The association between time-series milk ß-hydroxybutyrate dynamics and early reproductive performance of dairy cows.

The DeLaval Herd Navigator is an on-farm sensor system that measures on a frequent basis milk progesterone (P4) and ß-hydroxybutyrate (BHB) in individual cows to closely monitor reproductive performance and energy balance. This information provides the opportunity to investigate the dynamics of BHB measured in milk (mBHB) and study the association between mBHB and early reproductive performance. The objectives of the study were (1) to describe mBHB dynamics within the first 20 d in milk (DIM), and (2) to evaluate the association between mBHB dynamics and early reproductive performance at cow-level. Two-year time-series data from 4,133 dairy cows in 38 Dutch dairy farms were available for analysis. Data included information on mBHB, daily milk yield and the indicators of early reproductive performance, days from calving to resumption of cyclicity, days from calving to first estrus, and days from calving to first insemination. The following mBHB dynamic parameters were defined based on the first 20 DIM for each individual cow: average mBHB (AvgBHB), DIM when mBHB was for the first time ≥80 µmol/L (OnsetKeto), the total number of consecutive days a cow had mBHB concentration ≥80 µmol/L, and the number of measurements mBHB concentration was ≥80 µmol/L. Three Cox proportional hazard regression models with random herd effect were developed to evaluate the association between cow level mBHB dynamics and days from calving to resumption of cyclicity, first estrus, and first insemination. Results showed that the mean AvgBHB within 20 DIM among all cows was 73 µmol/L. The mean OnsetKeto within 20 DIM, was 8 DIM. Among all cows having hyperketolactia, 55.8% (1,350/2,419) had OnsetKeto in the first week of lactation. In total, 41.5% (1,714/4,133) of the cows did not have OnsetKeto in the first 20 DIM. An early onset of hyperketolactia was associated with delayed fertility events. Cows with higher AvgBHB have a prolonged time interval from calving to resumption of cyclicity and first estrus. Information on mBHB dynamics and the association with early reproductive performance provides insights that might be helpful to improve reproductive performance of individual dairy cows.

Homogeneity density scores of quarter milk in automatic milking systems.

Milk quality and clinical mastitis in dairy cows are monitored by detecting visually abnormal milk. A standardized method to evaluate clots in milk and studies of the incidence and dynamics of clots in milk at the quarter level are lacking. We validated a method to score clot density in quarter milk samples and describe the prevalence and dynamics of the density scores between consecutive samplings and periods in 4 farms with automatic milking systems. Using in-line filters, we collected quarter milk samples at each milking during 3 periods of 30 h each in each farm. Clot density was scored based on coverage of the filter area as 0 (negative), 1 (trace), 2 (mild), 3 (moderate), 4 (heavy), and 5 (very heavy). The score for a specific quarter and milking is referred to as the quarter milking score (QMS). Three assessors independently scored 902 images of filter samples with a Fleiss kappa value of 0.72. In total, 21,202 quarter milk samples from 5,398 milkings of 621 cows were collected. Of the quarter filter samples, 2.4% had visible clots, distributed as mild (1.4%), moderate (0.6%), heavy (0.3%), and very heavy (<0.1%, n = 8). Cases with a cow period sum of QMS ≥ 4, corresponding to 9.4% of all periods, harbored 86% and 94% of all QMS of 2 to 5 and 3 to 5, respectively. Of these cases, cows sampled in all 3 periods and clots in only 1 period had a quarter period sum score ≥ 1 in 1.8 different quarters in average. Corresponding numbers for the cows with clots or traces in 2 or 3 periods were 2.2 and 2.5 different quarters, respectively. A QMS of 2 to 5 in the preceding milking increased the chance of a QMS >1 in the following milking, with an average chance of 38%. The probability of a QMS > 1 increased with increasing previous QMS, a higher sum of QMS during the milking period, longer milking interval, and higher lactation number, but decreased with increasing days in milk. Our study showed that the method of clot-density scoring is feasible to perform and reproducible for investigating the occurrence and dynamics of clots in milk. Elevated clot-density scores clustered within certain cows and cow periods and appeared in new quarters of the cows over time. The low recurrence of QMS of 1 and 2 within quarters indicated that QMS 3 could be a reasonable threshold for detecting quarters with abnormal milk that require further attention.

