Bacteriological Survey of Fresh Minced Beef on Sale at Retail Outlets in Scotland in 2019: Three Foodborne Pathogens, Hygiene Process Indicators, and Phenotypic Antimicrobial Resistance.

ABSTRACT: The health and economic burden of foodborne illness is high, with approximately 2.4 million cases occurring annually in the United Kingdom. A survey to understand the baseline microbial quality and prevalence of food-related hazards of fresh beef mince on retail sale could inform risk assessment, management, and communication to ensure the safety of this commodity. In such a survey, a two-stage sampling design was used to reflect variations in population density and the market share of five categories of retail outlets in Scotland. From January to December 2019, 1,009 fresh minced beef samples were collected from 15 geographic areas. The microbial quality of each sample was assessed using aerobic colony count and Escherichia coli count. Samples were cultured for Campylobacter and Salmonella, and PCR was used to detect target genes (stx1 all variants, stx2 a to g, and rfbO157) for Shiga toxin-producing E. coli (STEC). The presence of viable E. coli O157 and STEC in samples with a positive PCR signal was confirmed via culture and isolation. Phenotypic antimicrobial sensitivity patterns of cultured pathogens and 100 E. coli isolates were determined, mostly via disk diffusion. The median aerobic colony count and E. coli counts were 6.4 × 105 (interquartile range, 6.9 × 104 to 9.6 × 106) and <10 CFU/g (interquartile range, <10 to 10) of minced beef, respectively. The prevalence was 0.1% (95% confidence interval [CI], 0 to 0.7%) for Campylobacter, 0.3% (95% CI, 0 to 1%) for Salmonella, 22% (95% CI, 20 to 25%) for PCR-positive STEC, and 4% (95% CI, 2 to 5%) for culture-positive STEC. The evidence for phenotypic antimicrobial resistance detected did not give cause for concern, mainly occurring in a few E. coli isolates as single nonsusceptibilities to first-line active substances. The low prevalence of pathogens and phenotypic antimicrobial resistance is encouraging, but ongoing consumer food safety education is necessary to mitigate the residual public health risk.

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

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

HbA1c determination from HemaSpot™ blood collection devices: comparison of home prepared dried blood spots with standard venous blood analysis.

AIM: To assess the clinical performance and patient acceptance of HemaSpot™ blood collection devices as an alternative blood collection method. METHODS: Adult men and women with any type of diabetes, routinely carrying out self-monitoring of blood glucose were recruited (n = 128). Participants provided a venous blood sample and prepared two HemaSpot dried blood spots, one at clinics and one at home. HbA1c analysis was by Tosoh G8 high-performance liquid chromatography. Participants also completed a questionnaire. RESULTS: Strong linear relationships been HbA1c levels in dried blood spots and venous blood were observed and a linear model was fitted to the data. Time between dried blood spot preparation and testing did not impact the model. Participants were accepting of the approach: 69.2% would use this system if available and 60.7% would be more likely to use this system than going to their general practitioner. CONCLUSIONS: The combination of a robust desiccating dried blood spot device, home sample preparation and return by post produces HbA1c data that support the use of a time-independent linear calibration of dried blood spot to venous blood HbA1c . A robust remote sample collection service would be valuable to people living with diabetes in urban areas who are working or house-bound as well as those living in remote or rural locations.

Strategies for screening young stock for antibodies - optimising numbers to test, cut-points, & predictive values for bovine viral diarrhoea virus.

The antibody seroprevalence of young stock can be a useful indicator of recent or current infection in a herd. We examine the factors that contribute to the assessment of herd exposure to disease, via spot testing for antibody, using bovine viral diarrhoea virus (BVDv) as an example. A statistical distribution of seroprevalences for BVDv in beef herds identified three groups of herds: low, intermediate and high within-herd BVDv antibody seroprevalence. We tested two assumptions -the intermediate seroprevalence group of herds is assumed to be negative for BVDv at the herd level and alternatively if this group is assumed to be positive. We found that: The herd-level sensitivity and specificity are sensitive to the assumption regarding the herds with intermediate seroprevalence. If an appropriate cut-point is chosen, reducing the sample size from ten to five does not produce a large drop in herd-level test performance. Increasing the cut-point may be valuable at the outset of an eradication programme. Increasing the sample size and decreasing the cut-point is advantageous towards the end of an eradication programme, to minimise the risk of positive herds being misclassified. The framework presented here illustrates how seroprevalence screening may be understood and assessed.

