Hygiene management practices and adenosine triphosphate luminometry of feeding equipment in preweaning calves on dairy farms in Quebec, Canada.

The objective of this study was to describe the cleaning practices currently used for preweaning calves on dairy farms in Quebec, Canada. In addition, contamination of feeding equipment for preweaning calves was described using ATP (expressed as relative light units, RLU), visual assessment, and bacteriological analysis. A questionnaire was administered on 50 commercial dairy farms in Quebec, Canada, regarding the self-reported cleaning protocol used for feeding equipment of preweaning calves. During the visit, a visual score was given to the feeding equipment available at the farm. Afterward, ATP luminometry measurements were obtained using Hygiene UltraSnap and MicroSnap swabs (Hygiene, Camarillo, CA), and the liquid rinsing technique for buckets, nipples, bottles, esophageal tube feeders (ET), the tube of automatic milk feeders (AMF), water samples, and milk replacer. An additional direct swabbing technique was performed on buckets and nipples. The fluid retrieved from the liquid rinsing technique was also used to determine the total bacterial count (TBC) and total coliform count. Based on the bacteriological analysis, optimal RLU cutoff values to determine contamination were obtained. The median (interquartile range) luminometer measurements using the UltraSnap and direct technique for buckets and nipples were 2,082 (348-7,410) and 3,462 (462-7,518) RLU, respectively; and, using the liquid technique for bottles, ET, AMF, water, and milk replacer were 43 (4-974), 15 (4-121), 301 (137-1,323), 190 (71-358), and 94 (38-218) RLU, respectively. Overall, for all equipment and both techniques used, higher RLU values were seen in UltraSnap samples compared with MicroSnap samples. Additionally, for buckets and nipples, higher RLU values were obtained for the direct swabbing method compared with the liquid sampling method for both swabs used. No differences in the level of contamination were seen between the different feeding equipment used within a farm. Overall, a higher correlation with bacteriological results was noticed for ATP luminometry compared with the visual score, with a high correlation for nipples and bottles using the UltraSnap and liquid technique. Based on the classification of "contaminated" (TBC ≥100,000 cfu/mL) or "not contaminated" (TBC <100,000 cfu/mL), optimal ATP luminometer cutoff values for buckets, nipples, bottles, AMF, water, and milk replacer were 798, 388, 469, 282, 1,432, and 93 RLU, respectively. No clear association was found between ATP measurements and the self-reported cleaning protocol. This study gave new insights into the current cleaning procedures and contamination of feeding equipment for preweaning calves on dairy farms in Quebec. In addition, ATP luminometry cutoff values could help benchmark farms regarding cleaning practices and provide customized advice, improving the overall hygiene management, and thus the health, of preweaning calves on dairy farms.

Standardization and validation of ATP luminometry as a diagnostic tool to assess the cleanliness of feeding equipment in preweaning calves.

The objective of this cross-sectional study was to standardize a reliable and repeatable swabbing technique using ATP luminometry (light emission proportional to the amount of ATP with result provided in relative light units [RLU]) to describe the cleanliness of various feeding equipment used for preweaning calves in dairy farms. A total of 7 Québec commercial dairy herds were selected conveniently. Following visual hygiene scoring, the cleanliness of every available piece of feeding equipment was assessed using direct surface swabbing for buckets and nipples with Hygiena UltraSnap swabs. A liquid rinsing technique was used for esophageal feeders, bottles, and automatic milk feeders (AMF) with UltraSnap, AquaSnap, and MicroSnap swabs. To validate direct swabbing technique of buckets, a stage within and between operators was realized, as well as a conventional bacterial culture. A total of 519 swab samples were obtained from 201 pieces of equipment. The median (interquartile range) contamination in RLU for a bottle, esophageal feeder, AMF, bucket and nipple was 2 (1;6), 2 (0;12), 52 (19;269), 886 (128;7,230) and 899 (142;6,928), respectively. The direct swabbing technique, which consists in swabbing directly the surface of an equipment, showed excellent correlation for intrarater reliability (intraclass correlation (ICC) = 0.93; 95% CI: 0.88-0.96). The interoperator (2 sessions with 3 different operators) reliability also showed high correlation (ICC = 0.88; 95% CI: 0.78-0.94 for the first session, and ICC = 0.89; 95% CI: 0.79-0.95 for the second session). Luminometer values were positively associated with the visual score of esophageal feeders, AMF and buckets. A positive correlation between bacterial culture and direct swabbing of buckets was also found for the UltraSnap (rs = 0.653; 95% CI: 0.283-0.873; P = 0.0003) and MicroSnap (rs = 0.569, 95% CI: 0.309-0.765; P = 0.002). This study describes a standardized and practical on-farm swabbing technique for assessing the hygienic status of feeding equipment by luminometry, which can be integrated in the investigation of preweaning dairy calves problems.

Reproductive performance of lactating dairy cows with elevated milk ß-hydroxybutyrate levels during first 6 weeks of lactation.

