Survival of the ovine footrot pathogen Dichelobacter nodosus in different soils.

Dichelobacter nodosus (D. nodosus) is the causative agent of footrot in sheep; one of the most important health and welfare issues of sheep worldwide. For control programmes to be effective, it is essential that the transmission cycle of D. nodosus is understood and bacterial reservoirs in the environment are better defined. This study evaluated the survival of D. nodosus in different soils using soil microcosms. Cultivation independent and dependent methods were used to detect D. nodosus over 40 days from seeding in soil. A D. nodosus specific probe was used for quantification by qPCR and viability was assessed by cell permeability to an intercalating dye, PMA, and by culture. Survival varied dramatically depending on soil type, matric potential (MP) and temperature. Our findings indicate that D. nodosus survival was higher at 5 °C compared with 25 °C in all soils and significantly longer at both temperatures in clay soil (>44% clay) compared with other soil types. Survival under all conditions was longer than 30 days for both culture independent and dependent methods, this is substantially longer than previous studies and, if this is an infectious dose, longer than the current recommendation of resting a field for 14 days to prevent onward infection.

Challenges and Opportunities of Epidemiological Studies to Reduce the Burden of Cancers in Young Adults.

There are >1.9 million survivors of adolescent and young adult cancers (AYA, diagnosed at ages 15-39) living in the U.S. today. Epidemiologic studies to address the cancer burden in this group have been a relatively recent focus of the research community. In this article, we discuss approaches and data resources for cancer epidemiology and health services research in the AYA population. We consider research that uses data from cancer registries, vital records, healthcare utilization, and surveys, and the accompanying challenges and opportunities of each. To illustrate the strengths of each data source, we present example research questions or areas that are aligned with these data sources and salient to AYAs. Integrating the respective strengths of cancer registry, vital records, healthcare data, and survey-based studies sets the foundation for innovative and impactful research on AYA cancer treatment and survivorship to inform a comprehensive understanding of diverse AYA needs and experiences.

Evaluating observer agreement of scoring systems for foot integrity and footrot lesions in sheep.

BACKGROUND: A scoring scale with five ordinal categories is used for visual diagnosis of footrot in sheep and to study its epidemiology and control. More recently a 4 point ordinal scale has been used by researchers to score foot integrity (wall and sole horn damage) in sheep. There is no information on observer agreement using either of these scales. Observer agreement for ordinal scores is usually estimated by single measure values such as weighted kappa or Kendall's coefficient of concordance which provide no information where the disagreement lies. Modeling techniques such as latent class models provide information on both observer bias and whether observers have different thresholds at which they change the score given. In this paper we use weighted kappa and located latent class modeling to explore observer agreement when scoring footrot lesions (using photographs and videos) and foot integrity (using post mortem specimens) in sheep. We used 3 observers and 80 photographs and videos and 80 feet respectively. RESULTS: Both footrot and foot integrity scoring scales were more consistent within observers than between. The weighted kappa values between observers for both footrot and integrity scoring scales ranged from moderate to substantial. There was disagreement between observers with both observer bias and different thresholds between score values. The between observer thresholds were different for scores 1 and 2 for footrot (using photographs and videos) and for all scores for integrity (both walls and soles). The within observer agreement was higher with weighted kappa values ranging from substantial to almost perfect. Within observer thresholds were also more consistent than between observer thresholds. Scoring using photographs was less variable than scoring using video clips or feet. CONCLUSIONS: Latent class modeling is a useful method for exploring components of disagreement within and between observers and this information could be used when developing a scoring system to improve reliability.

Differences in composition of interdigital skin microbiota predict sheep and feet that develop footrot.

Footrot has a major impact on health and productivity of sheep worldwide. The current paradigm for footrot pathogenesis is that physical damage to the interdigital skin (IDS) facilitates invasion of the essential pathogen Dichelobacter nodosus. The composition of the IDS microbiota is different in healthy and diseased feet, so an alternative hypothesis is that changes in the IDS microbiota facilitate footrot. We investigated the composition and diversity of the IDS microbiota of ten sheep, five that did develop footrot and five that did not (healthy) at weekly intervals for 20 weeks. The IDS microbiota was less diverse on sheep 2 + weeks before they developed footrot than on healthy sheep. This change could be explained by only seven of > 2000 bacterial taxa detected. The incubation period of footrot is 8-10 days, and there was a further reduction in microbial diversity on feet that developed footrot in that incubation period. We conclude that there are two stages of dysbiosis in footrot: the first predisposes sheep to footrot and the second occurs in feet during the incubation of footrot. These findings represent a step change in our understanding of the role of the IDS microbiota in footrot pathogenesis.

