A review of risk factors for bovine tuberculosis infection in cattle in the UK and Ireland.

Bovine tuberculosis (bTB) is an important disease of cattle caused by infection with Mycobacterium bovis, a pathogen that may be extremely difficult to eradicate in the presence of a true wildlife reservoir. Our objective was to identify and review relevant literature and provide a succinct summary of current knowledge of risk factors for transmission of infection of cattle. Search strings were developed to identify publications from electronic databases to February 2015. Abstracts of 4255 papers identified were reviewed by three reviewers to determine whether the entire article was likely to contain relevant information. Risk factors could be broadly grouped as follows: animal (including nutrition and genetics), herd (including bTB and testing history), environment, wildlife and social factors. Many risk factors are inter-related and study designs often do not enable differentiation between cause and consequence of infection. Despite differences in study design and location, some risk factors are consistently identified, e.g. herd size, bTB history, presence of infected wildlife, whereas the evidence for others is less consistent and coherent, e.g. nutrition, local cattle movements. We have identified knowledge gaps where further research may result in an improved understanding of bTB transmission dynamics. The application of targeted, multifactorial disease control regimens that address a range of risk factors simultaneously is likely to be a key to effective, evidence-informed control strategies.

Mycobacterium bovis infections in domesticated non-bovine mammalian species. Part 1: Review of epidemiology and laboratory submissions in Great Britain 2004-2010.

Mycobacterium bovis, the causative agent of bovine tuberculosis (bTB), can infect a broad range of mammalian species in addition to domestic and feral cattle and badgers. Since legislation introduced in 2006 in Great Britain requires animal keepers, meat inspectors and veterinarians to notify the authorities of suspect bTB lesions or the isolation of M. bovis in any mammal excluding humans, the organism has been increasingly identified in domestic species other than cattle. Although in most cases 'spill-over' hosts, these remain a potential source of infection for cattle, wildlife, and possibly humans. In this first part of a two-part review of M. bovis infections in non-bovine domestic species, current knowledge of the epidemiology of such infections is presented along with novel data relating to diagnostic submissions for mycobacterial culture between 2004 and 2010. Over this period M. bovis infection was identified in 116 cats, 7 dogs, 34 llamas, 133 alpacas, 35 goats, 24 sheep and 85 pigs and wild boar. The risk that such infections pose to the control of bTB, and as zoonoses, is discussed. In part two, the options available to diagnose bTB in these species, as well as the challenges posed to disease detection and control will be discussed in depth.

Mycobacterium bovis infections in domesticated non-bovine mammalian species. Part 2: A review of diagnostic methods.

Despite the large host range of Mycobacterium bovis, ante-mortem diagnostic tests for the infection mostly lack sensitivity/specificity and/or remain unvalidated in non-bovine species. The epidemiology and importance of M. bovis infection in these species are discussed in the first part of this two-part review. This second part focuses on the diagnostic options available to identify infected species such as sheep, goats, dogs, cats, and camelids, and highlights the significant challenges posed, both in establishing estimates of disease prevalence and in controlling infections in these species, in the absence of fully validated tests.

Factors associated with bovine tuberculosis confirmation rates in suspect lesions found in cattle at routine slaughter in Great Britain, 2003-2008.

Bovine tuberculosis (bTB) is one of the most complex and intractable animal health problems facing the British cattle industry today. The inspection of carcasses from cattle sent to slaughter is part of routine surveillance for bTB in Great Britain (GB). Tissue with suspect lesions from cattle from herds previously considered uninfected with bTB is sent to the Animal Health and Veterinary Laboratories Agency (AHVLA) for culture and histopathological examination for Mycobacterium bovis infection. In this study, risk factors for confirmation of infection in suspect bTB lesions found at routine slaughter of cattle from officially bTB-free (OTF) herds in GB were investigated. The study sample included the first record of a suspect lesion in a bovine from any OTF herd identified during post-mortem inspection between 2003 and 2008. There were 3663 submissions from 151 slaughterhouses of which 2470 (67.4%) were confirmed as culture positive for M. bovis. Logistic regression analysis with a random intercept for slaughterhouse was used to investigate relationships between bTB confirmation and animal and herd-level risk factors. Slaughterhouse of post mortem and the following factors related to bTB prevalence were significant predictors of confirmation probability: region of farm of origin of the animal, the testing interval for routine field surveillance for bTB on the farm, number of reactors in the last bTB incident on the farm within the last 4 years, if applicable, the animal's date of birth and the year of animal's slaughter. The modelled predicted population averaged probabilities for confirmation varied from 0.14 to 0.90 between slaughterhouses. Differences in the detection of cattle with bTB between British slaughterhouses warrant further study.

