The effect of flow-responsive pulsation on teat tissue condition and milking performance in Holstein dairy cows.

The objectives of this study were to assess the effect of a reduced liner-open phase applied through flow-responsive pulsation (FRP), as a method to provide supplemental stimulation, on teat tissue conditions and milking characteristics in dairy cows. In 2 switch-back trials, 156 Holstein cows milked 3 times daily were assigned to the FRP or conventional (CON) group in alternating sequences. Trial I lasted for 35 d and was split into 5 alternating 1-week periods of FRP and CON. The duration of Trial II was 84 d, consisting of 4 alternating 3-week periods of FRP and CON. Premilking udder preparation for both groups consisted of predipping, forestripping and wiping the teats. Upon milking unit attachment, the FRP cows were milked at a pulsation rate of 50 cycles/min and a pulsation ratio of 30:70 until the preset milk flow threshold of 0.5 kg/min was reached. When the threshold value of 0.5 kg/min was reached, the pulsation was automatically switched to milking mode, which consisted of a pulsation rate of 60 cycles/min and a pulsation ratio of 70:30. Cows in the CON group were milked by milking mode (pulsation rate, 60 cycles/min; pulsation ratio, 70:30) immediately after attachment of the milking unit. We assessed machine milking-induced short-term changes to the teat tissue by palpation and visual inspection during Trial I, and we assessed teat-end hyperkeratosis in Trial II. Electronic on-farm milk meters were used to assess milking characteristics [milk yield (kg/milking session), milking unit-on time (s), 2-min milk yield (kg), peak milk flow rate (kg/min), and duration of low milk flow rate (s)]. Generalized linear mixed models were used to analyze the effect of treatment on the outcome variables. The odds of machine milking-induced short-term changes to the teat tissue were lower for cows in the FRP group than for those in the CON group [odds ratio (95% confidence interval; 95% CI) = 0.41 (0.31-0.55)]. There were no meaningful differences in the odds of teat-end hyperkeratosis between the FRP and CON groups [odds ratio (95% CI) = 1.05 (0.38-2.89)]. The least squares means (95% CI) of milking characteristics in the FRP and CON groups were 14.3 (13.8-14.7) and 14.3 (13.8-14.7) kg for milk yield, respectively; 272 (264-281) and 270 (262-278) s for milking unit-on time, respectively; and 5.0 (4.8-5.1) and 4.9 (4.7-5.1) kg/min for peak milk flow rate, respectively. The FRP group had lower odds of bimodality than the CON group [odds ratio (95% CI) = 0.67 (0.61-0.74)]. In reference to CON, the odds ratios (95% CI) in FRP were 1.05 (0.76-1.46) for kick-off, and 1.02 (0.85-1.23) for milking unit reattachment. In this study, cows that were milked using FRP had lower odds of postmilking short-term changes to the teat tissue and lower odds of bimodal milk flow. We conclude that FRP may foster adequate teat stimulation in cows before the initiation of milk harvest and has the potential to improve teat tissue conditions.

The association between teat shape and bimodal milk ejection in Holstein dairy cows.

Our objectives were to investigate the association of teat shape with (1) bimodality (2) incremental milk flow rates (average flow rates during the first 15 s, 15-30 s, 30-60 s, and 60-120 s of milking), and (3) peak milk flow rate in Holstein dairy cows milked 3 times per day. In this prospective cohort study, we analyzed 220,928 milking observations that were collected from 2,520 cows from a single dairy herd over a period of 31 d. Teat shape was visually assessed and classified into 1 of 4 categories as follows: (1) triangular barrel and pointed teat end (TP), (2) square barrel and round teat end (SR), (3) square barrel, round teat end, and flat in the area of the teat orifice (SRF), and (4) square barrel and flat teat end (SF). Individual cow-level milk flow rates were obtained using electronic on-farm milk meters. We considered bimodality to be present if any of the incremental milk flow rates (flow rates during 15-30 s, 30-60 s, or 60-120 s) were lower than any of the previous rates (flow rates during the first 15 s, 15-30 s, or 30-60 s). The frequency distribution of cows with different teat shapes was as follows: TP, 96 (3.8%); SR, 1,751 (69.5%); SRF, 617 (24.5%); and SF, 56 (2.2%). A generalized linear mixed model revealed differences in the odds of bimodality among cows with different teat shapes. Compared with cows in category SR, the odds (95% confidence interval) of bimodality of cows in other categories were as follows: TP, 0.68 (0.48-0.97); SF, 1.96 (1.21-3.19); and SRF, 1.46 (1.23-1.72). General linear mixed models indicated an association between teat shape and all incremental milk flow rates, with higher milk flow rates being present for cows with teat shapes in the SRF and SF categories. The general linear mixed model for the dependent variable peak milk flow rate indicated an association between teat shape and peak milk flow rate. Least squares means and 95% confidence intervals were 5.1 (4.9-5.3) kg/min for TP, 5.8 (5.5-6.1) kg/min for SF, 5.6 (5.5-5.7) for SRF, and 5.3 (5.3-5.4) for SR, respectively. We conclude that in this study cohort, bimodality is more likely to occur in cows with flat teat ends compared with those with a round teat ends. We attribute this relationship to the difference in milking speed across cows with different teat shapes. Teat shape may serve as a useful phenotype to identify cows that are more likely to exhibit bimodality. Because our study population was unique, future studies considering different circumstances such as breed, milking system, and milking routine are needed before results can be extrapolated.

A retrospective cohort study investigating the association of individual quarter dry-off with udder health, milk production, and culling risk.