Although there is evidence that ketosis negatively affects fertility, the effect of late and early ketosis on the reproductive performance of lactating cows has not been systematically investigated. The aim of this study was to evaluate the association between time and amplitude of elevated milk BHB (EMB) occurring within 42 d in milk (DIM) and subsequent reproductive performance of lactating Holstein cows. The dairy herd information data of 30,413 cows with 2 test-day milk BHB recordings during early lactation periods 1 and 2 (5-14 and 15-42 DIM, respectively) assessed as negative (<0.15 mmol/L), suspect (0.15-0.19 mmol/L), or positive (≥0.2 mmol/L) for EMB were used in this study. Based on the time and amplitude of milk BHB, cows were grouped into 7 groups: (1) healthy cows negative in both periods 1 and 2 were classified as NEG; (2) suspect in period 1 and negative in period 2: EARLY_SUSP; (3) suspect in period 1 and suspect/positive in period 2: EARLY_SUSP_Pro; (4) positive in period 1 and negative in period 2: EARLY_POS; (5) positive in period 1 and suspect/positive in period 2: EARLY_POS_Pro; (6) negative in period 1 and suspect in period 2: LATE_SUSP; and (7) negative in period 1 and positive in period 2: LATE_POS. The overall prevalence of EMB within 42 DIM was 27.4%, with the highest prevalence being EARLY_SUSP (10.49%). Cows in EARLY_POS and EARLY_POS_Pro, but not other EMB categories, had a longer interval from calving to first service compared with NEG cows. For the reproductive parameters, first service to conception interval, days open and calving interval, cows in all EMB groups except EARLY_SUSP had longer intervals compared with NEG cows. These data indicate that there is a negative association between EMB within 42 d and reproductive performance after the voluntary waiting period. The intriguing findings of this study are the unaltered reproductive performance of EARLY_SUSP cows, and the negative association between late EMB and reproductive performance. Hence, monitoring and prevention of ketosis during the first 6 wk of lactation is necessary to optimize reproductive performance of lactating dairy cows.

Toward Precision Feeding Regarding Minerals: What Is the Current Practice in Commercial Dairy Herds in Québec, Canada?

This analysis is performed to obtain information on the current situation regarding phosphorus (P), cobalt (Co), copper (Cu), iron (Fe), manganese (Mn), and zinc (Zn) concentrations in cow diets of commercial dairy herds in Québec, Canada, and to compare them with National Research Council recommendations. Data are collected on 100 Holstein dairy herds in Québec, Canada, and 4430 cows were involved. Rations are analyzed for selected minerals and cow requirements relative to the recommendations were calculated. Median percentages of mineral recommendations fulfilled by forage were 55%, 196%, 54%, 776%, 181%, and 44% for P, Co, Cu, Fe, Mn, and Zn, respectively. Daily dietary concentrations of P, Cu, Mn, and Zn decreased as lactation progressed, whereas Co and Fe were stable throughout lactation. Phosphorus was the mineral fed the closest to the requirements, cows below 21 days in milk were even underfed by 11%. All studied trace minerals were fed in excess for the majority of cows. Cobalt was fed on average 480% above requirements regardless of the stage of lactation. For Cu, Fe, Mn, and Zn, rations for cows below 21 days in milk were fed 23% (95% confidence interval: 15-32), 930% (849-1019), 281% (251-314), and 35% (22-47) above the recommendations, respectively, and were closer to the requirements than after 21 days in milk. These results show that most nutritionists are aware that precision feeding regarding P is important to minimize detrimental environmental impacts of dairy production. However, some efforts should be made to limit trace mineral overfeeding to ensure environmental resiliency.

Fatty Acid Profiles from Routine Milk Recording as a Decision Tool for Body Weight Change of Dairy Cows after Calving.

Cows mobilize body reserves during early lactation, which is reflected in the milk fatty acid (FA) profile. Milk FA can be routinely predicted by Fourier-transform infrared (FTIR) spectroscopy, and be, thus, used to develop an early indicator for bodyweight change (BWC) in early lactating cows in commercial dairy farms. Cow records from 165 herds in Denmark between 2015 and 2017 were used with bodyweight (BW) records at each milking from floor scales in automatic milking systems. Milk FA in monthly test-day samples was predicted by FTIR. Predictions of BWC were based on a random forest model and included parity, stage of lactation, and test day milk production and components (fat, protein, and FA). Bodyweight loss was mainly explained by decreased short-chain FA (C4:0-C10:0) and increased C18:0 FA. The root mean square error (RMSE) of prediction after cross-validation was 1.79 g/kg of BW (R2 of 0.94). Model evaluation with previously unseen BWC records resulted in reduced prediction performance (RMSE of 2.33 g/kg of BW; R2 of 0.31). An early warning system may be implemented for cows with a large BW loss during early lactation based on milk FA profiles, but model performance should be improved, ideally by using the full FTIR milk spectra.