Long-term temporal trends and estimated transmission rates for Mycobacterium bovis infection in an undisturbed high-density badger (Meles meles) population.

We describe epidemiological trends in Mycobacterium bovis infection in an undisturbed wild badger (Meles meles) population. Data were derived from the capture, clinical sampling and serological testing of 1803 badgers over 9945 capture events spanning 24 years. Incidence and prevalence increased over time, exhibiting no simple relationship with host density. Potential explanations are presented for a marked increase in the frequency of positive serological test results. Transmission rates (R0) estimated from empirical data were consistent with modelled estimates and robust to changes in test sensitivity and the spatial extent of the population at risk. The risk of a positive culture or serological test result increased with badger age, and varied seasonally. Evidence consistent with progressive disease was found in cubs. This study demonstrates the value of long-term data and the repeated application of imperfect diagnostic tests as indices of infection to reveal epidemiological trends in M. bovis infection in badgers.

Tuberculin manufacturing source and breakdown incidence rate of bovine tuberculosis in British cattle, 2005-2009.

The single intradermal comparative cervical tuberculin (SICCT) test is the primary test used for surveillance for bovine tuberculosis (bTB) in cattle in Great Britain (GB). The tuberculin used can, with other factors, influence test accuracy. In this analysis, the detection of infected cattle in GB 2005-2009 was compared between SICCT tests using tuberculins manufacturered by different manufacturers. Higher rates of reactors (adjusted rate 209 vs 186 per 100,000 tests, P = 0.003) and herd bTB incidents (adjusted total breakdown rate 5.1 vs 4.5 per 100 herd-years at risk, P < 0.001) were detected using tuberculin manufactured at Weybridge compared with Lelystad. However, confirmation of infection in reactors by postmortem evidence was higher with Lelystad tuberculin (adjusted percent 44.1 vs 47.1, P = 0.018). The findings, overall, suggest slightly higher test sensitivity and lower test specificity associated with Weybridge tuberculin compared with Lelystad. Assuming effective adjustment for confounding, the overall impact of tuberculin manufacturing source (2007-2009), was calculated to range somewhere between 315 false positive breakdowns, and 1086 bTB breakdowns missed (624 confirmed) as a result of using Weybridge and Lelystad tuberculin, respectively. However, animals that tested negative to the SICCT were not slaughtered at the time of the tests, so definitive conclusions are not possible.

Geographical association between the genotype of bovine tuberculosis in found dead badgers and in cattle herds.

In a survey, 457 badgers that had been found dead in Wales were postmortem-examined, and samples were examined by histology and by extended culture (for up to 12 weeks). Mycobacterium bovis was cultured from 55 badgers (12.0 per cent), and the histology typical of M bovis infection was seen in a further six (1.3 per cent). The prevalence in badgers in each of 10 geographical areas varied between 0 and 26 per cent (P<0.001), and was associated with the incidence of confirmed M bovis infection in cattle herds in the same areas (P<0.01). In northern Wales, bTB was rare in both hosts. An infected badger was 12.3 times more likely to be within 5 km of a confirmed cattle bTB breakdown than an uninfected badger. The M bovis isolates from badgers belonged to one of four genotypes defined by spoligotype and variable number tandem repeat type. These genotypes were also found in 290 concurrent confirmed herd breakdowns, and tended to be similar to the genotypes in badgers in the same geographical areas. When badgers and cattle no more than 30 km apart were compared, the genotype diversity was greater in cattle than in badgers (P=0.016), suggesting that the movement of cattle plays a greater part in the spatial distribution of M bovis than the movement of badgers.

Herd-level risk factors of bovine tuberculosis in England and Wales after the 2001 foot-and-mouth disease epidemic.

