Paratuberculosis, caused by Mycobacterium avium subsp. paratuberculosis (Map), is a chronic incurable infection of intestinal tract of animals. Molecular characterization of Map isolates classifies them into two major groups, ‘Cattle’ or Type II and ‘Sheep’ or Type I/III with a different phenotype, epidemiology, virulence and pathogenesis. The aim of this study was to examine 192 Map ELISA-positive sheep and goats from Cyprus using faecal culture and genotype Map isolates using IS1311 PCR and restriction endonuclease analysis (IS1311 PCR-REA) with HinfI restriction enzyme. Map was isolated from only four (4.6%) faecal samples out of 88 sheep and 15 (14.4%) faecal samples out of 104 goats. Genotyping of the isolates using IS1311 PCR-REA revealed that sheep and goat populations on the island are infected primarily by ‘Sheep’ strains. Only three Map isolates from goats originated from one farm were characterized as ‘Cattle’ strains.

Bovine ephemeral fever (BEF) is an economically important vector-borne viral disease of cattle and buffalo. It has been reported from most of the world's tropical and subtropical regions. In the last few decades, outbreaks of BEF have occurred in Israel almost every other year. Several serological studies have demonstrated a wide range of wild animal species that are positive for BEF virus (BEFV) antibodies. However, the question of whether wild animals and domesticated species other than cattle also play an important role in the maintenance and transmission of BEFV in Israel remains. Here, we examined the prevalence of anti-BEFV antibodies in 942 samples collected from various wild, semi-captive and domesticated animal species during the years 2000–2009 using the serum neutralization (SN) method. SN test revealed the presence of BEFV-neutralizing antibodies in nine samples (0.96%), from three species: Bubalus bubalis (4/29, 13.79%), Gazella g. gazella (3/68, 4.44%) and Dama d. mesopotamica (2/296, 0.68%). All positive samples were collected in areas of earlier outbreaks. The low prevalence of positive animals and the solid correlation with prior outbreaks indicate that the tested species probably do not serve as virus reservoirs and may play only a minor role in the maintenance of BEFV in the Middle East.

We developed a model to quantify the effect of factors influencing the spatio-temporal distribution of foot-and-mouth disease (FMD) in Tanzania. The land area of Tanzania was divided into a regular grid of 20 km × 20 km cells and separate grids constructed for each of the 12-month periods between 2001 and 2006. For each year, a cell was classified as either FMD positive or negative dependent on an outbreak being recorded in any settlement within the cell boundaries. A Bayesian mixed-effects spatial model was developed to assess the association between the risk of FMD occurrence and distance to main roads, railway lines, wildlife parks, international borders and cattle density. Increases in the distance to main roads decreased the risk of FMD every year from 2001 to 2006 (ORs ranged from 0.43 to 0.97). Increases in the distance to railway lines and international borders were, in general, associated with a decreased risk of FMD (ORs ranged from 0.85 to 0.99). Increases in the distance from a national park decreased the risk of FMD in 2001 (OR 0.80; 95% CI 0.68–0.93) but had the opposite effect in 2004 (OR 1.06; 95% CI 1.01–1.12). Cattle population density was, in general, positively associated with the risk of FMD (ORs ranged from 1.01 to 1.30). The spatial distribution of high-risk areas was variable and corresponded to endemic (2001, 2002 and 2005) and epidemic (2003, 2004 and 2006) phases. Roads played a dominant role in both epidemiological situations; we hypothesize that roads are the main driver of FMD expansion in Tanzania. Our results suggest that FMD occurrence in Tanzania is more related to animal movement and human activity via communication networks than transboundary movements or contact with wildlife.

Bluetongue virus serotype 8 (BTV-8) was responsible for a large outbreak among European ruminant populations in 2006–2009. In spring 2008, a massive vaccination campaign was undertaken, leading to the progressive disappearance of the virus. During surveillance programmes in Western Europe in 2010–2011, a low but significant number of animals were found weakly positive using BTV-specific real-time RT-PCR, raising questions about a possible low level of virus circulation. An interference of the BTV-8 inactivated vaccine on the result of the real-time RT-PCR was also hypothesized. Several studies specifically addressed the potential association between a recent vaccination and BTV-8 RNA detection in the blood of sheep. Results were contradictory and cattles were not investigated. To enlighten this point, a large study was performed to determine the risks of detection of bluetongue vaccine-associated RNA in the blood and spleen of cattle using real-time RT-PCR. Overall, the results presented clearly demonstrate that vaccine viral RNA can reach the blood circulation in sufficient amounts to be detected by real-time RT-PCR in cattle. This BTV-8 vaccine RNA carriage appears as short lasting.

Encephalitozoon cuniculi is an obligate intracellular microsporidian that is the causal agent of encephalitozoonosis, an important and emerging disease in both humans and animals. Little is known about its occurrence in wildlife. In this study, serum samples from 793 wild rodents [178 bank voles (BV), 312 field voles (FV) and 303 wood mice (WM)], 96 foxes and 27 domestic cats from three study areas in the UK were tested for the presence of antibodies to E. cuniculi using a direct agglutination test (DAT). Seroprevalence in the wild rodents ranged from 1.00% to 10.67% depending on species (overall 5.31%) and was significantly higher in foxes [49.50% (50/96)]. None of the 27 cats sampled were found to be seropositive. This is the first report of seroprevalence to E. cuniculi in BV, FV, WM, foxes and cats in the UK and provides some evidence that foxes could act as sentinels for the presence of E. cuniculi in rodents. The study demonstrates that wildlife species could be significant reservoirs of infection for both domestic animals and humans.

Prevalence of the protozoan Perkinsus spp. in the gills of the pleasure oyster Crassostrea corteziensis from two estuaries in Nayarit, Mexico, was measured. The protozoan was identified by PCR amplification of the internal transcribed spacer (ITS) region of the rDNA of Perkinsus spp. The pathogen was found in 92% of oysters from Boca de Camichín and 77% of oysters from Pozo Chino. ITS sequences characterized from C. corteziensis showed 96–100% similarity to Perkinsus marinus. The most frequent ITS sequence (GenBank JQ266236) had 100% identity with the ITS locus of P. marinus from New Jersey, Maryland, South Carolina and Texas, and the second most frequent observed sequence (GenBank JQ266240) was 100% identical to ITS sequences of P. marinus from New Jersey, South Carolina, Louisiana, and Bahía Kino, Sonora, Mexico. The 14 sequences from the non-transcribed spacer (NTS) showed 98% similarity to P. marinus from Texas. The most frequent polymorphism identified was at nucleotide 446 of the ITS region; however, the NTS showed the highest nucleotide diversity, thereby suggesting that this region is suitable for genotype identification. Moreover, the most conserved ITS marker is better for species-specific diagnosis. Both the ITS and NTS sequences of P. marinus obtained from C. corteziensis were grouped in two clades, identifying two allelic variants of P. marinus.

Two Bangladeshi isolates of Newcastle disease virus (NDV), one from a chicken and one from a pigeon, were characterized in this study. Pathogenicity of the isolates was evaluated on the basis of intracerebral pathogenicity index (ICPI). Both the isolates were found to be of velogenic pathotype having ICPI of 1.83 and 1.51 for the chicken and pigeon isolate, respectively. Genotype of the isolates was determined by phylogenetic analysis based on partial F gene sequences. A 766-bp genome fragment spanning partial M and F gene was amplified by RT-PCR and sequenced. The first 354 bp of the coding region of F gene and corresponding deduced amino acid sequences (residues 1–118) of these two NDV isolates were aligned with that of other NDV strains retrieved from GenBank. A phylogenetic tree constructed from the alignment showed that the chicken isolate (BD-C162) belonged to the newly described genotype XIII and the pigeon isolate (BD-P01) to genotype VI. Both the chicken and pigeon isolates possessed a virulent-like fusion protein cleavage site ¹¹²RRQKRF¹¹⁷.

The potential control efficacy of Aeromonas phage PAS-1 was evaluated against Aeromonas salmonicida subsp. salmonicida infection in rainbow trout (Oncorhynchus mykiss) model in this study. The phage was co-cultured with the virulent A. salmonicida subsp. salmonicida strain AS05 that possesses the type III secretion system (TTSS) ascV gene, and efficient bacteriolytic activity was observed against the bacteria. The administration of PAS-1 in rainbow trout demonstrated that the phage was cleared from the fish within 200 h post-administration, and a temporal neutralizing activity against the phage was detected in the sera of phage-administrated fish. The administration of PAS-1 (multiplicity of infection: 10 000) in A. salmonicida subsp. salmonicida infected rainbow trout model showed notable protective effects, with increased survival rates and mean times to death. These results demonstrated that Aeromonas phage PAS-1 could be considered as an alternative biological control agent against A. salmonicida subsp. salmonicida infections in rainbow trout culture.

Peste des petits ruminants (PPR) is an endemic disease of small ruminants, and vaccination has been the method of control but outbreaks are continuously occurring in Pakistan. The following study presents a detailed investigation of an outbreak, suspected to be PPR, probably introduced by PPRV-infected sheep and goats from Sindh Province (north-west) to Punjab Province (central) of Pakistan during the flood relief campaign in 2011. A total of 70 serum samples from 28 different flocks were tested with competitive ELISA (H antibodies), which detected 24 (34.2%) samples positive for PPRV antibodies. Nasal swabs and faeces were tested with immunocapture ELISA (N antigen), which detected 18 (25.7%) samples positive for PPRV antigen. The RNA detected positive (n = 28, 40%) using real-time PCR was subjected to conventional PCR for the amplification of the fusion and nucleoprotein genes. Sequencing of both genes and subsequent phylogenetic analysis indicated the grouping of all the sequences to be in lineage IV along with other Asian isolates of PPRV. However, sequences of both genes were divided into two groups within lineage IV. One group of viruses clustered with previously characterized Pakistani isolates, whereas the other group was distinctly clustered with isolates from the Middle East or India. The sequence identity indicated the introduction of at least one population of PPRV from a different source and circulation in the local flocks of small ruminants, which emphasized the need to obtain health clearance certificate before movement of animals. The results of this study provide baseline data for the genetic characterization of different PPRV populations in Pakistan.

Emerging infectious diseases affect the health of animal and human populations, but the impact goes beyond health as it extends to political, economic, social and environmental domains, as well as inter-state relations. Deeper understanding of these impacts aids public health authorities in their duties of protection and improvement of the health of their communities, promotion of healthy practices and research on disease, injury and threat prevention and mitigation. This empirical essay gathers insights from Cambodia, Hong Kong and Indonesia as they attempt to design and implement control and surveillance systems against avian influenza – an infectious disease.

Rift Valley fever (RVF) is an emerging zoonotic mosquito-borne infectious disease that has been identified as a risk for spread to other continents and can cause mass livestock mortality. In equatorial Africa, outbreaks of RVF are associated with high rainfall, when vector populations are at their highest. It is, however, unclear how RVF virus persists during the inter-epidemic periods and between seasons. Understanding inter-epidemic persistence as well as the role of vectors and hosts is paramount to creating effective management programmes for RVF control. We created a mathematical model for the spread of RVF and used the model to explore different scenarios of persistence including vertical transmission and alternate wildlife hosts, with a case study on buffalo in Kruger National Park, South Africa. Our results suggest that RVF persistence is a delicate balance between numerous species of susceptible hosts, mosquito species, vertical transmission and environmental stochasticity. Further investigations should not focus on a single species, but should instead consider a myriad of susceptible host species when seeking to understand disease dynamics.

Development of point of concentration (POC) surveillance strategies for bovine tuberculosis (bTB) would facilitate global efforts to eradicate bTB. The interferon-gamma (IFNγ) assay can detect IFNγ responses to Mycobacterium bovis in blood collected at commencement of exsanguination (COE) of experimentally challenged cattle but has not been evaluated under field conditions. The current study was aimed at determining (i) whether blood collected at COE of cattle at slaughter, under field conditions, is practical to obtain and useful for identifying cattle as IFNγ positive for bTB, (ii) whether the results of the IFNγ assay obtained at COE reliably compare with results obtained from live animals in the field, and (iii) whether the identified animal(s) originated from bTB-infected or bTB-exposed herds. Cattle from three risk groups were used: the highest risk group consisted of 49 cattle from 3 bTB-infected herds; the medium risk group consisted of 24 cattle from a potentially exposed herd; and the lowest risk group consisted of 60 cattle from herds with no known history of bTB exposure. The IFNγ assay was performed on blood collected both before stunning and at COE of cattle at slaughter. An enhanced slaughter inspection for gross lesions consistent with bTB was performed on all cattle. In addition, lymph nodes were cultured for M. bovis for cattle that tested positive for bTB via the IFNγ assay and for most cattle that tested negative for bTB. Cattle, both with and without lesions consistent with bTB, were identified as positive for bTB by the IFNγ assay using blood collected at COE, but none of the positive cattle originated from the lowest risk group. The current study demonstrates that blood collected at COE of cattle is both a practical and moderately reliable sample for accessing bTB infection using the IFNγ assay.

Non-tuberculous mycobacteria (NTM) are widely distributed in the environment, particularly in wet soil, marshland, rivers or streams, but also are causative agents of a wide variety of infections in animals and humans. Little information is available regarding the NTM prevalence in wildlife and their effects or significance in the bovine tuberculosis (bTB) epidemiology and diagnosis. This research shows the most frequently NTM isolated in lymph nodes of wild boar (Sus scrofa) from southern Spain, relating the NTM presence with the individual characteristics, the management of animals and the possible misdiagnosis of Mycobacterium bovis in concurrent infections. A total of 219 NTM isolates were obtained from 1249 wild boar mandibular lymph nodes sampled between 2007 and 2011. All but 75 isolates were identified by the PCR-restriction analysis-hsp65, and a partial sequencing of the 16S rDNA was carried out to identify the rest of the isolates. Results showed that Mycobacterium chelonae was the most frequently isolated NTM specie (133 isolates, 60.7%), followed by Mycobacterium avium (24 isolates, 11%). No relation was found regarding sex, body condition and management, but M. chelonae was more frequently detected in adults, whereas M. avium was more prevalent in subadults. The high NTM prevalence observed in the studied wild boar populations could make difficult the bTB diagnostic.

Four-hundred and forty-two F4+ pathogenic Escherichia coli were isolated in a period of 10 years (2002–2011), from pigs that were suffering from diarrhoea belonging to Italian swine herds. The strains were analysed for their susceptibility to 12 antimicrobials using the disc diffusion method. During the study period, a statistically significant proportion of isolates resistant to enrofloxacin (14.5–89.3%), marbofloxacin (5.4–60.7%), flumequine (49.1–92.9%), danofloxacin (21.6–80%), florfenicol (9.8–64.3%), thiamphenicol (50–92%) and cefquinome (3.8–44%) was recorded. An increase in resistance (not statistically significant) to gentamicin (63.6–85.7%), apramycin (61.8–82.1%), trimethoprim-sulphamethoxazole (75–89.3%), tetracycline (97–100%) and erythromycin (92.4–100%) was also observed. Based on antimicrobial multiresistance, the strains were collected into three groups: I. resistant to 2–5 antimicrobials; II. resistant to 6–8 antimicrobials; III. resistant to 9–12 antimicrobials. The number of isolates belonging to the first group showed a statistically significant decrease (P < 0.05; R² = 0.896; r = −0.9608), while the isolates belonging to the second and third groups showed a statistically significant increase in resistance (P < 0.05; R² = 0.753; r = 0.8890 and P < 0.05; R² = 0.727; r = 0.8701, respectively) over the period of study. The results of this study suggest the need for continued monitoring of the development of resistance.

West Nile fever (WNF) is a viral disease of wild birds transmitted by mosquitoes. Humans and equids can also be affected and suffer from meningoencephalitis. In Tunisia, two outbreaks of WNF occurred in humans in 1997 and 2003; sporadic cases were reported on several other years. Small-scale serological surveys revealed the presence of antibodies against WN virus (WNV) in equid sera. However, clinical cases were never reported in equids, although their population is abundant in Tunisia. This study was achieved to characterize the nationwide serological status of WNV in Tunisian equids. In total, 1189 sera were collected in 2009 during a cross-sectional survey. Sera were tested for IgG antibodies, using ELISA and microneutralization tests. The estimated overall seroprevalence rate was 28%, 95% confidence interval [22; 34]. The highest rates were observed (i) in the north-eastern governorates (Jendouba, 74%), (ii) on the eastern coast (Monastir, 64%) and (iii) in the lowlands of Chott El Jerid and Chott el Gharsa (Kebili, 58%; Tozeur, 52%). Environmental risk factors were assessed, including various indicators of wetlands, wild avifauna, night temperature and chlorophyllous activity (normalized difference vegetation index: NDVI). Multimodel inference showed that lower distance to ornithological sites and wetlands, lower night-time temperature, and higher NDVI in late spring and late fall were associated with higher serological prevalence rate. The model-predicted nationwide map of WNF seroprevalence rate in Tunisian equids highlighted different areas with high seroprevalence probability. These findings are discussed in the perspective of implementing a better WNF surveillance system in Tunisia. This system might rely on (i) a longitudinal survey of sentinel birds in high-risk areas and time periods for WNV transmission, (ii) investigations of bird die-offs and (iii) syndromic surveillance of equine meningoencephalitis.

Canine parvovirus causes serious disease in dogs. Study of the genetic variation in emerging CPV strains is important for disease control strategy. The antigenic property of CPV is connected with specific amino acid changes, mainly in the capsid protein VP2. This study was carried out to characterize VP2 gene of CPV viruses from two provinces of China in 2011. The complete VP2 genes of the CPV-positive samples were amplified and sequenced. Genetic analysis based on the VP2 genes of CPV was conducted. All of the isolates screened and sequenced in this study were typed as CPV-2a except GS-K11 strain, which was typed as CPV-2b. Sequence comparison showed nucleotide identities of 98.8–100% among CPV strains, whereas the Aa similarities were 99.6–100%. Compared with the reference strains, there are three distinctive amino acid changes at VP2 gene residue 267, 324 and 440 of the strains isolated in this study. Of the 27 strains, fourteen (51.85%) had the 267 (Phe-Tyr) and 440 (Thr-Ala) substitution, all the 27 (100%) had 324 (Tyr-Ile) substitution. Phylogenetically, all of the strains isolated in this study formed a major monophyletic cluster together with one South Korean isolate, two Thailand isolates and four Chinese former isolates.

