Frequency of non-aureus staphylococci and mammaliicocci species isolated from quarter clinical mastitis: a six-year retrospective study.

Non-aureus staphylococci and mammaliicocci (NASM) are the most frequently isolated bacterial group from bovine milk samples. Most studies focus on subclinical mastitis caused by NASM, however NASM can cause clinical mastitis (CM) as well. We evaluated retrospective data from 6 years (2017-2022) to determine the species and frequency of NASM isolated from quarter bovine CM. The data comprised of microbiological results from quarter CM samples routinely submitted to Quality Milk Production Services (QMPS), Cornell University, NY, US, for microbial identification by MALDI-TOF MS. A total of 9,909 microbiological results from 410 dairy herds were evaluated. Our results showed that 29 distinct NASM species were identified, with the 8 most prevalent NASM species being Staphylococcus chromogenes, S. haemolyticus, S. simulans, S. epidermidis, S. sciuri (now Mammaliicoccus sciuri), S. agnetis/S. hyicus, S. borealis, and S. xylosus. The NASM distribution remained similar among seasons, but the frequency of NASM CM cases was higher during the summer. Our results showed different patterns of variations in the isolation frequency over time, depending on the bacterial species: increasing or decreasing trends, cyclic fluctuations, and except for S. borealis, a significant seasonality effect for our study's most prevalent NASM was observed. This study showed that S. chromogenes remains the most frequent (43%) NASM species identified from bovine CM, followed by S. haemolyticus (18%), and S. simulans (12%).

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.

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.

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.

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.

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.

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.

A randomized trial to study the effect of automatic cluster remover settings on milking performance, teat condition, and udder health.

The objectives were to study the effect of 2 different automatic cluster remover settings on (1) milking characteristics, (2) milk component yields, (3) teat tissue condition, and (4) udder health. In a randomized controlled field trial, Holstein cows (n = 689) from 1 commercial dairy farm with a thrice-daily milking schedule were allocated to 2 treatment groups. Treatment consisted of a cluster remover take-off milk flow threshold of 1.2 (ACR1.2) or 0.8 kg/min (ACR0.8) for 57 d. Milking characteristics (milk yield; and milking unit-on time, MUOT) were obtained with electronic on-farm milk meters. Composite milk samples were collected and analyzed for fat, protein, lactose, and somatic cell count. Machine-milking-induced short- and long-term changes to the teat tissue condition were assessed visually. General linear mixed models demonstrated differences in MUOT, whereas no meaningful differences in milk yield were detected. Milk yield (least squares means, 95% confidence interval) was 11.3 (10.9-11.8) and 11.3 (10.8-11.8) kg in groups ACR1.2 and ACR0.8, respectively. The effect of treatment on MUOT was modified by parity. Milking unit-on time in first-, second-, and ≥third-lactation cows, respectively, was 260.7 (252.0-269.4), 257.8 (247.4-268.1), and 260.2 (252.6-267.9) s in group ACR1.2; and 273.7 (264.9-282.5), 279.1 (269.4-288.8), and 295.7 (287.9-303.6) s in group ACR0.8. We detected no meaningful differences in milk component yields or linear somatic cell score. Least squares means in groups ACR1.2 and ACR0.8, respectively, were milk fat yield, 0.42 (0.40-0.44) and 0.42 (0.40-0.44) kg; milk protein yield, 0.36 (0.35-0.37) and 0.37 (0.36-0.37) kg; milk lactose yield, 0.61 (0.60-0.63) and 0.63 (0.61-0.64) kg, and linear somatic cell score, 1.9 (1.8-2.0) and 1.9 (1.8-2.0). A generalized linear mixed model revealed an effect of treatment on machine-milking-induced short-term changes. The odds of short-term changes to the teat tissue were lower for cows in group ACR1.2 [odds ratio (95% confidence interval) = 0.78 (0.63-0.96)]. No meaningful differences were detected in machine-milking-induced long-term changes between treatment groups. Increasing cluster remover take-off milk flow threshold from 0.8 to 1.2 kg/min decreased individual milking duration and alleviated machine-milking-induced short-term changes to the teat tissue without adversely affecting milking performance or somatic cell count. Future studies are warranted to investigate the effect on milk production and udder health over a whole lactation period.

Technical note: Development and evaluation of a standardized technique to assess blood perfusion in teats of dairy cows using power Doppler ultrasonography.