Individual quarter dry-off (QDO) has been increasingly employed as a strategy for managing cows with chronically elevated SCC and recurrent clinical mastitis. However, little knowledge is available on the effects of QDO on milk production, SCC, the risk of clinical mastitis, and the risk of removal from the herd. Therefore, this retrospective cohort study aimed to investigate these associations. Data from 471 dairy cows subjected to QDO were analyzed. The cows were housed on a 4,000-cow dairy farm with a thrice-daily milking schedule. The cows were grouped based on the reason for QDO: (1) cows detected with a nonlactating quarter at a fresh cow check (QFRESH); (2) cows with recurrent clinical mastitis (QMAST); (3) cows diagnosed with Staphylococcus aureus IMI (QSA); and (4) cows with chronic subclinical mastitis (QSCC). Additionally, we randomly selected herd mates at a ratio of 1:1 to serve as a control group (CON). Cows in the CON group were matched in terms of parity and stage of lactation. Generalized linear mixed models with an identical link were used to estimate milk yield and SCC at 1 test day before QDO (T-1) as well as 1, 2, and 3 (T1, T2, and T3) test days after QDO. All cows subjected to QDO exhibited a decrease in milk yield following QDO compared with their respective control groups. All QDO cows approached the yield of their control group by T3. In particular, the difference in milk yield between QMAST cows and their controls at T3 was less than the difference at T1. Cows in the QMAST and QSCC groups exhibited a decrease in their SCC following QDO. In particular, the SCC was significantly higher among QMAST cows than among their controls at T1, but this difference was no longer significant by T3. Proportional hazards regression models revealed that QDO was associated with clinical mastitis occurrence and removal from the herd. Compared with CON cows, the hazard ratio (95% CI) for clinical mastitis occurrence was 3.70 (1.65-8.28), 1.80 (1.31-2.47), and 2.27 (0.93-5.54) among QFRESH, QMAST, and QSA cows, respectively. The hazard ratio among QSCC cows was modified by the effect of parity. The hazard ratio (95% CI) for removal from the herd was higher among cows subjected to QDO than among CON cows (hazard ratio [95% CI] values of 2.30 [0.99-5.33], 3.27 [2.20-4.86], and 4.87 [1.81-13.12] for QFRESH, QMAST, and QSCC cows, respectively). We conclude that QDO can be a viable strategy for managing cows with recurrent clinical mastitis. However, the results for the cows that underwent QDO for other reasons are less clear, partially due to low statistical power. Therefore, future research should examine how to decrease the risks of clinical mastitis and removal from the herd among cows subjected to QDO.

The effect of 2 different premilking stimulation regimens, with and without a latency period, on teat tissue condition and milking performance in Holstein dairy cows.

The objectives of this study were to assess the effect of 2 different premilking stimulation regimens, with and without a latency period between tactile stimulation and the attachment of the milking unit, on the teat tissue condition and milking performance of dairy cows. In a randomized controlled crossover study, 145 Holstein cows milked 3 times daily were assigned to treatment (TRT) or control (CON) groups. Premilking udder preparation for the TRT group consisted of the application of a latency period resulting in a preparation lag time of 90 s. The only difference in the premilking udder preparation of the CON group was the absence of latency period; the milking unit was attached immediately after completion of the tactile stimulation. The average duration of total tactile stimulation in TRT and CON group was 8 ± 2 and 9 ± 2 s, respectively. The study lasted for 14 d and was split into 2 periods, each consisting of a 2-d adjustment period followed by 5 d of data collection. We assessed machine milking-induced short-term changes to the teat tissue by palpation and visual inspection postmilking. Electronic on-farm milk meters were used to assess milking characteristics (milk yield [kg/milking session], machine-on time [s], 2-min milk yield [kg], and duration of low milk flow rate [s]). Generalized linear mixed models were used to analyze the effect of treatment on the outcome variables. The odds of machine milking-induced short-term changes to the teat tissue were lower for cows that received a 90-s preparation lag time (TRT cows) compared with cows in the CON group (odds ratio [95% confidence interval; 95% CI] = 0.13 [0.08-0.20]). The least squares means (95% CI) values of cows in the TRT and CON groups were 15.4 (14.9-15.9) and 15.3 (14.8-15.8) kg, respectively, for milk yield, and 246 (239-253) and 253 (247-260) s for machine-on time. The 2-min milk yield was higher for the TRT compared with CON group cows at all the parity levels. The 2-min milk yields of animals in lactation 1, 2, and ≥3 were 5.7, 5.7, and 6.5 kg, respectively, in the TRT group and 4.6, 5.0, and 5.9 kg in the CON group. The TRT cows spent less time in low milk flow rate compared with CON cows at all parity levels. The durations of low milk flow rate of cows in lactation 1, 2, and ≥3 in the TRT group were 19, 17 and 13 s, respectively, and those in the CON group were 31, 22, and 15 s. In this study, cows that received a latency period, and thus were subjected to a 90-s preparation lag time had lower odds of exhibiting short-term changes to the teat tissue after machine milking, shorter machine-on time, higher 2-min milk yields, and lower durations of low milk flow rates. We conclude that consideration of latency period leading to a 90-s preparation lag time in the premilking stimulation regimen facilitated cows' milk-ejection reflex. This latency period can alleviate the adverse effects of vacuum-induced forces on teat tissue during machine milking, improve udder health, and promote animal well-being.

A randomized controlled trial to study the effects of an automated premilking stimulation system on milking performance, teat tissue condition, and udder health in Holstein dairy cows.