An empirical comparison of isolate-based and sample-based definitions of antimicrobial resistance and their effect on estimates of prevalence.

Antimicrobial resistance is primarily a problem in human medicine but there are unquantified links of transmission in both directions between animal and human populations. Quantitative assessment of the costs and benefits of reduced antimicrobial usage in livestock requires robust quantification of transmission of resistance between animals, the environment and the human population. This in turn requires appropriate measurement of resistance. To tackle this we selected two different methods for determining whether a sample is resistant - one based on screening a sample, the other on testing individual isolates. Our overall objective was to explore the differences arising from choice of measurement. A literature search demonstrated the widespread use of testing of individual isolates. The first aim of this study was to compare, quantitatively, sample level and isolate level screening. Cattle or sheep faecal samples (n=41) submitted for routine parasitology were tested for antimicrobial resistance in two ways: (1) "streak" direct culture onto plates containing the antimicrobial of interest; (2) determination of minimum inhibitory concentration (MIC) of 8-10 isolates per sample compared to published MIC thresholds. Two antibiotics (ampicillin and nalidixic acid) were tested. With ampicillin, direct culture resulted in more than double the number of resistant samples than the MIC method based on eight individual isolates. The second aim of this study was to demonstrate the utility of the observed relationship between these two measures of antimicrobial resistance to re-estimate the prevalence of antimicrobial resistance from a previous study, in which we had used "streak" cultures. Boot-strap methods were used to estimate the proportion of samples that would have tested resistant in the historic study, had we used the isolate-based MIC method instead. Our boot-strap results indicate that our estimates of prevalence of antimicrobial resistance would have been considerably lower in the historic study had the MIC method been used. Finally we conclude that there is no single way of defining a sample as resistant to an antimicrobial agent. The method used greatly affects the estimated prevalence of antimicrobial resistance in a sampled population of animals, thus potentially resulting in misleading results. Comparing methods on the same samples allows us to re-estimate the prevalence from other studies, had other methods for determining resistance been used. The results of this study highlight the importance of establishing what the most appropriate measure of antimicrobial resistance is, for the proposed purpose of the results.

British Escherichia coli O157 in Cattle Study (BECS): to determine the prevalence of E. coli O157 in herds with cattle destined for the food chain.

Escherichia coli O157 are zoonotic bacteria for which cattle are an important reservoir. Prevalence estimates for E. coli O157 in British cattle for human consumption are over 10 years old. A new baseline is needed to inform current human health risk. The British E. coli O157 in Cattle Study (BECS) ran between September 2014 and November 2015 on 270 farms across Scotland and England & Wales. This is the first study to be conducted contemporaneously across Great Britain, thus enabling comparison between Scotland and England & Wales. Herd-level prevalence estimates for E. coli O157 did not differ significantly for Scotland (0·236, 95% CI 0·166-0·325) and England & Wales (0·213, 95% CI 0·156-0·283) (P = 0·65). The majority of isolates were verocytotoxin positive. A higher proportion of samples from Scotland were in the super-shedder category, though there was no difference between the surveys in the likelihood of a positive farm having at least one super-shedder sample. E. coli O157 continues to be common in British beef cattle, reaffirming public health policy that contact with cattle and their environments is a potential infection source.

Difficulties arising from the variety of testing schemes used for bovine viral diarrhoea virus (BVDV).

Globally, the eradication of bovine viral diarrhoea virus (BVDV) is still in its infancy, but eradication has been, or is being, adopted by several countries or regions. Comparisons between countries' schemes allow others to assess best practice, and aggregating published results from eradication schemes provides greater statistical power when analysing data. Aggregating data requires that results derived from different testing schemes be calibrated against one another. The authors aimed to evaluate whether relationships between published BVDV test results could be created and present the outcome of a systematic literature review following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. The results are tabulated, providing a summary of papers where there is potential cross-calibration and a summary of the obstacles preventing such data aggregation. Although differences in measuring BVDV present barriers to academic progress, they may also affect progress within individual eradication schemes. The authors examined the time taken to retest following an initial antibody BVDV test in the Scottish eradication scheme. The authors demonstrate that retesting occurred quicker if the initial not negative test was from blood rather than milk samples. Such differences in the response of farmers/veterinarians to tests may be of interest to the design of future schemes.