Streptococcus suis (S. suis) can be classified into 33 serotypes based on the structure of capsular polysaccharides. Recent research indicated that a new surface protein designated as Sao (surface antigen one) reacts with 30 serotypes of convalescent-phase sera during S. suis infections, which makes Sao a good potential antigen for developing S. suis vaccines. The objectives of this study were to produce recombinant Sao-L protein (rSao-L) from a strain of S. suis serotype 2 by a prokaryotic expression system in bioreactors and to use rSao-L as the antigen for a S. suis vaccine in mouse and swine models. The antibody titres in mice and pigs immunized with rSao-L were significantly (P < 0.05) increased. After challenge with live S. suis serotype 1 bacteria, the anatomical lesions in pigs immunized with rSao-L were reduced by 60%. These data indicated that immunization with rSao-L can confer cross-serotype protection against S. suis. Moreover, percentages of CD8⁺ and CD4⁺/CD8⁺ double-positive T cells in immunized pigs were significantly higher than those of the control group (P < 0.01). Using bioreactors to produce rSao-L as the antigen for S. suis vaccines may broaden protective efficacy and reduce production costs.

Foot-and-mouth disease (FMD) is endemic in Eritrea and in most parts of Africa. To be able to control FMD using vaccination, information on the occurrence of various foot-and-mouth disease serotypes in Eritrea is needed. In this cross-sectional study, 212 sera samples were collected from FMD infected and recovered animals in Eritrea. These samples were tested for the presence of antibodies against FMD non-structural proteins (NSP) and neutralizing antibodies against six of the seven (all but SAT 3) serotypes of FMD virus (FMDV). Of these, 67.0% tested positive to non-structural protein antibodies in the FMD NS ELISA. By virus neutralization, FMDV serotype O antibodies were shown to be the most dominant (approximately 50%). Virus neutralization test results indicate that infection with serotype C and SAT 1 might have occurred, although there are no reports of isolation of these two serotypes. Because the samples were not randomly selected, further random serological surveillance in all age group animals is necessary both to estimate the prevalence of FMD in the country and to confirm the serological results with serotype C and SAT 1.

Q fever is a zoonosis occurring worldwide in livestock. Often neglected in differential diagnoses, Q fever can persist in herds causing financial losses in the long run. In ruminants, well-known manifestations of Q fever are abortion, stillbirth, delivery of weak offspring and premature delivery. In cattle, Q fever is frequently asymptomatic and/or under-reported. The use of new methodologies in veterinary clinical epidemiology is of prime importance to find accurate clinical indicators of exposure to C. burnetii at herd level. A retrospective randomly cross-sectional survey was conducted to estimate the seroprevalence of Q fever in southern Belgium by means of an ELISA test performed on the bulk tank milk (n = 206 cattle herds). At the same time, a questionnaire was accomplished allowing the investigation of presumptive clinical signs observed at herd level during the previous twelve months for dairy cows. A multivariate logistic regression analysis was used to identify abortion and irregular repeat breeding as two indicators associated with Q fever exposure in dairy herds. In addition, a bootstrapped quantile regression revealed that the average score of putative clinical signs related to Q fever was significantly more important in exposed versus non-exposed herds. A classification and regression tree (CART) analysis confirmed the importance of the average clinical score and the irregular repeat breeding as main splitters, considering or not each clinical sign separately. Considering herd clinical patterns, instead of taking each clinical sign separately, seems to be more useful to differentiate herds at risk of Q fever exposure.

A case–control study conducted during 2011 involved 90 randomly selected commercial layer farms infected with highly pathogenic avian influenza type A subtype H5N1 (HPAI) and 175 control farms randomly selected from within 5 km of infected farms. A questionnaire was designed to obtain information about potential risk factors for contracting HPAI and was administered to farm owners or managers. Logistic regression analyses were conducted to identify significant risk factors. A total of 20 of 43 risk factors for contracting HPAI were identified after univariable logistic regression analysis. A multivariable logistic regression model was derived by forward stepwise selection. Both unmatched and matched analyses were performed. The key risk factors identified were numbers of staff, frequency of veterinary visits, presence of village chickens roaming on the farm and staff trading birds. Aggregating these findings with those from other studies resulted in a list of 16 key risk factors identified in Bangladesh. Most of these related to biosecurity. It is considered feasible for Bangladesh to achieve a very low incidence of HPAI. Using the cumulative list of risk factors to enhance biosecurity pertaining to commercial farms would facilitate this objective.

A longitudinal study has been conducted in the provinces of Sindh, Punjab and Islamabad Capital Territory area, Pakistan, to evaluate the impact of foot-and-mouth disease on milk yield in a sample of farmers owning cattle and buffaloes. The sample consisted of 50 farms where the presence of foot-and-mouth disease (FMD) virus was initially suspected on the basis of clinical signs and subsequently confirmed through either a field test or laboratory confirmation. In each farm, the total number of clinical cases was registered, and clinically diseased milking cattle and buffaloes were followed up for the next 60 days from the onset of clinical signs and the amount of milk yield measured.

The average milk yield, estimated to be around 10 l per animal before the onset of FMD, decreased significantly in the 2 months following the onset of acute clinical disease. The loss of milk production in the 60 days following the onset of clinical signs was estimated to be around 220 and 201 l for cattle and buffaloes, respectively. Under the assumption that the administration of a good-quality vaccine matching circulating FMD strains could protect against clinical disease, the benefit/cost ratio for having all animals vaccinated in all 50 farms was estimated to be 5.7.

Recent European contingency plans envisage emergency vaccination as an animal-friendly control strategy for foot-and-mouth disease (FMD). Anti-viral drugs may be used as an alternative or complementary measure. We here demonstrate that the nucleoside analogue 2′-C-methylcytidine (2′CMC) protects severe combined immunodeficient (SCID) mice against lethal FMD virus infection. In brief, SCID mice were inoculated with serotype A FMD virus and treated for five consecutive days with 2′CMC. All 15 treated mice remained healthy until the end of the study at 14 days post-infection (dpi). At that time, viral RNA was no longer detected in 13 of 15 treated mice. All eight untreated mice suffered from an acute generalized disease and were euthanized for ethical reasons on average at 4 dpi. These results illustrate the potential of small molecules to control FMD.

Outbreaks of bluetongue disease have occurred in Spain six times and have been caused by the following serotypes of bluetongue virus (BTV), in chronological order: BTV10, BTV2, BTV4, BTV1 and BTV8. Serotypes BTV1, BTV2 and BTV4 may have entered the country in Culicoides transported by wind; BTV8 via infected animal movements; and BTV10 across the Portuguese border. The evolution of each serotype has been different: BTV1, BTV4 and BTV10 spread throughout mainland Spain; BTV2 did not spread from the Balearic Islands to the Iberian Peninsula; and BTV8 has proven very poor at spreading throughout mainland Spain. The significant economic impact of the disease has led authorities to adopt control and eradication measures, which have evolved as new diagnostic tools and vaccines have become available. This review describes BTV infection in Spain, and it focuses on the clinical disease produced by each serotype, the Culicoides species which were present at what time, the origin of the virus and the control measures adopted. In the field, it has proven necessary to vaccinate livestock against each new BTV serotype as it arrived. Therefore, future eradication strategies should focus on developing polyvalent vaccines and vaccines that allow the differentiation of infected and vaccinated animals. As of 1 January 2013, the Iberian Peninsula is considered a restricted area for BTV1, and a small zone in southern Spain is a restricted area for BTV4, which includes the little BTV8 restricted area. Serotypes BTV1 and BTV4 were detected in sentinel animals in January and November and in March 2012, respectively. The last BTV8 positive animal was detected in November 2010, which implies that in the coming months, Spain may be declared free of BTV8.

Giardia duodenalis is a common intestinal parasite in humans and a wide range of livestock species. It is a genetically heterogeneous parasite that has been characterized in seven distinct genetic assemblages or cryptic species, and molecular markers can be used to differentiate both animal-specific and potentially zoonotic genotypes. Little is known about G. duodenalis and the range of assemblages occurring in domestic livestock species in the UK. Here, we present data on the occurrence and molecular diversity of G. duodenalis detected in the faeces or large intestinal contents of cattle, sheep, pigs, goats and camelids from farms in the north-west of England. Both healthy and clinically diseased animals were included in the survey. The presence of Giardia spp. and assemblages was determined by sequencing of the small-subunit ribosomal RNA gene. The potential association of infection with various clinical and epidemiological parameters was studied in cattle using both univariate and multivariate analyses. Giardia spp. were detected in 127 (34.3%) of the 370 animals tested. G. duodenalis assemblage E was found to be predominant in cattle and sheep, followed by assemblage A. Mixed infections with assemblages A and E were also detected. Interestingly, some cattle, sheep and pigs were found to be infected with more unexpected assemblages (C, D, F). Pre-weaned calves were more likely to test positive than adult animals, but no association between the occurrence of overt intestinal disease and G. duodenalis infection was detected. The common occurrence of assemblage A and the finding of unusual assemblages in atypical hosts suggest that in future, a multilocus analysis should be used to confirm the actual diversity of G. duodenalis in livestock and the presence of potentially zoonotic genotypes. These data also suggest that there is a need to re-evaluate the clinical significance of G. duodenalis infection in livestock.

The aim of the study was to assess whether blood samples collected onto FTA^® cards could be used in combination with real-time PCR for the detection of African swine fever virus (ASFV) DNA in samples from resource-poor settings under the assumption that asymptomatically (sub-clinically) infected pigs may be present. Blood samples were collected from clinically healthy pigs from Mbeya Region, Tanzania. The blood samples were stored on FTA^® cards and analysed by real-time PCR assays in duplicate; three pigs had high levels of viral DNA (Ct values of 27–29), and three pigs had a low level of viral DNA (Ct 36–45). Four pigs were positive in one of the duplicate samples only, but clear products of the expected size were obtained when the reactions were analysed by gel electrophoresis. For comparison, blood samples from pigs experimentally infected with either a pathogenic (OURT T88/1) or a non-pathogenic (OURT T88/3) isolate of ASFV were collected, stored on FTA^® cards and analysed in the same way. The blood from pigs infected with the OURT T88/1 isolate showed high levels of viral DNA (Ct 22-33), whereas infection with non-pathogenic OURT T88/3 isolate resulted in only low levels of viral DNA (Ct 39) in samples collected at 10–14 days after inoculation.

Streptococcus agalactiae (Group B streptococcus, GBS) has emerged as an important pathogen that affects humans and animals, including aquatic species. In August 2011, a severe infectious disease affecting rabbits, which caused 42% mortality, occurred in Mianyang, Sichuan Province, China. The main clinical signs included acute respiratory distress syndrome, fever, paddling and convulsions. A Gram-positive, chain-forming coccus was isolated from the primary organs and tissues of diseased rabbits and then identified as S. agalactiae by morphology, biochemical and physiological characteristics, 16S rDNA and gyrB gene sequences analysis. All isolates of S. agalactiae showed a similar antibiotic susceptibility, which were sensitive to florfenicol, ampicillin,gentamicin and norfloxacin, as well as being resistant to penicillin, amoxicillin and tetracycline. To our knowledge, this is the first report on S. agalactiae natural infection in domestic rabbits.

Animal health surveillance programmes may change in response to altering requirements or perceived weaknesses but are seldom subjected to any formal evaluation to ensure that they provide valuable information in an efficient manner. The literature on the evaluation of animal health surveillance systems is sparse, and those that are published may be unstructured and therefore incomplete. To address this gap, we have developed SERVAL, a SuRveillance EVALuation framework, which is novel and aims to be generic and therefore suitable for the evaluation of any animal health surveillance system. The inclusion of socio-economic criteria ensures that economic evaluation is an integral part of this framework. SERVAL was developed with input from a technical workshop of international experts followed by a consultation process involving providers and users of surveillance and evaluation data. It has been applied to a range of case studies encompassing different surveillance and evaluation objectives. Here, we describe the development, structure and application of the SERVAL framework. We discuss users' experiences in applying SERVAL to evaluate animal health surveillance systems in Great Britain.

This study demonstrated the prevalence of Porcine circovirus type 2 (PCV2) among pig farms in Vietnam. Analyses of the genome, capsid protein and phylogeny classified all 30 Vietnamese PCV2 strains as the PCV2b genotype, belonging to the clusters of 1A, 1B, 1C and recombinant forms. Each viral genome was 1767 nucleotides long and shared 96.0–100% nucleotide sequence identity. The amino acid substitutions in the capsid protein of the Vietnamese PCV2 strains were in immunodominant regions, and the majority of strains (24/30) contained a lysine extension at the C-terminus. Bayesian phylogeographic analysis revealed epidemic links of the PCV2 recombinant cluster within and among countries, which supports a circulating recombinant form of PCV2. Further analysis by the Jameson–Wolf antigenic index indicated antigenic alterations at important sites in the capsid protein (sites 131–133) among the recombinant cluster and the other clusters of PCV2b.

Bluetongue (BT) was monitored in wildlife in France during two consecutive years corresponding to contrasting incidence rates in livestock: in 2008 at the peak of domestic outbreaks and in 2009 when very few outbreaks were observed. The disease status of 2 798 ruminants comprising 837 red deer (Cervus elaphus) was explored using ELISA test on serum and real-time RT-PCR test on blood or spleen. A large proportion of red deer were seropositive and positive to RT-PCR in 2008, but also in 2009 (seroprevalence: 47.1% and 24.3%), suggesting that red deer could maintain infection when domestic incidence was negligible. By contrast, low seroprevalence (<3%) and few RT-PCR positive results were observed in other wild ruminant species, which rather appeared thus as dead-end hosts. The risk factors of bluetongue circulation during the periods of high (2008) and low (2009) domestic incidence were explored in red deer using logistic mixed models. In this species, prevalence has been mainly influenced by the initial peak of BT in livestock, but also by environmental factor such as elevation and edge density between forest and pastures. Surprisingly, cattle density has a negative influence on prevalence in red deer, possibly due to the protective effect of cattle regarding midges' bites and/or to still unexplained factors dealing with the host/midge interface. To our knowledge, this study is the first attempt at measuring the effect of landscape and wildlife/domestic interface on BT prevalence in wildlife in Europe.

Newcastle disease is a contagious and often fatal disease, capable of affecting all species of birds. A velogenic Newcastle disease virus (vNDV) outbreak occurred in an Israeli zoo, in which Little owls (Athene noctua) and African penguins (Spheniscus demersus) were found positive for presence of NDV. Some of them have died. The diagnostic process included: post-mortem examination, histopathology, real-time RT-PCR assay, virus isolation, serology, intracerebral pathogenicity index and phylogenetic analysis. A vNDV was diagnosed and found to be closely related to isolates from vNDV outbreaks that occurred in commercial poultry flocks during 2011. All isolates were classified as lineage 5d.

Switzerland is currently porcine reproductive and respiratory syndrome virus (PRRSV) free, but semen imports from PRRSV-infected European countries are increasing. As the virus can be transmitted via semen, for example, when a free boar stud becomes infected, and the risk of its import in terms of PRRSV introduction is unknown, the annual probability to accidentally import the virus into Switzerland was estimated in a risk assessment. A quantitative stochastic model was set up with data comprised by import figures of 2010, interviews with boar stud owners and expert opinion. It resulted in an annual median number of 0.18 imported ejaculates (= imported semen doses from one collection from one donor) from PRRSV-infected boars. Hence, one infected ejaculate would be imported every 6 years and infect a mean of 10 sows. These results suggest that under current circumstances, there is a substantial risk of PRRSV introduction into Switzerland via imported boar semen and that measures to enhance safety of imports should be taken. The time from infection of a previously negative boar stud to its detection had the highest impact on the number of imported ‘positive’ ejaculates. Therefore, emphasis should be placed on PRRSV monitoring protocols in boar studs. Results indicated that a substantial increase in safety could only be achieved with much tighter sampling protocols than currently performed. Generally, the model could easily be customized for other applications like other countries or regions or even sow farms that want to estimate their risk when purchasing semen from a particular boar stud.

Porcine brucellosis is a disease caused by Brucella suis, which is characterized by reproductive disorders in pigs. The number of cases of swine brucellosis has risen in many European countries, likely because of the presence of a wild reservoir of B. suis in wild boar. This study aimed at evaluating factors that may influence the probability of infection with Brucella spp. in wild boar and at assessing the impact of a previous contact with Brucella spp. on reproductive parameters of wild boar. Two hundred and four wild boar living in Extremadura (south-western Spain) were studied. The presence of anti-Brucella antibodies was determined using an indirect ELISA, while the presence of living bacteria in genital organs was evaluated through microbiological cultures. Sex, age, density of wild boar in summer and presence of outdoor pigs were selected as possible risk factors for being seropositive for Brucella spp. in wild boar. In addition, reproductive parameters such as breeding status or potential fertility in females and testis weight in males were estimated and related to the presence of anti-Brucella antibodies. A total of 121 animals were seropositive, resulting in a prevalence of 59.3% (95% CI). In addition, seven isolates of B. suis biovar 2 were obtained. Wild boar density in summer, as well as age and sex, was proposed as factors to explain the probability of Brucella seroconversion, although wild boar density in summer was the key factor. Current measures of reproductive parameters were not influenced by a previous contact with Brucella spp. Isolation of B. suis confirms that wild boar could represent a risk to domestic pig health in the study area. Wild boar density seems to have a great influence in the probability of infections with B. suis and suggests that density management could be useful to control Brucella infection in wild boar.

