Metabolic Clusters of Early-Lactating Dairy Cows Based on Blood ß-hydroxybutyrate Trajectories and Predicted from Milk Compounds.

High-yielding dairy cows encounter metabolic challenges in early lactation. Typically, ß-hydroxybutyrate (BHB), measured at a specific time point is employed to diagnose the metabolic status of cows based on a predetermined threshold. However, in early lactation, BHB is highly dynamic, and there is high interindividual variability in its time profile. This could limit the effectiveness of the single measurement and threshold-based diagnosis probably contributing to the disparities in reports linking metabolic status with productive and reproductive outcomes. This research delves into the examination of the trajectories of BHB to unveil inter-cow variations and identify latent metabolic groups. We compiled a data set from 2 observational studies involving a total of 195 lactations from multiparous Holstein Friesian cows. The data set encompasses measurements of BHB, NEFA, and insulin from blood samples collected at 3, 6, 9, and 21 d in milk (DIM), along with weekly determinations of milk composition and fatty acids (FA) proportions in milk fat. In both experiments, milk yield (MY) and feed intake were recorded daily during the first month of lactation. We explored interindividual and intraindividual variations in metabolic responses using the trajectories of blood BHB and evaluated the presence of distinct metabolic groups based on such variations. For this purpose, we employed the growth mixture model (GMM), a trajectory clustering technique. Our findings unveil novel insights into the diverse metabolic responses among cows, encompassing both trajectory patterns and the magnitude of blood BHB concentrations. Specifically, we identified 3 latent metabolic groups: the "QuiBHB" cluster (≈10%) exhibited a higher initial BHB concentration than other clusters, peaking on d 9 (average maximum BHB of 2.4 mM) and then declining by d 21; the "SloBHB" cluster (≈23%) started with a lower BHB concentration, gradually increasing until d 9, and at the highest BHB concentration at d 21 (1.6 mM serum BHB at the end of the experimental period); and the "LoBHB" cluster (≈67%) began with the lowest serum BHB concentration (serum BHB <0.75 mM), remaining relatively stable throughout the sampling period. Notably, the 3 metabolic groups exhibited significant physiological disparities, evident in blood NEFA and insulin concentrations. The QuiBHB and SloBHB cows exhibited higher NEFA and lower insulin concentrations as compared with the LoBHB cows. Interestingly, these metabolic differences extended to MY and DMI during the first month of lactation. The elevated BHB concentrations observed in QuiBHB cows were linked with lower DMI and MY as compared with SloBHB and LoBHB cows. Accordingly, these animals were considered metabolically impaired. Conversely, SloBHB cows displayed higher MY along with increased DMI, and thus the elevated BHB might be indicative of an adaptive response for these cows. The QuiBHB cows also displayed higher proportions of unsaturated FA (UFA), monounsaturated FA (MUFA), and total C18:1 FA in milk during the first week of lactation. Prediction of the QuiBHB cows using these FA and test day variables resulted in moderate predictive accuracy (ROCAUC > 0.7). Given the limited sample size for the development of prediction models, and the variation in DIM among samples in the same week, the result is indicative of the predictive potential of the model and room for model optimization. In summary, distinct metabolic groups of cows could be identified based on the trajectories of blood BHB in early lactation.

Comparison of 3 mathematical models to estimate lactation performance in dairy cows.

Milk yield dynamics and production performance reflect how dairy cows cope with their environment. To optimize farm management, time series of individual cow milk yield have been studied in the context of precision livestock farming, and many mathematical models have been proposed to translate raw data into useful information for the stakeholders of the dairy chain. To gain better insights on the topic, this study aimed at comparing 3 recent methods that allow one to estimate individual cow potential lactation performance, using daily data recorded by the automatic milking systems of 14 dairy farms (7 Holstein, 7 Italian Simmental) from Belgium, the Netherlands, and Italy. An iterative Wood model (IW), a perturbed lactation model (PLM), and a quantile regression (QR) were compared in terms of estimated total unperturbed (i.e., expected) milk production and estimated total milk loss (relative to unperturbed yield). The IW and PLM can also be used to identify perturbations of the lactation curve and were thus compared in this regard. The outcome of this study may help a given end-user in choosing the most appropriate method according to their specific requirements. If there is a specific interest in the post-peak lactation phase, IW can be the best option. If one wants to accurately describe the perturbations of the lactation curve, PLM can be the most suitable method. If there is need for a fast and easy approach on a very large dataset, QR can be the choice. Finally, as an example of application, PLM was used to analyze the effect of cow parity, calving season, and breed on their estimated lactation performance.