Replicating disease spread in empirical cattle networks by adjusting the probability of infection in random networks.

Comparisons between mass-action or "random" network models and empirical networks have produced mixed results. Here we seek to discover whether a simulated disease spread through randomly constructed networks can be coerced to model the spread in empirical networks by altering a single disease parameter - the probability of infection. A stochastic model for disease spread through herds of cattle is utilised to model the passage of an SEIR (susceptible-latent-infected-resistant) through five networks. The first network is an empirical network of recorded contacts, from four datasets available, and the other four networks are constructed from randomly distributed contacts based on increasing amounts of information from the recorded network. A numerical study on adjusting the value of the probability of infection was conducted for the four random network models. We found that relative percentage reductions in the probability of infection, between 5.6% and 39.4% in the random network models, produced results that most closely mirrored the results from the empirical contact networks. In all cases tested, to reduce the differences between the two models, required a reduction in the probability of infection in the random network.

Associations between bovine viral diarrhoea virus (BVDV) seropositivity and performance indicators in beef suckler and dairy herds.

Data from 255 Scottish beef suckler herds and 189 Scottish dairy herds surveyed as part of national bovine viral diarrhoea virus (BVDV) prevalence studies from October 2006 to May 2008 were examined retrospectively to determine the relationship between serological status and key performance indicators derived from national cattle movement records. On average, calf mortality rates were 1.35 percentage points higher in seropositive beef herds and 3.05 percentage points higher in seropositive dairy herds than in negative control herds. Seropositive beef herds were also more likely to show increases in calf mortality rates and culling rates between successive years. There were no discernible effects of BVDV on the average age at first calving or calving interval for either herd type. Accompanying questionnaire data revealed that only 27% of beef farmers and 25% of dairy farmers with seropositive herds thought their cattle were affected by BVDV, which suggests that the clinical effects of exposure may be inapparent under field conditions or masked by other causes of reproductive failure and culling. Beef farmers were significantly more likely to perceive a problem when their herd experienced acute changes in calf mortality rates, culling rates, and calving intervals between successive years. However, only 35% of these perceived positive herds were actually seropositive for BVDV. These findings emphasize both the importance of routinely screening herds to determine their true infection status and the potential for using national cattle movement records to identify herds that may be experiencing outbreaks from BVDV or other infectious diseases that impact herd performance.

Relative associations of cattle movements, local spread, and biosecurity with bovine viral diarrhoea virus (BVDV) seropositivity in beef and dairy herds.

The success of bovine viral diarrhoea virus (BVDV) eradication campaigns can be undermined by spread through local transmission pathways and poor farmer compliance with biosecurity recommendations. This work combines recent survey data with cattle movement data to explore the issues likely to impact on the success of BVDV control in Scotland. In this analysis, data from 249 beef suckler herds and 185 dairy herds in Scotland were studied retrospectively to determine the relative influence of cattle movements, local spread, and biosecurity on BVDV seropositivity. Multivariable logistic regression models revealed that cattle movement risk factors had approximately 3 times greater explanatory power than risk factors for local spread amongst beef suckler herds, but approximately the same explanatory power as risk factors for local spread amongst dairy herds. These findings are most likely related to differences in cattle husbandry practices and suggest that where financial prioritization is required, focusing on reducing movement-based risk is likely to be of greatest benefit when applied to beef suckler herds. The reported use of biosecurity measures such as purchasing cattle from BVDV accredited herds only, performing diagnostic screening at the time of sale, implementing isolation periods for purchased cattle, and installing double fencing on shared field boundaries had minimal impact on the risk of beef or dairy herds being seropositive for BVDV. Only 28% of beef farmers and 24% of dairy farmers with seropositive herds recognized that their cattle were affected by BVDV and those that did perceive a problem were no less likely to sell animals as replacement breeding stock and no more likely to implement biosecurity measures against local spread than farmers with no perceived problems. In relation to the current legislative framework for BVDV control in Scotland, these findings emphasize the importance of requiring infected herds take appropriate biosecurity measures to prevent further disease transmission and conducting adequate follow-up to ensure that biosecurity measures are being implemented correctly in the field.