An 8-month-old crossbred ewe, normal upon physical examination, was humanely euthanized for tissue collection. After approximately 3 weeks in tissue culture, fungi began budding out of cells obtained from the choroid plexus. After an additional 3 weeks, budding was observed in kidney cell cultures and eventually in monocyte cultures as well. Serum from the lamb was submitted to the Veterinary Diagnostic Laboratory at Colorado State University for fungal diagnosis and was found negative for Aspergillus, Blastomyces, Coccidioidomycosis and Histoplasmosis. DNA was isolated from fungi collected from tissue culture supernatants and used in a set of pan-fungal PCR assays with DNA from Candida acting as a positive control. PCR products were sequenced and BLAST analysis performed. The unknown fungal sequence aligned with 100% identity to Rhodotorula minuta an emerging opportunistic pathogen. Samples were submitted to The Fungal Testing Laboratory at The University of Texas Health Science Center at San Antonio for additional validation. We believe this to be the first report of Rhodotorula fungemia in a sheep in the United States.

In December 2008, bird species in two geographically distant holdings were found positive for H5 viruses following the annual Avian influenza serological screening in Belgium. The virological tests performed identified in one holding a low-pathogenic avian influenza (LPAI) virus subtype H5N2, and a H5 LPAI virus was identified by real-time PCR and direct sequencing at the second holding. The first farm was an outdoor mixed holding housing ornamental birds and poultry (n = 6000) and the second a free-range geese breeding farm (n = 1500). No clinical signs or mortalities were reported. Control measures defined by Council Directive 2005/94/EC were followed, including notification to the European Commission via the Animal Disease Notification System and to the World Organization for Animal Health, and poultry were killed, while ornamental bird species were quarantined. Partial sequencing of the H5N2 virus haemagglutinin and neuraminidase N2 gene sequences revealed a close homology to some recent LPAI isolates identified from wild birds in Germany and Italy and from wild birds in Eurasia and Africa, respectively. It is noteworthy that, these two holdings were already H5 positive based on HI test results carried out during the previous serological screening; however, no virus was detected at that time. To have a better understanding of the potential ‘silent’ circulation of the H5N2 isolate in the field, experimental infections of chickens and turkeys were performed. The low excretion detected might in part explain viral persistence not associated with spread between gallinaceous birds in the same holding, indicating that the H5N2 LPAI isolate was not fully adapted to these two poultry species. Our results highlighted limitations to only using serological screening for the early detection of LPAI in an ‘at-risk farm’, suggesting that virological and serological monitoring tests be applied simultaneously as a means of testing animals in ‘at-risk farms’.

The epidemiological situation of foot-and-mouth disease virus (FMDV) is uncertain in Nigeria, where the disease is endemic, and the majority of outbreaks are unreported. Control measures for FMD in Nigeria are not being implemented due to the absence of locally produced vaccines and an official ban on vaccine importation. This study summarizes the findings of a 3-year study aimed at quantifying the seroprevalence of FMD, its distribution in susceptible species and the genetic diversity of FMDV isolated from the Plateau State of Nigeria. A 29% FMD prevalence was estimated using 3ABC enzyme-linked immunosorbent assay (3ABC ELISA). Farms with suspected FMD nearby, with contact with wildlife, that used drugs or FMD vaccines or with >100 animals, and animals of large ruminant species and in pastures other than nomadic grazing were significantly (P < 0.05) associated with FMD. Antibodies against five FMDV serotypes, (A, O, SAT1, SAT2 and SAT3) were detected by the virus neutralization test (VNT) at various titres (<100–>800) from all tested sera from most parts of the region. This is probably the first report of the presence of FMDV SAT3 in Nigeria. Further studies to investigate the potential probable presence and prevalence of SAT 3 virus in Nigeria are required. Tissue samples collected from clinical animals were positive for FMDV. Virus isolates were sequenced and confirmed as serotype A. All of the isolates showed marked genetic homogeneity with >99% genetic identity in the VP1 region and were most closely related to a previously described virus collected from Cameroon in 2000. This study provides knowledge on the epidemiological situation of FMD in Plateau State, Nigeria, and will probably help to develop effective control and preventive strategies for the disease in Nigeria and other countries in the West African subregion.

Understanding viral transmission is an important factor for the effective prevention one of the most devastating swine diseases, porcine reproductive and respiratory syndrome. Focusing on molecular epidemiology of type 1 PRRSV, this study analysed a large ORF5 dataset collected worldwide from 1991 to 2012 using a coalescent-based Bayesian Markov chain Monte Carlo approach. The results suggested that the virus diversified into unique subpopulations in Russia & Belarus and Italy approximately 100 years ago. Previously unreported consecutive diffusions of the virus were identified, which showed that some countries, such as Spain and Germany, acted as distribution sources to some extent. This study also provided statistical evidence for the existence of an ORF5-based phylogeographical structure of type 1 PRRSV, in which the virus tended to cluster by geographical locations more tightly than expected by chance. In contrast to this tight geographical structure, the evolution of the ORF5 gene, based on mapping of non-synonymous/synonymous substitutions, was best described by a non-homogeneous process that could be implicated as a mechanism for viral immune evasion.

Sixteen sheep and 18 cattle were followed up during 1 year to estimate the duration of immunity induced by inactivated bluetongue virus serotype 8 (BTV-8) vaccines (sheep and cattle) and a bluetongue virus serotype 1 (BTV-1) vaccine (cattle) under field conditions using cELISA and seroneutralization test (SNT). Four sheep never seroconverted. Those that seroconverted were all seronegative by BTV-8 SNT at the date of last sampling [378 days post-vaccination (dpv)]. Eight sheep were still positive by competitive ELISA (cELISA) 378 dpv. All the cattle seroconverted. At the end of the study, eight and 11 cattle were still positive by BTV-8 SNT and cELISA, respectively (335 dpv); and nine were still positive by BTV-1 SNT (301 dpv).

A prospective cohort study of avian influenza infection in poultry flocks was carried out in the Mekong River Delta of Viet Nam between December 2008 and April 2010. Our objectives were to (i) estimate the prevalence and incidence of avian influenza virus infection and (ii) assess the efficacy of H5N1 vaccination programmes as indicated by the presence of H5 antibody in vaccinated and unvaccinated poultry. Real-time PCR and H5 multiplex assays were used to detect the antigen of avian influenza viruses from swab samples. The haemagglutination inhibition test was used to detect H5 antibody. A total of 17 968 swab and 14 878 blood samples were collected from 5476 birds over the study period. The overall incidence rate of influenza type A virus infection was 5 (95% CI 4–7) positive birds per 100 bird-months at risk. The overall incidence rate of H5 virus infection was 0.2 (95% CI 0.1–0.5) positive birds per 100 bird-months at risk. Fifty (95% CI 48–52) birds per 100 tested birds were H5 HI positive in the unvaccinated group compared with 71 (95% CI 69–73) birds per 100 in the vaccinated group. Influenza type A and H5 viruses were circulating in village poultry throughout the study period with no recorded signs of clinical disease. This implies that interventions need to be carried out continuously throughout the year rather than only focusing on the established high-risk periods. Broiler ducks had an incidence rate of influenza H5 virus infection approximately four times greater than that of layer ducks and in-contact species. We conclude that broiler ducks are likely to be the main entry route for H5 virus into poultry flocks in the MRD. Control efforts would benefit from understanding why there is a difference between villages in H5 incidence and developing strategies to provide greater protection to broiler ducks.

In this study, we undertook the genomic characterization of 54 pseudorabies virus (PRV) strains isolated in Italy during 1984–2010. The characterization was based on partial sequencing of the UL44 (gC) and US8 (gE) genes; 44 strains (38 for gene gE and 36 for gC) were isolated on pig farms; 9 originated from dogs and 1 from cattle. These porcine PRV strains, which were closely related to those isolated in Europe and America in the last 20 years, and the bovine strain bovine/It/2441/1992 belong to cluster B in both phylogenetic trees. Six porcine strains that do not belong to cluster B are related in both gE and gC phylogenetic trees to the ‘old’ porcine PRV strains isolated in the 1970s and 1980s. In the last two decades, the presence of these strains in domestic pig populations has been reduced drastically, whereas they are prevalent in wild boar. The two remaining strains have an interesting genomic profile, characterized by the gC gene being closely related to the old porcine PRV strains, and the gE gene being similar to that of recently isolated strains. Three strains originating from working dogs on pig farms are located in cluster B in both phylogenetic trees. Five strains isolated from hunting dogs have a high degree of correlation with PRV strains circulating in wild boar. The last isolate has a gC gene similar to that in the two porcine strains mentioned previously, and the gE gene is correlated with the strains isolated from hunting dogs. These results provide interesting insight into the genomic characterization of PRV strains and reveal a clear differentiation between the strains isolated from hunting dogs that are related to the wild boar strains and those originating from domestic pigs.

Peste des petits ruminants virus (PPRV) causes one of the most contagious and highly infectious respiratory diseases in sheep and goats known as peste des petits ruminants (PPR). Reports of outbreaks of PPR in captive and wild small ruminants have extended the known spectrum of susceptible species to include antelopes. Phylogenetic analysis of nucleoprotein and fusion genes indicates that all PPRVs isolated from wild ungulate outbreaks belong to lineage IV. While it is clear that a number of wildlife species are susceptible to infection, the role of wildlife in the epidemiology of PPR remains uncertain. The available information about the occurrence of disease in free-ranging wildlife is mainly derived from surveys based on serological evidence. Data on the genetic nature of circulating PPRV strains are scarce. Given the scope of PPR in wild ungulates that are widespread in many countries, current disease surveillance efforts are inadequate and warrant additional investment. This is crucial because domestic and wild ruminants mingle together at several points, allowing inter-species transmission of PPRV. There is no reason to believe that PPRV circulates in wild animals and acts as a potential source of virus for domestic species. Irrespective of the possibility of wild small ruminants as the reservoir of PPRV, concerns about the role of susceptible species of antelopes need to be addressed, due to the fact that the disease can pose a serious threat to the survival of endangered species of wild ruminants on the one hand and could act as a constraint to the global eradication of PPR on the other hand. In this review, knowledge gained through research or surveillance on the sustainability of PPRV in wild ruminants is discussed.

Vaccination is considered as an important tool to control foot-and-mouth disease (FMD). A good quality vaccine containing relevant serotypes and matching strains is a pre-requisite for vaccination to be effective. The present study investigated the quality of different brands of FMD vaccine available in Pakistan, including three locally produced and two imported products. All the vaccines were found free of bacterial or fungal contamination. No adverse effects were noted in suckling mice and buffalo calves inoculated with the vaccines, showing that the vaccines were sterile and safe. The humoral immune response to the FMD vaccines was determined in buffalo calves for 234 days post-vaccination. Very low humoral immune responses against FMD serotypes O, A and Asia 1 viruses were detected to the locally produced vaccines. The imported vaccines, however, elicited a higher antibody response which persisted for a long period in one of the 2 vaccines. The present study highlights the need of assessing an independent vaccine quality control of finished FMD vaccine products.

A serological survey to detect Schmallenberg virus (SBV)-specific antibodies by ELISA was organized in the Belgian sheep population to study the seroprevalence at the end of the epidemic. One thousand eighty-two sheep samples which were collected from 83 herds all over Belgium between November 2011 and April 2012 were tested. The overall within-herd seroprevalence and the intraclass correlation coefficient were estimated at 84.31% (95% CI: 84.19–84.43) and 0.34, respectively. The overall between-herd seroprevalence was 98.03% (95% CI: 97.86–98.18). A spatial cluster analysis identified a cluster of six farms with significantly lower within-herd seroprevalence in the south of Belgium compared with the rest of the population (P = 0.04). It was shown that seroprevalence was associated to flock density and that the latter explained the presence of the spatial cluster. Additionally, 142 goat samples from eight different herds were tested for SBV-specific antibodies. The within-herd seroprevalence in goats was estimated at 40.68% (95% CI: 23.57–60.4%). The results of the current study provided evidence that almost every Belgian sheep herd has been in contact with SBV during 2011 and should be taken into consideration as part of comprehensive SBV surveillance and control strategies.

Influenza A viruses are common causes of respiratory disease in pigs and can be transmitted among multiple host species, including humans. The current lack of published information on infection dynamics of influenza viruses within swine herds hinders the ability to make informed animal health, biosecurity and surveillance programme decisions. The objectives of this serial cross-sectional study were to describe the infection dynamics of influenza virus in a two-site swine system by estimating the prevalence of influenza virus in animal subpopulations at the swine breeding herd and describing the temporal pattern of infection in a selected cohort of growing pigs weaned from the breeding herd. Nasal swab and blood samples were collected at approximately 30-day intervals from the swine breeding herd (Site 1) known to be infected with pandemic 2009 H1N1 influenza virus. Sows, gilts and neonatal pigs were sampled at each sampling event, and samples were tested for influenza virus genome using matrix gene RRT-PCR. Influenza virus was detected in neonatal pigs, but was not detected in sow or gilt populations via RRT-PCR. A virus genetically similar to that detected in the neonatal pig population at Site 1 was also detected at the wean-to-finish site (Site 2), presumably following transportation of infected weaned pigs. Longitudinal sampling of nasal swabs and oral fluids revealed that influenza virus persisted in the growing pigs at Site 2 for at least 69 days. The occurrence of influenza virus in neonatal pigs, but not breeding females, at Site 1 emphasizes the potential for virus maintenance in this dynamic subpopulation, the importance of including this subpopulation in surveillance programmes and the potential transport of influenza virus between sites via the movement of weaned pigs.

Zoonotic agents such as Brucella spp., Salmonella spp., Toxoplasma gondii and Trichinella spp., all considered high-risk zoonotic pathogens by the European Food Safety Agency (EFSA), may cause no symptoms of infection in free-range pigs yet still have a significant public health impact. A serological survey was therefore performed to determine the history of occurrence of these pathogens in such pigs in southern Spain. A total of 709 serum samples were collected at abattoir from pigs from 79 farms and analysed for specific antibodies against the above pathogens using commercially available ELISA kits. Encysted Trichinella spp. larvae were also sought following the artificial digestion method of diaphragm pillar muscle. The results showed Salmonella spp. to be widely distributed among the sampled herds [73.42%, 95% confidence interval (CI₉₅) 65.6–81.78] and Toxoplasma gondii to be present in over half (58.23%, CI₉₅ 47.33–69.07). The seroprevalence of Brucella spp. was very low (3.8%, CI₉₅ 0.18–7.42), and antibodies against Trichinella spp. were not detected. No encysted Trichinella spp. larvae were microscopically detected.

Between late February and May 2012, a preliminary anonym survey was conducted among sheep farmers in south of Belgium in order to contribute to future estimations of the economic losses caused by Schmallenberg virus (SBV). Based on clinical signs consistent with SBV infection, this survey involved 13 meat sheep flocks considered as positive flocks with subsequent SBV detection by RT-qPCR [SBV-positive flocks (PF); total of 961 animals], and 13 meat sheep flocks considered as negative flocks (NF; total of 331 animals). These preliminary results indicated several significant characteristics that were more present in PF than in NF. These include an increased rate of abortions (6.7% in PF versus 3.2% in NF), of lambs born at term but presenting malformations (10.1% in PF versus 2.0% in NF) and of dystocia (10.1% in PF versus 3.4% in NF). Lamb mortality during the first week of life was reported more frequently in PF (8 of 13 PF, 61.5%) than in NF (1 of 13 NF, 7.7%). In PF, the observed prolificacy rate was 2-fold lower (93%) than expected (186%). The implementation of a survey at larger scale, including a high number of breeders, is necessary to allow a more detailed analysis of the SBV impact in the sheep sector.

Early disease detection and efficient methods of proving disease freedom can substantially improve the response to incursions of important transboundary animal diseases in previously free regions. We used a spatially explicit, stochastic disease spread model to simulate the spread of classical swine fever in wild pigs in a remote region of northern Australia and to assess the performance of disease surveillance strategies to detect infection at different time points and to delineate the size of the resulting outbreak. Although disease would likely be detected, simple random sampling was suboptimal. Radial and leapfrog sampling improved the effectiveness of surveillance at various stages of the simulated disease incursion. This work indicates that at earlier stages, radial sampling can reduce epidemic length and achieve faster outbreak delineation and control, but at later stages leapfrog sampling will outperform radial sampling in relation to supporting faster disease control with a less-extensive outbreak area. Due to the complexity of wildlife population dynamics and group behaviour, a targeted approach to surveillance needs to be implemented for the efficient use of resources and time. Using a more situation-based surveillance approach and accounting for disease distribution and the time period over which an epidemic has occurred is the best way to approach the selection of an appropriate surveillance strategy.

It is known that lumpy skin disease virus (LSDV) can be shed in bull semen following infection and also that artificial insemination (AI) poses a biosecurity risk. However, it is not known whether the use of LSDV infected semen in AI poses a biosecurity risk. The aim of this study was to investigate whether LSDV, transmitted through semen, can infect cows and their embryos. Two controlled trials were performed simultaneously. Eleven young beef heifers, naïve to LSDV, were synchronized using an OvSynch protocol and inseminated on Day 0 with fresh semen spiked with a field strain of LSDV on day 0. Six of the heifers were superovulated on Day 1 using pregnant mare serum gonadotropin, and embryos were flushed from these heifers on Day 6. Blood and serum samples were collected from Day 4 until Day 27 to determine the presence of LSDV by PCR and virus isolation, and the presence of antibodies against LSDV by SNT. The first clinical signs of LSD were noticed on Day 10, followed by severe generalized LSD in three heifers and mild LSD in two more heifers. Two heifers were humanely euthanized due to severe unresponsive stranguria. LSDV was detected by PCR, virus isolation or electron microscopy in blood, embryos and organs of experimentally infected animals; and eight heifers had seroconverted by Day 27. Two control animals were not affected. This is the first report of experimental seminal transmission of LSDV in cattle.