Milk yield residuals and their link with the metabolic status of dairy cows in the transition period.

The transition period is one of the most challenging periods in the lactation cycle of high-yielding dairy cows. It is commonly known to be associated with diminished animal welfare and economic performance of dairy farms. The development of data-driven health monitoring tools based on on-farm available milk yield development has shown potential in identifying health-perturbing events. As proof of principle, we explored the association of these milk yield residuals with the metabolic status of cows during the transition period. Over 2 yr, 117 transition periods from 99 multiparous Holstein-Friesian cows were monitored intensively. Pre- and postpartum dry matter intake was measured and blood samples were taken at regular intervals to determine ß-hydroxybutyrate, nonesterified fatty acids (NEFA), insulin, glucose, fructosamine, and IGF1 concentrations. The expected milk yield in the current transition period was predicted with 2 previously developed models (nextMILK and SLMYP) using low-frequency test-day (TD) data and high-frequency milk meter (MM) data from the animal's previous lactation, respectively. The expected milk yield was subtracted from the actual production to calculate the milk yield residuals in the transition period (MRT) for both TD and MM data, yielding MRTTD and MRTMM. When the MRT is negative, the realized milk yield is lower than the predicted milk yield, in contrast, when positive, the realized milk yield exceeded the predicted milk yield. First, blood plasma analytes, dry matter intake, and MRT were compared between clinically diseased and nonclinically diseased transitions. MRTTD and MRTMM, postpartum dry matter intake and IGF1 were significantly lower for clinically diseased versus nonclinically diseased transitions, whereas ß-hydroxybutyrate and NEFA concentrations were significantly higher. Next, linear models were used to link the MRTTD and MRTMM of the nonclinically diseased cows with the dry matter intake measurements and blood plasma analytes. After variable selection, a final model was constructed for MRTTD and MRTMM, resulting in an adjusted R2 of 0.47 and 0.73, respectively. While both final models were not identical the retained variables were similar and yielded comparable importance and direction. In summary, the most informative variables in these linear models were the dry matter intake postpartum and the lactation number. Moreover, in both models, lower and thus also more negative MRT were linked with lower dry matter intake and increasing lactation number. In the case of an increasing dry matter intake, MRTTD was positively associated with NEFA concentrations. Furthermore, IGF1, glucose, and insulin explained a significant part of the MRT. Results of the present study suggest that milk yield residuals at the start of a new lactation are indicative of the health and metabolic status of transitioning dairy cows in support of the development of a health monitoring tool. Future field studies including a higher number of cows from multiple herds are needed to validate these findings.

Sensor-based behavioral patterns can identify heat-sensitive lactating dairy cows.

Heat stress impairs the health and performance of dairy cows, yet only a few studies have investigated the diversity of cattle behavioral responses to heat waves. This research was conducted on an Italian Holstein dairy farm equipped with precision livestock farming sensors to assess potential different behavioral patterns of the animals. Three heat waves, defined as at least five consecutive days with mean daily temperature-humidity index higher than 72, were recorded in the farm area during the summer of 2021. Individual daily milk yield data of 102 cows were used to identify "heat-sensitive" animals, meaning the cows that, under a given heat wave, experienced a milk yield drop that was not linked with other health events (e.g., mastitis). Milk yield drops were detected as perturbations of the lactation curve estimated by iteratively using Wood's equation. Individual daily minutes of lying, chewing, and activity were retrieved from ear-tag-based accelerometer sensors. Semi-parametric generalized estimating equations models were used to assess behavioral deviations of heat-sensitive cows from the herd means under heat stress conditions. Heat waves were associated with an overall increase in the herd's chewing and activity times, along with an overall decrease of lying time. Heat-sensitive cows spent approximately 15 min/days more chewing and performing activities (p < 0.05). The findings of this research suggest that the information provided by high-frequency sensor data could assist farmers in identifying cows for which personalized interventions to alleviate heat stress are needed.

