive, from 1 test-positive animal to 2, the PPV of serological results could be increased without substantial loss in NPV. The herd sensitivity based on (1) bulk milk sampling, (2) fecal sampling of all animals, and (3) bulk milk sampling and individual sera from 20 animals within each age group was 0.53, 0.88, and 0.95, respectively. In low-prevalence regions, this sensitivity was enough to verify a high probability of freedom (>0.99), as the probability of infection in such Swedish regions has been shown to be 0.01. For herds with a higher prior probability of infection, repeated sampling (2-9 sampling occasions) was needed to reach the same level of confidence. Analysis of surveillance data indicated that boot swabs can be used to replace the standard fecal sampling presently used in Sweden. It was also confirmed that the individual specificity of the tests used for serological testing of Swedish calves is high (0.99). The results can form a basis for fit-for-purpose testing strategies (e.g., surveillance or prepurchase testing).

Serological testing of Schmallenberg virus in Swedish wild cervids from 2012 to 2016.

BACKGROUND: Schmallenberg virus (SBV) first emerged in Europe in 2011, and in Sweden in late 2012. The virus was still circulating in parts of Europe in 2015. In recent testing, the virus has not been detected in Swedish domestic animals, indicating that it is no longer circulating in Sweden. It is not known if the virus has circulated and is still circulating in Swedish wild cervid populations and whether wildlife can act as virus reservoirs. The aim of this study was to investigate whether SBV has circulated, and is still circulating among wild cervids in Sweden. RESULTS: Ninety-two sera from moose (Alces alces, n = 22), red deer (Cervus elaphus, n = 15), fallow deer (Dama dama, n = 44), and roe deer (Capreolus capreolus, n = 11) were collected and analyzed for antibodies against SBV. The sampling occurred in the southern and middle part of Sweden during three time periods: 1) before the vector season in 2012, 2) after the vector season in 2012, and 3) after the vector season in 2015. Animals from periods 1 and 2 were of varying ages, whereas animals collected in period 3 were born after the vector season 2013. Animals from period 1 (n = 15) and 3 (n = 47) were seronegative, but, 53% (16 of 30) of animals from period 2 were seropositive, determined by SBV competitive ELISA. Samples from period 2 were additionally analyzed for SBV-neutralizing antibodies. Such antibodies were detected in 16/16 SBV-N-antibody-positive, 3/12 negative and 2/2 doubtful sera. The two tests were in accordance at SBV-neutralizing antibody titers of 1:32 or higher. CONCLUSION: Our results show that SBV circulated among wild cervids during the vector season of 2012. Three years later, no SBV-antibodies were detected in animals born after the vector season 2013. The likely absence of SBV circulation in Sweden, in contrast to other parts of Europe, might be explained by the annual occurrence of a vector-free season due to climate conditions. Interpretations are limited by the small sample-size, but the results suggest that the SBV competitive ELISA has high specificity but might have slightly lower sensitivity compared to a seroneutralization assay, when using samples from wild cervids.

Disease Risk Analysis and Post-Release Health Surveillance for a Reintroduction Programme: the Pool Frog Pelophylax lessonae.

There are risks from disease in undertaking wild animal reintroduction programmes. Methods of disease risk analysis have been advocated to assess and mitigate these risks, and post-release health and disease surveillance can be used to assess the effectiveness of the disease risk analysis, but results for a reintroduction programme have not to date been recorded. We carried out a disease risk analysis for the reintroduction of pool frogs (Pelophylax lessonae) to England, using information gained from the literature and from diagnostic testing of Swedish pool frogs and native amphibians. Ranavirus and Batrachochytrium dendrobatidis were considered high-risk disease threats for pool frogs at the destination site. Quarantine was used to manage risks from disease due to these two agents at the reintroduction site: the quarantine barrier surrounded the reintroduced pool frogs. Post-release health surveillance was carried out through regular health examinations of amphibians in the field at the reintroduction site and collection and examination of dead amphibians. No significant health or disease problems were detected, but the detection rate of dead amphibians was very low. Methods to detect a higher proportion of dead reintroduced animals and closely related species are required to better assess the effects of reintroduction on health and disease.