Social behaviour and transmission of lameness in a flock of ewes and lambs.

Introduction: Sheep have heterogenous social connections that influence transmission of some infectious diseases. Footrot is one of the top five globally important diseases of sheep, it is caused by Dichelobacter nodosus and transmits between sheep when infectious feet contaminate surfaces, e.g., pasture. Surfaces remain infectious for a few minutes to a few days, depending on surface moisture levels. Susceptible sheep in close social contact with infectious sheep might be at risk of becoming infected because they are likely to step onto infectious footprints, particularly dams and lambs, as they cluster together. Methods: High resolution proximity sensors were deployed on 40 ewes and their 54 lambs aged 5-27 days, in a flock with endemic footrot in Devon, UK for 13 days. Sheep locomotion was scored daily by using a 0-6 integer scale. Sheep were defined lame when their locomotion score (LS) was ≥2, and a case of lameness was defined as LS ≥2 for ≥2 days. Results: Thirty-two sheep (19 ewes, 9 single, and 4 twin lambs) became lame during the study, while 14 (5 ewes, 5 single, and 4 twin lambs) were lame initially. These 46 sheep were from 29 family groups, 14 families had >1 lame sheep, and transmission from ewes to lambs was bidirectional. At least 15% of new cases of footrot were from within family transmission; the occurrence of lameness was higher in single than twin lambs. At least 4% of transmission was due to close contact across the flock. Most close contact occurred within families. Single and twin lambs spent 1.5 and 0.9 hours/day with their dams, respectively, and twin lambs spent 3.7 hours/day together. Non-family sheep spent only 0.03 hours/day in contact. Lame single lambs and ewes spent less time with non-family sheep, and lame twin lambs spent less time with family sheep. Discussion: We conclude that most transmission of lameness is not attributable to close contact. However, in ewes with young lambs, some transmission occurs within families and is likely due to time spent in close contact, since single lambs spent more time with their dam than twin lambs and were more likely to become lame.

Impact of Strain Variation of Dichelobacter nodosus on Disease Severity and Presence in Sheep Flocks in England.

AprV2 and aprB2 are variants of the apr gene of Dichelobacter nodosus, the cause of footrot in sheep. They are putative markers for severe and mild disease expression. The aim of our study was to investigate the distribution of aprV2 and aprB2 in flocks with and without footrot. Our hypotheses were that both strains are present in endemically affected flocks, with aprB2 and aprV2 associated with mild and virulent phenotypes respectively but that D. nodosus is not present in flocks without footrot. Alternatively, aprB2 persists in flocks without footrot. Despite extensive searching over 3 years only three flocks of sheep without footrot were identified. D. nodosus was not detected in these three flocks. In one further flock, only mild interdigital dermatitis was observed, and only aprB2 was detected. Twenty-four flocks with endemic footrot of all severities were sampled on three occasions and all were positive for D. nodosus and the aprV2 variant; aprB2 was detected in only 11 of these flocks. AprB2 was detected as a co-infection with aprV2 in the 22% of samples positive for aprB2 and was more likely in mild footrot phenotypes than severe. Dichelobacter nodosus serogroups were not associated with footrot phenotype. We conclude that D. nodosus, even aprB2 strains, do not persist in flocks in the absence of footrot. Our results support the hypothesis that aprB2 is associated with mild footrot phenotypes. Finally, we conclude that given the small number of flocks without footrot that were identified, footrot is highly endemic in English sheep flocks.

A Pilot Study to Investigate the Feasibility of a Multiple Locus Variable Number Tandem Repeat Analysis to Understand the Epidemiology of Dichelobacter nodosus in Ovine Footrot.