The objectives were to examine the effect of an automated premilking stimulation (APS) by means of a high pulsation frequency (300 cycles/min) without a reduction of the vacuum in the pulsation chamber or claw piece on (1) milking performance, (2) teat tissue condition, and (3) udder health in dairy cows. In a randomized controlled field study, Holstein cows (n = 427) from 1 commercial dairy farm with a milking schedule of 3 times per day were assigned to treatment and control groups over a 90-d period. Treatments consisted of a maximum of 80 s (APS80) or 99 s (APS99) of mechanical stimulation at a pulsation rate of 300 pulses per minute and a ratio of 25:75 (no reduction of the pulsation chamber or milking vacuum). Cows in the control group (CON) received traditional premilking stimulation by means of manual forestripping for 8 s. Milking characteristics were documented with on-farm milk meters. Short- and long-term changes in teat tissue condition induced by machine milking were assessed visually on a weekly basis. Composite milk samples were analyzed once per month to determine somatic cell count. Generalized linear mixed models were used to study the effect of the treatment on the outcome variables. We observed no meaningful differences in milk yield or milking unit-on time. Least squares means and their 95% confidence intervals (95% CI) for cows in the APS80, APS99, and CON groups were 13.5 (13.1-14.0), 13.2 (12.8-13.7), and 13.2 (12.8-13.7) kg for milk yield and 222 (213-231), 219 (210-228), and 223 (214-232) s for milking unit-on time, respectively. The effect of treatment on bimodality was modified by milk yield such that the odds of bimodality increased in the treatment groups with increasing milk yield. Compared with cows in the CON group, the odds ratios (95% CI) of bimodality were 1.08 (0.62-1.89) in the APS80 group and 0.89 (0.55-1.42) in the APS99 group at a milk yield of 11 kg and 2.0 (1.24-3.22) in the APS80 group and 2.08 (1.29-3.35) in the APS99 group at a milk yield of 16 kg. We observed differences in short- and long-term changes in teat tissue condition between the treatment and control groups. Compared with cows in the CON group, the odds (95% CI) of short-term changes were 1.87 (1.35-2.58) for the APS80 group and 1.49 (1.08-2.07) for the APS99 group, and the odds of long-term changes were 1.52 (1.24-1.85) for cows in the APS80 group and 1.59 (1.31-1.94) for cows in the APS99 group. The least squares means (95% CI) for somatic cell counts (log10-transformed) were 4.74 (4.68-4.81) for the APS80 group, 4.77 (4.71-4.83) for the APS99 group, and 4.79 (4.73-4.86) for the CON group. We conclude that the APS system tested here had no negative effects on milk yield or milking unit-on time. However, differences in bimodality and teat tissue condition suggest that the APS system did not provide sufficient stimulation to facilitate a gentle milk harvest and adversely affects teat tissue condition.

Genomic Characterization of Mycoplasma arginini Isolated from a Housefly on a Dairy Farm and Comparison with Isolates from Bovine Milk and Lung Tissue.

Mycoplasma mastitis can be highly contagious, unresponsive to treatment, and cause severe economic problems in affected herds. Notable routes of Mycoplasma spp. transmissions are contaminated milking equipment and animal contact through respiratory secretions. Only a few studies report the environment as a possible source of infection. Our group studied the presence of pathogens in houseflies (Musca domestica) in a New York State dairy in the United States. Among others, a Mycoplasma spp. was found in the gut of a housefly captured in the sick pen and identified as M. arginini. Here, we characterized its genome and investigated its relatedness with eight isolates from milk, one isolate from lung tissue collected in the same dairy, and five other dairies in New York State. We applied whole-genome sequencing and phylogenetic analysis based on the sequences of the 16S rRNA gene and 76 conserved proteins. We also assessed an in silico virulence profile by considering a panel of 94 putative virulence genes. As a result of the genome analysis, the housefly M. arginini isolate was highly similar to the milk isolates; interestingly, the similarity was highest with M. arginini isolated from milk on the same dairy farm where the housefly was captured. The housefly and milk M. arginini isolates possessed 54 of the 94 pathogenicity genes considered. Our data support the hypothesis that houseflies are carriers of Mycoplasma spp. and can be considered within the possible roots of environmental transmission of infection in dairy cows. Nevertheless, M. arginini pathogenicity will need to be investigated with dedicated studies. IMPORTANCE It is critical to control the spread of bovine mastitis caused by Mycoplasma spp., as this disease can be highly contagious and have a severe economic impact on affected dairies. A better understanding of possible transmission routes is crucial for infection control and prevention. Based on our data, the composite milk isolates are genetically similar to the housefly isolate. This provides evidence that the same Mycoplasma species found in milk and associated with mastitis can also be isolated from houseflies captured in the dairy environment.

Multivariable time series classification for clinical mastitis detection and prediction in automated milking systems.

In this study, we developed a machine learning framework to detect clinical mastitis (CM) at the current milking (i.e., the same milking) and predict CM at the next milking (i.e., one milking before CM occurrence) at the quarter level. Time series quarter-level milking data were extracted from an automated milking system (AMS). For both CM detection and prediction, the best classification performance was obtained from the decision tree-based ensemble models. Moreover, applying models on a data set containing data from the current milking and past 9 milkings before the current milking showed the best accuracy for detecting CM; modeling with a data set containing data from the current milking and past 7 milkings before the current milking yielded the best results for predicting CM. The models combined with oversampling methods resulted in specificity of 95 and 93% for CM detection and prediction, respectively, with the same sensitivity (82%) for both scenarios; when lowering specificity to 80 to 83%, undersampling techniques facilitated models to increase sensitivity to 95%. We propose a feasible machine learning framework to identify CM in a timely manner using imbalanced data from an AMS, which could provide useful information for farmers to manage the negative effects of CM.

A randomized controlled trial assessing the effect of intermittent and abrupt cessation of milking to end lactation on the well-being and intramammary infection risk of dairy cows.