Prediction of first test day milk yield using historical records in dairy cows.

The transition between two lactations remains one of the most critical periods during the productive life of dairy cows. In this study, we aimed to develop a model that predicts the milk yield of dairy cows from test day milk yield data collected in the previous lactation. In the past, data routinely collected in the context of herd improvement programmes on dairy farms have been used to provide insights in the health status of animals or for genetic evaluations. Typically, only data from the current lactation is used, comparing expected (i.e., unperturbed) with realised milk yields. This approach cannot be used to monitor the transition period due to the lack of unperturbed milk yields at the start of a lactation. For multiparous cows, an opportunity lies in the use of data from the previous lactation to predict the expected production of the next one. We developed a methodology to predict the first test day milk yield after calving using information from the previous lactation. To this end, three random forest models (nextMILKFULL, nextMILKPH, and nextMILKP) were trained with three different feature sets to forecast the milk yield on the first test day of the next lactation. To evaluate the added value of using a machine-learning approach against simple models based on contemporary animals or production in the previous lactation, we compared the nextMILK models with four benchmark models. The nextMILK models had an RMSE ranging from 6.08 to 6.24 kg of milk. In conclusion, the nextMILK models had a better prediction performance compared to the benchmark models. Application-wise, the proposed methodology could be part of a monitoring tool tailored towards the transition period. Future research should focus on validation of the developed methodology within such tool.

Milk losses and dynamics during perturbations in dairy cows differ with parity and lactation stage.

Milk yield dynamics during perturbations reflect how cows respond to challenges. This study investigated the characteristics of 62,406 perturbations from 16,604 lactation curves of dairy cows milked with an automated milking system at 50 Belgian, Dutch, and English farms. The unperturbed lactation curve representing the theoretical milk yield dynamics was estimated with an iterative procedure fitting a model on the daily milk yield data that was not part of a perturbation. Perturbations were defined as periods of at least 5 d of negative residuals having at least 1 day that the total daily milk production was below 80% of the estimated unperturbed lactation curve. Every perturbation was characterized and split in a development and a recovery phase. Based hereon, we calculated both the characteristics of the perturbation as a whole, and the duration, slopes, and milk losses in the phases separately. A 2-way ANOVA followed by a pairwise comparison of group means was carried out to detect differences between these characteristics in different lactation stages (early, mid-early, mid-late, and late) and parities (first, second, and third or higher). On average, 3.8 ± 1.9 (mean ± standard deviation) perturbations were detected per lactation in the first 305 d after calving, corresponding to an estimated 92.1 ± 135.8 kg of milk loss. Only 1% of the lactations had no perturbations. On average, 2.3 kg of milk was lost per day in the development phase, while the recovery phase corresponded to an average increase in milk production of 1.5 kg/d, and these phases lasted an average of 10.1 and 11.6 d, respectively. Perturbation characteristics were significantly different across parity and lactation stage groups, and early and mid-early perturbations in higher parities were found to be more severe with faster development rates, slower recovery rates, and higher milk losses. The method to characterize perturbations can be used for precision phenotyping purposes that look into the response of cows to challenges or that monitor applications (e.g., to evaluate the development and recovery of diseases and how these are affected by preventive actions or treatments).

Productive life span and resilience rank can be predicted from on-farm first-parity sensor time series but not using a common equation across farms.