Ample research has described the assessment of dimensional changes for different teat traits, whereas diagnostic techniques to reliably assess blood circulation in teats of dairy cows are limited. Here, we describe the development and evaluation of a scanning technique to quantify blood flow in teats of dairy cows using power Doppler ultrasonography. In 2 consecutive trials, 384 teat scans [trial 1, n = 256 (sagittal plane, n = 128; transverse plane, n = 128); trial 2, n = 128 (transverse plane)] from 16 cows were obtained by the same 2 operators. Perfusion intensity from single images (trial 1) and video images (trial 2) were assessed using a commercially available software program. Intraclass correlation coefficients (ICC) and concordance correlation coefficients (CCC) were used to assess interoperator reproducibility (agreement between measurements performed by different operators) and intraoperator repeatability (agreement between measurements performed by the same operator). In trial 1, interoperator ICC and CCC indicated poor agreement (ICC ≤0.26, CCC ≤0.26). Intraoperator ICC and CCC demonstrated poor agreement between duplicate measurements within operators (ICC ≤0.19, CCC ≤0.19). Modifications after trial 1 included (1) a different ultrasound device, (2) analysis of video clips rather than single images, (3) restriction to 1 sectional plane (i.e., transverse), and (4) a scanning sequence such that repeated scans within operators were measured one after another. Through these modifications, intraoperator repeatability in trial 2 yielded fair to good agreement, with intraoperator ICC and CCC over both operators ranging from 0.44 to 0.70 and from 0.57 to 0.69, respectively, whereas interoperator ICC and CCC showed poor agreement (ICC = 0.35, CCC = 0.34). We conclude that repeatable measurements of blood perfusion intensity of teats in dairy cows can be attained with power Doppler ultrasonography. Power Doppler ultrasonography is a suitable tool to quantify slow flow in small vessels and may be an acceptable diagnostic technique to assess changes in blood circulation that result from machine milking in teats of dairy cows, although further research is necessary to validate this hypothesis.

Machine milking-induced changes in teat canal dimensions as assessed by ultrasonography.

Changes in teat canal diameter as induced by machine milking are thought to be related to teat canal penetrability and susceptibility to new intramammary infections. The objective of this study was to describe the effects of machine milking on teat canal dimensions as assessed by ultrasonography and to evaluate postmilking changes of teat canal dimensions throughout an 8-h milking interval. Ultrasonographic images were taken of the left front and right hind teats of 80 Holstein cows milked 3 times per day. Imaging occurred before attachment of the milking unit after completion of premilking udder preparation (t-1), immediately after removal of the milking unit (t0), and then every hour after milking until 8 h had elapsed (t1 to t8). Teat canal length and teat canal diameter (at the proximal, middle, and distal regions) were measured. General linear mixed model analysis indicated differences in the relative change of teat canal length (compared with t-1) with least squares means (LSM) and 95% confidence interval (95% CI) of 11.1% (8.7-13.4%) after machine milking (t0). Teat canal length decreased throughout the 8-h milking interval but remained elevated relative to t-1. Teat canal diameter at the proximal and middle regions increased near the end of the milking interval, whereas that at the distal region did not change meaningfully during the 8 h of observation. Our findings suggest that the teat canal and its surrounding tissue is in a state of near-constant remodeling in cows milked 3 times daily. Future work is warranted to study the association between ultrasonographically assessed changes of teat canal dimensions after machine milking and teat canal penetrability to mastitis-causing pathogens.

An observational study investigating the association of ultrasonographically assessed machine milking-induced changes in teat condition and teat-end shape in dairy cows.

Mechanical forces during machine milking induce changes in teat condition which can be differentiated into short-term and long-term changes. Machine milking-induced short-term changes in teat condition (STC) are defined as tissue responses to a single milking and have been associated with the risk of new intramammary infection. Albeit, their association with teat characteristics, such as teat-end shape, has not been investigated by rigorous methods. The primary objective was to determine the association of STC, as measured by ultrasonography, with teat-end shape. The second objective was to describe possible differences in the recovery time of teat tissue after machine milking among teats with different teat-end shapes. Holstein cows (n=128) were enrolled in an observational study, housed in free-stall pens with sand bedding and milked three times a day. Ultrasonography of the left front and right hind teat was performed after teat preparation before milking (t-1), immediately after milking (t 0) and 1, 3, 5 and 7 h after milking (t 1, t 3, t 5, t 7). The teat tissue parameters measured from ultrasound scans were teat canal length, teat-end diameter, teat-end diameter at the midpoint between the distal and proximal end of the teat canal, teat wall thickness, and teat cistern width. Teat-end shape was assessed visually and classified into three categories: pointed, flat and round. Multivariable linear regression analyses showed differences in the relative change of teat tissue parameters (compared with t-1) at t 0 among teats with different teat-end shapes, with most parameters showing the largest change for round teats. The premilking values were reached (recovery time) after 7 h in teats with a pointed teat-end shape, whereas recovery time was greater than 7 h in teats with flat and round teat-end shapes. Under the same liner and milking machine conditions, teats with a round teat-end shape had the most severe short-term changes. The results of this observational study indicated that teat-end shape may be one of the factors that contribute to the severity of STC.

