DNA carryover in milk samples from routine milk recording used for PCR-based diagnosis of bovine Staphylococcus aureus mastitis.

Real-time PCR techniques are increasingly used to detect udder pathogens from milk samples collected non-aseptically at routine milk recording. The objectives of this study were (1) to estimate the statistical associations between cycle threshold (Ct) values for Staphylococcus aureus in non-aseptically collected composite samples taken at routine milk recording from cows milked consecutively with the same milking unit and milk meter; and (2) to formulate practical and plausible guidelines for understanding the diagnostic implications of PCR testing for Staph. aureus intramammary infection at routine milk recording. The study included 4 herds with conventional milking parlors and repeatedly low Ct-values for Staph. aureus (representing a high DNA load) in bulk tank milk. Composite milk samples were collected from all cows at all milking units during routine milk recording using the Tru-Test electronic milk meter (Tru-Test Group, Auckland, New Zealand) and analyzed using the PathoProof PCR (Thermo Fisher Scientific, Vantaa, Finland) assay. Milking clock times were retrieved at each milk meter to establish the milking order of the cows at each unit. A multinomial logistic regression was applied to estimate the association between Ct-values from cows milked consecutively with the same milking unit and milk meter. The following groups were selected based on Ct-values: (1) 0-31.3, (2) 31.4-33.9, (3) 34.0-37, (4) 37.1-39.9, and (5) 40 (negative result). The association between groups from cows milked consecutively with the same milking unit and milk meter was statistically significant. Approximately 60% of cows were in Ct group 5 if the antecedent cow was also in Ct group 5, but only 20% of cows were in Ct group 5 if the antecedent cow was in Ct group 1. The probability of cows being in Ct group 1 was not markedly influenced by the group of the antecedent cow. Statistical relationships in the intermediate range gave a plausible indication of a dose-response relationship. Carryover of bacterial DNA via the milking unit and milk meter is very likely to affect PCR results for Staph. aureus. Therefore, information about milking order must be considered in mastitis control efforts. We suggest a practical interpretation of PCR results: cows with a Ct-value <32 can be labeled "very likely to be infected with Staph. aureus," but cows with Ct-values of >37 and 32-37 can be labeled "very likely to be negative for Staph. aureus" and "uncertain Staph. aureus status," respectively.

A quantitative screening method to detect rater-introduced bias in clinical ratings.

BACKGROUND: We suggest a 'screening test' to examine large data files with clinical ratings for the occurrence of rater-introduced bias prior to using the data for quantitative analyses. The test is based on a statistical model in which a well-standardized interval-scale outcome (for example, milk yield) is related to clinical ratings (for example, body condition scores) obtained from multiple contexts (for example, dairy herds). FINDINGS: 84,968 calvings from 279 herds, with subsequent body condition scores performed by 117 veterinarians within the first 21 days postpartum were analyzed with a multilevel random coefficient regression model. The model included an independent variable, where body condition score was centered within veterinarian. This is a so-called comparison effect to describe possible rater-introduced bias in the body condition scores. A highly significant comparison effect was found for second and older parities, indicating occurrence of possible rater-introduced bias in this large multi-herd data file. CONCLUSIONS: A within-group centering technique (the comparison effect) appeared to be useful for discriminating between biased and unbiased clinical scores. In some cases, this test for bias should prevent further analysis of the data and divert the focus of study to the calibration of raters or alternative study designs.

Veterinary decision making in relation to metritis--a qualitative approach to understand the background for variation and bias in veterinary medical records.

BACKGROUND: Results of analyses based on veterinary records of animal disease may be prone to variation and bias, because data collection for these registers relies on different observers in different settings as well as different treatment criteria. Understanding the human influence on data collection and the decisions related to this process may help veterinary and agricultural scientists motivate observers (veterinarians and farmers) to work more systematically, which may improve data quality. This study investigates qualitative relations between two types of records: 1) 'diagnostic data' as recordings of metritis scores and 2) 'intervention data' as recordings of medical treatment for metritis and the potential influence on quality of the data. METHODS: The study is based on observations in veterinary dairy practice combined with semi-structured research interviews of veterinarians working within a herd health concept where metritis diagnosis was described in detail. The observations and interviews were analysed by qualitative research methods to describe differences in the veterinarians' perceptions of metritis diagnosis (scores) and their own decisions related to diagnosis, treatment, and recording. RESULTS: The analysis demonstrates how data quality can be affected during the diagnostic procedures, as interaction occurs between diagnostics and decisions about medical treatments. Important findings were when scores lacked consistency within and between observers (variation) and when scores were adjusted to the treatment decision already made by the veterinarian (bias). The study further demonstrates that veterinarians made their decisions at 3 different levels of focus (cow, farm, population). Data quality was influenced by the veterinarians' perceptions of collection procedures, decision making and their different motivations to collect data systematically. CONCLUSION: Both variation and bias were introduced into the data because of veterinarians' different perceptions of and motivations for decision making. Acknowledgement of these findings by researchers, educational institutions and veterinarians in practice may stimulate an effort to improve the quality of field data, as well as raise awareness about the importance of including knowledge about human perceptions when interpreting studies based on field data. Both recognitions may increase the usefulness of both within-herd and between-herd epidemiological analyses.