Bayesian estimation of qPCR and bacterial culture accuracy for detection of bovine coagulase-negative staphylococci from milk and teat apex at different test cut-off points.

AIM: To primarily estimate the sensitivity (Se) and specificity (Sp) of the commercially available Mastit4 quantitative PCR (qPCR) assay and bacterial culture (BC) for diagnosis of intramammary infections (IMI) and teat apex colonization (TAC) with coagulase-negative staphylococci (CNS) at different cut-offs for qPCR cycle threshold values using Bayesian latent class analysis. A secondary objective was to evaluate two cut-offs of BC for diagnosis of IMI and TAC with CNS. METHODS AND RESULTS: We randomly selected 13-20 cows with subclinical mastitis from eight dairy herds. Teat skin samples and aseptically collected foremilk samples were collected from the right hindquarters (n = 149) for BC and qPCR analysis. The Se of qPCR was always higher than BCSe in diagnosis of IMI, however; the Sp of BC was higher than qPCRSp . BCSe and BCSp showed no substantial difference between the tested BC cut-offs. In contrast to IMI, estimates of BC and qPCR in diagnosing TAC were different. BCSe was higher than qPCRSe at all tested cut-offs, however; qPCRSp was higher than BCSp . CONCLUSION: The overall performance of qPCR is higher than BC in the diagnosis of IMI; however, the performance of BC is better than qPCR in diagnosis of TAC. The qPCR and BC are valid diagnostics for bovine IMI with CNS. However, for TAC, both techniques require further investigation to reduce the uncertainty of the true status of the quarter and teat skin. SIGNIFICANCE AND IMPACT OF THE STUDY: We reported, for the first time, the diagnostic performance of new mastitis technology (Mastit4 PCR) and culture for detection of CNS in milk and nonmilk samples in dairy herds with automatic milking systems. Our findings will improve the interpretation of the test results of culture and qPCR assay and subsequently, will strengthen the control of IMI with CNS in dairy cows.

An update on environmental mastitis: Challenging perceptions.

Environmental mastitis is the most common and costly form of mastitis in modern dairy herds where contagious transmission of intramammary pathogens is controlled through implementation of standard mastitis prevention programmes. Environmental mastitis can be caused by a wide range of bacterial species, and binary classification of species as contagious or environmental is misleading, particularly for Staphylococcus aureus, Streptococcus uberis and other streptococcal species, including Streptococcus agalactiae. Bovine faeces, the indoor environment and used pasture are major sources of mastitis pathogens, including Escherichia coli and S. uberis. A faeco-oral transmission cycle may perpetuate and amplify the presence of such pathogens, including Klebsiella pneumoniae and S. agalactiae. Because of societal pressure to reduce reliance on antimicrobials as tools for mastitis control, management of environmental mastitis will increasingly need to be based on prevention. This requires a reduction in environmental exposure through bedding, pasture and pre-milking management and enhancement of the host response to bacterial challenge. Efficacious vaccines are available to reduce the impact of coliform mastitis, but vaccine development for gram-positive mastitis has not progressed beyond the "promising" stage for decades. Improved diagnostic tools to identify causative agents and transmission patterns may contribute to targeted use of antimicrobials and intervention measures. The most important tool for improved uptake of known mastitis prevention measures is communication. Development of better technical or biological tools for management of environmental mastitis must be accompanied by development of appropriate incentives and communication strategies for farmers and veterinarians, who may be confronted with government-mandated antimicrobial use targets if voluntary reduction is not implemented.

Stoichiometric control of the density of states in PbS colloidal quantum dot solids.