Revisiting the phylogeography and demography of European badgers (Meles meles) based on broad sampling, multiple markers and simulations.

Although the phylogeography of European mammals has been extensively investigated since the 1990s, many studies were limited in terms of sampling distribution, the number of molecular markers used and the analytical techniques employed, frequently leading to incomplete postglacial recolonisation scenarios. The broad-scale genetic structure of the European badger (Meles meles) is of interest as it may result from historic restriction to glacial refugia and/or recent anthropogenic impact. However, previous studies were based mostly on samples from western Europe, making it difficult to draw robust conclusions about the location of refugia, patterns of postglacial expansion and recent demography. In the present study, continent-wide sampling and analyses with multiple markers provided evidence for two glacial refugia (Iberia and southeast Europe) that contributed to the genetic variation observed in badgers in Europe today. Approximate Bayesian computation provided support for a colonisation of Scandinavia from both Iberian and southeastern refugia. In the whole of Europe, we observed a decline in genetic diversity with increasing latitude, suggesting that the reduced diversity in the peripheral populations resulted from a postglacial expansion processes. Although MSVAR v.1.3 also provided evidence for recent genetic bottlenecks in some of these peripheral populations, the simulations performed to estimate the method's power to correctly infer the past demography of our empirical populations suggested that the timing and severity of bottlenecks could not be established with certainty. We urge caution against trying to relate demographic declines inferred using MSVAR with particular historic or climatological events.

Investigations and actions taken during 2011 due to the first finding of Echinococcus multilocularis in Sweden.

Echinococcus multilocularis is a parasite that can cause alveolar echinococcosis disease. After the first positive finding of E. multilocularis in Sweden in 2011, a consulting group with representatives from relevant authorities was summoned. In this group, all relevant information was shared, strategies for information dissemination and any actions to be taken due to the finding of E. multilocularis were discussed and decided. The present paper describes the actions taken during 2011 and the results thereof, including surveillance in animals, risk assessment for humans to become infected and recommendations given to the public. Further discussion about whether the parasite was introduced, and if so, how, as well as possible future development of the infection in animals and humans in Sweden and future actions are included.

First detection of Echinococcus multilocularis in Sweden, February to March 2011.

Surveillance for the fox tapeworm, Echinococcus multilocularis, has been carried out in Sweden since 2000, with about 300 red foxes analysed annually. We report the first finding of E. multilocularis in Sweden, in a fox shot in December 2010 in the south-west of the country. A second infected fox shot in the same location was detected in March 2011. This paper describes the national monitoring programme and the ongoing work to estimate the prevalence and spread of the infection.

Possible means of introduction of bluetongue virus serotype 8 (BTV-8) to Sweden in August 2008: comparison of results from two models for atmospheric transport of the Culicoides vector.

In September 2008, bluetongue virus serotype 8 (BTV-8) infection was detected for the first time in Sweden, in a dairy herd on the west coast. Two different previously published operational atmospheric dispersion models indicate that midges from infected regions in Europe are likely to have reached Sweden by atmospheric transport during an estimated infection window. Both models indicated that the likely dates for the incursion of midges were overnight on August 6 to 7 and August 14 to 15; however, the less constrained model indicated a number of additional possible dates. The distribution of infected herds detected by active surveillance coincides with the regions that were indicated by the models to have been reached by midges from regions in Denmark and Germany with infected herds. It is likely that several points of introduction of infected midges occurred, possibly on different occasions. No alternative routes for introduction of the infection to Sweden were identified, supporting the theory that BTV-8 was introduced by infected midges carried by the wind.

Reproductive maturation in the male Eurasian lynx (Lynx lynx): a study on 55 reproductive organs collected from carcasses during 2002-2005.