OBJECTIVES: We present the results of a 2005 case-control study of bovine tuberculosis (bTB) breakdowns in English and Welsh herds. The herd management, farming practices, and environmental factors of 401 matched pairs of case and control herds were investigated to provide a picture of herd-level risk factors in areas of varying bTB incidence. METHODS: A global conditional logistic regression model, with region-specific variants, was used to compare case herds that had experienced a confirmed bTB breakdown to contemporaneous control herds matched on region, herd type, herd size, and parish testing interval. RESULTS: Contacts with cattle from contiguous herds and sourcing cattle from herds with a recent history of bTB were associated with an increased risk in both the global and regional analyses. Operating a farm over several premises, providing cattle feed inside the housing, and the presence of badgers were also identified as significantly associated with an increased bTB risk. CONCLUSIONS: Steps taken to minimize cattle contacts with neighboring herds and altering trading practices could have the potential to reduce the size of the bTB epidemic. In principle, limiting the interactions between cattle and wildlife may also be useful; however this study did not highlight any specific measures to implement.

Recurrence of bovine tuberculosis breakdowns in Great Britain: risk factors and prediction.

Bovine tuberculosis (bTB) is an important economic disease worldwide with implications for both animal and human health. In Great Britain the number of herds that test positive for bTB, termed "breakdowns", has increased over the last two decades. Despite more intensive testing during a breakdown, around 23% of breakdowns recur within 12 months of the previous breakdown ending, and around 38% within 24 months. These "recurrent" breakdowns may be important for onward transmission of infection. Detailed case-control data were analysed to identify factors associated with recurrence within 12 months. The model predicted 83% of all recurrent breakdowns, with a positive predictive value (PPV) of 44%. A further model, restricted to data currently available nationally, was not sufficient to predict recurrence reliably; at a sensitivity of 72-76%, the PPV was 33-36%, when validated on independent data. Factors identified to be associated with recurrence are consistent with previous studies; namely, the number of reactors, a recent bTB history in the herd and a lack of association with the confirmation status of the initial breakdown. These variables are indicative of a higher level of infection or residual infection, and could be useful in the future development of predictive models for bTB recurrence.

Predicting prolonged bovine tuberculosis breakdowns in Great Britain as an aid to control.

Bovine tuberculosis (bTB) is an important notifiable disease in cattle in Great Britain (GB), and is subject to statutory control measures. Despite this, disease incidence has increased since the mid-1980s, and around 30% of herd breakdowns continue for more than 240 days. This is twice the shortest possible time for confirmed breakdowns to test clear from infection (≈120 days), and four times the shortest possible time for unconfirmed breakdowns (≈60 days). These "prolonged" breakdowns consume substantial resources and may act as an ongoing source of infection. It is not clear why some breakdowns become prolonged. Existing detailed case-control data have been re-analysed to determine risk factors for breakdowns lasting longer than 240 days, the strongest of which was the confirmation status of the breakdown: OR 12.6 (95%CI: 6.7-25.4). A further model restricted to data available early on in a breakdown for all breakdowns nationally, can predict 82-84% of prolonged breakdowns with a positive predictive value of 44-49% when validated using existing national datasets over a 4-year period. Identification of prolonged breakdowns at an earlier stage could help to target bTB controls in GB.

Evidence of genetic resistance of cattle to infection with Mycobacterium bovis.

Anecdotal evidence points to genetic variation in resistance of cattle to infection with Mycobacterium bovis, the causative agent of bovine tuberculosis (BTB), and published experimental evidence in deer and cattle suggests significant genetic variation in resistance and reactivity to diagnostic tests. However, such genetic variation has not been properly quantified in the United Kingdom dairy cattle population; it is possible that it exists and may be a factor influencing the occurrence of BTB. Using models based on the outcome of the process of diagnosis (ultimate fate models) and on the outcome of a single stage of diagnosis (continuation ratio models, herd test-date models), this study shows that there is heritable variation in individual cow susceptibility to BTB, and that selection for milk yield is unlikely to have contributed to the current epidemic. Results demonstrate that genetics could play an important role in controlling BTB by reducing both the incidence and the severity of herd breakdowns.

Comparison of a standard and a detailed postmortem protocol for detecting Mycobacterium bovis in badgers.