Rapid, evidence-based decision-making is critical during a disease outbreak response; however, compliance by stakeholders is necessary to ensure that such decisions are effective – especially if the response depends on voluntary action. This mixed method study evaluated technical policy decision-making processes during the 2007 outbreak of equine influenza in Australia by identifying and analysing the stakeholder network involved and the factors driving policy decision-making. The study started with a review of the outbreak literature and published policy documents. This identified six policy issues regarding policy modifications or differing interpretations by different state agencies. Data on factors influencing the decision-making process for these six issues and on stakeholder interaction were collected using a pre-tested, semi-structured questionnaire. Face-to-face interviews were conducted with 24 individuals representing 12 industry and government organizations. Quantitative data were analysed using social network analysis. Qualitative data were coded and patterns matched to test a pre-determined general theory using a method called theory-oriented process-tracing. Results revealed that technical policy decisions were framed by social, political, financial, strategic and operational considerations. Industry stakeholders had influence through formal pre-existing channels, yet specific gaps in stakeholder interaction were overcome by reactive alliances formed during the outbreak response but outside the established system. Overall, the crisis management system and response were seen as positive, and 75–100% of individuals interviewed were supportive of, had interest in and considered the outcome as good for the majority of policy decisions, yet only 46–75% of those interviewed considered that they had influence on these decisions. Training to increase awareness and knowledge of emergency animal diseases (EADs) and response systems will improve stakeholder participation in emergency disease management and preparedness for future EAD incursions.

Free-grazing ducks (FGD) have been associated with highly pathogenic avian influenza (HPAI) H5N1 outbreaks and may be a viral reservoir. In July–August 2010, we assessed influenza exposure of Thai FGD and risk factors thereof. Serum from 6254 ducks was analysed with enzyme-linked immunosorbent assay (ELISA) to detect antibodies to influenza A nucleoprotein (NP), and haemagglutinin H5 protein. Eighty-five per cent (5305 ducks) were seropositive for influenza A. Of the NP-seropositive sera tested with H5 assays (n = 1423), 553 (39%) were H5 ELISA positive and 57 (4%) suspect. Twelve per cent (74 of 610) of H5 ELISA-positive/suspect ducks had H5 titres ≥ 1 : 20 by haemagglutination inhibition. Risk factors for influenza A seropositivity include older age, poultry contact, flock visitors and older purchase age. Study flocks had H5 virus exposure as recently as March 2010, but no HPAI H5N1 outbreaks have been identified in Thailand since 2008, highlighting a need for rigorous FGD surveillance.

We implemented a questionnaire-based methodology targeting veterinary field practitioners to evaluate clinical and economic impact of Schmallenberg virus in Belgium. First suspicious cases were detected as soon as July 2011. The mean cost for individual symptomatic treatment was 65 or 107 Euros, in case of fatal outcome or apparent recovery, respectively.

The objective of this study was to develop two indirect enzyme-linked immunosorbent assays (iELISAs) for detection of serum antibodies against classical vaccine strain of porcine reproductive and respiratory syndrome virus (PRRSV) and highly pathogenic PRRSV (HP-PRRSV). To detect the common antibodies against classical and HP-PRRSV, the coating antigen used in the iELISA (designated iELISA-180) was the antigen of Nsp2-180, the 180aa at amino terminal of Nsp2. To detect the different antibodies against classical and HP-PRRSV, the coating antigen in the second iELISA (designated iELISA-D29) was Nsp2-D29, the deleted 29aa in Nsp2 of HP-PRRSV. The antigen concentration and serum dilutions were optimized using a draughtboard titration. The cut-off values of 0.361 at OD_450nm for the iELISA-180 and 0.27 at OD_450nm for the iELISA-D29 were determined by testing a panel of 120 classical PRRSV positive and 198 PRRSV negative pig serum samples, which generated the specificity of 97.1% and 96.7%, the sensitivity of 96.9% and 96.3% for iELISA-180 and iELISA-D29, respectively. The agreements between the Western blot and iELISA-180 and iELISA-D29 were 98%, 96.7%, respectively. The developed iELISAs can be used to differentiate serologically HP-PRRSV from the vaccinated or classical PRRSV in clinical serum samples.

Aujeszky's disease (AD) causes significant economic losses in the Spanish pig sector due to import trade restrictions imposed by disease-free countries. Most regions of Spain have achieved ‘low AD prevalence’ status as a result of an intensive national AD eradication programme involving vaccination and other measures. However, to achieve AD-free status that would eliminate trade restrictions, vaccination must be stopped. For this final stage of eradication, up to date and reliable estimates of the risk of AD reintroduction are essential. Here, we propose an approach based on spatio-temporal scan statistics that the assesses risk of AD reintroduction in a disease-free territory by analysing the two most frequent risk pathways: movement of live domestic pigs and contact with wildlife reservoirs. The approach is illustrated using the case of Navarre, one of the first Spanish regions which plan to stop vaccination. Moreover, direct contacts among pig farms in Navarre were used to evaluate the potential spread of AD in the event of reintroduction. Areas at highest risk of AD reintroduction were in the southern part of the region during the second half of the year through pig movements and in the western and east-central parts of Navarre through contact with wild boars. Northern Navarre, despite having the highest density of pig farms, seems to be at low risk of AD reintroduction. Analysing the network of pig movements within Navarre revealed distinct northern and southern compartments that may be used in preventive compartmentalization strategies to reduce potential risk of AD re-infection in the scenario without vaccination. The approach described here may be extended to other regions and may be useful for guiding risk-based measures that reduce the risk of AD re-infection in a more cost-effective manner. Such analysis in Spain may allow authorities to stop vaccination in the safest possible way.

Salmonella enterica is a zoonotic agent of worldwide importance found in a wide range of wild hosts. However, its prevalence in many popular game species has never been assessed. Iberian ibex (Capra pyrenaica) is the main game caprinae of the Iberian Peninsula and around two thousand individuals are hunted every year for trophy or for home consumption. In this work, 313 Iberian ibexes from the Ports de Tortosa i Beseit National Game Reserve (NE Spain) were tested for Salmonella enterica in faeces, and anti microbial susceptibility was determined. The exact location of shooting or capture was recorded with a GPS device to study the links of Salmonella infection with cattle presence and human proximity. Additionally, samples were taken from cattle grazing inside this reserve (n = 73). Only three Iberian ibexes (0.96%, 95% CI 0.2–2.8) were positive to Salmonella (serotype Enteritidis, Bardo and 35:r:z35), while prevalence was moderate in cattle: 21.92% (95% CI 13.10–33.14, serotype Meleagridis, Anatum, Kedougou and Othmarschen). All isolates were susceptible to the anti microbial agents tested. Moreover, a case of fatal septicaemic salmonellosis in an 11-year-old male Iberian ibex is described where Salmonella enterica serotype Enteritidis was isolated from the lung, liver and spleen samples. The low prevalence of Salmonella in Iberian ibex and the lack of shared serotypes suggest no association to cattle. Despite this, game meat aimed for human consumption should be examined, and it is strongly recommended that hunters and game keepers manipulate animals and carcasses under maximal hygienic conditions to avoid environmental contamination and human contagion.

Papillomavirus (PV) are double-stranded DNA viruses that can cause both benignant and malignant tumours in mammals. Twelve genotypes of bovine papillomavirus (BPV1–12) have been identified so far. The presence of BPV1 and 2 has been found in the body fluids of cattle and horses. The aim of this study is to investigate the presence of BPV DNA and the expression of viral genes in the blood and sperm cells of healthy horses using PCR and RT-PCR. BPV-1 or 2 was detected in 14 of 70 blood samples (20%) and in 11 of 31 semen samples (35%). In five of fourteen blood samples, the E5 expression tested positive, while no blood sample was positive for L1 expression. Four of 11 (36%) semen cell samples proved to be positive for E5 expression, while no gene expression in L1 could be detected. This is the first study that shows BPV1 gene expression in the blood and semen of healthy horses. Our data illustrate the need for a better understanding of the presence of BPV in non-epithelial tissues of horses and their role in the vertical and horizontal transmission of these viruses.

A combined epidemiological–economic modelling approach was used to analyse strategies for highly pathogenic avian influenza (HPAI) control for the Netherlands. The modelling framework used was InterSpread Plus (ISP), a spatially based, stochastic and dynamic simulation model. A total of eight control strategies were analysed, including pre-emptive depopulation and vaccination strategies. The analysis was carried out for three different regions in the Netherlands: high-, medium- and low-density areas (HDA, MDA and LDA, respectively). The analysis included the veterinary impact (e.g. number of infected premises and duration), but was particularly focused on the impact on direct costs (DC) and direct consequential costs. The efficient set of control strategies for HDA and MDA included strategies based on either pre-emptive depopulation only or combined vaccination and pre-emptive depopulation: D2 (pre-emptive depopulation within a radius of 2 km), RV3 + D1 (ring vaccination within a radius of 3 km and additional pre-emptive depopulation within a radius of 1 km) and PV + D1 (preventive vaccination in non-affected HDAs and pre-emptive depopulation within a radius of 1 km in the affected HDA). Although control solely based on depopulation in most cases showed to be effective for LDA, pre-emptive depopulation showed to have an additional advantage in these areas, that is, prevention of ‘virus jumps’ to other areas. The pros and cons of the efficient control strategies were discussed, for example, public perception and risk of export restrictions. It was concluded that for the Netherlands control of HPAI preferably should be carried out using strategies including pre-emptive depopulation with or without vaccination. Particularly, the short- and long-term implications on export, that is, indirect consequential costs (ICC) and aftermath costs of these strategies, should be analysed further.

Seven clinical cases of cutaneous papillomatosis in yaks were studied in Arunachal Pradesh, India. Sporadic, single or a chain of multiple varying size warts appeared around the eyes or on the body. Predominant site of warts was around eyes. Histopathologically, these cases were diagnosed as fibropapilloma. It was confirmed by the detection of BPV-1 and BPV-2 or their mixed infection by PCR and sequencing. Quantitative SYBR Green real-time PCR detected comparatively lower viral DNA copy number in cutaneous warts (CWs). Cases of CWs and its causative agent as bovine papillomavirus (BPVs) are reported for the first time in yaks.

The aim of the present study was to identify potential risk factors for the occurrence of enzootic pneumonia (EP) in herds situated in a region of high pig density, where a majority of herds is endemically infected with Mycoplasma hyopneumoniae. Between 2006 and 2010, overall 100 herds were enrolled in a case–control study. Data were collected through personal interview with the farmers, clinical examination of pigs and their environments, and serological testing for M. hyopneumoniae, swine influenza virus and porcine reproductive and respiratory syndrome virus. There were 40 case herds (coughing index high, seroprevalence high) with a mean coughing index of 4.3 and a seroprevalence of 86.6%. There were two control groups. Control group I consisted of 25 herds (coughing index low, seroprevalence low) with mean values of 0.7 and 11.2%, and 35 herds were allocated to control group II (coughing index low, seroprevalence high) where the mean coughing index was 0.9 and seroprevalence 86.3%. Case herds and control II herds had an increased age of piglets at weaning compared to control I herds. Any contact between fattening pigs of different age during restocking of compartments increased the risk for the occurrence of EP in a herd. Finally, farms that use living animals for the exposure to gilts during the acclimatization and farms that had increased number of weaned piglets per sow and year were less likely to test positive for M. hyopneumoniae and less likely to develop clinical symptoms of EP in fattening pigs.

Pestiviruses, a genetically and antigenically highly diverse group, include one of the most historically significant swine pathogens, that is, classical swine fever virus. In Australia, investigations into swine outbreaks characterized by neonatal mortality, stillbirths and mummified foetuses resulted in the discovery of a new pestivirus, Bungowannah virus. This finding raised the possibility that Bungowannah virus, or a variant thereof, was circulating in swine herds elsewhere in the World. If so, it raised the possibility of a pestivirus emerging as a new swine disease with unknown consequences for animal health and food safety. Thus, we developed three specific qRT-PCR assays to evaluate tissue samples from undiagnosed cases of abortion or respiratory disease for evidence of Bungowannah virus. Examination of 64 samples collected between the Fall of 2007 and Spring of 2010 tested negative for all three genes examined. We conclude that Bungowannah-like pestivirus is unlikely to be present in swine in the upper Midwestern USA.

Mycobacterium bovis, the causative agent of tuberculosis in animals, has a broad host range, including humans. Historically, public health concerns prompted programs to eradicate tuberculosis from cattle in many nations. Eradication efforts decreased the prevalence of bovine tuberculosis; nevertheless, some countries encountered significant obstacles, not least of which was a wildlife reservoir of M. bovis. Efforts to decrease the size of the affected wildlife populations have neither eliminated disease nor eliminated transmission to cattle. Consequently, the use of a vaccine for wildlife is being explored. The vaccine most studied is M. bovis BCG, an attenuated live vaccine, first developed 100 years ago. The most efficient and effective means of vaccinating wildlife will be an oral vaccine. White-tailed deer in Michigan, USA, constitute a reservoir of M. bovis. White-tailed deer are a popular game species, and as such, represent a food animal to many hunters. BCG persistence in deer tissues could result in human exposure to BCG. Although non-pathogenic, BCG exposure could induce false-positive skin test results, confounding the central component of public health surveillance for TB. The objective of the present study in white-tailed deer was to evaluate persistence of lipid-encapsulated BCG and a liquid suspension of BCG after oral administration at two different dosages. Vaccine was not recovered at any time after oral consumption of a bait containing a single dose (1 × 10⁸ CFU) of lipid-encapsulated BCG. However, persistence was consistent in deer consuming 10 lipid-encapsulated baits (1 × 10⁹ CFU), with BCG recovered from at least one deer at 1, 3, 6, 9 and 12 months after consumption. Persistence of up to 9 months was seen in deer vaccinated with orally with a liquid suspension. Persistence of BCG was limited to lymphoid tissue and never found in samples of muscle collected at each time point. Although the risk of exposure to hunters is low, BCG persistence should be considered prior to field use in white-tailed deer.

In January 2010, the United Kingdom notified cases of equine infectious anaemia (EIA) in two horses introduced from Belgium. The animals came from one assembly centre in Romania and had transited through Belgium with 16 other horses. Nine of them, bought by a Belgian horse breeder, were investigated in Belgium and revealed one additional EIA-positive animal. Afterwards, the Belgian Federal Agency for the Safety of the Food Chain (FASFC) organized a serological EIA survey of the horses introduced into Belgium from Romania between 2007 and 2009. Among the 95 horses identified, six additional serological positive cases were found that had been introduced into Belgium in 2008 (n = 4) and in 2009 (n = 2). The survey was extended to the horses in contact with the positive cases, but all contact animals were negative, indicating the absence of transmission. Virological examination performed on tissue samples collected from two seropositive animals demonstrated the presence of viral DNA of EIA virus. Phylogenetic analysis based on the sequences of EIA virus gag gene clustered the Belgian isolates with Romanian strains isolated in 2009. The presumption of a common Belgian origin could be rejected.

Thirteen outbreaks of foot-and-mouth disease (FMD) were reported in pigs and cattle in Korea between 8 April and 4 June 2010. The FMD virus (FMDV) isolates were of serotype O, indicating that they were related to the virus strains of the Southeast Asia topotype that are circulating in East Asian countries. Animals carrying the viruses were identified by reverse transcriptase-polymerase chain reaction (RT-PCR) during a 29-day period between 8 April and 6 May, 2010. Prior to this outbreak, these FMDVs had not been detected in Korea and may therefore have been introduced from neighbouring countries into Ganghwa Island and subsequently spread inland to other areas, including Gimpo, Chungju and Cheongyang. Tests conducted to lift restrictions on animal movements lead to detection of two additional FMD-positive farms. Through appropriate responses, including swift diagnoses and culling policies, Korea was able to quickly regain its recognition as being free of FMD, without vaccination, by the World Organization for Animal Health (OIE) on 27 September 2010.

Japanese encephalitis (JE) is a disease that threatens both human and animal populations in Asian countries, and the causative agent of JE, Japanese encephalitis virus (JEV), has recently changed from genotype III (GIII) to genotype I (GI). However, a test for the rapid differentiation of GI and GIII JEV is still unavailable, especially one that can be used for mosquito-based surveillance. We have designed GI- and GIII-specific primer sets for the rapid detection and differentiation of GI and GIII JEV by multiplex reverse transcriptase-polymerase chain reaction (multiplex RT-PCR). The GI-specific and GIII-specific primer sets were able to specifically amplify the target gene from GI and GIII JEV, respectively. The limitations of detection were 0.00225 and 0.225 pfu for the GI-specific and GIII-specific primers, respectively. Using a mixture of GI-specific and GIII-specific primers, the multiplex RT-PCR was able to specifically detect and differentiate GI and GIII JEV. The multiplex RT-PCR was able to successfully differentiate GI and GIII virus in JEV-infected mosquitoes. Thus, a sensitive and specific multiplex RT-PCR system for the rapid detection and differentiation of GI and GIII JEV has been developed, and this test is likely to be valuable when carrying out mosquito-based JEV surveillance.

Echinococcus multilocularis is a cestode parasites that frequently occurs in the red fox (Vulpes vulpes), which is the main definitive host in Central Europe. The parasite may infect humans as accidental intermediate hosts and cause alveolar echinococcosis. In the German federal state of Saxony-Anhalt, the occurrence of E. multilocularis in red foxes as a possible source of infection for humans was studied from 1998 to 2010. A significant shift in the geographical centroid of the occurrence of E. multilocularis from a long-known highly endemic area in the southwest of the state towards the north-northeast (3.2 km/year) was found. The overall prevalence in the state increased significantly from 13.6% (1998–2005) to 23.4% (2006–2010). No autochthonous cases of alveolar echinococcosis have been reported to date in Saxony-Anhalt, but this might change in the near future with the spread and increasing biomass of the parasite.