Colloidal quantum dots, and nanostructured semiconductors in general, carry the promise of overcoming the limitations of classical materials in chemical and physical properties and in processability. However, sufficient control of electronic properties, such as carrier concentration and carrier mobility, has not been achieved until now, limiting their application. In bulk semiconductors, modifications of electronic properties are obtained by alloying or doping, an approach that is not viable for structures in which the surface is dominant. The electronic properties of PbS colloidal quantum dot films are fine-tuned by adjusting their stoichiometry, using the large surface area of the nanoscale building blocks. We achieve an improvement of more than two orders of magnitude in the hole mobility, from below 10-3 to above 0.1 cm2/V⋅s, by substituting the iodide ligands with sulfide while keeping the electron mobility stable (~1 cm2/V⋅s). This approach is not possible in bulk semiconductors, and the developed method will likely contribute to the improvement of solar cell efficiencies through better carrier extraction and to the realization of complex (opto)electronic devices.

Molecular epidemiology and strain-specific characteristics of Streptococcus agalactiae at the herd and cow level.

Host-adaptation of Streptococcus agalactiae subpopulations has been described whereby strains that are commonly associated with asymptomatic carriage or disease in people differ phenotypically and genotypically from those causing mastitis in dairy cattle. Based on multilocus sequence typing (MLST), the most common strains in dairy herds in Denmark belong to sequence types (ST) that are also frequently found in people. The aim of this study was to describe epidemiological and diagnostic characteristics of such strains in relation to bovine mastitis. Among 1,199 cattle from 6 herds, cow-level prevalence of S. agalactiae was estimated to be 27.4% based on PCR and 7.8% based on bacteriological culture. Quarter-level prevalence was estimated at 2.8% based on bacteriological culture. Per herd, between 2 and 26 isolates were characterized by pulsed-field gel electrophoresis (PFGE) and MLST. Within each herd, a single PFGE type and ST predominated, consistent with a contagious mode of transmission or point source infection within herds. Evidence of within-herd evolution of S. agalactiae was detected with both typing methods, although ST belonged to a single clonal complex (CC) per herd. Detection of CC23 (3 herds) was associated with significantly lower approximate count (colony-forming units) at the quarter level and significantly lower cycle threshold value at the cow level than detection of CC1 (2 herds) or CC19 (1 herd), indicating a lower bacterial load in CC23 infections. Median values for the number of infected quarters and somatic cell count (SCC) were numerically but not significantly lower for cows infected with CC23 than for cows with CC1 or CC19. For all CC, an SCC threshold of 200,000 cells/mL was an unreliable indicator of infection status, and prescreening of animals based on SCC as part of S. agalactiae detection and eradication campaigns should be discouraged.

Experienced and inexperienced observers achieved relatively high within-observer agreement on video mobility scoring of dairy cows.

Assessment of lameness prevalence and severity requires visual evaluation of thelocomotion of a cow. Welfare schemes including locomotion assessments are increasingly being adopted, and more farmers and their veterinarians might implement a locomotion-scoring routine together. However, high within-observer agreement is a prerequisite for obtaining valid mobility scorings, and within-observer agreement cannot be estimated in a barn, because the gait of cows is dynamic and may change between 2 occasions. The objective of this study was to estimate the within-observer agreement according to the observers' educational background and experience with cattle, based on video recordings with very diverse types of gait. Groups of farmers, bovine veterinarians, first- and fourth-year veterinary students, researchers, and cattle-inexperienced sensory assessors evaluated mobility using a 5-point mobility score system developed specifically for walking cows (n=102 observers). The evaluation sessions were similar for all groups, lasted 75 min, and were organized as follows: introduction, test A, short training session, break, and test B. In total, video recordings of 22 cows were displayed twice in a random order (11 cows in each test × 2 replicates). Data were analyzed applying kappa coefficient, logistic regression, and testing for random effects of observers. The crude estimates of 95% confidence interval for weighted kappa in test A and B ranged, respectively, from 0.76 to 0.80 and 0.70 to 0.75. When adjusting for the fixed effects of video sample and gait scoring preferences, the probability of assigning the same mobility score twice to the same cow varied from 55% (sensory assessors) to 72% (fourth-year veterinary students). The random effect of the individual observers was negligible. That is, in general observers could categorize the mobility characteristics of cows quite well. Observers who preferred to assess the attributes back arch or the overall mobility score (based on uneven gait) had the highest agreement, respectively, 69 or 68%. The training session seemed insufficient to improve agreement. Nonetheless, even novice observers were able to achieve perfect agreement up to 60% of the 22 scorings with merely the experience obtained during the study (introduction and training session). The relatively small differences between groups, together with a high agreement, demonstrate that the new system is easy to follow compared with previously described scoring systems. The mobility score achieves sufficiently high within-observer repeatability to allow between-observer agreement estimates, which are reliable compared with other more-complex scoring systems. Consequently, the new scoring scale seems feasible for on-farm applications as a tool to monitor mobility within and between cows, for communication between farmers and veterinarians with diverse educational background, and for lamenessbenchmarking of herds.