The objectives were to compare the effects of an intermittent milking schedule with a thrice daily milking schedule during the final week of lactation on the well-being, udder health, milk production, and risk of culling of dairy cows. We hypothesized that cows subjected to an intermittent milking schedule would experience less udder engorgement and pain, lower concentrations of fecal glucocorticoid metabolites (11,17-dioxoandrostanes; 11,17-DOA concentration) after dry-off, lower risk of an intramammary infection during the dry period, higher milk production and lower somatic cell count in the subsequent lactation, and lower culling risk compared with herd mates milked 3 times daily and dried off by abrupt cessation. In a randomized controlled field study, Holstein cows (n = 398) with a thrice daily milking schedule were assigned to treatment and control groups. The treatment consisted of an intermittent milking schedule for 7 d before dry-off (gradual cessation of milking, GRAD). Gradual-cessation cows were milked once daily until the day of dry-off, whereas cows in the control group (abrupt cessation of milking, APT) were milked 3 times daily until the day of dry-off. Udder firmness and pain responses of the udder 3 d after dry-off, as well as the percentage change in fecal 11,17-DOA concentration (3 d after dry-off compared with the dry-off day), were used to assess the well-being of the animals. Compared with cows in the GRAD group, the odds [95% confidence interval (CI)] of udder firmness were 1.55 (0.99-2.42) for cows in the APT group, and the odds of a pain response were 1.48 (0.89-2.44) for cows in the APT group. The least squares means (95% CI) of the percentage change in 11,17-DOA concentration were 129.3% (111.1-150.4) for the APT group and 113.6% (97.5-132.4) for the GRAD group. Quarter-level culture results from the periods before dry-off and after calving were compared, to assess the likelihoods of microbiological cure and new infection. Cows in the APT group had lower odds of a new intramammary infection in the dry period [odds ratio, 95% CI: 0.63 (0.37-1.05)], whereas we observed no meaningful differences in the microbiological cure likelihood among groups. The least squares means (95% CI) for somatic cell counts (log10-transformed) were 4.9 (4.8-5.0) in the APT group and 4.9 (4.8-5.0) in the GRAD group. The odds (95% CI) of clinical mastitis in the first 30 d postcalving were 1.32 (0.53-3.30) in the APT group compared with the GRAD group. We observed no meaningful differences in milk production at the first test date postcalving or the culling risk among groups. We conclude that the gradual-cessation protocol tested herein failed to significantly improve animal well-being, udder health, milk production, and survival in the tested study cohort. However, the observed differences in udder firmness, as well as the numerical differences in udder pain and the percentage change in fecal 11,17-DOA concentrations suggest that this line of research may be useful. Future research is needed to develop drying-off strategies that are appropriate for lowering milk production at the end of the lactation and improve animal well-being without compromising udder health.

Comparison of 2 types of milk flow meters for detecting bimodality in dairy cows.

The primary objective of this observational study was to investigate whether incremental milk flow rates (in the 0-15 s, 15-30 s, 30-60 s, and 60-120 s intervals) from electronic on-farm milk flow meters can be used to detect bimodal milk flow curves in dairy cows compared with the use of a portable milk flow meter. Our second objective was to study the concordance between an electronic on-farm milk flow meter and a portable milk flow meter for assessing the 2-min milk yield and total milk yield. In this cross-sectional study, data from 92 milking observations from individual cows were analyzed. We collected data on incremental milk flow rates, the 2-min milk yield, and the total milk yield simultaneously with an on-farm milk flow meter and a portable milk flow meter. Bimodality detected by the on-farm milk flow meter was defined as lower milk flow rates during any of the 15-30 s, 30-60 s, and 60-120 s intervals compared with the previous intervals (0-15 s, 15-30 s, and 30-60 s). Bimodality according to the portable milk flow meter (BIMLC) was observed through automatic detection. κ statistics indicated good agreement between bimodality detected by the on-farm milk flow meter and BIMLC [κ (95% confidence interval): 0.69 (0.49-0.90)]. Using BIMLC as the gold standard, diagnostic test statistics for bimodality detected by the on-farm milk flow meter indicated moderate performance for sensitivity [0.73 (0.54-0.86)] as well as high performance for positive predictive value [0.83 (0.63-0.93)], specificity [0.94 (0.85-0.98)], and negative predictive value [0.90 (0.81-0.95)]. Receiver operating characteristic curve analyses revealed that the 30-60 s milk flow rate was the variable that best predicted BIMLC, yielding an area under the curve of 0.89. Pearson correlation coefficients (r) revealed a very strong correlation between the 2 devices for both the 2-min milk yield [0.97 (0.96-0.98)] and total milk yield [r (95% confidence interval), 0.97 (0.96-0.98)]. Additionally, intraclass correlation coefficients (ICC) and concordance correlation coefficients (CCC) indicated excellent agreement between the 2 devices for the 2-min milk yield [ICC, 0.97 (0.96-0.98); CCC, 0.94 (0.92-0.96)] and total milk yield [ICC, 0.97 (0.96-0.98); CCC, 0.97 (0.95-0.98)]. Therefore, we concluded that electronic on-farm milk flow meters that measure incremental milk flow rates can be used to detect bimodality in dairy cows and that on-farm milk flow meters facilitate precise measurements of the 2-min milk yield and total milk yield.

Prospective cohort study of the relationship between milking machine liner slip, milking performance, and cow characteristics.