Dichelobacter nodosus is the essential pathogen in ovine footrot, an important cause of lameness in sheep that reduces productivity and welfare. The aim of this study was to investigate the feasibility of using multiple locus variable number tandem repeat analysis (MLVA) developed to investigate isolates to understand the molecular epidemiology of Dichelobacter nodosus in ovine footrot by investigation of communities of strains. MLVA sensitivity was improved by optimizing PCR conditions to 100% specificity for D. nodosus. The improved MLVA scheme was used to investigate non-cultured DNA purified from swabs (swab DNA) and cultured DNA from isolates (isolate DNA) from 152 foot and 38 gingival swab samples from 10 sheep sampled on four occasions in a longitudinal study. Isolate DNA was obtained from 6/152 (3.9%) feet and 5/6 yielded complete MLVA profiles, three strains were detected. Two of the three isolate strains were also detected in isolate DNA from 2 gingival crevice cultures. Complete MLVA profiles were obtained from swab DNA from 39 (25.7%) feet. There were 22 D. nodosus community types that were comprised of 7 single strain and 15 multi-strain communities. Six community types were detected more than once and three of these were detected on the same four sheep and the same two feet over time. There were a minimum of 17 and a maximum of 25 strain types of D. nodosus in the study. The three isolate strain types were also the most frequently detected strain types in swab DNA. We conclude that the MLVA from swab DNA detects the same strains as culture, is much more sensitive and can be used to describe and differentiate communities and strains on sheep, feet and over time. It is therefore a sensitive molecular tool to study D. nodosus strains directly from DNA without culture.

Serogroups of Dichelobacter nodosus, the cause of footrot in sheep, are randomly distributed across England.

We present the largest and most representative study of the serological diversity of Dichelobacter nodosus in England. D. nodosus causes footrot and is one of the top five globally important diseases of sheep. The commercial vaccine, containing nine serogroups, has low efficacy compared with bivalent vaccines. Our aim was to investigate the prevalence and distribution of serogroups of D. nodosus in England to elucidate whether a bivalent vaccine could protect the national flock. Farmers from 164 flocks submitted eight interdigital swabs from eight, preferably diseased, sheep. All serogroups, A-I, were detected by PCR in 687/1150 D. nodosus positive swabs, with a prevalence of 2.6-69.3% of positive swabs per serogroup. There was a median of two serogroups per flock (range 0-6). Serogroups were randomly distributed between, but clustered within, flocks, with 50 combinations of serogroups across flocks. H and B were the most prevalent serogroups, present in > 60% of flocks separately but in only 27% flocks together. Consequently, a bivalent vaccine targeting these two serogroups would protect 27% of flocks fully (if only H and B present) and partially, if more serogroups were present in the flock. We conclude that one bivalent vaccine would not protect the national flock against footrot and, with 50 combinations of serogroups in flocks, flock-specific vaccines are necessary.

A cross sectional study of the prevalence, risk factors and population attributable fractions for limb and body lesions in lactating sows on commercial farms in England.

BACKGROUND: Lesions on sows' limbs and bodies are an abnormality that might impact on their welfare. The prevalence of and risks for limb and body lesions on lactating sows on commercial English pig farms were investigated using direct observation of the sows and their housing. RESULTS: The prevalence of lesions on the limbs and body were 93% (260/279) and 20% (57/288) respectively. The prevalence of limb and body lesions was significantly lower in outdoor-housed sows compared with indoor-housed sows. Indoor-housed sows had an increased risk of wounds (OR 6.8), calluses (OR 8.8) and capped hock (OR 3.8) on their limbs when housed on fully slatted floors compared with solid concrete floors. In addition, there was an increased risk of bursitis (OR 2.7), capped hock (OR 2.3) and shoulder lesions (OR 4.8) in sows that were unwilling to rise to their feet. There was a decreased risk of shoulder lesions (OR 0.3) and lesions elsewhere on the body (OR 0.2) in sows with more than 20 cm between their tail and the back of the crate compared with sows with less than 10 cm. CONCLUSION: The sample of outdoor housed sows in this study had the lowest prevalence of limb and body lesions. In lactating sows housed indoors there was a general trend for an increased risk of limb and body lesions in sows housed on slatted floors compared with those housed on solid concrete floors with bedding. Sows that were less responsive to human presence and sows that had the least space to move within their crates had an additional increased risk of lesions.

A cross sectional study of prevalence, risk factors, population attributable fractions and pathology for foot and limb lesions in preweaning piglets on commercial farms in England.