The microbiome of Escherichia coli and culture-negative nonsevere clinical mastitis: Characterization and associations with linear score and milk production.

Culture-negative and Escherichia coli cases are uncommonly treated in pathogen-based protocols for nonsevere mastitis. High-throughput 16S rRNA sequencing might reveal the presence of other pathogens and can provide information on microbial diversity. The objective was to explore the milk microbiome at the time of the mastitis event (enrollment) and its association with survival in the herd, milk production, and postevent linear score (LS) for cows with clinical mastitis characterized as negative or E. coli by culture. Fifty E. coli-positive and 35 culture-negative samples from cases were enrolled. No cases were treated with antimicrobials. All E. coli-positive quarters were characterized as transient; microbiological culture of samples taken 15 d postmastitis were negative for this organism. However, a difference in α-diversity (Shannon index) was present between enrollment and follow-up samples (3.8 vs. 5.1). When α-diversity was explored for enrollment E. coli samples, no relationship was observed between the Shannon indices of these samples and postmastitis LS. Alpha-diversity of the enrollment samples was lower for E. coli-positive cows that subsequently had greater losses in milk production. This difference was explained by a greater relative abundance of the family Enterobacteriaceae (67.8 vs. 38.4%) for cows that dropped in production. Analysis of composition of the microbiome identified one phylum, Proteobacteria, that differed between E. coli-positive cows that dropped in production and those that did not. Evaluation of ß -diversity found no statistical relationship between postmastitis LS and the microbiome. When evaluating α- and ß-diversities and composition of the microbiomes for culture-negative quarters, no associations were found for milk production changes and postmastitis LS. Three cows did not remain in the herd, limiting the ability to analyze survival. The findings suggest that a contributing factor to negative outcomes in E. coli-positive cows is relative abundance of this pathogen, and that no single or collective group of bacterial families is associated with milk production changes or postmastitis LS in culture-negative quarters. Although additional studies should be performed, the absence of associations between outcomes explored and microbial profiles in this study suggests that we are not missing opportunities by not treating nonsevere E. coli or culture-negative mastitis cases.

Short communication: Teat-end shape and udder-level milking characteristics and their associations with machine milking-induced changes in teat tissue condition.

Machine milking-induced changes in teat tissue condition, such as congestion and edema, have been associated with teat canal openness, penetrability, and thus susceptibility to new intramammary infections and diminished animal well-being. The primary objective of this study was to investigate the association of teat-end shape and machine milking-induced short-term changes in teat tissue condition. Our secondary objective was to study the association of udder-level milking characteristics and short-term changes. Data from 125 Holstein cows were analyzed in a longitudinal prospective cohort study. Cows were housed in freestall pens with sand bedding, fed a total mixed ration, and milked 3 times per day. Teat-end shape was classified into 3 categories: pointed, flat, and round. Udder-level milking characteristics were obtained from electronic on-farm milk meters. Short-term changes in teat tissue condition after machine milking were assessed visually. Multivariable generalized mixed model analysis showed an association between teat-end shape and short-term changes. Compared with teats with round teat-end shape, the odds ratio (95% confidence interval) for short-term changes was 1.68 (0.53-5.31) and 0.03 (0.004-0.19) in teats with pointed and flat teat-end shape, respectively. There was an association between milking characteristics and short-term changes such that higher milk flow rate during the first 15 s of milking decreased the likelihood of short-term changes. The adjusted probability of short-term changes for a milking observation of a mid-lactation cow and an average first 15-s milk flow rate of 0.5 and 1.5 kg/min was 53.0% (42.8-63.8) and 32.9% (15.2-57.3), respectively. Our results suggest that teat-end shape may be one of the risk factors that contribute to machine milking-induced short-term changes. Milking characteristics (e.g., first 15-s milk flow rate) may have the potential as a measure to indirectly monitor teat tissue changes associated with machine milking on a daily basis, though further research is needed to validate this hypothesis and to establish thresholds that could serve as on-farm guidelines.