We conducted a prospective cohort study to investigate the associations of machine milking liner slip with (1) milking performance and (2) cow characteristics. Parlor data including milk flow characteristics and data on the occurrence of milking machine liner slips from a 4,000-cow dairy with a thrice-daily milking schedule were obtained with electronic on-farm milk meters over a 2-mo period. We analyzed data from a total of 686,330 milking observations. A multivariable general linear mixed model revealed no association between liner slip and milking unit on time. Least squares means (95% confidence intervals, 95% CI) were 237 (235-238) s for milking observations with and without a liner slip. We observed statistically significant differences in average milk flow rate; however, these were biologically irrelevant. Least squares means were 3.40 (3.37-3.42) kg/min for a milking observation with and 3.42 (3.40-3.44) kg/min without a liner slip. A multivariable generalized linear mixed model showed an association between liner slip and cow characteristics. Compared with late-lactation cows, the odds ratios (OR, 95% CI) of occurrence of a liner slip were 2.03 (1.59-2.59) in early lactation cows and 1.26 (0.97-1.64) in cows from 101 to 200 days in milk. Presence of a nonlactating quarter increased the odds of liner slip occurrence [OR, 95% CI: 10.35 (8.02-13.35)]. Bimodal milking observations had higher odds of occurrence of a liner slip compared with milking observations with a unimodal milk flow curve [OR, 95% CI: 1.05 (1.005-1.09)]. A 1-kg increase in 2-min milk yield increased the odds of a liner slip [OR, 95% CI: 1.26 (1.25-1.28)]. We conclude that, in the study cohort presented herein, the negative effect of liner slips on milking performance can be diminished. The identified cow characteristics could offer unique opportunities to identify and manage cows at increased risk of liner slips.

Pathogens associated with houseflies from different areas within a New York State dairy.

Houseflies (Musca domestica) are nonbiting muscoids of importance because they can be mechanical vectors of many kinds of pathogens such as bacteria, protozoa, viruses, and helminth eggs. This study aimed to evaluate the bacterial communities associated with houseflies captured in 3 different areas on a dairy farm located in New York State. Variations in the bacterial community were also evaluated based on the flies' sex and external or internal location where the bacteria were isolated. A total of 101 flies were collected: 27 flies from the sick pen, 42 from calf hutches, and 32 from the milking parlor. A total of 485 organisms were isolated, 233 (48.0%) from 53 female flies and 252 (52.0%) from 48 male flies. Most (74%) bacteria were found in the internal parts of the flies, with only 26% isolated from the external surfaces. The number of isolates detected per fly ranged between 1 and 11. A total of 392 bacteria were identified at the species level. We isolated 26 species reported to be bovine contagious or environmental mastitis pathogens. Within the group of organisms considered contagious, we isolated Staphylococcus aureus and Mycoplasma arginini. This was the first time that a Mycoplasma species was isolated from houseflies. We identified 5 organisms considered foodborne pathogens that affect human health: Salmonella spp., Escherichia coli, Staph. aureus, Bacillus cereus, and Bacillus subtilis. Four of the organisms isolated in this study were also linked with milk spoilage, including Pseudomonas aeruginosa, Bacillus cereus, Bacillus licheniformis, and Paenibacillus lactis. This study confirmed that houseflies carry a high bacterial diversity, including organisms associated with animal infections, organisms that could be a concern for public health, or organisms that could negatively affect milk quality.

Development and evaluation of a standardized technique to assess teat skin temperature of dairy cows using infrared thermography.

Much research has been done to develop methods to assess dimensional teat traits in dairy cows. In contrast, diagnostic techniques to reliably assess the circulatory system of teats are limited. Infrared thermography facilitates measurements of skin temperature and could be used to detect physiological and pathological changes to the teat tissue associated with machine milking, as temperature reflects the underlying blood circulation and tissue metabolism. Our objective was to develop and evaluate a scanning technique to quantify teat skin temperature in dairy cows using infrared thermography. Using a portable thermography camera, 2 operators obtained duplicate scans of both hind teats from 20 Holstein cows, resulting in 80 thermographic images (20 cows × 2 operators × 2 images). Average teat skin temperatures at the proximal, middle, and distal teat aspects were determined. We used Pearson correlation coefficients (r), intraclass correlation coefficients (ICC), and concordance correlation coefficients (CCC) to assess interoperator reproducibility (i.e., agreement between measurements performed by different operators) and intraoperator repeatability (i.e., agreement between measurements performed by the same operator). Pearson correlation coefficients revealed a very strong correlation for measurements at the proximal, middle, and distal aspects of the teat, respectively, between operators (r ≥0.95) and duplicate scans (r ≥0.94) within operators. Intraclass correlation coefficients and CCC indicated excellent interoperator reproducibility (ICC ≥0.95, CCC ≥0.95) and excellent intraoperator repeatability (ICC ≥0.94, CCC ≥0.94), respectively, for measurements at all 3 aspects. Least squares means (95% confidence interval) for average teat skin temperatures at the proximal, middle, and distal teat aspects, respectively, were 33.2 (32.6-33.8), 32.4 (31.5-33.2), and 30.9 (29.8-32.0) °C for operator 1, and 33.2 (32.6-33.8), 32.4 (31.6-33.3), and 31.0 (29.9-32.0) °C for operator 2. Average temperatures between duplicate scans within operators at the proximal, middle, and distal aspects, respectively, were 33.3 (32.7-33.9), 32.5 (31.7-33.3), and 31.0 (29.9-32.1) °C for the first scan and 33.2 (32.6-33.8), 32.3 (31.5-33.1), and 30.8 (29.7-31.9) °C for the second scan. We conclude that infrared thermography facilitates precise measurements of skin temperatures of cows' hind teats.

Delta thermal radiomics: An application in dairy cow teats.