BACKGROUND: In a cross sectional study of 88 indoor and outdoor English pig farms, the prevalence of foot and limb lesions in 2843 preweaning piglets aged 1-4 weeks from 304 litters was recorded. The environmental risks for the prevalence of lesions and population attributable fractions were calculated. The risks for lesions in piglets were compared with those for limb and body lesions in their mothers. A small number of piglets with each type of lesion were examined post mortem to elucidate the pathology of the clinical lesions observed. RESULTS: The prevalence of sole bruising, sole erosion, skin abrasion and swollen joints or claws in 2843 piglets was 49.4% (1404), 15.5% (441), 43.6% (1240) and 4.7% (143) respectively. The prevalence of all foot and limb lesions was higher in indoor housed piglets than in outdoor housed piglets. The prevalence of sole bruising (OR 0.3) and skin abrasion (OR 0.6) decreased with each week of age from 1-4 weeks, but there was no significant association between piglet age and the prevalence of sole erosion or swollen joints and claws. There was an increased prevalence of sole bruising (OR 3.0) and swollen joints or claws (OR 3.0) and a decreased prevalence of skin abrasion (OR 0.3, piglets

A four year longitudinal sero-epidemiological study of bovine herpesvirus type-1 (BHV-1) in adult cattle in 107 unvaccinated herds in south west England.

BACKGROUND: Bovine herpesvirus type-1 (BHV-1) is an important pathogen of cattle that presents with a variety of clinical signs, including the upper respiratory tract infection infectious bovine rhinotracheitis (IBR). A seroepidemiological study of BHV-1 antibodies was conducted in England from 2002 - 2004: 29,782 blood samples were taken from 15,736 cattle from 114 herds which were visited on up to three occasions. Antibody concentration was measured using a commercial ELISA. Farm management information was collected using an interview questionnaire, and herd size and cattle movements were obtained from the cattle tuberculosis testing database and the British Cattle Movement Service. Hierarchical statistical models were used to investigate associations between cattle and herd variables and the continuous outcome percentage positive (PP) values from the ELISA test in unvaccinated herds. RESULTS: There were 7 vaccinated herds, all with at least one seropositive bovine. In unvaccinated herds 83.2% had at least one BHV-1 seropositive bovine, and the mean cattle and herd BHV-1 seroprevalence were 42.5% and 43.1% respectively. There were positive associations between PP value, age, herd size, presence of dairy cattle. Adult cattle in herds with grower cattle had lower PP values than those in herds without grower cattle. Purchased cattle had significantly lower PP values than homebred cattle, whereas cattle in herds that were totally restocked after the foot-and-mouth epidemic in 2001 had significantly higher PP values than those in continuously stocked herds. Samples taken in spring and summer had significantly lower PP values than those taken in winter, whereas those taken in autumn had significantly higher PP values than those taken in winter. The risks estimated from a logistic regression model with a binary outcome (seropositive yes/no) were similar. CONCLUSION: The prevalence of BHV-1 seropositivity in cattle and herds has increased since the 1970s. Although the study population prevalence of BHV-1 was temporally stable during study period, the associations between serological status and cattle age, herd size, herd type, presence of young stock and restocked versus continuously stocked herds indicate that there is heterogeneity between herds and so potential for further spread of BHV-1 within and between herds.

Gene mutations and genomic rearrangements in the mouse as a result of transposon mobilization from chromosomal concatemers.

Previous studies of the Sleeping Beauty (SB) transposon system, as an insertional mutagen in the germline of mice, have used reverse genetic approaches. These studies have led to its proposed use for regional saturation mutagenesis by taking a forward-genetic approach. Thus, we used the SB system to mutate a region of mouse Chromosome 11 in a forward-genetic screen for recessive lethal and viable phenotypes. This work represents the first reported use of an insertional mutagen in a phenotype-driven approach. The phenotype-driven approach was successful in both recovering visible and behavioral mutants, including dominant limb and recessive behavioral phenotypes, and allowing for the rapid identification of candidate gene disruptions. In addition, a high frequency of recessive lethal mutations arose as a result of genomic rearrangements near the site of transposition, resulting from transposon mobilization. The results suggest that the SB system could be used in a forward-genetic approach to recover interesting phenotypes, but that local chromosomal rearrangements should be anticipated in conjunction with single-copy, local transposon insertions in chromosomes. Additionally, these mice may serve as a model for chromosome rearrangements caused by transposable elements during the evolution of vertebrate genomes.