Data on reproductive physiology from the Eurasian lynx (Lynx lynx) are still scarce. The lynx is protected under Swedish hunting legislation. All lynx that are found dead or that are culled at hunting are to be sent to the Swedish National Veterinary Institute. In this study, we examined reproductive organs from 55 male lynx collected during the years 2002-2005. Age, body weight, testicular weight and volume, production of spermatozoa, and sperm viability were evaluated. The majority of the animals (39) had been killed in February and March, which is during the hunting season. The ages varied between 6 months and 17 years, body weight between 3.6 and 25.5 kg, and mean testes weight between 0.16 and 3.16 g. The gonadosomatic index was low compared with other species (approximately 0.02% in mature males). Mean testes weight differed significantly between males <12 months of age and all other age groups but did not differ between males of 18-23 months and older males. Spermatozoa could be collected but had lost most of their viability. Seven of 10 males of 18-23 months were fertile, as defined by the production of spermatozoa while no males < or =15 months of age were fertile. Adherence of the prepuce to the penis and absence of penile spines were associated with immaturity. The results indicate that most males are fertile during the reproductive season of their second year.

Unilateral failure of development of mandibular premolars and molars in an Eastern Grey kangaroo (Macropus giganteus) and its effects on molar progression.

An adult male Eastern Grey kangaroo from a wildlife reserve near Melbourne was submitted for necropsy examination and was discovered to have abnormal dentition. There was no evidence that any premolars or molars had ever been present on the right mandible, whilst the incisors were normal. The age of the kangaroo was estimated to be 1 year 9 months using the right maxillary molars and 2 years 4 months old using the contralateral side, presumably due to the asymmetry of the dental arcades. 'Lumpy jaw', a common periodontal disease of kangaroos, from which Bacteroides sp was cultured, was present on the base of the vertical ramus of the left mandible. Complete unilateral absence of premolar and molar teeth in the mandible of a kangaroo has not been described. This condition affected molar progression in both sets of maxillary molars.

Molecular characterization of Echinococcus isolates of cervid origin from Finland and Sweden.

The species Echinococcus granulosus is made up of several genotypic strain groups, whose taxonomical classification is still undetermined. Genotypes in the cervid-wolf life-cycle are poorly known, especially in Europe. In this study, 33 Echinococcus isolates from cervids from Finland and Sweden were characterized using mitochondrial ND1 gene sequencing. In addition, phylogenetic analysis of E. granulosus strains using the mitochondrial ATP6, ND1, ND3 and CO1 genes was performed using maximum likelihood, neighbour-joining and maximum parsimony methods. The Finnish and Swedish cervid isolates were found to represent the genotype G10. In the phylogenetic analyses, the camel (G6), pig (G7), cervid (G8) and Fennoscandian cervid (G10) strains clustered in a well-supported monophyletic group. This group differed clearly from the common sheep (G1) and horse (G4, 'E. equinus') strains, but was closely related to the cattle strain (G5, 'E. ortleppi'). Our results support the previous studies suggesting that the genotypes G6-10 should be separated from the species E. granulosus sensu stricto. However, additional morphological studies are needed, and the relationship to the cattle strain ('E. ortleppi') should be thoroughly evaluated before a final decision of the taxonomical status of the G6-10 group can be made.

Trichinella pseudospiralis foci in Sweden.

In Sweden, the prevalence of Trichinella infection in domestic pigs has greatly decreased since the 1970s, with no reports in the past 4 years. However, infected wild animals continue to be found. The objective of the present study was to identify the species of Trichinella present in animals of Sweden, so as to contribute to the knowledge on the distribution area and hosts useful for the prevention and control of this zoonosis. In the period 1985-2003, Trichinella larvae were detected in the muscles of 81/1800 (4.5%) red foxes (Vulpes vulpes), 1/6 (16.7%) arctic fox (Alopex lagopus), 1/7 (14.3%) wolf (Canis lupus), 10/200 (5.0%) lynxes (Lynx lynx), 4/8000 (0.05%) wild boars (Sus scrofa), and 27/66 x 10(6) (0.000041%) domestic pigs. All four Trichinella species previously found in Europe were detected (Trichinella spiralis, T. nativa, T. britovi and T. pseudospiralis). The non-encapsulated species T. pseudospiralis was detected in three wild boars from Holo (Stockholm area) and in one lynx from Froso (Ostersund area), suggesting that this species is widespread in Sweden. These findings are consistent with those of a study from Finland, both for the unexpected presence of T. pseudospiralis infection and the presence of the same four Trichinella species, suggesting that this epidemiological situation is present in the entire Scandinavian region. The widespread diffusion of T. pseudospiralis in the Scandinavian region is also important in terms of it potential impact on public health, given that human infection can occur and the difficulties to detect it by the trichinelloscopic examination.