The influence of empirical contact networks on modelling diseases in cattle.

We present two stochastic models of the passage of an SEIR (susceptible-latent-infected-resistant) disease through herds of cattle. One model is based on a contact network constructed via continuously recorded interaction data from two herds of cattle, the other, a matching network constructed using the principles of mass-action mixing. The recorded contact data were produced by attaching proximity data loggers to two separate herds of cattle during two separate recording periods. The network constructed using the principles of mass-action mixing uses the same number of contacts as the recorded network but distributes them randomly amongst the animals. The recorded networks had a greater number of repeated contacts, lower closeness and clustering scores and greater average path length than the mass-action networks. A lower proportion of simulations of the recorded network produce any disease spread when compared to those simulations of the mass-action network and, of those that did, fewer infected animals were predicted. For all parameter values tested, within the sensitivity analysis, similar differences were found between the recorded and mass-action network models.

Prevalence of antibodies to bovine viral diarrhoea virus in bulk tank milk and associated risk factors in Scottish dairy herds.

Bulk tank milk samples were collected from 374 dairy farms in Scotland in 2007/2008 along with questionnaire data relating to the management of the farm. Milk samples were tested for antibodies to bovine viral diarrhoea virus (BVDV) using a commercially available (Svanova) kit and percentage positivity scores calculated according to the manufacturer's guidelines. There were 220 farms that did not routinely vaccinate for bovine viral diarrhoea (BVD), and these were distributed according to the Swedish BVD eradication classes as 12.7 per cent, 22.3 per cent, 44.5 per cent and 20.5 per cent for Classes 0, 1, 2 and 3, respectively. A more sophisticated statistical method (finite mixture modelling) which does not depend on arbitrary thresholds and categories suggested a 73 per cent prevalence of herds with high mean levels of antibodies. Risk factor analysis suggested that routine vaccination for BVD, suspicion of BVD, housing of pregnant cows with calves, total number of cows and the proportion of cows that were dry were all associated with increased BVDV antibodies in bulk milk. The inclusion of BVD within the farm's health plan was associated with decreased BVDV antibodies in the bulk milk.

Does circumpatellar electrocautery improve the outcome after total knee replacement?: a prospective, randomised, blinded controlled trial.

The incidence of anterior knee pain following total knee replacement (TKR) is reported to be as high as 49%. The source of the pain is poorly understood but the soft tissues around the patella have been implicated. In theory circumferential electrocautery denervates the patella thereby reducing efferent pain signals. However, there is mixed evidence that this practice translates into improved outcomes. We aimed to investigate the clinical effect of intra-operative circumpatellar electrocautery in patients undergoing TKR using the LCS mobile bearing or Kinemax fixed bearing TKR. A total of 200 patients were randomised to receive either circumpatellar electrocautery (diathermy) or not (control). Patients were assessed by visual analogue scale (VAS) for anterior knee pain and Oxford knee score (OKS) pre-operatively and three months, six months and one year post-operatively. Patients and assessors were blinded. There were 91 patients in the diathermy group and 94 in the control. The mean VAS improvement at one year was 3.9 in both groups (control; -10 to 6, diathermy; -9 to 8, p < 0.001 in both cases, paired, two-tailed t-test). There was no significant difference in VAS between the groups at any other time. The mean OKS improvement was 17.7 points (0 to 34) in the intervention group and 16.6 (0 to 42) points in the control (p = 0.36). There was no significant difference between the two groups in OKS at any other time. We found no relevant effect of patellar electrocautery on either VAS anterior knee pain or OKS for patients undergoing LCS and Kinemax TKR.

Modelling the effects of previous infection and re-infection on the costs of bovine viral diarrhoea outbreaks in beef herds.