Increase in the flock prevalence of lameness in ewes is associated with a reduction in farmers using evidence-based management of prompt treatment: A longitudinal observational study of 154 English sheep flocks 2013-2015.

Since 2006, farmers in England have received new recommendations on best practice to manage lameness in sheep through a range of knowledge exchange activities. The adoption of each recommendation varied, but in 2013 approximately 50% of farmers reported treating all lame sheep within 3 days of onset of lameness (prompt treatment), 41% did not practice routine foot trimming, 50% culled sheep that had been lame and 14% vaccinated against footrot; all recommended best practices. The aim of this study was to investigate the prevalence of lameness in ewes in England from 2013 to 2015 and to identify changes in practice to manage lameness between 2013 and 2015 and the population attributable fraction for these managements. A longitudinal study with a cohort of 154 English sheep farmers was run for three years, farmers completed questionnaires on lameness in their flock for the previous 12 months in 2013, 2014 and 2015. The geometric mean prevalence of lameness in ewes was 4.1% in 2015, significantly higher than 3.3% and 3.2% for the same 128 farmers who provided data in both 2013 and 2014. Between 2013 and 2015 there was a significant reduction in farmers practising prompt treatment (50.6%-28.6%) but an increase in not practising routine foot trimming (40.9%-79.2%), culling sheep that had been lame (49.4%-81.8%), and vaccinating against footrot (14.3%-29.2%). Not practising prompt treatment, ≥5% of sheep feet bleeding during routine foot trimming, vaccinating ewes for <6 years or not vaccinating at all, and other flocks mixing with the flock, were associated with a significantly higher flock prevalence of lameness. Culling sheep that had been lame was not associated with prevalence of lameness. The population attributable fractions (PAFs) for not vaccinating for>5 years, not treating lame sheep promptly, ≥5% of sheep feet bleeding during routine foot trimming, and mixing of flocks were 34.5%, 25.3%, 2.9% and 2.4%. In 2013, when 50% of farmers used prompt treatment, the PAF for not using prompt treatment was only 13.3%. We conclude that the change in practice by these farmers towards flock-level managements and a reduction in individual prompt treatment of lame sheep negatively impacted the prevalence of lameness in sheep. This change occurred despite the evidence that prompt treatment of lame sheep is highly effective at reducing the prevalence of lameness in sheep flocks and is an example of cognitive dissonance.

Sites of persistence of Fusobacterium necrophorum and Dichelobacter nodosus: a paradigm shift in understanding the epidemiology of footrot in sheep.

Sites of persistence of bacterial pathogens contribute to disease dynamics of bacterial diseases. Footrot is a globally important bacterial disease that reduces health and productivity of sheep. It is caused by Dichelobacter nodosus, a pathogen apparently highly specialised for feet, while Fusobacterium necrophorum, a secondary pathogen in footrot is reportedly ubiquitous on pasture. Two prospective longitudinal studies were conducted to investigate the persistence of D. nodosus and F. necrophorum in sheep feet, mouths and faeces, and in soil. Molecular tools were used to detect species, strains and communities. In contrast to the existing paradigm, F. necrophorum persisted on footrot diseased feet, and in mouths and faeces; different strains were detected in feet and mouths. D. nodosus persisted in soil and on diseased, but not healthy, feet; similar strains were detected on both healthy and diseased feet of diseased sheep. We conclude that D. nodosus and F. necrophorum depend on sheep for persistence but use different strategies to persist and spread between sheep within and between flocks. Elimination of F. necrophorum would be challenging due to faecal shedding. In contrast D. nodosus could be eliminated if all footrot-affected sheep were removed and fade out of D. nodosus occurred in the environment before re-infection of a foot.

Porcine reproductive and respiratory syndrome virus (PRRSV) in GB pig herds: farm characteristics associated with heterogeneity in seroprevalence.