A dairy cow's lifetime resilience and her ability to recalve gain importance on dairy farms, as they affect all aspects of the sustainability of the dairy industry. Many modern farms today have milk meters and activity sensors that accurately measure yield and activity at a high frequency for monitoring purposes. We hypothesized that these same sensors can be used for precision phenotyping of complex traits such as lifetime resilience or productive life span. The objective of this study was to investigate whether lifetime resilience and productive life span of dairy cows can be predicted using sensor-derived proxies of first-parity sensor data. We used a data set from 27 Belgian and British dairy farms with an automated milking system containing at least 5 yr of successive measurements. All of these farms had milk meter data available, and 13 of these farms were also equipped with activity sensors. This subset was used to investigate the added value of activity meters to improve the model's prediction accuracy. To rank cows for lifetime resilience, a score was attributed to each cow based on her number of calvings, her 305-d milk yield, her age at first calving, her calving intervals, and the DIM at the moment of culling, taking her entire lifetime into account. Next, this lifetime resilience score was used to rank the cows within their herd, resulting in a lifetime resilience ranking. Based on this ranking, cows were classified in a low (last third), moderate (middle third), or high (first third) resilience category within farm. In total, 45 biologically sound sensor features were defined from the time series data, including measures of variability, lactation curve shape, milk yield perturbations, activity spikes indicating estrous events, and activity dynamics representing health events (e.g., drops in daily activity). These features, calculated on first-lactation data, were used to predict the lifetime resilience rank and, thus, to predict the classification within the herd (low, moderate, or high). Using a specific linear regression model progressively including features stepwise selected at farm level (cutoff P-value of 0.2), classification performances were between 35.9 and 70.0% (46.7 ± 8.0, mean ± SD) for milk yield features only, and between 46.7 and 84.0% (55.5 ± 12.1, mean ± SD) for lactation and activity features together. This is, respectively, 13.7 and 22.2% higher than what random classification would give. Moreover, using these individual farm models, only 3.5 and 2.3% of cows were classified high when they were actually low, or vice versa, whereas respectively 91.8 and 94.1% of wrongly classified animals were predicted in an adjacent category. The sensor features retained in the prediction equation of the individual farms differed across farms, which demonstrates the variability in culling and management strategies across farms and within farms over time. This lack of a common model structure across farms suggests the need to consider local (and evidence-based) culling management rules when developing decision support tools for dairy farms. With this study we showed the potential of precision phenotyping of complex traits based on biologically meaningful features derived from readily available sensor data. We conclude that first-lactation milk and activity sensor data have the potential to predict cows' lifetime resilience rankings within farms but that consistency between farms is currently lacking.

Competitive inhibition assay for the detection of progesterone in dairy milk using a fiber optic SPR biosensor.

Analytical methods that are often used for the quantification of progesterone in bovine milk include immunoassays and chromatographic techniques. Depending on the selected method, the main disadvantages are the cost, time-to-result, labor intensity and usability as an automated at-line device. This paper reports for the first time on a robust and practical method to quantify small molecules, such as progesterone, in complex biological samples using an automated fiber optic surface plasmon resonance (FO-SPR) biosensor. A FO-SPR competitive inhibition assay was developed to determine biologically relevant concentrations of progesterone in bovine milk (1-10 ng/mL), after optimizing the immobilization of progesterone-bovine serum albumin (P4-BSA) conjugate, the specific detection with anti-progesterone antibody and the signal amplification with goat anti-mouse gold nanoparticles (GAM-Au NPs). The progesterone was detected in a bovine milk sample with minimal sample preparation, namely ½ dilution of the sample. Furthermore, the developed bioassay was benchmarked against a commercially available ELISA, showing excellent agreement (R2 = 0.95). Therefore, it is concluded that the automated FO-SPR platform can combine the advantages of the different existing methods for quantification of progesterone: sensitivity, accuracy, cost, time-to-result and ease-of-use.

Visible and near-infrared bulk optical properties of raw milk.