A previously published model was re-employed to examine the potential impact of different epidemiological circumstances on output losses due to bovine viral diarrhoea virus (BVDV) infection in typical British hill cow-calf enterprises. The average annuity equivalent of unchecked losses from 100 simulated 10-year disease scenarios ranged from almost pound0/cow to approximately pound40/cow. Significant differences were found under certain circumstances, depending on the initial disease status of the herd, the initial source of virus, the probability and source of further infection, the probability of virus transmission within the herd and herd size. For naïve herds, losses depended only on the risk of incursion. In most other circumstances, the losses could be mitigated if the annual risk of incursion was <0.3 and risk of within herd transmission was extremely low. Greater understanding of the interaction between these risk factors and management actions are required so that total costs of BVDV infection can be minimised under different circumstances.

A model of the relationship between the epidemiology of Johne's disease and the environment in suckler-beef herds.

A non-predictive, dynamic and stochastic herd-level simulation model of an outbreak of Johne's in a suckler-beef herd is reported. Importantly, the model incorporates, with a simple method, the environment as the primary source of infection, reflecting the consensual understanding of the disease. The model also takes into account the density of the infectious agent in the environment. A sensitivity analysis suggests that the model is highly and equally sensitive to certain parameters (probability of infection in the presence of one unit of bacterial density, infectious area and bacterial shedding rate). Mathematical reasons for this similarity in sensitivity are presented. Compared to many other diseases, data for Johne's are scarce. Therefore models of Johne's outbreaks including this one cannot be predictive or easily validated. The qualitative results: (a) demonstrate the modelled effect of inclusion of infection via the environment; (b) suggest management factors that could be tested by experimentation or observation. Estimates for the rate of transmission, arising from the model output, are similar to published empirical estimates. The results of future empirical research should aid scientific understanding of the disease, help validate this model and might bring economic benefits through improved management.

Modelling BVD at herd level compared with individual animal level.

We assess two closely related stochastic models of BVD in a beef herd for their utility. One is an individual (animal) based model and the other a herd based model.

Assessing economic and social pressure for the control of bovine viral diarrhoea virus.

The objective of this paper is to present a preliminary assessment of variation in the economic impact of bovine viral diarrhoea virus (BVDV) at dairy farm level between a sample of nations within the EU and hence assess differences in pressure to respond to this disease that may be impeding progress in control and hence restricting collective benefits from healthier livestock. We used a questionnaire to obtain national average values of key epidemiological and economic parameters for a typical dairy farm from BVDV experts in the countries concerned. These parameters were converted into assessments of economic impact using a computer simulation model. Uncontrolled output losses for a BVDV-naïve herd with virus introduced in year 1 of a 10-year epidemic represented 22, 7, 8, 5, 8 and 20% of the BVDV-free annuity for the UK, Northern Portugal, Holland, Norway, Italy and Germany, respectively. Differences between countries will be widened by differences in the risk of acquiring BVDV. These will be much reduced in countries, such as Norway that have a national BVDV eradication programme. Farmers in such countries can therefore justify spending much less on maintaining BVDV-free status than BVDV-free farms in other countries. This result illustrates the paradox that in countries where BVDV prevalence is high, farmers have least to gain from unilateral BVDV eradication because of the high cost of maintaining freedom from the disease. We discuss this issue in the light of increasing recognition at international level of the importance of BVDV control.

Optimal risk management versus willingness to pay for BVDV control options.

Previous work has shown that the least-cost BVD-control option for cow-calf herds in Scotland was not necessarily the risk-minimising option. Thus, assessing BVDV prevention measures must account for risk reduction as well as decision-makers' attitude towards risks. We therefore describe a method to do this using a hypothetical example. Data for this analysis were generated using a simulation model of BVDV transmission in a typical Scottish cow-calf herd over a 10-year period. Herd infection/re-infection was adjusted to reflect the expected risk of infection/re-infection and the use of either biosecurity or vaccination strategies at various levels of effectiveness. The level of risk-free financial return that maximises farmers' utility of wealth was significantly affected by the assumed effectiveness of the control strategy. More importantly, it was observed that utility maximisation of wealth as a business objective is not an optimal solution in terms of animal welfare when dealing with a BVDV outbreak.