BACKGROUND: The between- and within-herd variability of porcine reproductive and respiratory syndrome virus (PRRSV) antibodies were investigated in a cross-sectional study of 103 British pig herds conducted 2003-2004. Fifty pigs from each farm were tested for anti-PRRSV antibodies using ELISA. A binomial logistic model was used to investigate management risks for farms with and without pigs with PRRSV antibodies and multilevel statistical models were used to investigate variability in pigs' log ELISA IRPC (relative index x 100) in positive herds. RESULTS: Thirty-five herds (34.0%) were seronegative, 41 (39.8%) were seropositive and 27 (26.2%) were vaccinated. Herds were more likely to be seronegative if they had < 250 sows (OR 3.86 (95% CI 1.46, 10.19)) and if the nearest pig herd was > or = 2 miles away (OR 3.42 (95% CI 1.29, 9.12)). The mean log IRPC in seropositive herds was 3.02 (range, 0.83 - 5.58). Sixteen seropositive herds had only seropositive adult pigs. In these herds, pigs had -0.06 (95% CI -0.10, -0.01) lower log IRPC for every mile increase in distance to the nearest pig unit, and -0.56 (95% CI -1.02, -0.10) lower log IRPC when quarantine facilities were present. For 25 herds with seropositive young stock and adults, lower log IRPC were associated with isolating purchased stock for > or = 6 days (coefficient -0.46, 95% CI -0.81, -0.11), requesting > or = 48 hours 'pig-free time' from humans (coefficient -0.44, 95% CI -0.79, -0.10) and purchasing gilts (coefficient -0.61, 95% CI -0.92, -0.29). CONCLUSION: These patterns are consistent with PRRSV failing to persist indefinitely on some infected farms, with fadeout more likely in smaller herds with little/no reintroduction of infectious stock. Persistence of infection may be associated with large herds in pig-dense regions with repeated reintroduction.

A four year longitudinal sero-epidemiology study of Neospora caninum in adult cattle from 114 cattle herds in south west England: associations with age, herd and dam-offspring pairs.

BACKGROUND: Neosporosis caused by the protozoan parasite Neospora caninum, is an economically important cause of abortion, stillbirth, low milk yield, reduced weight gain and premature culling in cattle. Consequently, a seroepidemiological study of N. caninum antibodies was conducted in England with 29,782 samples of blood taken from 15,736 cattle from 114 herds visited on three occasions at yearly intervals. Herds were categorised into lower (< 10%) and higher (> or = 10%) median herd seroprevalence. Hierarchical models were run to investigate associations between the sample to positive (S/P) ratio and herd and cattle factors. RESULTS: Ninety-four percent of herds had at least one seropositive cow; 12.9% of adult cattle had at least one seropositive test. Approximately 90% of herds were seropositive at all visits; 9 herds (8%) changed serological status between visits. The median N. caninum seroprevalence in positive herds was 10% (range 0.4% to 58.8%). There was a positive association between the serostatus of offspring and dams that were ever seropositive. In the hierarchical model of low seroprevalence herds there was no significant association between S/P ratio and cattle age. There was a significantly lower S/P ratio in cattle in herds that were totally restocked after the foot-and-mouth epidemic of 2001 compared with those from continuously stocked herds and cattle purchased into these herds had a higher S/P ratio than homebred cattle. In the model of high seroprevalence herds the S/P ratio increased with cattle age, but was not associated with restocking or cattle origin. CONCLUSION: There were no strong temporal changes in herd seroprevalence of N. caninum but 90% of herds had some seropositive cattle over this time period. Vertical transmission from seropositive dams appeared to occur in all herds. In herds with a high seroprevalence the increasing S/P ratio in 2-4 year old cattle is suggestive of exposure to N. caninum: horizontal transmission between adult cattle, infection from a local source or recrudescence and abortions. Between-herd movements of infected cattle enhance the spread of N. caninum, particularly into low seroprevalence herds. Some restocked herds had little exposure to N. caninum, while in others infection had spread in the time since restocking.

Development and validation of a multiple locus variable number tandem repeat analysis (MLVA) scheme for Fusobacterium necrophorum.