We describe a novel approach for analyzing thermal images by way of radiomics (i.e., thermal radiomics) and how it can be used to monitor short-term temperature changes of dairy cow hind teats; that is, delta thermal radiomics. The heat generated from metabolic activities and blood-flow patterns can be visualized using thermal radiography of the skin surface. The hind teats from 25 dairy cows were imaged with a digital thermal camera and the images were converted to medical images (DICOM format) by mapping the multi-channel colorized thermal image to a monochromatic image whose intensities represent temperature. The 50 teats (left and right hind) were then manually segmented by 2 investigators. Radiomics analysis, which is a common method of extracting semantic and nonsemantic image biomarkers from medical images for machine learning, was performed. To evaluate whether this approach can detect pre- and postmilking differences, 18 cows were imaged before and after milking, the teats were manually segmented, and radiomic calculations were performed. Student's t-test was used to provide an estimate of the likelihood of whether postmilking thermal image biomarkers are the same as premilking thermal image biomarkers, and Cohen's d was used to evaluate the size of the effect (d > 1.2). To evaluate uncertainties from manual segmentation, the Dice similarity score (DS) between the 2 investigators' segments was computed. The average DS (95% confidence limit) was 0.952 (0.913-0.982) when comparing the 2 investigators' segmentations. There was no significant difference in DS when comparing the left and right segmented teats, suggesting that teats can be segmented consistently. No differences (d < 0.36) were observed when comparing image biomarkers from one investigator's segments with the other's, suggesting that image biomarkers computed from one investigator's segmentation of teats are not likely to differ from those computed from the other investigator. When comparing image biomarkers before and after milking, 109 image biomarkers were analyzed, and 17 image biomarkers were simultaneously significant and exhibited effect size. Thus, delta thermal radiomics offers a noninvasive and quantitative method of monitoring skin temperature changes in humans and animals after an intervention. The advantage of this approach is that it can reveal both perceptible and imperceptible surface temperature features that may be useful for detecting and managing dairy teat health.

Risk factors for delayed milk ejection in Holstein dairy cows milked 3 times per day.

Delayed milk ejection, manifested most often as bimodal milk flow, occurs when the cisternal milk fraction is removed before the alveolar milk reaches the gland cistern. It is thought to be a consequence of not meeting cows' physiological needs, due to insufficient premilking teat stimulation, inadequate timing of milking unit attachment, or both. It has been associated with decreased milking efficiency, reduced milk yield, and impaired teat and udder health. Traditionally, portable electronic milk meters have been used to assess the presence of delayed milk ejection in dairy cows. By contrast, incremental milk flow rates from on-farm milk meters and their suitability as a measure to assess delayed milk ejection have not been studied by rigorous methods. The objectives were (1) to describe a protocol for identification of cows with chronically delayed milk ejection (CDME) and (2) to investigate risk factors for CDME using incremental milk flow rates obtained from automated on-farm milk meters. In a retrospective case control study, milk flow data from a 4,300-cow dairy with a thrice-daily milking schedule were obtained over a 1-wk period. Incremental milk flow rates (0-15 s, 15-30 s, 30-60 s, and 60-120 s) were used to identify cows with delayed milk ejection. Cases of CDME were defined as presence of delayed milk ejection at all 21 milking observations. Cows that had no delayed milk ejection at any of the same 21 milking observations were included as controls. A total of 171 cases and 393 controls were included in the study based on these criteria. A logistic regression model was used to evaluate associations of the following risk factors with CDME: parity (1, 2, ≥3), stage of lactation (<100, 101-200, >200 DIM), presence of a nonlactating quarter, milk somatic cell count, average daily milk production, and health and management events. Parity and CDME were associated such that compared with cows in their third or greater lactation, the odds (95% confidence intervals, 95% CI) of CDME were 1.27 (0.71-2.25) for cows in their first and 4.77 (2.47-9.22) for animals in their second lactation. The odds of CDME increased with increasing stage of lactation, with an odds ratio of 0.20 (0.11-0.36) for early and 0.28 (0.15-0.52) for mid-lactation animals, respectively, compared with late lactation cows. A 1-kg increase in average daily milk production was associated with decreased odds of CDME [odds ratio (95% CI): 0.89 (0.87-0.92)]. A lameness event during the study period increased the odds of CDME [odds ratio (95% CI): 8.04 (1.20-53.83)], as did a vaccination event 1 wk before the study period [odds ratio (95% CI): 4.07 (0.99-16.71)]. This study confirmed associations between CDME and previously reported risk factors and identified several previously less rigorously investigated health and management events that could be associated with CDME. Incremental milk flow rates from individual cows serve as an automated tool to evaluate milk flow dynamics. This information could be used to improve individual premilking udder preparation to meet the animal's physiological requirements, improve teat and udder health, and enhance parlor efficiency.

Short communication: Comparative performance of 3 on-farm culture systems for detection of mastitis pathogens interpreted by trained and untrained observers.