The implementation of optical sensor technology to monitor the milk quality on dairy farms and milk processing plants would support the early detection of altering production processes. Basic visible and near-infrared spectroscopy is already widely used to measure the composition of agricultural and food products. However, to obtain maximal performance, the design of such optical sensors should be optimized with regard to the optical properties of the samples to be measured. Therefore, the aim of this study was to determine the visible and near-infrared bulk absorption coefficient, bulk scattering coefficient, and scattering anisotropy spectra for a diverse set of raw milk samples originating from individual cow milkings, representing the milk variability present on dairy farms. Accordingly, this database of bulk optical properties can be used in future simulation studies to efficiently optimize and validate the design of an optical milk quality sensor. In a next step of the current study, the relation between the obtained bulk optical properties and milk quality properties was analyzed in detail. The bulk absorption coefficient spectra were found to mainly contain information on the water, fat, and casein content, whereas the bulk scattering coefficient spectra were found to be primarily influenced by the quantity and the size of the fat globules. Moreover, a strong positive correlation (r ≥ 0.975) was found between the fat content in raw milk and the measured bulk scattering coefficients in the 1,300 to 1,400 nm wavelength range. Relative to the bulk scattering coefficient, the variability on the scattering anisotropy factor was found to be limited. This is because the milk scattering anisotropy is nearly independent of the fat globule and casein micelle quantity, while it is mainly determined by the size of the fat globules. As this study shows high correlations between the sample's bulk optical properties and the milk composition and fat globule size, a sensor that allows for robust separation between the absorption and scattering properties would enable accurate prediction of the raw milk quality parameters.

Visible and near-infrared spectroscopic analysis of raw milk for cow health monitoring: reflectance or transmittance?

The composition of produced milk has great value for the dairy farmer. It determines the economic value of the milk and provides valuable information about the metabolism of the corresponding cow. Therefore, online measurement of milk components during milking 2 or more times per day would provide knowledge about the current health and nutritional status of each cow individually. This information provides a solid basis for optimizing cow management. The potential of visible and near-infrared (Vis/NIR) spectroscopy for predicting the fat, crude protein, lactose, and urea content of raw milk online during milking was, therefore, investigated in this study. Two measurement modes (reflectance and transmittance) and different wavelength ranges for Vis/NIR spectroscopy were evaluated and their ability to measure the milk composition online was compared. The Vis/NIR reflectance measurements allowed for very accurate monitoring of the fat and crude protein content in raw milk (R(2)>0.95), but resulted in poor lactose predictions (R(2)<0.75). In contrast, Vis/NIR transmittance spectra of the milk samples gave accurate fat and crude protein predictions (R(2)>0.90) and useful lactose predictions (R(2)=0.88). Neither Vis/NIR reflectance nor transmittance spectroscopy lead to an acceptable prediction of the milk urea content. Transmittance spectroscopy can thus be used to predict the 3 major milk components, but with lower accuracy for fat and crude protein than the reflectance mode. Moreover, the small sample thickness (1mm) required for NIR transmittance measurement considerably complicates its online use.

Pressure distribution at the teat-liner and teat-calf interfaces.

During milking, the teat is loaded because of a combination of vacuum and pressure of the collapsing liner. It is assumed that pressure concentrations tend to cause teat-end injuries and hyperkeratosis. The pressure distribution on the bovine teat was measured to test the hypothesis that the pressures of the collapsed liner are unevenly distributed over the teat. With the aid of a pressure-sensitive sensor (approximately 2 gauge points/cm(2)), the pressures at the teat-liner and the teat-calf interfaces were measured at 100 Hz. Pressure distribution over the surface of an artificial teat was measured with 7 different liners, 1 liner at 3 different vacuum levels, and a suckling calf. One cow was equipped with a sensor at a teat during a milking with one of the liners. Conventional round liners concentrated the load over 2 sites at the teat end. Some liners (softer material, reduced tension, smaller barrel, reduced mouthpiece depth) distributed the compressive load over a larger area of the teat. Although all liners distributed the highest pressures at the teat end, some liner designs showed a 25% reduction at the site of interest at vacuum of 44 kPa. The calf forced milk flow by a combination of suckling and overpressure in the teat cistern caused by the tongue. While the calf was swallowing, teat pressure was reduced because of a decrease in vacuum. Moreover, the calf did not load the teat end, probably because the teat canal would be closed and the milk would not flow. The method of using a pressure sensor to analyze teat loading at the teat-liner and the teat-calf interfaces showed potential and is a first step toward developing a natural milking technique.