Fusobacterium necrophorum is associated with various diseases in humans and animals. Reservoirs (sites where the pathogen persists in the absence of disease) of F. necrophorum are believed to be present in healthy individuals e.g. tonsillar epithelium, or their environment e.g. soil, but for most diseases the reservoir sites are unknown. Strain typing of F. necrophorum would facilitate linking specific reservoirs with a specific disease. The aim of this study was to develop multiple locus variable number tandem repeat analysis (MLVA) as a strain typing technique for F. necrophorum, and to test the use of this scheme to analyse both isolates and mixed communities of bacteria. Seventy-three tandem repeat regions were identified in the F. necrophorum genome; three of these loci were suitable and developed as a MLVA scheme. The MLVA scheme was sensitive, specific, and discriminatory for both isolates and communities of F. necrophorum. The MLVA scheme strain typed 46/52F. necrophorum isolates including isolates of both subspecies and from different countries, host species and sample sites within host. There were 12 unique MLVA strain types that clustered by subspecies. The MLVA scheme characterised the F. necrophorum community in DNA from 32/49 foot- and 28/33 mouth swabs from sheep. There were 17 community types in total. In 31/32 foot swabs, single strains of F. necrophorum were detected while in the 28 mouth swabs there were up to a maximum of 8 strains of F. necrophorum detected. The results demonstrate the potential for this method to elucidate reservoirs of F. necrophorum.

Development and validation of an ELISA to detect antibodies to Corynebacterium pseudotuberculosis in ovine sera.

Several enzyme-linked immunosorbent assays (ELISAs) have been developed for the detection of antibodies to Corynebacterium pseudotuberculosis, the causative agent of caseous lymphadenitis (CLA). However, none are commercially available in the UK. It was therefore necessary to develop a new, economic ELISA for use in a research project studying the epidemiology of CLA in UK sheep. The ELISA with its diagnostic qualities is presented. The ELISA was developed using sonicated C. pseudotuberculosis and optimised to detect total antibody or IgG class antibody in serum. Receiver operating characteristic (ROC) curves were obtained and the area under the ROC curve was used to compare the sensitivity and specificity of the two ELISAs. Both versions of the ELISA were evaluated on a panel of 150 positive reference sera and 103 negative reference sera. Using the test at 100% specificity, the sensitivity of detection of total antibody was 71% (95% confidence interval 63-78%), and the sensitivity of detection of IgG antibody to C. pseudotuberculosis was 83% (76-89%), which compares favourably with other reported ELISA tests for CLA in sheep. The sensitivity of the IgG antibody assay may be higher because of the greater affinity of IgG class antibodies compared with the IgM antibodies also detected by the total antibody ELISA. The results of ROC analysis indicated that the IgG isotype ELISA was more accurate than the total antibody ELISA. The efficiency of the test was greatest when serum samples were run in a dilution series than when any single serum dilution was used. The ELISA is considered to be suitable for application in field studies of CLA in UK sheep.

Cost-benefit analysis of management practices for ewes lame with footrot.

The aim of this study was to investigate the cost-benefit of different strategies to treat and control ovine footrot. In November 2006, 162 sheep farmers in England responded to a survey on prevalence and management of lameness. The costs of lameness per ewe per year (PEPY) were calculated for 116 flocks. Linear regression was used to model the overall cost of lameness PEPY by management method. Associations between farmer satisfaction and time and money spent managing lameness were investigated. The median prevalence of lameness was 5% (inter-quartile range, IQR, 4-10%). The overall cost of lameness PEPY in flocks with ≥10% lameness was UK £6.35 versus £3.90 for flocks with <5% lameness. Parenteral antibiotic treatment was associated with a significantly lower overall cost of lameness by £0.79 PEPY. Routine foot trimming and foot bathing were associated with significantly higher overall costs of lameness PEPY of £2.96 and £0.90, respectively. Farmers satisfied with time managing lameness spent significantly less time (1.46 h PEPY) than unsatisfied farmers (1.90 h PEPY). Farmers satisfied with money spent managing lameness had significantly lower treatment (£2.94 PEPY) and overall (£5.00 PEPY) costs than dissatisfied farmers (£5.50 and £7.60 PEPY, respectively). If the farmers in this study adopted best practice of parenteral antibiotic treatment with no routine foot trimming, and minimised foot bathing to treatment/prevention of interdigital dermatitis, the financial benefits would be approximately £4.65 PEPY. If these costs are similar on other farms the management changes would lead to significant economic benefits for the sheep industry.