Evaluation of the new differential somatic cell count parameter as a rapid and inexpensive supplementary tool for udder health management through regular milk recording.

Mastitis, particularly in its subclinical form, is the costliest disease in milk production causing substantial financial losses to the dairy industry, impairing animal welfare, and one of the main reasons for treating dairy cows with antimicrobials. Somatic cell count (SCC) is broadly used as an indicator for mastitis or intramammary infection (IMI) and is the basis for udder health management programmes, e.g., through monthly dairy herd improvement (DHI) testing. While SCC shows the total number of cells in milk, the new Differential SCC (DSCC) shows also the combined proportion of polymorphonuclear neutrophils (PMN) and lymphocytes as a percentage of the total SCC. In this study, we investigated the test characteristics of DSCC as a new supplementary indicator for mastitis screening. We collaborated with 11 herds totalling 969 dairy cows and collected metered DHI samples once a month over four months. The IMI status was assessed through analysis of aseptic composite hand-stripped samples using culture and followed by species identification using MALDI-ToF. The pathogens detected were categorised as 'no', 'minor', 'major', or 'other' pathogens. The results of our study showed that the DSCC parameter was significantly associated with the IMI status and the cow's parity but not with days in milk or test-day milk weight. On the other hand, SCC was associated with all these four factors. DSCC counts were significantly higher in samples of cows with IMI caused by major pathogens as compared to cows with no IMI or IMI by minor or other pathogens. SCC alone, DSCC alone, and the combination of DSCC and SCC were further compared based on test characteristics using exemplary cut-offs. For example, working with a cut-off of 200,000 cells/ for SCC alone compared to working with the combination of DSCC of 65 % and/or 200,000 cells/mL to classify cows as infected by major pathogens, the sensitivity increased from 78 % to 92 % and the specificity decreased from 87 % to 66 %. With the combination, the positive predictive value changed from 52 % to 34 %, and the negative predictive value stayed at the same level (96 % vs 98 %). In summary, our study provides first insights on test characteristics of the DSCC parameter used in combination with the well-established SCC for monitoring udder health using DHI testing. This combination opens up the possibility to further improve udder health monitoring programmes (e.g., improved identification of IMI caused by major pathogens) but more work on the subject is needed.

Milk ß-hydroxybutyrate concentration measured by Fourier-transform infrared and flow-injection analyses from samples taken at different times relative to milking.

Analysis of milk BHB concentration by Fourier-transform infrared (FTIR) spectrometry more frequently than regular milk testing could help dairy producers in decision making, particularly if it would be possible to use small hand-stripped samples (hereinafter simply called samples) taken between dairy herd improvement (DHI) test-samples analysed using DHI algorithms. The aim of this Research Communication was to evaluate milk BHB concentration and the prevalence of elevated milk BHB concentration analysed by FTIR spectrometry compared with flow-injection analysis (SKALAR) from samples taken at different times relative to the milking. A total of 293 early-lactation cows in 44 commercial dairy herds were involved in the study. Herds were visited once during the morning milking when a routine DHI test-sample was obtained using in-line milk samplers. Additional milk samples were taken by hand stripping as follows: (1) Just before connecting the milking machine; (2) immediately after removing the milking machine; (3) 3 h after milking and (4) 6 h after milking. Milk samples were analysed for BHB concentration by FTIR and SKALAR, the latter being the reference method. Milk BHB concentration from samples taken before milking was different between FTIR and SKALAR whereas no difference was noted for other sampling times, although milk BHB concentration rose as time after milking increased. Except for DHI test-samples for which prevalence was not different between analysis methods, prevalence of elevated milk BHB concentration (≥0.15 mmol/l) was greater for FTIR analysis. However, no difference in prevalence was observed between SKALAR and FTIR when using a threshold of ≥0.20 mmol/l. In summary, hand-stripped milk samples taken any time after removing the milking machine until 6 h after the milking can be recommended for FTIR analysis of elevated milk BHB concentration prevalence provided a threshold of 0.20 mmol/l is used.

Calculations of apparent ruminal synthesis and intestinal absorption of biotin in dairy cows as influenced by the extraction method.

Biotin is present in nature either free or as biocytin, which is only degraded under the action of a specific enzyme: biotinidase. This enzyme is not included in analytical assays generally used. A method for sample preparation using biotinidase was developed in our laboratory before analysis by ELISA. Three cows equipped with duodenal and ileal cannulae were used to compare the effects of methods of sample preparation on calculations of apparent ruminal synthesis and intestinal absorption of biotin. There was no apparent ruminal synthesis of biotin, no matter whether free or total biotin was measured (p = 0.84). Results also suggested that rumen microbes cannot utilize nor degrade biocytin present in the feed. Estimates of apparent intestinal absorption were influenced by the sample preparation method (p = 0.002). Analysis of free biotin caused an artefact, suggesting intestinal synthesis of this vitamin; whereas determination of total biotin concentrations showed that absorption was taking place in the small intestine.