In case of a classical swine fever outbreak in the European Union (EU), its control is based upon the culling of swine on infected farms, movement restrictions in the protection and surveillance zones, and contact tracing. Additionally, preventive culling may be carried out. Emergency vaccination and rapid PCR testing are discussed as alternatives to avoid this measure. An outbreak of classical swine fever and the success of its control are influenced by different factors. Using a spatial and temporal Monte-Carlo simulation model the control strategies ‘Restriction Zone’, ‘Traditional Control’, ‘Emergency Vaccination’, ‘Test To Slaughter’, ‘Test To Control’ and ‘Vaccination in conjunction with Rapid Testing’ were compared under various conditions. Farm density, compliance with movement restrictions and delay in the establishment of an emergency vaccination were analysed as influencing factors. It was found that all these factors had a significant influence on the number of infected and culled farms. In a low-density region, the basic measures are sufficient to control an epidemic, provided strict compliance with movement restrictions is adhered to. In a high-density region, additional measures are necessary. They can compensate non-strict compliance with movement restriction to a certain extent. In the high-density region, ‘Emergency Vaccination’ and ‘Vaccination in conjunction with Rapid Testing’ reached the same level of infected farms as ‘Traditional Control’, independent of the value of compliance with movement restrictions. However, in the case of an emergency vaccination, an early start to the vaccination campaign is essential for successful disease control.

A qualitative import risk assessment was undertaken to assess the likelihood of introduction and establishment of viral haemorrhagic septicaemia virus (VHSV) genotype 1a in England and Wales (E&W), via the processing of imported rainbow trout (Oncorhynchus mykiss) carcasses from continental Europe. The likelihood was estimated for one import from an infected farm. Four main routes by which susceptible populations could be exposed to VHSV via processing waste were considered: (i) run-off from solid waste to watercourses, (ii) contamination of birds or rodents with VHSV by scavenging solid waste, (iii) discharge of liquid waste to mains drainage, and (iv) discharge of liquid waste directly to watercourses. Data on the biophysical characteristics of VHSV, its epidemiology, fish processing practices and waste management were collected. Likelihoods for each step of the four pathways were estimated. Pathway 4 (discharge of liquid waste to a watercourse) was judged as the most likely to result in infection of susceptible individuals. Levels of virus entering the aquatic environment via pathways 1–3 were judged to be many times lower than pathway 4 due mainly to the treatment of solid waste (pathways 1 and 2) and high levels of dilution (pathways 1, 2 and 3). Thirty-four trout farms process fish, of which seven have imported carcasses for processing. Compared with other processing facilities, on-farm processing results in a higher likelihood of VHSV exposure and establishment via all four pathways. Data availability was an issue; the analysis was particularly constrained by a lack of data on the prevalence of VHSV in Europe, volume of trade of carcasses into the UK and processing practices in E&W. It was concluded that the threat of VHSV introduction into E&W could be reduced by treatment of liquid effluent from processing plants and by sourcing carcasses for on-farm processing only from approved VHSV free areas.

The central European lineage of Usutu virus was isolated from a blackbird (Turdus merula), which was found dead in the city of Brno, Czech Republic, in 2011. The virus RNA was detected in two other dead blackbirds in Brno during 2012.

African horse sickness (AHS) is associated with high morbidity and mortality in equids, especially horses. A retrospective analysis was carried out concerning 737 AHS outbreaks that occurred during 2007–2010 in Ethiopia. A total of ten outbreaks were investigated in the study period. All four forms of the disease (pulmonary, cardiac, horse sickness fever and the combined form) were observed, with the cardiac form being the most prevalent. Multiple African horse sickness virus serotypes (AHSV-2, AHSV-4, AHSV-6, AHSV-8 and AHSV-9) were detected by molecular methods (type-specific real-time RT-PCR assays), and fourteen isolates were derived from blood and tissue samples collected during 2009–2010. This is the first report of AHSV-4, AHSV-6 or AHSV-8 in Ethiopia.

Porcine circovirus 2 (PCV2) is a common virus in pig population and is associated with the postweaning multisystemic wasting disease (PMWS). In this study, it was developed and evaluated the single-tube nested PCR (STNPCR) method for the detection of PCV2 DNA. PCV2 reference controls and swine tissue samples were used, and primers were selected for targeting specific regions of the viral genome. In comparison of the methods, STNPCR was 10 times more sensitive than conventional PCR and showed the same sensitivity to nested PCR (NPCR), but with reduction in the risk of cross-contamination. In clinical application, 55 tissue samples were analysed by conventional PCR and resulted in 67% (37/55) of positive reactions, while the NPCR and STNPCR were able to identify the presence of viral DNA in 100% (55/55) of the samples. The high sensitivity combined with the elimination of cross-contamination makes the STNPCR method suitable for the epidemiological studies of PCV2 and can aid in the diagnosis of PMWS.

Economic analysis of control strategies for contagious diseases is a necessity in the development of contingency plans. Economic impacts arising from epidemics such as highly pathogenic avian influenza (HPAI) consist of direct costs (DC), direct consequential costs (DCC), indirect consequential costs (ICC) and aftermath costs (AC). Epidemiological models to support economic analysis need to provide adequate outputs for these critical economic parameters. Of particular importance for DCC, ICC and AC is the spatial production structure of a region. Spatial simulation models are therefore particularly suited for economic analysis; however, they often require a large number of parameters. The aims of this study are (i) to provide an economic rationale of epidemiological modelling in general, (ii) to provide a transparent description of the parameterization of a spatially based epidemiological model for the analysis of HPAI control in the Netherlands and (iii) to discuss the validity and usefulness of this model for subsequent economic analysis. In the model, HPAI virus transmission occurs via local spread and animal movements. Control mechanisms include surveillance and tracing, movement restrictions and depopulation. Sensitivity analysis of key parameters indicated that the epidemiological outputs with the largest influence on the economic impacts (i.e. epidemic duration and number of farms in the movement restriction zone) were more robust than less influential indicators (i.e. number of infected farms). Economically relevant outputs for strategy comparison were most sensitive to the relative role of the different transmission parameters. The default simulation and results of the sensitivity analysis were consistent with the general outcomes of known HPAI models. Comparison was, however, limited due to the absence of some economically relevant outputs. It was concluded that the model creates economically relevant, adequate and credible output for subsequent use in economic analysis. A detailed economic analysis is presented in a subsequent article.

This paper analyses the potential gains and the main challenges for increased cross-border collaboration in the control of highly contagious livestock diseases in regions with cross-border reliance on production and consumption of livestock commodities. The aim of this intensification of cross-border collaboration is to retain the economic advantages of cross-border trade in livestock and livestock commodities while maintaining a low risk of highly contagious livestock diseases. From these two foci, possibilities for future policy making with respect to highly contagious livestock diseases are discussed: peacetime cross-border cooperation to improve the cost-effectiveness of routine veterinary measures and crisis time cross-border harmonization of current disease control strategies. A general disease management framework was used to describe the way in which these two fields are related to and affect the epidemiological system and, consequently, how they impact the stakeholders. In addition to this framework, the importance of a good understanding of influencing factors, that is, the production structure of livestock, was stressed because these factors are important determinants of the frequency and magnitude of highly contagious livestock diseases and their economic impact. The use of the suggested integrated approach was illustrated for the extended cross-border region of the Netherlands and Germany, that is, North Rhine Westphalia and Lower Saxony. For this region, current difficulties in cross-border trade in livestock and livestock commodities and possibilities for future cross-border collaboration were examined. The concepts and ideas presented in this paper should foster future development of cross-border collaboration in animal health control.

Bovine brucellosis is a zoonotic disease spread worldwide. The infection in cattle is predominantly caused by Brucella abortus and is usually detected in pregnant females through abortions. The disease is endemic in Argentina; however, infection in humans is underestimated and often not reported. The prevalence of bovine brucellosis in countries bordering Argentina is quite variable: 0.04% in Uruguay, 10.20% in the north and 0.06% in the south of Brazil, 0.2% in Chile, 3.15% in Paraguay and 2.27% in Bolivia. In 1999, the Argentine National Control and Eradication Program was implemented. Its strategies include identification of vaccinated animals, compulsory vaccination with B. abortus S19 of 100% of 3- to 8-month-old females, negative serological tests before animal movements and categorization of farms in terms of their brucellosis status. The epidemiological surveillance in milk is performed through the milk ring test and the indirect ELISA. The result of a national brucellosis survey performed in 2004 indicates that 12.4% (95% CI: 10.89–14.0) of Argentine beef farms are seropositive to Brucella and that the apparent prevalence in cattle is 2.10% (95% CI: 1.90–2.40). The official serological diagnostic tests are as follows: buffered plate antigen test, as screening, serum agglutination test, 2-mercaptoethanol and fluorescence polarization assay, competitive ELISA, as confirmatory tests, and complement fixation test, as definitive test. Santa Fe and a district in Córdoba have ‘Outstanding Plans’. Tierra del Fuego is a ‘Zone free from bovine brucellosis’. One question arising when studying the Argentine situation is why the disease remains endemic if good regulations exist to control and eradicate it. In future, some different aspects might be evaluated to understand it, and further studies should be performed to prioritize, select and refine control strategies.

In commercial swine populations, influenza is an important component of the porcine respiratory disease complex (PRDC) and a pathogen with major economic impact. Previously, a commercial blocking ELISA (FlockChek^™ Avian Influenza Virus MultiS-Screen^® Antibody Test Kit, IDEXX Laboratories, Inc., Westbrook, ME, USA) designed to detect influenza A nucleoprotein (NP) antibodies in avian serum was shown to accurately detect NP antibodies in swine serum. The purpose of this study was to determine whether this assay could detect NP antibodies in swine oral fluid samples. Initially, the procedure for performing the NP-blocking ELISA on oral fluid was modified from the serum testing protocol by changing sample dilution, sample volume, incubation time and incubation temperature. The detection of NP antibody was then evaluated using pen-based oral fluid samples (n = 182) from pigs inoculated with either influenza A virus subtype H1N1 or H3N2 under experimental conditions and followed for 42 days post inoculation (DPI). NP antibodies in oral fluid were detected from DPI 7 to 42 in all inoculated groups, that is, the mean sample-to-negative (S/N) ratio of influenza-inoculated pigs was significantly different (P < 0.0001) from uninoculated controls (unvaccinated or vaccinated-uninoculated groups) through this period. Oral fluid versus serum S/N ratios from the same pen showed a correlation of 0.796 (Pearson's correlation coefficient, P < 0.0001). The results showed that oral fluid samples from influenza virus-infected pigs contained detectable levels of NP antibodies for ≥42 DPI. Future research will be required to determine whether this approach could be used to monitor the circulation of influenza virus in commercial pig populations.

Several strategies for eradicating Pseudorabies virus (Aujeszky's disease) in Chiang-Mai and Lampoon Provinces, Thailand, were compared using a computer simulation model, the North American Animal Disease Spread Model (NAADSM). The duration of the outbreak, the number of affected herds and the number of destroyed herds were compared during these simulated outbreaks. Depopulation, zoning for restricted movement and improved detection and vaccination strategies were assessed. The most effective strategies to eradicate Pseudorabies as per the findings from this study are applying depopulation strategies with MOVEMENT RESTRICTIONS in 3-, 8- and 16-km ZONES surrounding infected herds and enhancing the eradication with vaccination campaign on 16-km radius surrounding infected herds.

Dynamic mathematical modelling and stochastic simulation of disease–host systems for the purpose of epidemiological analysis offer great opportunities for testing hypotheses, especially when field experiments are impractical or when there is a need to evaluate multiple experimental scenarios. This, combined with the ever increasing computer power available to researchers, has contributed to the development of many mathematical models for epidemic simulations, such as the individual-based model (IBM). Nevertheless, few of these models undergo extensive validation and proper assessment of intrinsic variability. The Ontario rabies model (ORM) will be used here to exemplify some advantages of appropriate model behaviour validation and to illustrate the use of a simple geometric procedure for testing directional bias in distributed stochastic dynamic model of spread of diseases. Results were obtained through the comparison of 10 000 epizootics resulting from 100 epidemic simulations started using 100 distinct base populations. The analysis results demonstrated a significant directional bias in epidemic dispersion, which prompted further verification of the model code and the identification of a coding error, which was then corrected. Subsequent testing of the corrected code showed that the directional bias could no longer be detected. These results illustrate the importance of proper validation and the importance of sufficient knowledge of the model behaviour to ensure the results will not confound the objectives of the end-users.

Porcine reproductive and respiratory syndrome virus (PRRSV) is the causative agent of PRRS, the most significant infectious disease currently affecting swine industry worldwide. In the United States alone, the economic losses caused by PRRS amount to more than 560 million US dollars every year. Due to immune evasion strategies and the antigenic heterogeneity of the virus, current commercial PRRSV vaccines (killed-virus and modified-live vaccines) are of unsatisfactory efficacy, especially against heterologous infection. Continuous efforts have been devoted to develop better PRRSV vaccines. Experimental PRRSV vaccines, including live attenuated vaccines, recombinant vectors expressing PRRSV viral proteins, DNA vaccines and plant-made subunit vaccines, have been developed. However, the genetic and antigenic heterogeneity of the virus limits the value of almost all of the PRRSV vaccines tested. Developing a universal vaccine that can provide broad protection against circulating PRRSV strains has become a major challenge for current vaccine development. This paper reviews current status of PRRSV vaccine development and discusses strategies to develop a universal PRRSV vaccine.

Bulk-tank milk (BTM) samples are frequently used to evaluate the health status of dairy livestock. A large-scale investigation carried out in BTM samples from dairy cattle herds from a Q fever-endemic region in Northern Spain revealed a high degree of exposure to Coxiella burnetii. This study was aimed at assessing the value of BTM samples analysis as an indicator of the C. burnetii status in dairy cattle herds. Three herds with BTM samples positive for C. burnetii by ELISA and PCR were selected, and blood, faeces and individual milk and BTM samples were analysed by serology and PCR. In spite of the high antibodies titres found in BTM samples, only one of the three farms presented an active infection by C. burnetii, as revealed by the presence of bacterial DNA in vaginal mucus and in environmental samples collected in the calving area, a seroprevalence around 40% in heifers and the seroconversion rate observed in cows. Results obtained indicated that the analysis of BTM samples is a good epidemiological tool at the population level that can be used to discriminate between seropositive and seronegative herds, but at the herd level, additional tests are necessary to evaluate whether Q fever is a potential problem in the farm. When Q fever is suspected in a cattle herd, sera from a small group of 1- to 3-year-old animals need to be analysed to investigate recent contact with C. burnetii.

Control of bovine viral diarrhoea (BVD) in Belgium is currently implemented on a voluntary basis at herd level and mainly relies on detection and culling of persistently infected (PI) animals. The present field study was conducted during the winter of 2010/2011 to assess the performances of diagnostic assays used in the testing scheme for BVD as proposed by the two Belgian regional laboratories. Individual blood samples were collected from 4972 animals, and individual samples from the same herd were pooled (maximum of 30 individual samples per pool) and screened for the presence of Bovine Viral Diarrhoea Virus (BVDV)-specific RNA using a commercial real-time RT-PCR test (ADIAGENE). Individual samples from positive pools were then tested in parallel with the same RT-PCR test and with an antigen-capture ELISA test (IDEXX) to detect viremic animals. This study demonstrated that individual results differed according to the type of assay used (P < 0.001): 140 animals (2.8%) were positive by RT-PCR and 72 (1.4%) by antigen-ELISA. A second blood sample was taken 40 days later from 74 PCR positive animals to detect persistent viremia: 17 (23%) of these were still PCR positive and considered to be PI and the 57 that no longer tested positive were assumed to be transiently infected (TI) animals. All PI animals were positive also by antigen-ELISA at both time points. Among TI animals, 10 (16%) were positive by antigen-ELISA at the first but none at the second sampling. A highly significant difference in cycle threshold (C_t) values obtained by RT-PCR was observed between PI and TI animals. ROC analysis was performed to establish thresholds to confirm with high probability that an animal is PI, based on the result of RT-PCR test performed on a single individual blood sample.

Leptospirosis is a zoonotic disease of global importance and one of the notifiable diseases in Sri Lanka. Recent studies on human leptospirosis have suggested that the cattle could be one of the important reservoirs for human infection in the country. However, there is a dearth of local information on bovine leptospirosis, including its implications for human transmission. Thus, this study attempted to determine the carrier status of pathogenic Leptospira spp in cattle in Sri Lanka. A total of 164 cattle kidney samples were collected from the meat inspection hall in Colombo city during routine inspection procedures conducted by the municipal veterinary surgeons. The DNA was extracted and subjected to nested PCR for the detection of leptospiral flaB gene. Amplicons were sequenced, and phylogenic distances were calculated. Of 164 samples, 20 (12.2%) were positive for flaB-PCR. Sequenced amplicons revealed that Leptospira species were deduced to L. borgpetersenii (10/20, 50%), L. kirschneri (7/20, 35%) and L. interrogans (3/20, 15%). The results indicate that a high proportion of the sampled cattle harbour a variety of pathogenic Leptospira spp, which can serve as important reservoirs for human disease.

The Mediterranean basin is a biodiversity hotspot; it has historically had a large human presence that has shaped ecosystems for millennia. As the cradle of many civilizations, the area was one of the main theatres for transitions that punctuated both human and pathogen histories, which are intimately linked. Today we are living through another great historical transition summarized in the expression ‘global changes’. In this context, we are witnessing a rise in the emergence of pathogens widely associated with aforementioned global changes. The Mediterranean basin might be especially vulnerable to this phenomenon due to the acute consequences global changes will have in this key intercontinental interface region. In addition, Arab revolutions and European economic crisis are creating both sanitary issues and presenting new opportunities to improve infectious disease control and prevention in the region. The aim of this review is to identify the impacts that ongoing changes might have on the risk of infectious disease emergence in the Mediterranean basin. We focussed on three key domains undergoing transformations: (i) resources, namely safe drinking water and animal products, (ii) socio-economic factors including health inequalities within countries and poor sanitary conditions linked to ongoing conflicts and (iii) movements of people and goods that are reshaped by current changes and are intimately linked to the risk of disease proliferation. Building on recent examples, we try to identify upcoming challenges and discuss ways to meet them in the light of existing international human and veterinary health guidelines and their possible improvements.