Prevalence and severity of foot lesions in Danish Holstein heifers through first lactation.

Several studies have shown that foot lesions and clinical lameness occur before first calving and develop further during the lactation period. Lameness may cause production losses, but the relationship between foot lesions, particularly in the claw horn, and lameness in heifers is unclear. The objectives of this study were to describe the development of and evaluate the relationship between lameness and foot lesions in Danish Holsteins before and after first calving. In a longitudinal study, 147 heifers were examined for lameness and foot lesions 2-5 times over an 18-month period. Lameness was assessed by means of a visual locomotion score and foot lesion severity was recorded. The prevalence of a locomotion score 3 was 25% before calving, and 90% at approximately 250 days in milk (DIM). Prevalence of moderate to severe sole haemorrhage (SH) was 27% before calving and 56% at 250 DIM, and that of moderate to severe white line lesion (WLL) 44% before calving with a peak of 70% at 200 DIM. There was one case of white line abscess but SH was seen throughout the entire study period. Digital dermatitis (DD) was prevalent prior to first calving (15%) and peaked at 39% at 0-100 DIM. Heel horn erosion (HHE) occurred in almost all cows (93-100%) and was strongly correlated with DD (r=0.51). The correlation coefficient between SH and WLL was also high (0.42). The relatively high correlations between WLL and both DD and HHE were more surprising (0.38 and 0.35, respectively), those between SH and both DD and HHE were moderate (around 0.18). Interdigital dermatitis was significantly correlated with both HHE and DD, but completely unrelated to SH and WLL. The overall average locomotion score increased by about one-half of a score unit from 1 month prior to calving until 250 DIM, with a large difference between herds, although this was unsurprising as cows may alter their locomotion pattern with management factors (e.g. floor properties). DD and WLL were both associated with a locomotion score > or = 3 but of the cows with severe WLL there was no clear association between a locomotion score > or = 3 and DD. The highest locomotion scores occurred among cows with DD but without WLL.

A mixed methods inquiry: How dairy farmers perceive the value(s) of their involvement in an intensive dairy herd health management program.

BACKGROUND: Research has been scarce when it comes to the motivational and behavioral sides of farmers' expectations related to dairy herd health management programs. The objectives of this study were to explore farmers' expectations related to participation in a health management program by: 1) identifying important ambitions, goals and subjective well-being among farmers, 2) submitting those data to a quantitative analysis thereby characterizing perspective(s) of value added by health management programs among farmers; and 3) to characterize perceptions of farmers' goals among veterinarians. METHODS: The subject was initially explored by means of literature, interviews and discussions with farmers, herd health management consultants and researchers to provide an understanding (a concourse) of the research entity. The concourse was then broken down into 46 statements. Sixteen Danish dairy farmers and 18 veterinarians associated with one large nationwide veterinary practice were asked to rank the 46 statements that defined the concourse. Next, a principal component analysis was applied to identify correlated statements and thus families of perspectives between respondents. Q-methodology was utilized to represent each of the statements by one row and each respondent by one column in the matrix. A subset of the farmers participated in a series of semi-structured interviews to face validate the concourse and to discuss subjects like animal welfare, veterinarians' competences as experienced by the farmers and time constraints in the farmers' everyday life. RESULTS: Farmers' views could be described by four families of perspectives: Teamwork, Animal welfare, Knowledge dissemination, and Production. Veterinarians believed that farmers' primary focus was on production and profit, however, farmers' valued teamwork and animal welfare more. CONCLUSION: The veterinarians in this study appear to focus too much on financial performance and increased production when compared to most of the participating farmers' expectations. On the other hand veterinarians did not focus enough on the major products, which farmers really wanted to buy, i.e. teamwork and animal welfare. Consequently, disciplines like sociology, economics and marketing may offer new methodological approaches to veterinarians as these disciplines have understood that accounting for individual differences is central to motivate change, i.e. 'know thy customer'.