Lameness detection challenges in automated milking systems addressed with partial least squares discriminant analysis.

Lameness causes decreased animal welfare and leads to higher production costs. This study explored data from an automatic milking system (AMS) to model on-farm gait scoring from a commercial farm. A total of 88 cows were gait scored once per week, for 2 5-wk periods. Eighty variables retrieved from AMS were summarized week-wise and used to predict 2 defined classes: nonlame and clinically lame cows. Variables were represented with 2 transformations of the week summarized variables, using 2-wk data blocks before gait scoring, totaling 320 variables (2 × 2 × 80). The reference gait scoring error was estimated in the first week of the study and was, on average, 15%. Two partial least squares discriminant analysis models were fitted to parity 1 and parity 2 groups, respectively, to assign the lameness class according to the predicted probability of being lame (score 3 or 4/4) or not lame (score 1/4). Both models achieved sensitivity and specificity values around 80%, both in calibration and cross-validation. At the optimum values in the receiver operating characteristic curve, the false-positive rate was 28% in the parity 1 model, whereas in the parity 2 model it was about half (16%), which makes it more suitable for practical application; the model error rates were, 23 and 19%, respectively. Based on data registered automatically from one AMS farm, we were able to discriminate nonlame and lame cows, where partial least squares discriminant analysis achieved similar performance to the reference method.

Effect of presampling procedures on real-time PCR used for diagnosis of intramammary infections with Staphylococcus aureus in dairy cows at routine milk recordings.

Aseptic procedures for milk sample collection are considered crucial for bacterial culture to avoid misdiagnosis and subsequently unnecessary treatment or culling. The objective of this field study was to investigate the effect of presampling procedures on the PCR-positivity at cycle threshold value ≤ 37 of real-time PCR assay to detect Staphylococcus aureus from composite milk samples at routine milk recordings while accounting for known cow-level risk factors. A total of 1,199 dairy cows from 6 herds with conventional milking parlors were sampled and tested by PCR in 2011. Following the farmers' routine premilking preparations, 624 cows of the 1,199 cows were randomly selected for bacterial culture preceded by presampling procedures. These procedures were: cleaning of udder teats, removing the first streams of milk, and 70% alcohol teat disinfection. Data on parity, somatic cell counts, days in milk, daily milk yield, fat %, and protein % were extracted from the Danish Cattle Database, whereas energy-corrected milk was calculated based on the latter 3. The within-herd prevalence of intramammary infections with Staph. aureus was 31%, ranging from 16 to 48% in the 6 herds. Univariable analysis showed that the presampling procedures, somatic cell counts, energy-corrected milk, and days in milk were significantly associated with PCR-positivity, whereas parity was not significant. A multivariable model with herd as random effect showed that presampling procedures decreased the chance of being PCR-positive to 0.75 (95% CI; 0.58-0.97) compared with cows where the presampling procedures were not carried out. In conclusion, presampling procedures decreased the cow's chance of being PCR-positive to Staph. aureus. Presampling procedures appeared to improve the specificity of PCR for detection of Staph. aureus by reducing false positives through destruction of Staph. aureus bacteria colonizing or contaminating the teat skin, orifice, and canal. Random herd effects accounted for only 8.9%, indicating that the cluster effect due to herd management on the PCR positivity to Staph. aureus was virtually negligible.

Short communication: scoring of digital dermatitis during milking as an alternative to scoring in a hoof trimming chute.