The 2003 outbreak of Highly pathogenic avian influenza (HPAI) A(H7N7) in the Netherlands, Belgium and Germany resulted in significant genetic diversification that proved informative for tracing transmission events. Building on previous investigations on the Dutch outbreak, we focused on the potential transnational transmissions between the Netherlands and Belgium. Although no clear epidemiological links could be identified from the tracing data, the transmission network based on concatenated HA-NA-PB2 sequences supports at least three independent introductions from the Netherlands to Belgium and suggests one possible introduction form Belgium back to the Netherlands. Two introductions in the Belgian province of Limburg occurred from nearby farms in the Dutch province of Limburg. One introduction resulted in three secondary infected farms, while a second introduction did not cause secondary infections. The third introduction into Belgium occurred in the north of the Antwerp province, very close to the national border, and originated from the North of the Dutch province Brabant (long distance transmission, >65 km). The virus spread to two additional Belgian farms, one of which may be the source of a secondarily infected farm in the Netherlands. One infected turkey farm in the province of Antwerp (Westmalle) was geographically close to the latter introduction, but genetically clustered with the first introduction event in the Limburg province. Epidemiological tracing data could neither confirm nor exclude whether this outbreak was a result from long distance contacts within Belgium or whether this farm presented a fourth independent transboundary introduction. These multiple transnational transmissions of HPAI in spite of reinforced biosecurity measures and trade restrictions illustrate the importance of international cooperation, legislation and standardization of tools to combat transboundary diseases.

Vampire bat rabies causes significant impacts within its endemic range in Mexico. These impacts include livestock mortality, animal testing costs, post-exposure prophylaxis costs, and human mortality risk. Mitigation of the impacts can be achieved by vaccinating livestock and controlling vampire bat populations. A benefit-cost analysis was performed to examine the economic efficiency of these methods of mitigation, and Monte Carlo simulations were used to examine the impact that uncertainty has on the analysis. We found that livestock vaccination is efficient, with benefits being over six times higher than costs. However, bat control is inefficient because benefits are very unlikely to exceed costs. It is concluded that when these mitigation methods are judged by the metric of economic efficiency, livestock vaccination is desirable but bat control is not.

Pakistan is at an initial stage for progressive control of foot and mouth disease (FMD). Understanding the risk factors for introduction, spread and persistence of the infection is important to design an evidence-based disease control programme. A rapid appraisal method was adopted, and a convenient sample of twenty commercial dairy farmers was interviewed. The following were considered to contribute in secondary transmission of infection: (i) intermediaries and service providers [animal health workers, animal traders and transporters, raw milk collectors, persons who remove skin of dead animals], (ii) places where animals come in close contact [livestock markets, animal fairs, communal grazing pastures, routes in villages where livestock move, watering points, animal transport vehicles], (iii) use of bulls immediately after recovery from FMD infection, (iv) range land/desert livestock production, (v) small holder sheep and goat production, (vi) purchase of replacement stock and fodder from infected locations. This article reveals contacts within and between villages, some of which may act as routes of transmission of FMD. The study suggests the need for zoosanitary education of the livestock keepers.

Newcastle disease (ND), caused by virulent strains of avian paramyxovirus type 1 (APMV-1), is considered throughout the world as one of the most important animal diseases. For over three decades now, there has been a continuing panzootic caused by a variant virulent APMV-1 strain, so-called pigeon paramyxovirus type 1 (PPMV-1), primarily in racing pigeons, which has also spread to wild birds and poultry. PPMV-1 isolations have been made in Great Britain every year since 1983. In this study, we have completed a comparative phylogenetic analysis based on a 374 nucleotide section of the fusion protein gene of 63 isolates of PPMV-1 that were isolated over a 26-year period; 43 of these were sequenced for this study. Phylogenetic analysis of these sequences revealed that all were closely related and placed in the genetic sublineage 4b (VIb), subdivision 4biif.

Acute infections with bovine viral diarrhoea virus (BVDV), a major pathogen of cattle, are often asymptomatic or produce only mild clinical symptoms. However, they may play an important role in the bovine respiratory disease complex by exerting a marked immunosuppressive effect, as a result of the death of the immunocompetent cell populations involved in controlling innate and adaptive immune responses, together with a marked reduction of both cytokine expression and co-stimulatory molecule synthesis. Although experimental research and field studies have shown that acute BVDV infection enhances susceptibility to secondary infection, the precise mechanism involved in BVDV-induced immunosuppression remains unclear. The present study is aimed at measuring a range of blood parameters in a single group of fourteen calves infected with non-cytopathic BVDV-1. Focus has been put on those related to the cell-mediated immune response just as leucocyte populations and lymphocyte subpopulations, serum concentrations of cytokines (IL-1β, TNF-α, IFN-γ, IL-12, IL-4 and IL-10) and acute phase proteins [haptoglobin, serum amyloid A (SAA), fibrinogen and albumin], as well as BVDV-specific antibodies and viremia. After non-cytopathic BVDV-1 infection, clinical signs intensity was never more than moderate coinciding with the presence of viremia and leucocyte and lymphocyte depletion. An early increase in TNF-α, IFN-γ and IL-12 levels in contrast to IL-1β was observed in line with a raise in haptoglobin and SAA levels on the latest days of the study. As regards IL-4 levels, no evidence was found of any changes. However, a slight increase in IL-10 was observed, matching up the TNF-α decline during the acute phase response. These findings would help to increase our knowledge of the immune mechanisms involved in acute infection with non-cytopathic BVDV-1 strains, suggesting the existence of a clear tendency towards a type 1 immune response, thereby enhancing resistance against viral infections.

The recent outbreak caused by Schmallenberg virus, which affected sheep, goats and cattle in Europe, highlighted the importance of having a robust surveillance plan capable of monitoring abortions and malformations in the livestock offspring. In this context, bluetongue viruses (BTVs) represented and represent one of the major threats to the European livestock industry. Aiming to improve the understanding on BTV cross placental transmission and serotype involvement, in this retrospective study foetal spleens and/or brains of 663 ovines, 429 bovines, 155 goats and 17 buffaloes were tested for the presence of BTV by virus isolation. BTV vaccine strains were isolated from 31 foetuses (2.4%; 95% CI: 1.7–3.4%): 24 (3.6%; 95% CI: 2.4–5.3%) from ovine foetal tissues; 6 (1.4%; 95% CI: 0.6–3.0%) from bovine foetal tissues and 1 (0.6%; 95% CI: 0.2–3.5%) from the spleen of a caprine foetus. All foetuses were from animals vaccinated with either BTV-2 or BTV-2, and BTV-9 modified live vaccines (MLVs) produced by Onderstepoort Biological Products (OBP), South Africa. Among the 31 isolated vaccine strains, serotype 9 (n = 28) was more frequently isolated (P < 0.05) than serotype 2 (n = 3). In two cases infectious vaccine strains were found in the foetal tissues 2 months after the vaccine administration. Other pathogens known to be causative agents of abortion in ruminants were not detected nor isolated. This study demonstrates, for the first time, that BTV-2 and BTV-9 vaccine strains are able to cross the placental barrier of sheep, cattle and goats. BTV-2 and BTV-9 vaccine strains are able to infect foetuses and cause abortions or malformations depending on the period of pregnancy at the time of vaccination.

An outbreak of suspected malignant catarrhal fever (MCF) was investigated by molecular and histopathological assays. Of the 70 Holstein beef calf herds, 14 were affected by multiple clinical signs suggestive of MCF infection. These beef calves were housed next to sheep flocks. In the complete blood count, the 14 affected calves had severe anaemia with leucopaenia, lymphopaenia and neutropaenia. Upon PCR amplification using a hemi-nested PCR assay for the detection of the Ovine herpesvirus 2 (OvHV-2), bovine tissue samples from the mesenteric lymph nodes and spleen and ovine blood samples were shown to be positive with the expected PCR bands amplified. Direct sequencing of the hemi-nested PCR product confirmed the identity of the causative virus as OvHV-2. The histopathological findings confirmed the clinical and laboratory diagnosis of MCF. Collective clinical, PCR and histopathological data confirmed the identity of this outbreak to be a sheep-associated malignant catarrhal fever (SA-MCF).

We quantified the between-village transmission rate, β (the rate of transmission of H5N1 HPAI virus per effective contact), and the reproductive number, R_e (the average number of outbreaks caused by one infectious village during its entire infectious period), of H5N1 highly pathogenic avian influenza (HPAI) virus in Nigeria using outbreak data collected between December 2005 and July 2008. We classified the outbreaks into two phases to assess the effectiveness of the control measures implemented. Phase 1 (December 2005–October 2006) represents the period when the Federal Government of Nigeria managed the HPAI surveillance and response measures, while Phase 2 (November 2006–July 2008) represents the time during which the Nigeria Avian Influenza Control and Human Pandemic Preparedness project (NAICP), funded by a World Bank credit of US$ 50 million, had taken over the management of most of the interventions. We used a total of 204 outbreaks from 176 villages that occurred in 78 local government areas of 25 states. The compartmental susceptible-infectious model was used as the analytical tool. Means and 95% percentile confidence intervals were obtained using bootstrapping techniques. The overall mean β (assuming a duration of infectiousness, T, of 12 days) was 0.07/day (95% percentile confidence interval: 0.06–0.09). The first and second phases of the epidemic had comparable β estimates of 0.06/day (0.04–0.09) and 0.08/day (0.06–0.10), respectively. The R_e of the virus associated with these β and T estimates was 0.9 (0.7–1.1); the first and second phases of the epidemic had R_e of 0.84 (0.5–1.2) and 0.9 (0.6–1.2), respectively. We conclude that the intervention measures implemented in the second phase of the epidemic had comparable effects to those implemented during the first phase and that the R_e of the epidemic was low, indicating that the Nigeria H5N1 HPAI epidemic was unstable.

The major policy for eradication of classical swine fever (CSF) in South Korea has focused on the implementation of compulsory vaccination of the susceptible pig population. A vaccine strain of CSF virus, the LOM strain, is used to maintain high herd seroconversion, a practice complementary to the ‘stamping-out policy’ and restriction of animal movement during disease outbreaks. To survey for the prevalence of CSF in domestic pigs in South Korea over the past 13 years (1999–2011), we tested 4 193 782 and 1 162 645 samples for antibodies and antigens, respectively. Whereas seropositivity for CSF antibodies has been maintained at over 95% in the mainland, in Jeju Island, where no-vaccination has been administered since 1999, seroprevalence has been below 1% during the last 3 years of study (2009-2011). The highest number of outbreaks in South Korea occurred in 2002 and 2003; since then, outbreaks have decreased each year, with the last CSF outbreak recorded in 2009. No outbreaks have occurred during the past 3 years, and a high level of herd immunity has been maintained in the mainland pig population for 8 years; therefore, South Korea could now switch to a no-vaccination policy throughout the country. However, the constant threat of the re-emergence of the disease in the susceptible pig population should be the main consideration in planning and carrying out the last phase of the CSF eradication process.

To gain further insight into the immunopathogenesis of bovine tuberculosis (bTB), the cytokine and chemokine expression of cattle experimentally infected with Mycobacterium bovis was analysed in TB granulomas, using immunohistochemistry (IHC) and laser capture microdissection (LCM) followed by qPCR. Immunohistochemistry was conducted for cell types using labelling for CD68, CD3, CD4, CD8, WC1 and CD79a and for the cytokines IFN-γ, TNF-α and TGF-β as well as inducible form of nitric oxide synthase (iNOS). qPCR was conducted for mRNA expression of IFN-γ, TNF-α, TGF-β, IL-17A, IL-22, IL-2, granzyme A and the chemokines CXCL9 and CXCL10. Early stages of granuloma were primarily comprised of epithelioid MΦs expressing high levels of IFN-γ and iNOS, with significantly upregulated expression of CXCL9 and CXCL10 when compared with control tissue. These chemokines displayed a trend of decreasing mRNA expression as lesion progressed, suggesting a higher level of importance during the early stages of the immune response to mycobacterial infection. IL-22 levels showed a strong trend of decrease through granuloma development, and IL-17A was shown to be upregulated, supporting its investigation as a potential biomarker of bTB. The use of LCM and qPCR may prove especially useful for the study of IL-17A as previous attempts to analyse its expression using IHC and in situ hybridization proved unsuccessful.

Bovine vaccinia (BV), a zoonosis caused by Vaccinia virus (VACV), affects dairy cattle and milkers, causing economic, veterinary and human health impacts. Despite such impacts, there are no experimental studies about the pathogenesis of BV in cows to assess whether there is a systemic spread of the virus and whether there are different ways of VACV shedding. Trying to answer some of these questions, a study was proposed using experimental inoculation of VACV in cows. All experimentally infected cows developed lesions compatible with VACV infection in cattle. Two of the six animals presented VACV DNA in blood and faecal samples, starting at the 2nd and the 3rd day post-infection (d.p.i.), respectively, and lasting until the 36th d.p.i., in an intermittent way. This study provides new evidence that VACV can be detected in blood and faeces of infected cows, suggesting that BV could be a systemic disease, and also bringing new information about the epidemiology and pathogenesis of BV.

Hepatitis E virus (HEV) causes an important public health disease in many developing countries and is also endemic in some industrialized countries. In addition to humans, strains of HEV have been genetically identified from pig, chicken, rat, mongoose, deer, rabbit and fish. While the genotypes 1 and 2 HEV are restricted to humans, the genotypes 3 and 4 HEV are zoonotic and infect humans and other animal species. As a part of our ongoing efforts to search for potential animal reservoirs for HEV, we tested goats from Virginia for evidence of HEV infection and showed that 16% (13/80) of goat sera from Virginia herds were positive for IgG anti-HEV. Importantly, we demonstrated that neutralizing antibodies to HEV were present in selected IgG anti-HEV positive goat sera. Subsequently, in an attempt to genetically identify the HEV-related agent from goats, we conducted a prospective study in a closed goat herd with known anti-HEV seropositivity and monitored a total of 11 kids from the time of birth until 14 weeks of age for evidence of HEV infection. Seroconversion to IgG anti-HEV was detected in seven of the 11 kids, although repeated attempts to detect HEV RNA by a broad-spectrum nested RT-PCR from the faecal and serum samples of the goats that had seroconverted were unsuccessful. In addition, we also attempted to experimentally infect laboratory goats with three well-characterized mammalian strains of HEV but with no success. The results indicate that a HEV-related agent is circulating and maintained in the goat population in Virginia and that the goat HEV is likely genetically very divergent from the known HEV strains.

Sweden experienced its first outbreak of bluetongue virus (BTV) infection beginning in September 2008. Mandatory vaccination with an inactivated vaccine (BTVPUR Alsap8; Merial, Lyon, France) began 2 days after bluetongue was confirmed in the country. The aim of this study was to investigate whether the goal of 80% seroconversion by the susceptible population within the vaccination area was met during the initial phase of the Swedish vaccination campaign and whether there were discrepancies between subpopulations. Milk or blood samples were collected from 274 cattle randomly selected from the vaccinated population. Blood samples were also collected from ten ewes on each of 28 randomly selected vaccinated herds. The vaccination campaign in Sweden may be regarded as successful, as measured by apparent seroprevalence in the vaccinated population. The overall apparent seroprevalence was 77%, and in cattle, which constituted the majority of the susceptible population, the apparent seroprevalence was 82%. Factors that influenced the titres after vaccination were as follows: (i) the time span between vaccination and sampling and (ii) the age of the animals.

Ambient water temperature is a key factor controlling the distribution and impact of disease in fish populations, and optimum temperature ranges have been characterised for the establishment of a number important aquatic diseases exotic to the UK. This study presents a simple regression method to approximate daily average surface water temperature in lakes of 0.5–15 ha in size across the UK using 5 km² gridded daily average air temperatures provided by the UK Meteorological Office. A Geographic information system (GIS) is used to present thematic maps of relative risk scores established for each grid cell based on the mean number of days per year that water temperature satisfied optimal criteria for the establishment of two economically important pathogens of cyprinid fish (koi herpesvirus (KHV) and spring viraemia of carp virus (SVCV)) and the distribution and density of fish populations susceptible to these viruses. High-density susceptible populations broadly overlap the areas where the temperature profiles are optimal for KHV (central and south-east England); however, few fish populations occur in areas where temperature profiles are most likely to result in the establishment of spring viremia of carp (SVC) (namely northern England and Scotland). The highest grid-cell risk scores for KHV and SVC were 7 and 6, respectively, out of a maximum score of 14. The proportion of grid cells containing susceptible populations with risk scores of 5 or more was 37% and 5% for KHV and SVC, respectively. This work demonstrates a risk-based approach to inform surveillance for exotic pathogens in aquatic animal health management, allowing efficient use of resources directed towards higher risk animals and geographic areas for early disease detection. The methodology could be used to examine the change in distribution of high-risk areas for both exotic and endemic fish diseases under different climate change scenarios.

One of the most challenging aspects of foot-and-mouth disease (FMD) control is the high genetic variability of the FMD virus (FMDV). In endemic settings such as the Indian subcontinent, this variability has resulted in the emergence of pandemic strains that have spread widely and caused devastating outbreaks in disease-free areas. In countries trying to control and eradicate FMD using vaccination strategies, the constantly evolving and wide diversity of field FMDV strains is an obstacle for identifying vaccine strains that are successful in conferring protection against infection with field viruses. Consequently, quantitative knowledge on the factors that are associated with variability of the FMDV is prerequisite for preventing and controlling FMD in the Indian subcontinent. A hierarchical linear model was used to assess the association between time, space, host species and the genetic variability of serotype O FMDV using viruses collected in Pakistan from 2005 to 2011. Significant (P < 0.05) amino acid and nucleotide variations were associated with spatial distance, but not with differences in host species, which is consistent with the frequent multi-species infection of this serotype O FMDV. Results from this study will contribute to the understanding of FMDV variability and to the design of FMD control strategies in Pakistan. Viruses sequenced here also provide the earliest reported isolate from the Pan Asia II^ANT-10 sublineage, which has caused several outbreaks in the Middle East and spread into Europe (Bulgaria) and Africa (Libya).