A standard postmortem protocol, consisting of gross pathology, culture for mycobacteria and limited selective histopathology, was used in the randomised badger culling trial in Great Britain to detect Mycobacterium bovis infection. This standard protocol was compared with a more detailed protocol in which more tissues were examined grossly, more tissues were cultured, more culture slopes were seeded, the culture period was extended and tissues were examined routinely by histopathology. The standard protocol was more sensitive in badgers with gross visible lesions than in badgers with no gross visible lesions. When applied to the study population of badgers, the overall sensitivity of the standard protocol relative to the more detailed protocol was estimated to be 54.6 per cent (95 per cent confidence interval 44.9 to 69.8 per cent). Badgers with tuberculosis (tb) detected by the standard protocol had a mean of 7.6 tissues with microscopic lesions suspicious of tb. The additional badgers detected by the detailed protocol had a mean of 4.4 tissues with microscopic lesions suspicious of tb.

The prevalence, distribution and severity of detectable pathological lesions in badgers naturally infected with Mycobacterium bovis.

The Randomized Badger Culling Trial (RBCT) began in 1998 to determine the impact of badger culling in controlling bovine tuberculosis in cattle. A total of 1166 badgers (14% of total) proactively culled during the RBCT were found to be tuberculous, offering a unique opportunity to study the pathology caused by Mycobacterium bovis in a large sample of badgers. Of these, 39% of adults (approximately 6% of all adults culled) had visible lesions (detectable at necropsy) of bovine tuberculosis; cubs had a lower prevalence of infection (9%) but a higher percentage of tuberculous cubs (55.5%) had visible lesions. Only approximately 1% of adult badgers had extensive, severe pathology. Tuberculous badgers with recorded bite wounds (approximately 5%) had a higher prevalence of visible lesions and a different distribution of lesions, suggesting transmission via bite wounds. However, the predominance of lesions in the respiratory tract indicates that most transmission occurs by the respiratory route.

Bovine tuberculosis infection in wild mammals in the South-West region of England: a survey of prevalence and a semi-quantitative assessment of the relative risks to cattle.

In the United Kingdom, badgers are implicated in the transmission of Mycobacterium bovis to cattle, but little information is available on the potential role of other wild mammals. This paper presents the results of the largest systematic UK survey of M. bovis infection in other wild mammals. Mammal carcasses (4715) from throughout the South-West region of England were subjected to a systematic post mortem examination, microbiological culture of tissues and spoligotyping of isolates. Infection was confirmed in fox, stoat, polecat, common shrew, yellow-necked mouse, wood mouse, field vole, grey squirrel, roe deer, red deer, fallow deer and muntjac. Prevalence in deer may have been underestimated because the majority were incomplete carcasses, which reduced the likelihood of detecting infection. Infected cases were found in Wiltshire, Somerset, Devon and Cornwall, Gloucestershire and Herefordshire. Lesions were found in a high proportion of spoligotype-positive fallow, red and roe deer, and a single fox, stoat and muntjac. M. bovis spoligotypes occurred in a similar frequency of occurrence to that in cattle and badgers. Data on prevalence, pathology, abundance and ecology of wild mammals was integrated in a semi-quantitative risk assessment of the likelihood of transmission to cattle relative to badgers. Although most species presented a relatively low risk, higher values and uncertainty associated with muntjac, roe, red and in particular fallow deer, suggest they require further investigation. The results suggest that deer should be considered as potential, although probably localised, sources of infection for cattle.

Ante mortem diagnosis of tuberculosis in cattle: a review of the tuberculin tests, gamma-interferon assay and other ancillary diagnostic techniques.