A mixed methods inquiry into the validity of data.

BACKGROUND: Research in herd health management solely using a quantitative approach may present major challenges to the interpretation of the results, because the humans involved may have responded to their observations based on previous experiences and own beliefs. This challenge can be met through increased awareness and dialogue between researchers and farmers or other stakeholders about the background for data collection related to management and changes in management. By integrating quantitative and qualitative research methods in a mixed methods research approach, the researchers will improve their understanding of this potential bias of the observed data and farms, which will enable them to obtain more useful results of quantitative analyses. CASE DESCRIPTION: An example is used to illustrate the potentials of combining quantitative and qualitative approaches to herd health related data analyses. The example is based on two studies on bovine metritis. The first study was a quantitative observational study of risk factors for metritis in Danish dairy cows based on data from the Danish Cattle Database. The other study was a semi-structured interview study involving 20 practicing veterinarians with the aim to gain insight into veterinarians' decision making when collecting and processing data related to metritis. DISCUSSION AND EVALUATION: The relations between risk factors and metritis in the first project supported the findings in several other quantitative observational studies; however, the herd incidence risk was highly skewed. There may be simple practical reasons for this, e.g. underreporting and differences in the veterinarians' decision making. Additionally, the interviews in the second project identified several problems with correctness and validity of data regarding the occurrence of metritis because of differences regarding case definitions and thresholds for treatments between veterinarians. CONCLUSION: Studies where associations between specific herd health management routines and disease outcome variables are drawn based purely on quantitative observational studies may benefit greatly by adding a qualitative perspective to the quantitative approach as illustrated and discussed in this article. The combined approach requires, besides skills and interdisciplinary collaboration, also openness, reflection and scepticism from the involved scientists, but the benefits may be extended to various contexts both in advisory service and science.

Resistance to penicillin of Staphylococcus aureus isolates from cows with high somatic cell counts in organic and conventional dairy herds in Denmark.

BACKGROUND: Quarter milk samples from cows with high risk of intramammary infection were examined to determine the prevalence of Staphylococcus aureus (SA) and penicillin resistant SA (SAr) in conventional and organic dairy herds and herds converting to organic farming in a combined longitudinal and cross-sectional study. METHODS: 20 conventional herds, 18 organic herds that converted before 1995, and 19 herds converting to organic farming in 1999 or 2000 were included in the study. Herds converting to organic farming were sampled three times one year apart; the other herds were sampled once. Risk of infection was estimated based on somatic cell count, milk production, breed, age and lactation stage. RESULTS: The high-risk cows represented about 49 % of the cows in the herds. The overall prevalence of SA and SAr among these cows was 29% (95% confidence interval: 24%-34%) and 4% (95% confidence interval: 2%-5%) respectively. The prevalence of penicillin resistance among SA infected cows was 12% (95% confidence interval: 6%-19%) when calculated from the first herd visits. No statistically significant differences were observed in the prevalence of SAr or the proportion of isolates resistant to penicillin between herd groups. CONCLUSION: The proportion of isolates resistant to penicillin was low compared to studies in other countries except Norway and Sweden. Based on the low prevalence of penicillin resistance of SA, penicillin should still be the first choice of antimicrobial agent for treatment of bovine intramammary infection in Denmark.

The Mycobacterium avium subsp. paratuberculosis ELISA response by parity and stage of lactation.

Two cross-sectional studies were carried out to determine the enzyme-linked immunosorbent assay (ELISA) response to Mycobacterium avium subsp. paratuberculosis (Map) by cow characteristics and stage of lactation. One of the studies (referred to as "milk-group") used milk samples from all lactating cows (n=7994) in 108 Danish dairy herds. The other study (referred to as "serum-group") used serum samples collected from all cows (n=5323) in a subset of 72 herds from the 108 herds. These samples were analysed using a similar ELISA for detection of antibodies. The results from the ELISAs were interpreted with two cut-off values as the optimal cut-off value is not known, and as several levels are recommended to be used in practice. The results showed that the probability of being ELISA-positive was two to three times lower for cows in parity 1 relative to cows in other parities using both milk and serum ELISA. At the beginning of the lactation the probability of being positive was highest in the milk ELISA. In the serum ELISA the odds of being positive was highest at the end of lactation. The findings are important in the interpretation of ELISA results at cow level with a subsequent tentative diagnosis and correction for parity and stage of lactation should be considered when providing a diagnosis of paratuberculosis. Some issues related to the pathogenesis are also discussed.