Movements of animals and animal products are one of the most important ways of disease introduction and spread between regions and countries. Maybe one of the most complex animal species in terms of diversity of uses, nature and extent of movements are equidae, for which animal movement records are usually not available. The study presented here is the first characterization of a complete and reliable network of equidae movements in Castile and Leon, which is one of the most important equidae production regions of Spain. Social network analysis and space–time cluster analysis were used to describe the contact patterns of the equidae network and to identify the most important premises, areas and time periods for potential disease introduction or spread into the region. The studied network was complex, with very heterogeneous types of premises and diverse nature and extent of the movements compared with other livestock species, which have important implications for prevention and control of equidae diseases. Centrality measures revealed that production and reproduction farms and centres of livestock competition were the most important type of premises in the studied network. Cluster analyses allowed to identify seventeen significant spatio-temporal clusters of premises at high risk of dispatching or receiving equidae, which formed four interconnected compartments. These clusters were mainly located in the north-west region and in the second part of the year. The results of this study may be useful to design risk-based surveillance and control programmes of equidae diseases and increase the speed of detection and control of potential secondary outbreaks in future epidemics. Consequently, these results will help to minimize the great economic and sanitary impact of equidae diseases. The analytical approach used here may be easily extended to characterize the equidae movement patterns in other countries and regions of the world.

This study assessed whether recently weaned piglets with maternally derived antibodies were able to generate infectious influenza aerosols. Three groups of piglets were assembled based on the vaccination status of the dam. Sows were either non-vaccinated (CTRL) or vaccinated with the same (VAC-HOM) strain or a different (VAC-HET) strain to the one used for challenge. Piglets acquired the maternally derived antibodies by directly suckling colostrum from their respective dams. At weaning, pigs were challenged with influenza virus by direct contact with an infected pig (seeder pig) and clinical signs evaluated. Air samples, collected using a liquid cyclonic air collector, and individual nasal swabs were collected daily for 10 days from each group and tested by matrix real-time reverse transcriptase polymerase chain reaction (RRT-PCR) assay. Virus isolation and titration were attempted for air samples on Madin–Darby canine kidney cells. All individual pigs from both VAC-HET and CTRL groups tested positive during the study but only one pig in the VAC-HOM group was positive by nasal swab RRT-PCR. Influenza virus could not be detected or isolated from air samples from the VAC-HOM group. Influenza A virus was isolated from 3.2% and 6.4% air samples from both the VAC-HET and CTRL groups, respectively. Positive RRT-PCR air samples were only detected in VAC-HET and CTRL groups on day 7 post-exposure. Overall, this study provides evidence that recently weaned pigs with maternally derived immunity without obvious clinical signs of influenza infection can generate influenza infectious aerosols which is relevant to the transmission and the ecology of influenza virus in pigs.

The objective of this study was to determine the association between phenotypic resistance, genotypic resistance and virulence genes of Escherichia coli isolates in Jiangsu province, China and Punjab province Pakistan. A total of 62 E. coli isolates were characterized for phenotypic resistance, genotypic resistance and virulence factor genes. The anti-microbial resistance phenotype and genotypes in relation to virulence factor genes were assessed by statistical analysis. Of 20 tested virulence genes, twelve were found and eight were not found in any isolates. sitA and TspE4C2 were the most prevalent virulence genes. Of the 13 anti-microbial agents tested, resistance to ampicillin, sulphonamide and tetracycline was the most frequent. All isolates were multiresistant, and 74% were resistant to trimethoprim and sulphamethaxazole. Phenotypically, tetracycline-, cefotaxime- and trimethoprim-resistant isolates had increased virulence factors as compared with susceptible isolates. Genotypically, resistant genes Tem, ctx-M, Tet, Sul 1, dhfr1, Cat2 and flo-R showed the association with the virulence genes. Almost all classes of anti-microbial-resistant genes have a high association with virulence. Resistant isolates have more virulent genes than the susceptible isolates.

Porcine reproductive and respiratory syndrome virus (PRRSV) can persist in different organs of infected pigs, which suggests a failure in the immune response. Antigen-presenting cells (APCs) play a pivotal role in the induction of effective T- and B-cell responses. In this study, we investigated the changes in the different APC subpopulations and T- and B-cell counts in the tonsil, retropharyngeal and mediastinal lymph nodes of pigs experimentally infected with a European PRRSV field isolate. Our results demonstrated that the expression of S100, SWC3, HLA-DR molecule and CD3 was diminished in the studied organs throughout the study, observing a significant negative correlation between viral antigen and HLA-DR expression in both retropharyngeal and mediastinal lymph nodes. In contrast, λ-light chains showed an increase during the study. Taking all into account, after PRRSV infection, no enhancement in the number of APCs and T cells was observed, suggesting an impairment of the immune function which may allow the persistence of PRRSV into the organism.

Dabbling ducks, particularly Mallards (Anas platyrhynchos) have been frequently and consistently reported to play a pivotal role as a reservoir of low pathogenic avian influenza viruses (AIV). From October 2006 to November 2008, hand-raised Mallard ducks kept at a pond in an avifaunistically rich area of Southern Germany served as sentinel birds in the AIV surveillance programme in Germany. The pond was regularly visited by several species of dabbling ducks. A flock of sentinel birds, consisting of the same 16 individual birds during the whole study period, was regularly tested virologically and serologically for AIV infections. Swab samples were screened by RT-qPCR and, if positive, virus was isolated in embryonated chicken eggs. Serum samples were tested by the use of competitive ELISA and hemagglutinin inhibition (HI) assay. Sequences of full-length hemagglutinin (HA) and neuraminidase (NA) genes were phylogenetically analysed. Four episodes of infections with Eurasian-type AIV occurred in August (H6N8), October/November (H3N2, H2N3) 2007, in January (H3N2) and September (H3N8) 2008. The HA and NA genes of the H3N2 viruses of October 2007 and January 2008 were almost identical rendering the possibility of a re-introduction of that virus from the environment of the sentinel flock highly likely. The HA of the H3N8 virus of September 2008 belonged to a different cluster. As a correlate of the humoral immune response, titres of nucleocapsid protein-specific antibodies fluctuated in correlation with the course of AIV infection episodes. However, no specific systemic response of hemagglutination inhibiting antibodies could be demonstrated even if homologous viral antigens were used. Besides being useful as early indicators for the circulation of influenza viruses in a specific region, the sentinel ducks also contributed to gaining insights into the ecobiology of AIV infection in aquatic wild birds.

Tick-borne pathogens can spread easily through the movements of infested birds. An important example is viruses that pose a threat to humans and that are carried in Hyalomma ticks that move from Africa into south-western Europe. This study evaluates the probability of arrival of migrating birds from Africa into Spain and the environmental suitability of different regions of Spain for the survival of tick stages introduced by these birds. This evaluation produced a spatial risk index measuring the probability that foreign tick populations will survive in the target area. Periods of highest risk were observed for large areas of Spain, from the second fortnight of April to the second fortnight of May. Although birds may arrive as early as January and massive migrations may take place in March, the environmental suitability for Hyalomma marginatum ticks is low in these periods and high mortality of the spread stages (nymphs) is expected. This study introduces new methods of objective analysis based on spatial and process-driven models for both ticks and hosts and critically evaluates the usefulness of spatial spreading methods for assessing the risk of tick-borne pathogens.

African swine fever virus (ASFV) causes one of the most dreaded transboundary animal diseases (TADs) in Suidae. African swine fever (ASF) often causes high rates of morbidity and mortality, which can reach 100% in domestic swine. To date, serological diagnosis has the drawback of not being able to differentiate variants of this virus. Previous studies have identified the 22 genotypes based on sequence variation in the C-terminal region of the p72 gene, which has become the standard for categorizing ASFVs. This article describes a genotyping assay developed using a segment of PCR-amplified genomic DNA of approximately 450 bp, which encompasses the C-terminal end of the p72 gene. Complementary paired DNA probes of 15 or 17 bp in length, which are identical except for a single nucleotide polymorphism (SNP) in the central position, were designed to either individually or in combination differentiate between the 22 genotypes. The assay was developed using xMAP technology; probes were covalently linked to microspheres, hybridized to PCR product, labelled with a reporter and read in the Luminex 200 analyzer. Characterization of the sample was performed by comparing fluorescence of the paired SNP probes, that is, the probe with higher fluorescence in a complementary pair identified the SNP that a particular sample possessed. In the final assay, a total of 52 probes were employed, 24 SNP pairs and 4 for general detection. One or more samples from each of the 22 genotypes were tested. The assay was able to detect and distinguish all 22 genotypes. This novel assay provides a powerful novel tool for the simultaneous rapid diagnosis and genotypic differentiation of ASF.

Horizon scanning techniques can be developed to identify novel routes and sources for the emergence of viruses in the medium to long term. Central to horizon scanning is prediction of the complex scenarios through which viruses could emerge before they occur. One approach involves ‘spidergrams’ in which complex scenarios are generated by combining factors randomly selected from different categories of events. Spidergrams provide a framework for how different factors could interact, irrespective of the virus, and also enable testing of combinations not previously considered but which would be ‘tested’ in nature by a virus. The emergence of viruses through new routes is often related to changes, for example, in environmental and social factors, and the Internet will undoubtedly be used to identify long-term trends for consideration. In addition, online games may provide horizon scanners with suggestions for new routes and strategies that could be used by emerging viruses.

The pathogenesis of highly pathogenic porcine reproductive and respiratory syndrome virus (PRRSV) strain (HuN4) is poorly understood. Therefore, highly pathogenic PRRSV strain (HuN4) and its derivative strain (HuN4-F112) (obtained by propagation in MARC145 cells for 112 passages) were inoculated into a total of 48 PRRSV-sero-negative pigs (age: 4–5 weeks) by the intranasal route. Virological, pathological and in situ hybridization analyses were performed. The results exhibited that pigs infected with HuN4 showed a loss of appetite, decrease in body weight, raised body temperature and respiratory symptoms, along with interstitial pneumonia lesions. In the HuN4 group, multifocal interstitial pneumonia with macrophage infiltration was found in the lung. The lesions in the lymph node were characterized by collapsed follicles, depletion of germinal centres and reduction in lymphocytes. Perivascular cuffing and glial nodules were observed in the brains of some pigs. By comparison, the HuN4-F112 group had milder lesions. PRRSV was detected in macrophages, alveolar epithelial cells and vascular endothelial cells in the tonsil and lymph nodes. The PRRSV amounts in the pigs infected with HuN4 were 10⁵–10⁹ copies/ml in the blood and 10¹⁰–10¹¹ copies/g in the lung tissues, whereas the virus amounts with HuN4-F112 were 10^2.15–10^3.13 copies/ml in the blood and 10^3.0–10^3.6 copies/g in the lung. Our results demonstrate that the PRRS HuN4 virus infects alveolar epithelial cells, macrophages and vascular endothelial cells causing diffuse alveolar damage and lymph node necrosis. Its higher pathogenicity compared with HuN4-F112 virus may be explained in part by higher replication rate in the previously mentioned organs.

In recent years, several animal disease epidemics have occurred within the European Union (EU). At the 4th Annual Meeting of the EPIZONE network (7–10 June 2010, St. Malo, France), an interactive session was run to elicit the opinions of delegates on a pre-defined list of epidemic threats to the EU. Responses from over 190 delegates, to questions relating to impact and likelihood, were used to rank six virus groups with respect to their perceived threat now (2010) and in 2020. The combined opinions of all delegates suggested that, from the pre-selected list of virus groups, foot-and-mouth disease and influenza are currently of most concern. Delegates thought that influenza would be less of a threat and zoonotic arboviruses would be more of a threat in 2020. Although the virus group rankings should not be taken as definitive, the results could be used in conjunction with experimental and field data, by scientists, policy-makers and stakeholders when assessing and managing risks associated with these virus groups.

In 2010, Coxiella burnetii was identified at a high prevalence in the placentas of Northern fur seals (Callorhinus ursinus) collected at a single rookery on St. Paul Island Alaska; an area of the United States where the agent was not known to be present. As contamination was hypothesized as a potential cause of false positives, but nothing was known about environmental C. burnetii in the region, an environmental survey was conducted to look for the prevalence and distribution of the organism on the island. While environmental prevalence was low, two strains of the organism were identified using PCR targeting the COM1 and IS1111 genes. The two strains are consistent with the organism that has been increasingly identified in marine mammals as well as a strain type more commonly found in terrestrial environments and associated with disease in humans and terrestrial animals. Further work is needed to elucidate information regarding the ecology of this organism in this region, particularly in association with the coastal environment.

The aim of this study was to estimate the incidence of mycobacterial infections in cats in Great Britain (GB). This was performed using the proxy measure of feline tissue samples submitted to diagnostic laboratories in GB that were found to have histopathological changes typical of mycobacterial infection (‘MYC’). Sixteen primary diagnostic laboratories were asked for information on the number of feline samples submitted in 2009, the number with MYC, the number undergoing Ziehl–Neelsen (ZN) staining and, for comparison, the number diagnosed with lymphoma.

Eight laboratories provided full data for the whole year: 11 782 samples; lymphoma 3.2% (mean, 95% CI: 2.89, 3.5), MYC 1.16% (0.98; 1.37) and ZN-positive 0.31% (0.22; 0.43). Data on 1569 samples from seven laboratories that provided partial data on samples for the whole year revealed similar results, although all changes were more frequent: lymphoma 5.42% (4.35; 6.66), MYC 2.36% (1.66; 3.23) and ZN-positive 0.77% (0.40; 1.33). One laboratory only provided data for part of the year (4.5 months), reporting all three types of histopathology less frequently: 18 232 samples; lymphoma 0.2% (0.18; 0.32), MYC 0.07% (0.04; 0.12) and ZN-positive 0.05% (0.02; 0.09). The reasons for low reporting rates in this high-throughput laboratory are unclear.

In total, 187 samples were reported as having MYC. Five Reference laboratories were also contacted, reporting 174 feline tissue submissions in 2009, with mycobacteria being cultured from 90.

The study shows that MYC are frequently reported in tissue samples from cats in GB, being reported in ∼1% of samples, with confirmation as ZN-positive in ∼0.3%. Lymphoma is recognized as a common disease in cats, being seen in ∼3% of samples in this study. When compared against MYC, lymphoma was reported only twice as frequently. This confirms that far from being rare, clinically significant mycobacterial infections occur commonly in cats in GB.

Attacks against livestock and poultry using biological agents constitute a subtype of agroterrorism. These attacks are defined as the intentional introduction of an animal infectious disease to strike fear in people, damage a nation’s economy and/or threaten social stability. Livestock bioterrorism is considered attractive to terrorists because biological agents for use against livestock or poultry are more readily available and difficult to monitor than biological agents for use against humans. In addition, an attack on animal husbandry can have enormous economic consequences, even without human casualties. Animal husbandry is vulnerable to livestock-targeted bioterrorism because it is nearly impossible to secure all livestock animals, and compared with humans, livestock are less well-guarded targets. Furthermore, anti-livestock biological weapons are relatively easy to employ, and a significant effect can be produced with only a small amount of infectious material. The livestock sector is presently very vulnerable to bioterrorism as a result of large-scale husbandry methods and weaknesses in the systems used to detect disease outbreaks, which could aggravate the consequences of livestock-targeted bioterrorism. Thus, terrorism against livestock and poultry cannot be thought of as either a ‘low-probability’ or ‘low-consequence’ incident. This review provides an overview of methods to prevent livestock-targeted bioterrorism and respond to terrorism involving the deliberate introduction of a pathogen-targeting livestock and poultry.

Beginning in 2006, point infection control operations and aerial distribution of oral rabies vaccines along the US border were performed in Quebec, Canada, to control the potential spread of raccoon rabies. A benefit-cost analysis assessed the economic efficiency of this rabies control programme into the future. In this study, a mathematical simulation model was used to determine the potential spread of raccoon rabies from the 2006 index case, and incidence rates of human post-exposure prophylaxis (PEP), animal testing and human exposure investigations were calculated. Benefits were calculated as the potential savings from reduced numbers of human PEP, animal testing and human exposure investigations owing to control, which ranged from $47 million to $53 million. Programme cost scenarios were based on projections of total expenditures, which ranged from $33 million to $49 million. Economic efficiency was indicated for approximately half of the modelled scenarios, with the greatest benefit-cost ratios resulting from reduced future programme costs.

Foot and mouth disease (FMD) is an endemic transboundary disease in the Mekong region, and FMD records of reports to animal health authorities in Lao PDR between 2009 and 2011 were reviewed. FMD outbreaks occurred in 2 of 3 years in eight districts in three of the eight northern Lao PDR provinces, locations suggested as FMD ‘hotspots’. The relatively higher risk of recurrence of FMD in these districts was likely due to the presence of a dense large ruminant population, extensive animal trading including transboundary movements and ineffective animal movement controls. As an understanding of the epidemiology of FMD in these ‘hotspots’ may offer insights into improved FMD control in the region, a study of an outbreak of FMD occurring in early 2010 following failure to vaccinate was conducted in the endemic ‘hotspot’ area of Paek district in Xiengkhoung province where in early 2009, a major outbreak of FMD in the district had been prevented in two villages by vaccination. The 2010 outbreak included collection of tissue samples 1 week after the onset of FMD that confirmed infection with FMD virus serotype O (Myanmar topotype) in a population of 239 large ruminants, comprising 167 cattle and 72 buffalo. A survey by interview of 30 farmers conducted in July 2010 documented high morbidity in cattle and buffalo (>90%) and identified disease risk factors, including increased trading of animals at the end of the rice harvest, plus several failures of biosecurity. In late 2010 and early 2011, a total of 40 and 72 serum samples were collected from large ruminants prior to and post-FMD vaccination respectively and tested by LPB-ELISA. Antibodies were present in the pre-vaccination samples attributable to previous exposure to FMD virus and significantly rising post-vaccination titres indicated likely temporary protection against future FMDV infection. It was concluded that to provide sufficient control of FMD in this ‘hotspot’, regular vaccination, particularly prior to the peak risk period in December-February, plus improved farmer knowledge of disease transmission risk and biosecurity, is required. Although low rural education standards and language barriers because of multiple ethnic groups pose a challenge for the successful delivery of extension programmes in northern Lao PDR, training to improve disease recognition and reporting plus village-level biosecurity practices is considered important in FMD ‘hotspots’ if sustainable regional initiatives directed at FMD control are to be achieved.