The early, preclinical stages of bovine TB can be detected in live animals by the use of tests of cellular immunity (the skin, gamma-interferon and lymphocyte transformation tests). Tests of humoral (antibody) immunity, Mycobacterium bovis PCR probes on early tissue cultures or live cattle specimens, and tests based on "electronic nose" technology have been developed more recently. The key measure of diagnostic test accuracy is the relationship between sensitivity and specificity, which determines the false-positive and false-negative proportions. None of the tests currently available for the diagnosis of bovine TB allow a perfectly accurate determination of the M. bovis infection status of cattle. Although various factors can reduce the sensitivity and specificity of the skin tests, these remain the primary ante mortem diagnostic tools for TB in cattle, providing a cost-effective and reliable means of screening entire cattle populations. Despite the inescapable limitations of existing diagnostic tests, bovine TB has been effectively eradicated from many developed countries and regions with the implementation of sound programmes of regular tuberculin skin testing and removal of reactors, coupled with slaughterhouse surveillance for undetected infections, repeat testing and culling of infected herds, cattle movement restrictions to prevent introduction of infected animals and occasional slaughter of entire herds with intractable breakdowns. This is likely to remain the mainstay of bovine TB control programmes for the foreseeable future. Additionally, newer ancillary in vitro diagnostic assays are now available to TB control programme managers to supplement the skin tests in defined circumstances according to the specific disease situation in each country or region. The strategic deployment of ancillary in vitro tests alongside the primary skin tests has enhanced the detection of M. bovis-infected cattle and reduced the number of animals slaughtered as false positives.

The status of Mycobacterium bovis infection in UK wild mammals: a review.

Bovine tuberculosis caused by Mycobacterium bovis is a zoonotic infection with a wide range of mammalian hosts. In parts of the UK M. bovis infection in cattle is a persistent problem. The European badger (Meles meles) is implicated in the transmission of M. bovis to cattle, and is widely believed to constitute the most important reservoir of infection in UK wildlife. However, few studies have been carried out on the status of M. bovis infection in other UK mammals. In this review we present information on the incidence and pathology of M. bovis infection in UK wild mammals from both published and previously unpublished sources. Although the evidence does not support the existence of a significant self-maintaining reservoir of infection in any wild mammal other than the badger, there is a clear lack of sufficient data to rule out the involvement of other species. In the light of this and the dynamic nature of epidemiological patterns, further surveillance for M. bovis infection in UK wild mammals, using modern methods of diagnosis, is essential.

Value of existing serological tests for identifying badgers that shed Mycobacterium bovis.

In the UK there has been a sharp rise in the incidence of bovine tuberculosis since the early 1990s and the badger has been identified as an important wildlife reservoir for this infection. Infected badgers can excrete Mycobacterium bovis, putting other badgers and cattle at risk of becoming infected. Vaccination has been proposed as an approach to reducing the excretion of M. bovis by tuberculous badgers. In order to evaluate the efficacy of a badger vaccine it will be necessary to accurately determine the number of badgers excreting M. bovis without removing them for post-mortem evaluation. The existing live tests for tuberculosis in the badger (culture, indirect ELISA, Western blot) have not been assessed for their ability to detect badgers excreting M. bovis. Over the past 18 years, badgers from 31 social groups have been trapped and sampled in a study area of the Cotswold escarpment. We have examined the serological responses of 128 badgers trapped between 1985 and 1998 from social groups where M. bovis infection was endemic. These responses were compared with culture from faeces, urine, tracheal aspirates and bite wound swabs taken from these animals while alive. ELISA was found to be more sensitive than Western blot in detecting badgers excreting M. bovis. The majority of culture-positive badgers excreted M. bovis intermittently over the period of study. As a result, there was only a 27.5% chance of sampling a badger for culture when it was excreting M. bovis. In contrast, a positive ELISA result correctly predicted 68.2% of badgers with a history of excreting M. bovis. In the absence of alternative live tests for the badger, the Brock Test indirect ELISA appears to be more valuable than culture for measuring the effect of vaccination on reducing the number of badgers at risk of transmitting tuberculosis.

Screening badgers (Meles meles) for Mycobacterium bovis infection by using multiple applications of an ELISA.

The current indirect ELISA used to evaluate whether live badgers are infected with Mycobacterium bovis has a low sensitivity (40.7 per cent), but a relatively high specificity (94.3 per cent). The low sensitivity of the test makes the diagnosis unreliable, but its sensitivity can be increased by using multiple tests. Two multiple testing procedures (involving up to three sequential tests) were investigated. A procedure in which two positive results were required from three tests before an animal was declared positive resulted in a lower sensitivity, but a higher specificity than the single test (38 and 98 per cent respectively). A more rigorous procedure, in which only one positive result was required from three tests, resulted in a marked increase in sensitivity but a slight reduction in specificity (79.5 and 83.1 per cent respectively) when compared to the single test.