Surgical implantation of radiotelemetry transmitters in European badgers (Meles meles).

The suitabilities of two subcutaneous and two intraabdominal surgical approaches were evaluated for implantation of telemetry transmitters in the European badger (Meles meles). Two transmitters, one for heart rate and the other for body temperature, were needed in each badger. Five wild badgers were trapped, housed in an outdoor pen, anesthetized, and surgically implanted with one or two transmitters per procedure. A total of 16 transmitters were implanted, 10 subcutaneously and six intraabdominally, and each badger had up to three procedures performed. Six subcutaneous transmitters were placed over the chest wall and four on the dorsal aspect of the neck. Three intraabdominal transmitters were placed free in the abdomen and three were fixed to the abdominal wall. Eight of the subcutaneous transmitters were later damaged or dislodged, probably by mechanical friction, injuries from fighting, or wound licking. The intraabdominal implantation procedures were easier and faster to perform compared with subcutaneous procedures. Two of the free intraabdominal transmitters became thinly encapsulated, with some omental adhesions. The fixed intraabdominal approach for transmitter implantation is preferable in badgers.

Conduction Block Analgesia in the Mandible: A comparative investigation of the techniques of Fischer and Gow-Gates.

An investigation has been made in order to compare two different intraoral methods for mandibular block analgesia. The first method (according to Fischer) anesthetizes the inferior alveolar and lingual nerves just above the level of the mandibular foramen and the other method (according to Gow-Gates) anesthetizes the mandibular nerve just after leaving the ovale foramen. The latter method should, with only one injection, give complete analgesia within the entire distribution zone of the sensory components of the mandibular nerve.The investigation has been performed on 12 test subjects, who each have achieved mandibular block analgesia according to the two methods mentioned above. 1.8 ml lidocaine-adrenaline (20 mg per ml + 12.5 mug per ml) has been used and the analgesic effect has been tested in the pulps and in the vestibular and lingual mucosa at different time intervals. The investigation shows that the onset of analgesia and complete analgesia was more rapid with the first method than with Gow-Gates method.Furthermore Gow-Gates' method shows no superiority in anesthetizing the long buccal nerve without separate injection for this nerve.However, regarding the analgesic effect only, there has been no statistically significant difference between the two methods with this small number of test subjects.

Conduction block analgesia in the mandible. A comparative investigation of the techniques of Fischer and Gow-Gates.

An investigation has been made in order to compare two different intraoral methods for mandibular block analgesia. The first method (according to Fischer) anesthetizes the inferior alveolar and lingual nerves just above the level of the mandibular foramen and the other method (according to Gow-Gates) anesthetizes the mandibular nerve just after leaving the oval foramen. The latter method should, with only one injection, give complete analgesia within the entire distribution zone of the sensory components of the mandibular nerve. The investigation has been performed on 12 test subjects, who each have achieved mandibular block analgesia according to the two methods mentioned above. 1.8 ml lidocaine-adrenaline (20 mg per ml + 12.5 microgram per ml) has been used and the analgesic effect has been tested in the pulps and in the vestibular and lingual mucosa at different time intervals. The investigation shows that the onset of analgesia and complete analgesia was more rapid with the first method than with Gow-Gates' method. Furthermore Gow-Gates' method shows no superiority in anesthetizing the long buccal nerve without separate injection for this nerve. However, regarding the analgesic effect only, there has been no statistically significant difference between the two methods with this small number of test subjects.