Digital dermatitis is a serious problem in dairy production in many countries. In many settings, it is important to evaluate the digital dermatitis status of individual cows or an entire dairy herd. Such an evaluation has traditionally been done in a hoof trimming chute. An evaluation in the milking parlor can take place without disturbing the cows to a large extent, it can be done using less labor compared with an evaluation in a hoof trimming chute, and is cheaper than using a chute. The objective was to evaluate the sensitivity and specificity of a rapid screening method for digital dermatitis in the milking parlor, without using any specialized tools and taking approximately 15 s/cow. All lactating cows in 3 commercial Danish dairy herds were included. Cows were first scored for the presence of digital dermatitis during milking and the next day all cows were scored during hoof trimming. A 6-point nominal scoring system based on a visual inspection of the digital dermatitis lesions was used. For the analysis, the scores were dichotomized (digital dermatitis positive or digital dermatitis negative). Additionally, lesions were classified as small (diameter 2 cm). Sensitivity and specificity were calculated using observations from the hoof trimming chute as the "gold standard" and observations during milking as the diagnostic test. Relatively large variation was found between herds with an overall sensitivity of 0.65 (95% confidence interval: 0.59 to 0.72) and a specificity of 0.84 (0.81 to 0.87). The sensitivity increased to 0.69 (0.62 to 0.76), when only large lesions were assessed. The method has several advantages compared with evaluation in a chute and may be a useful tool in the daily hoof health management in dairy herds.

Danish stable schools for experiential common learning in groups of organic dairy farmers.

The farmer field school (FFS) is a concept for farmers' learning, knowledge exchange, and empowerment that has been developed and used in developing countries. In Denmark, a research project focusing on explicit non-antibiotic strategies involves farmers who have actively expressed an interest in phasing out antibiotics from their herds through promotion of animal health. One way of reaching this goal was to form participatory focused farmer groups in an FFS approach, which was adapted to Danish conditions and named "stable schools." Four stable schools were established and went through a 1-yr cycle with 2 visits at each of the 5 or 6 farms connected to each group. A facilitator was connected to each group whose role was to write the meeting agenda together with the host farmer, direct the meeting, and write the minutes to send to the group members after the meeting. Through group focus interviews and individual semistructured qualitative interviews of all participants, the approach of the farmers' goal-directed work toward a common goal was judged to be very valuable and fruitful and based on a common learning process. Complex farming situations were the focus of all groups and in this context, problems were identified and solutions proposed based on each farmer's individual goals. In this article, we describe the experiences of 4 stable school groups (each comprising farmers and a facilitator), and the common process of building a concept that is suitable for Danish organic dairy farming.

Development and daily management of an explicit strategy of nonuse of antimicrobial drugs in twelve Danish organic dairy herds.

Promotion of animal health and well-being at the individual animal and herd level is an important goal in organic farming. At the same time, chemical products affecting the natural balance among living organisms are prohibited in all areas of the organic farm. From an animal welfare point of view, however, no animal must suffer. Therefore, veterinary drugs are allowed under the European Union's regulations for organic farming, despite the fact that they are powerful cell toxins affecting both pathogenic and necessary bacteria, and as such in organic terminology, are regarded as "chemical" or "artificial" products. In this article, we present and discuss interviews with 12 Danish organic dairy producers who claim that minimized use or nonuse of antimicrobial drugs is an explicit goal. The dairy producers were at different levels with regard to reduced antimicrobial treatment. An explicit strategy of no antimicrobial treatments is based primarily on a long-term effort to improve herd health, and secondarily, on finding alternative treatments for diseased animals. Improved hygiene, outdoor access, use of nursing cows, and blinding of chronic mastitis quarters were the main techniques in developing a strategy of not using antimicrobial treatments in the herd by dairy producers. Producers' perception of disease changed from something unavoidable to a disturbing break in the daily rhythm that often could have been avoided. Change toward a nonantimicrobial strategy was gradual and stepwise. All dairy producers in this study desired to preserve the possibility of using antimicrobial drugs in emergencies.

A stochastic model simulating pathogen-specific mastitis control in a dairy herd.