In the Netherlands, outbreaks of Aujeszky’s Disease (AD) are controlled by vaccination and movement restriction zones (MRZ). Although this strategy avoids the socio-ethical concerns associated with pre-emptive slaughter, it can easily result in animal welfare problems and negative economic consequences. These arise because movement restrictions result in surpluses of live (vaccinated) piglets on farms. The aim is to provide insight into the development of these surpluses and its impact and to describe how measures that allow early transportation of pigs under certain conditions and to specific destinations (channelling) could reduce these problems. For the analysis, a deterministic simulation model was developed, which calculates surpluses of piglets at multiplier farms during AD outbreaks. This is performed on a weekly basis for two areas (with and without piglet surplus), three outbreak durations (minimum, moderate and long) and three strategies for movement restrictions (strict, transports within the MRZ allowed and transports outside the MRZ allowed). The results show that in case of complete movement restrictions, surpluses of piglets varying in age and vaccination status will quickly arise. These surpluses are larger for longer epidemics and can become as large as 180–340 thousand piglets (45–75% of weekly domestic production) for moderate and long epidemics, respectively. Implementation of channelling strategies that allow earlier transportation within the MRZ can reduce surpluses by about 50% to 100–150 thousand piglets maximum. Strategies that also allow transportation outside the MRZ can reduce surpluses even further to below 100 thousand piglets. It was concluded that channelling of live piglets during AD outbreaks results in a drastic reduction of problems with accommodating ready-for-transport piglets. Moreover, it reduces shortages during movement restrictions and peak supply immediately after removing the restrictions. Channelling could therefore be an important instrument to reduce the economic and animal welfare impacts of containment measures.

Semen samples from 88 reproductively mature bulls were screened to detect the presence of Brucella spp. by polymerase chain reaction. Twenty-seven samples were found to be positive, underscoring the importance of researching brucellosis in males and the need for greater care in the selection of sperm-donating bulls for semen centres.

The E2 genes of 73 classical swine fever virus (CSFV) originated from CSF suspected cases in different regions of China were genetically characterized and compared with reference CSF viruses. All Chinese viruses that characterized were segregated into two major groups and subdivided into four subgroups. Most of isolates (61.6%) belonged to group 2 and were further divided into three subgroups: subgroup 2.1, 2.2 and 2.3. Subgroup 2.1 was the largest subgroup which contained 46.6% of isolates, whiles subgroup 2.3 was the smallest subgroup which contained only one isolate (1.4%). The remaining 38.4% of isolates were classified into subgroup 1.1 within group 1. However, no group 3 and subgroups 1.2 and 1.3 viruses were found in this study. This study has provided epidemiological information useful for assessing the virus origin and establishing a national prevention and control strategy against the disease.

Bovine cysticercosis (BC) is an important disease because of its zoonotic nature. There is a significant variation in the prevalence of BC in different countries, ranging from <0.01% to more than 20%. In this study, we followed the changes of BC prevalence in Israel during the last four decades and examined its association with import of live cattle. During 1973–2007, 629 549 cattle were subjected to post-mortem inspection conducted in ‘Marbek’ slaughterhouse located in the south of Israel. A specific comparison was made between the prevalence of BC in local and imported cattle during 2003–2007. Of 629 549 cattle, 2568 were infected with Cysticercus bovis (0.4%). From 1980, there was a gradual decrease in the prevalence of BC (R² = 0.53) with exceptional peaks. Moreover, from 1973 to 1998, only 4% of the documented cases appeared in outbreaks as opposed to 38% after 1998 when mass importation of live cattle to Israel was initiated. All of these late outbreak cases appeared in imported cattle of which 95% originated from Australia. During the years 2002–2007, importation from Australia was found as a significant risk factor for infection with BC, with prevalence in these cattle reaching 1.8% in 2006. The time from importation to BC detection suggests that infection occurred either in Australia or during the transport into Israel. We conclude that despite a reduction in the prevalence of BC as a result of a possible improvement in sanitary conditions at the farms, meticulous meat inspection is still essential in Israel and possibly in other developed countries exporting and importing live cattle.

Tembusu virus (TMUV) infection in ducks, geese and house sparrows was reported in China. To confirm the emergence of TMUV in humans, we investigated TMUV as a possible infection in duck industry workers in Shandong, China. Of 132 serum samples tested, 95 (71.9%) had TMUV antibodies. In oral swabs detection, 63 (47.7%) samples were positive for TMUV RNA. Nucleotide sequences of 277 bp coding the partial NS3 protein showed more than 99.5% identity with other duck TMUV strains, which can cause severe egg drop in ducks. These findings contribute to the realization that TMUV may be overlooked as a zoonotic transmission in China.

Foot-and-mouth disease (FMD) is endemic in India and causes severe economic loss. Status of FMD in the country for five fiscal years is presented. Outbreaks were more in number in 2007–2008 than 2010–2011. Three serotypes of FMD virus (O, A and Asia1) are prevalent. Serotype O was responsible for 80% of the confirmed outbreaks/cases, whereas Asia1 and A caused 12% and 8%, respectively. Geographical region-wise assessment indicated varying prevalence rate in different regions viz; 43% in Eastern region, 31.5% in Southern region, 11.6% in North-eastern region, 5% Central region, 4.4% Western region and 4% in Northern region. Highest number of outbreaks/cases was recorded in the month of September and lowest in June. Emergence and re-emergence of different genotypes/lineages within the serotypes were evident in real-time investigation carried out from time to time. Continues antigenic divergence in serotype A resulted in change in the vaccine strain in 2009. As on date, all genetic diversity within the serotypes is well tolerated by the vaccine strains. Unrestricted animal movements in the country play a major role in the spread of FMD.

Glanders or farcy, caused by Burkholderia mallei, is an infectious and zoonotic disease of solipeds. Horses, donkeys and mules are the only known natural reservoir of B. mallei. Although glanders has been eradicated from most countries, it has regained the status of a re-emerging disease because of the numerous recent outbreaks. Pre-symptomatic or carrier animals are the potential source of infection for the healthy equine population and play a crucial role in the spreading of the infectious agent. Glanders is characterized by ulcerating nodular lesions of the skin and mucous membrane. Generalized symptoms include fever, malaise, depression, cough, anorexia and weight loss. Burkholderia mallei can invade its host through mucous membranes, gastrointestinal tract and the integument. Its virulence mechanisms and pathogenesis are not yet completely understood. A major problem when using serological tests for diagnosing glanders is the occurrence of false-positive and false-negative results leading to difficulties in international trade with equids and to the spread of glanders to disease-free regions. Moreover, poor tests critically result in poor control of disease. These tests are not only incapable of discriminating between B. mallei and B. pseudomallei antibodies, they are also unable to differentiate between malleinized and naturally infected animals. Combined use of both serological and molecular detection methods increases the detection rate of glanders. Countermeasures against glanders include early detection of disease in susceptible animals, stringent quarantine measures, testing and safe destruction of infected carcasses, adequate compensation to the animal owners, disinfection of infected premises and awareness about glanders and the zoonotic implications through veterinary extension services. An account of the clinical picture and successful experimental therapy of spontaneous equine glanders is also given.

Outbreaks of highly pathogenic avian influenza (HPAI) H5N1 occurred in Nigeria between December 2005 and July 2008. We describe temporal and spatial characteristics of these outbreaks at State and Local Government Area (LGA) levels. A total of 25 of 37 States (67.6%; Exact 95% CI: 50.2–82.0%) and 81 of 774 LGAs (10.5%; Exact 95% CI: 8.4–12.8%) were affected by HPAI outbreaks over the period from 2005 to 2008. The incidence risk of HPAI outbreak occurrence at the State level was 5.6% (0.7–18.7%) for 2005, 50.0% (30.7–69.4%) for 2006, 54.5% (29.9–80.3%) for 2007 and 0% for 2008. Only very few LGAs experienced HPAI outbreaks within the affected States. The incidence risk of HPAI outbreak occurrence on a LGA level was 0.3% (0.0–0.9%) for 2005, 6.6% (4.9–8.6%) for 2006, 4.2% (2.9–6.0%) for 2007 and 0% for 2008. The mean period between farmers noticing HPAI outbreaks and reporting them to veterinary authorities, and between reporting HPAI outbreaks and the depopulation of infected premises, was for both 4.5 days; both periods also had medians of 1 day. We have estimated the spatially smoothed incidence risk for the whole outbreak period and identified the existence of a large corridor in the western part of Nigeria and a smaller corridor in south-eastern part, where the risk of HPAI occurrence was lower than in the rest of the country. The effect of HPAI control policies on the outbreaks patterns are discussed, as well as possible reasons why HPAI did not become endemic in Nigeria.

Forty-one cattle from seven Belgian farms and two French farms confirmed as infected with bluetongue virus serotype 8 (BTV-8) were monitored from the onset of clinical signs to describe the disease pattern and estimate the duration of blood RT-qPCR and competitiveELISA positivity under field conditions. On each visit, blood samples were taken, and a standardized clinical form was filled in for each animal. A clinical score was calculated for every week until the end of clinical signs. A classification and regression tree (CART) analysis was conducted to determine the most important clinical signs every week for the first 7weeks. The highest scores were recorded within 2 weeks of clinical onset. The first recorded clinical signs were quite obviously visible (lethargy, conjunctivitis, lesions of nasal mucosa, nasal discharge). Skin lesions, a drop in milk production and weight loss appeared later in the course of the disease. A biphasic pattern regarding nasal lesions was noticed: the first peak concerned mainly congestive and ulcerative lesions, whereas the second peak mainly concerned crusty lesions. The median time estimated by survival analysis to obtain negative RT-qPCR results from the onset of clinical signs was 195 days (range 166–213 days) in the 23 cattle included in the analysis. Serological results remained strongly positive until the end of the study. These results should ensure more accurate detection of an emerging infectious disease and are of prime importance in improving the modelling of BTV-8 persistence in Europe.

A loop-mediated isothermal amplification (LAMP) assay was developed to detect Borrelia burgdorferi s. l. in ticks, which is a pathogen that causes Lyme disease. Cross-reactions with Chlamydia psittaci, Mycoplasma mycoides subsp. capri and some tick-borne pathogens were excluded. Analytical sensitivity of LAMP showed its detection limit was from 0.02 to 0.2 pg of DNA in detection of the reference samples at 65°C for 40 min. The performance of LAMP was assessed by testing 110 samples from susceptible tick species and comparing the results with conventional and nested PCR tests previously described. The results demonstrated that LAMP was significantly more sensitive than the conventional PCR (32.7% versus 15.5%, P < 0.05) and slightly more sensitive, although not significantly so, than nested PCR (32.7% versus 26.4%, P > 0.05). The assay was used to analyse a total of 1052 ticks collected from eight provinces in China. The results showed that the infection rates of B. burgdorferi s. l. varied from 12.5% to 88.9% across the different geographical sites. Selected positive samples were subjected to sequencing and sequence analysis for conformation of the accuracy of the assay. Here we report a highly sensitive, specific and easy diagnostic assay based on LAMP technology. These data indicate that LAMP is a useful approach for detecting B. burgdorferi s. l. in field-collected ticks and has the potential as an alternative tool for the ecological and epidemiological surveillance of Lyme disease.

This work is an example of cooperation between veterinary and human medicine being fully complementary and at the same time, indispensable to improve our knowledge on animal chlamydiosis. This study investigated the existence of ocular chlamydiae and determined the prevalence of its presence, chlamydiosis, in asymptomatic and diseased farm animals and adjacent humans. Data were obtained by the omp2 gene family Chlamydiaceae-specific PCR. Two hundred cattle, buffaloes, sheep and goats and 44 human specimens were also examined. Conjunctival swabs from both the eyes were collected from all animals and humans using cotton swabs. Samples were tested for chlamydiae by Vero cells tissue culture, chicken embryo, modified Gimenez staining, direct fluorescein-conjugated monoclonal antibody staining (FA), immunoperoxidase, CFT and PCR. The PCR-RFLP revealed that Chlamydophila psittaci demonstrated in the conjunctival samples of cattle (68% asymptomatic and 88% diseased), of buffalo (68% asymptomatic and 72% diseased), of sheep (68% asymptomatic and 80% diseased), of goat (76% asymptomatic and 92% diseased) and of humans (77% asymptomatic and 82% diseased). The Cp. psittaci was the only chlamydiae demonstrated in all of the ocular conjunctival samples, which confirms the prevalence of Cp. psittaci in this population of animals and adjacent humans. Statistically, the animal species factor was calculated and was found to be of no significance. Yet, there appeared to be a significant difference in the percentage of animal that tested positive using the different methods. Detection of Cp. psittaci in most samples confirms the prevalence of Cp. psittaci in this population of animals and adjacent humans.

In vitro studies have demonstrated that bluetongue virus (BTV)-induced vasoactive mediators could contribute to the endothelial cells dysfunction and increased vascular permeability responsible of lesions characteristic of bluetongue (BT) like oedema, haemorrhages and ischaemic necrosis in different tissues. However, few in vivo studies have been carried out to clarify the causes of these lesions. The aim of this study was to elucidate in vivo the pathogenetic mechanisms involved in the appearance of vascular lesions in different organs during BT. For this purpose, tissue samples from goats naturally infected with bluetongue virus serotype 1 (BTV-1) were taken for histopathological and immunohistochemical studies to determine the potential role of proinflammatory cytokines (tumour necrosis factor alpha, TNFα and interleukin one alpha, IL-1α) in the increased vascular permeability and their relationship with the presence of virus. Gross and histopathological examination revealed the presence of vascular damage leading to generalized oedema and haemorrhages. Immunohistochemical studies displayed that endothelial injury may have been due to the direct pathogenic effect of BTV infection on endothelial cells or may be a response to inflammatory mediators released by virus-infected endothelial cells and, possibly, other cell types such as monocytes/macrophages. These preliminary results of what appears to be the first in vivo study of tissue damage in small BT-infected ruminants suggest a direct link between the appearance of vascular changes and the presence of BTV-induced vasoactive cytokines.

Bluetongue virus (BTV) is an economically important pathogen of ruminants that is the aetiological agent of the haemorrhagic disease bluetongue. Bluetongue virus is biologically transmitted by Culicoides biting midges (Diptera: Ceratopogonidae), and long-range dispersal of infected vector species contributes substantially to the rapid spread of the virus. The range of semi-passive flights of infected Culicoides on prevailing winds has been inferred to reach several hundred kilometres in a single night over water bodies. In this study, an atmospheric dispersion model was parameterized to simulate Culicoides flight activity based on dedicated entomological data sets collected in the UK. Five outbreaks of BTV in Europe were used to evaluate the model for use as an early warning tool and for retrospective analyses of BTV incursions. In each case, the generated predictions were consistent with epidemiological observations confirming its reliability for use in disease outbreak management. Furthermore, the model aided policy makers to predict, contain and eradicate BTV outbreaks in the UK during 2007 and 2008.

Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) infection is characterized by persisting in lungs and lymphoid tissue, resulting in systemic lymphoid depletion. The aim of this study was to correlate the histological changes, viral antigen expression and apoptosis phenomena in tonsil, medial retropharyngeal and mediastinal lymph nodes of 12 pigs inoculated with a type 2 PRRSV isolate (Chilean strain 2402). Apoptosis phenomena were observed mainly in lymphocytes and secondly in macrophages of lymph nodes and tonsils of inoculated animals, showing a peak of both apoptotic cells and viral antigen expression at the end of the study (21 dpi). However, the number of apoptotic cells was higher than the number of PRRSV-positive cells at the end of the study. This finding together with the location of apoptotic cells and PRRSV-positive cells in different structures of lymphoid organs supports the hypothesis that PRRSV-positive macrophages might modulate the apoptosis phenomena in other cells, mainly lymphocytes, by means of an indirect mechanism. Furthermore, apoptotic cells were detected both in B- and T-cell areas of lymphoid organs, suggesting that apoptosis phenomena may play a role in the impairment of the host immune response during PRRS.

Bovine spongiform encephalopathy (BSE), popularly known as ‘mad cow disease’, led to an epidemic in Europe that peaked in the mid-1990s. Its impact on developing countries, such as Nigeria, has not been fully established as information on livestock and surveillance has eluded those in charge of this task. The BSE risk to Nigeria’s cattle population currently remains undetermined, which has resulted in international trade restrictions on commodities from the cattle population. This is mainly because of a lack of updated BSE risk assessments and disease surveillance data. To evaluate the feasibility of BSE surveillance in Nigeria, we carried out a pilot study targeting cattle that were presented for emergency or casualty slaughter. In total, 1551 cattle of local breeds, aged 24 months and above were clinically examined. Ataxia, recumbency and other neurological signs were topmost on our list of criteria. A total of 96 cattle, which correspond to 6.2%, presented clinical signs that supported a suspect of BSE. The caudal brainstem tissues of these animals were collected post-mortem and analysed for the disease-specific form of the prion protein using a rapid test approved by the International Animal Health Organization (OIE). None of the samples were positive for BSE. Although our findings do not exclude the presence of BSE in Nigeria, they do demonstrate that targeted sampling of clinically suspected cases of BSE is feasible in developing countries. In addition, these findings point to the possibility of implementing clinical monitoring schemes for BSE and potentially other diseases with grave economic and public health consequences.