On-farm culture (OFC) systems facilitate pathogen-based mastitis management and can facilitate antimicrobial stewardship on dairy farms. Interpretation of the results, however, may present a challenge for those with limited microbiology experience. Here, we compared results of 3 OFC systems interpreted by trained and untrained observers against results of a standard laboratory reference method (aerobic culture and mass spectrometry). Milk samples (280 quarter and 60 composite) were selected from submissions for routine diagnostic testing to Quality Milk Production Services (Cornell University, Ithaca, NY) between August 2017 and January 2018. Samples were cultured simultaneously using the standard laboratory reference method and 3 commercially available OFC systems that varied in detail of pathogen identification (provided in parentheses) as follows: (1) Minnesota Easy Culture System II Bi-plate (University of Minnesota Laboratory for Udder Health, St. Paul; gram-positive, gram-negative), (2) Minnesota Easy Culture System II Tri-plate (gram-positive, gram-negative, some genus level), and (3) FERA Diagnostics and Biologicals AccuMast plate (Ithaca, NY; genus level, some species level). After 18 to 24 h of incubation, OFC plates were interpreted by 1 trained observer (>10 yr of experience in milk microbiology) and 6 untrained observers with no previous milk microbiology training, using only the manufacturers' instructions for guidance. Strength of agreement (κ) between observer groups and the reference method was determined for the available outcomes of each system. Interpreted by the trained observer, agreement was moderate for identifying gram-positive organisms (Bi-plate, κ = 0.56) and substantial for Streptococcus spp. (Tri-plate, κ = 0.64, AccuMast κ = 0.61). Interpretation by untrained observers resulted in fair agreement (κ = 0.29-0.37) for these organisms. Moderate agreement (κ = 0.43-0.59) was found across all 3 OFC for the identification of gram-negative organisms (Bi-plate), non-aureus staphylococci (Tri-plate and AccuMast), Lactococcus spp., and Enterococcus spp. (AccuMast) when interpreted by the trained observer, and fair to moderate agreement was found (κ = 0.31-0.53) among untrained observers. Across all 3 OFC, agreement was almost perfect (κ = 0.80-0.89) for Staphylococcus aureus for the trained observer, and moderate to substantial (κ = 0.56-0.61) for untrained observers. We concluded that all 3 OFC appeared suitable to support pathogen-based mastitis management when operated by trained observers. Training beyond the instruction manual is a prerequisite to make OFC systems useful for pathogen-based mastitis management.

Mycoplasma species isolated from bovine milk collected from US dairy herds between 2016 and 2019.

Determining the species of mycoplasma isolated from culture-positive milk samples is important for understanding the clinical significance of their detection. Between August 2016 and December 2019, 214,518 milk samples from 2,757 dairy herds were submitted to Quality Milk Production Services (QMPS) at Cornell University for mycoplasma culture. From these samples, 3,728 collected from 204 herds were culture positive. Based on the request of herd managers, owners, or veterinarians, 889 isolates from 98 herds were subjected to molecular identification by PCR and amplicon sequencing. The largest proportion of the identified isolates were from New York State (78.1%), while the others came from the eastern United States (17.8%), Texas (2.0%), and New Mexico (2.1%). As expected, Mycoplasma spp. were the most common (855 isolates, 96.2%) and Acholeplasma spp. accounted for the remainder (34 isolates, 3.8%). Mycoplasma bovis was the most prevalent Mycoplasma species (75.1%), followed by M. bovigenitalium (6.5%), M. canadense (5.9%), M. alkalescens (5%), M. arginini (1.7%), M. californicum (0.1%), and M. primatum (0.1%). A portion of the isolates were confirmed as Mycoplasma spp. other than M. bovis but were not identified at the species level (16 isolates, 1.8%) because further information was not requested by the manager, owner, or veterinarian. Mycoplasma bovis was the only species identified in 59 of the 98 herds. However, more than 1 Mycoplasma sp. was identified in 29 herds, suggesting that herd infection with 2 or more mycoplasmas is not uncommon. Moreover, a Mycoplasma sp. other than M. bovis was the only species identified in 8 herds. From the subset of 889 mycoplasma culture-positive isolates from 98 herds, we determined that over a third of the herds had either more than 1 Mycoplasma sp. or a Mycoplasma sp. other than M. bovis detected in their milk samples. In conclusion, we observed that M. bovis is the most common pathogenic Mycoplasma species found in mastitic milk, but other Mycoplasma species are not uncommon. Our results suggest that it is critical to test milk samples for mycoplasmas using diagnostic tests able to identify both the genus and the species.

Feasibility of the use of deep learning classification of teat-end condition in Holstein cattle.

Infections with pathogenic bacteria entering the mammary gland through the teat canal are the most common cause of mastitis in dairy cows; therefore, sustaining the integrity of the teat canal and its adjacent tissues is critical to resist infection. The ability to monitor teat tissue condition is a key prerequisite for udder health management in dairy cows. However, to date, routine assessment of teat condition is limited to cow-side visual inspection, making the evaluation a time-consuming and expensive process. Here, we demonstrate a digital teat-end condition assessment by way of deep learning. A total of 398 digital images from dairy cows' udders were collected on 2 commercial farms using a digital camera. The degree of teat-end hyperkeratosis was scored using a 4-point scale. A deep learning network from a transfer learning approach (GoogLeNet; Google Inc., Mountain View, CA) was developed to predict the teat-end condition from the digital images. Teat-end images were split into training (70%) and validation (15%) data sets to develop the network, and then evaluated on the remaining test (15%) data set. The areas under the receiver operator characteristic curves on the test data set for classification scores of normal, smooth, rough, and very rough were 0.778 (0.716-0.833), 0.542 (0.459-0.608), 0.863 (0.788-0.906), and 0.920 (0.803-0.986), respectively. We found that image-based teat-end scoring by way of deep learning is possible and, coupled with improvements in image acquisition and processing, this method can be used to assess teat-end condition in a systematic and efficient manner.

The effect of 2 different premilking stimulation regimens, with and without manual forestripping, on teat tissue condition and milking performance in Holstein dairy cows milked 3 times daily.