The objective of this study was to develop a model simulating mastitis control in dairy herds and to investigate how sensitive the model is when varying the effect parameters according to the uncertainty. The model simulates 9 pathogen-specific mastitis types, each of which can be subclinical or clinical. The clinical cases can be 1 of 4 severities defined according to the effect of the mastitis case: mild, moderate, severe, and permanent effect. The risk factors include lactation stage, parity, yield level, previous diseases, season, and contagious spread of the infection from herd mates. Occurrence of mastitis is modeled to have direct effects on feed intake, body weight, milk yield, somatic cell count in the milk, subsequent mastitis cases within the cow and in herd mates, voluntary and involuntary culling, mortality, and milk withdrawal. Thirty-five scenarios were simulated to study model behavior and model sensitivity. The consequences per cow/yr of mastitis in the default simulated herd included 0.42 clinical mastitis occurrences, 0.56 subclinical mastitis occurrences, loss of 385-kg milk yield, a 1.3% reduced feed intake, 61-kg milk withdrawal and 146 in reduced economic net return. Based on scenarios demonstrating model behavior and sensitivity analysis, the model appears to produce valid consequences of mastitis control strategies. Representation of the effect of subclinical mastitis and of variation in mastitis severity was concluded in this study to be important when modeling mastitis economics in a dairy herd. The model offers the opportunity to study the long-term herd specific effects of a wide range of control strategies against mastitis.

Cow-related risk factors for milk leakage.

Milk leakage in dairy cows is a symptom of impaired teat sphincter function. Milk leakage is related to an increased risk of mastitis in heifers and cows, and causes hygiene problems. The aim of our study was to assess whether teat shape, condition of teat orifice, and peak milk flow rate are risk factors for milk leakage. We conducted a longitudinal observational study in 15 German dairy farms in which cows were maintained in loose housing. The farms were visited monthly at 2 consecutive milkings. During the evening milking, milk flow curves were measured with the LactoCorder. Milk leakage was recorded during the subsequent morning milking, when cows entered the milking parlor. Immediately after detachment of the milking cluster, teat shape, teat end shape, and condition of the teat orifice of cows were assessed between 9 and 100 d in milk (DIM) and during late lactation (>250 DIM). Data from 1600 cows were analyzed. Milk leakage was treated as the binary response variable in a logistic regression model with herd as a random effect. Primiparous cows with high peak milk flow and teat canal protrusion were at greater risk of milk leakage. High peak milk flow rate, short teats, teat canal protrusion, inverted teat ends, and early lactation increased the risk of milk leakage in multiparous cows. Random herd effects accounted for only 10% of the total variation, indicating that the impact of management or other herd-level factors on the occurrence of milk leakage is virtually negligible for practical purposes.

Systematic clinical examinations for identification of latent udder health types in Danish dairy herds.

A cross-sectional study was conducted to explore the applicability of systematic clinical examinations of udders as an additional tool for the evaluation of udder health status on dairy farms. During 2000, each of the 16 dairy farms was visited 5 times; 20 cows per farm were chosen at random at each visit for clinical udder examination immediately after milking. The clinical examination included both pathological and morphological variables. One examination per cow was included in the analysis (n = 707 cows). Principal component analysis (PCA) was performed in 3 steps. First, 19 variables characterizing udder and teats were analyzed (PCA 1). Second, the variables parity and stage of lactation were included (PCA 2). Finally, somatic cell count (SCC) and milk yield (PCA 3) were included. The PCA resulted in 4 components that explained 30% of the variation of the data: 1) small udder, 2) distressed udder, 3) mastitis udder, and 4) soiled udder. Variables with high positive correlation to the "small udder" were small udder shape, short teats, and first parity. Impaired teat surface, hard udder texture, and a long udder shape were related to the "distressed udder." The "mastitis udder" was characterized by the clinical variables asymmetry between front quarters, asymmetry between hind quarters, knotty tissue, and acute clinical mastitis. Reduced milk yield and high SCC were related to the "mastitis udder," whereas low SCC was related to the "small udder." The "soiled udder" was related to early lactation. Including this information in the assessment of udder health may be of substantial value for data analysis in farms with suspected under-reporting of clinical mastitis.