The objectives were to study the effect of 2 different premilking stimulation regimens, with and without manual forestripping, on teat tissue condition and milking characteristics in dairy cows. In a randomized controlled crossover study, 130 Holstein cows milked 3 times daily were assigned to treatment and control groups. Premilking udder preparation for the treatment group consisted of: (1) predipping with 1% iodine, (2) sequential forestripping of 3 streams of milk per quarter, (3) wiping of teats, and (4) attachment of the milking unit. Premilking udder preparation for the control group was identical except that the forestripping step was omitted. The mean tactile stimulation durations were 16 s and 7 s for the treatment and control group, respectively. The time spent from first tactile stimulus (either forestripping or wiping of teats) to milking unit attachment was kept consistent at 90 s for both groups. The study lasted for 14 d with 2 periods, each consisting of a 2-d adjustment time followed by 5 d of data collection. Machine milking-induced short-term changes to the teat tissue were assessed by palpation and visually. The following milking characteristics were assessed with electronic on-farm milk meters: milk yield (MY), milking unit-on time (MUOT), 2-min MY (2MIN), and time spent in low milk flow rate (LMF). Generalized linear mixed models were used to describe the effect of treatment on the outcome variables. The odds of machine milking-induced short-term changes to the teat tissue were lower for cows that received forestripping compared with cows that were not forestripped (odds ratio = 0.31; 95% confidence interval = 0.22-0.42). Least squares means (95% confidence interval) for cows that were forestripped and animals that were not forestripped, respectively, were 12.7 (12.2-13.2) and 12.7 (12.2-13.2) kg for MY and 6.1 (5.8-6.4) and 5.6 (5.3-5.9) kg for 2MIN. There was an interaction between treatment and MY for LMF. Time spent in LMF for cows that were forestripped and received no forestripping, respectively, were 18 (17-20) and 24 (23-26) s for a MY level of 10 kg; and 13 (12-14) and 15 (14-16) s for a MY level of 15 kg. The effect of treatment on MUOT was modified by parity. Milking unit-on times for animals in first, second and third or greater lactation, respectively, were 230 (219-243), 249 (236-262), and 260 (249-272) s for cows that were forestripped, and 245 (232-258), 252 (239-266), and 268 (257-281) s for cows that received no forestripping. In this study, cows that were forestripped had shorter MUOT, higher 2MIN, lower LMF, and lower odds of exhibiting changes to the teat tissue after machine milking. We conclude that wiping of teats during premilking udder preparation alone and omitting forestripping of teats without compensating for the loss in stimulation time may not provide sufficient tactile stimulation to elicit the cows' maximum physiological milk-ejection capacity. This can aggravate the adverse effects of vacuum-induced forces on teat tissue during machine milking, diminish animal well-being, and possibly affect udder health.

Technical note: A digital technique and platform for assessing dairy cow teat-end condition.

Because infections with pathogenic bacteria entering the mammary gland through the teat canal are the most common cause of mastitis in dairy cows, sustaining the integrity of the teat canal and its adjacent tissues is critical to resist infection. The ability to monitor teat tissue condition is therefore a key prerequisite for udder health management in dairy cows. However, to date, routine assessment of teat-end condition is limited to cow-side visual inspection, making the evaluation a time-consuming and expensive process. Here, we illustrate and demonstrate a method for assessing teat-end condition of dairy cows through digital images and software. A digital workflow has been designed where images of dairy cow teats are obtained and processed to display individual teats, and the cow and teat images are labeled and displayed through a graphical user interface. The interface then allows an evaluator to assess quarter- and cow-level teat-end condition and store the results for review and future analysis. The digital workflow permits several advantages such as the ability to perform remote teat-end condition assessments, and assess inter- and intrarater variability of teat-end condition scoring. We demonstrate the image-based teat-end condition assessment of 194 dairy cows that also had cow-side teat-end condition assessments by 2 expert evaluators. Weighted Cohen's kappa statistic (κ) was computed to measure the evaluators' concordance of categorical scores of quarter- and cow-level assessments when using cow-side and image-based assessments. Substantial agreement (0.61 ≤ κ ≤ 0.80) was observed between an evaluator's cow-side and image-based assessments at the quarter and cow level. Moderate agreement (0.41 ≤ κ ≤ 0.60) was observed between evaluators when using image-based assessments at the quarter and cow level. Near perfect agreement (κ = 0.89, 95% confidence interval 0.78-1.00) was observed between evaluators when using cow-side assessments at the quarter level, and substantial agreement (κ = 0.66, 95% confidence interval 0.53-0.79) was observed when using cow-side assessments at the cow level. This suggests that image-based teat-end condition classification is possible, and coupled with improvements in image acquisition and image processing, this method can be used to assess teat-end condition in a systematic and convenient manner.

Blood perfusion of teat tissue in dairy cows: Changes associated with pre-milking stimulation and machine milking.

Mechanical forces during machine milking of dairy cows evoke circulatory impairment of the teat tissue that may affect the teats' defense mechanisms against mastitis pathogens. Ample research describes dimensional changes of different teat traits after machine milking, whereas reports that describe changes in blood circulation of dairy cows' teats are limited. Therefore, the objectives of this study were to (1) describe changes in teat blood circulation that occur after pre-milking teat stimulation and machine milking and (2) study the effect of 2 different milking liners on machine milking-induced changes in teat blood flow. In a randomized trial, Holstein dairy cows were stratified by parity, stage of lactation, and average daily milk yield during the previous week, and allocated to 1 of 2 treatment groups. Treatment consisted of 1 milking observation with either a round or multisided concave milking liner. Teat scans were taken of the left front and the right hind teats using power Doppler ultrasonography. Imaging occurred before pre-milking udder preparation (T1), after completion of pre-milking udder preparation but before milking-unit attachment (T2), and immediately after unit detachment (T3). Perfusion intensity measurements from teat scans were performed with a commercially available software program. Data from 109 cows were analyzed. A general linear mixed model showed differences in perfusion intensity between time points. Least squares means (95% confidence intervals) for T1, T2, and T3, respectively, were 0.035% (0.026-0.047), 0.124% (0.093-0.164), and 0.095% (0.073-0.124). Conversely, no statistically significant differences between treatment groups were observed. We conclude that teat blood circulation is subjected to several influences, including inherent circulatory regulation mechanisms, as well as extrinsic factors such as machine milking. Future research is warranted to decipher the magnitude of their influence and to further our understanding of how these changes relate to the susceptibility to intramammary infection and milking performance.