Use of a culture-independent on-farm algorithm to guide the use of selective dry-cow antibiotic therapy.

An algorithm using only computer-based records to guide selective dry-cow therapy was evaluated at a New York State dairy farm via a randomized field trial. DairyComp 305 (Valley Ag Software, Tulare, CA) and Dairy Herd Improvement Association test-day data were used to identify cows as low risk (cows that might not benefit from dry-cow antibiotics) or high risk (cows that will likely benefit). Low-risk cows were those that had all of the following: somatic cell count (SCC) ≤200,000 cells/mL at last test, an average SCC ≤200,000 cells/mL over the last 3 tests, no signs of clinical mastitis at dry-off, and no more than 1 clinical mastitis event in the current lactation. Low-risk cows were randomly assigned to receive intramammary antibiotics and external teat sealant (ABXTS) or external teat sealant only (TS) at dry-off. Using pre-dry-off and postcalving quarter-level culture results, low-risk quarters were assessed for microbiological cure risk and new infection risk. Groups were also assessed for differences in first-test milk yield and linear scores, individual milk weights for the first 30 d, and culling and mastitis events before 30 d in milk. A total of 304 cows and 1,040 quarters in the ABXTS group and 307 cows and 1,058 quarters in the TS group were enrolled. Among cows to be dried, the proportion of cows that met low-risk criteria was 64% (n = 611/953). Of cultures eligible for bacteriological cure analysis (n = 171), 93% of ABXTS cured, whereas 88% of TS cured. Of the non-cures, 95% were contributed by the minor pathogens coagulase-negative staphylococci (n = 19/20). These organisms also accounted for 57.5% of new infections (n = 77/134). We found no statistical differences between treatment groups for new infection risk (TS = 7.3% quarters experiencing new infections; ABXTS = 5.5%), milk production (ABXTS = 40.5 kg; TS = 41.2 kg), linear scores (ABXTS = 2.5; TS = 2.7), culling events (ABXTS, n = 18; TS, n = 15), or clinical mastitis events (ABXTS, n = 9; TS, n = 5). Results suggest that the algorithm used decreased dry-cow antibiotic use by approximately 60% without adversely affecting production or health outcomes.

Technical note: Development and evaluation of a standard operating procedure for ultrasound-based measurements of teat canal dimensions in dairy cows.

The objectives were to (1) develop a standard operating procedure (SOP) for ultrasound-based measurements of teat canal dimensions and (2) determine the precision of ultrasound-based measurements of teat canal dimensions in dairy cows. Teat scans (n = 64) of the right hind and left front teats from 16 cows were obtained with a portable ultrasound device before and after machine milking. Teat dimensions measured were teat canal length (TCL), teat canal diameter at the proximal end of the teat canal (TCDPROX), teat canal diameter at the midpoint between the proximal and distal ends of the teat canal (TCDMID), teat canal diameter at the distal end of the teat canal (TCDDIS), teat end diameter at the midpoint between the proximal and distal ends of the teat canal (TMD), and teat canal cross-sectional area (TCLAREA). An SOP was developed, reviewed, and modified by 3 operators. Measurements were performed by the same 3 operators using an open source software program. To assess the effect of implemented modifications and the precision of ultrasound-based measurements, concordance correlation coefficients (CCC) were calculated to determine interoperator reproducibility and intraoperator repeatability. Through modifications in the SOP, interoperator CCC increased from 0.45 to 0.86 for TCL, from 0.14 to 0.66 for TCDPROX, from 0.24 to 0.66 for TCDMID, from 0.06 to 0.56 for TCDDIS, from 0.64 to 0.91 for TMD, and from 0.17 to 0.64 for TCLAREA. Intraoperator CCC over all operators were 0.91 for TCL, 0.73 for TCDPROX, 0.80 for TCDMID, 0.69 for TCDDIS, 0.94 for TMD, and 0.75 for TCLAREA. Our results indicate that measurements of teat canal dimensions can be conducted with satisfactory precision when following an SOP. Development and strict implementation of an SOP has the potential to decrease inadvertent variability of ultrasound-based measurements among operators for ultrasonographic assessment of teat dimensions.

Clinical outcome comparison of immediate blanket treatment versus a delayed pathogen-based treatment protocol for clinical mastitis in a New York dairy herd.

The purpose was to compare immediate intramammary antimicrobial treatment of all cases of clinical mastitis with a selective treatment protocol based on 24-h culture results. The study was conducted at a 3,500-cow commercial farm in New York. Using a randomized design, mild to moderate clinical mastitis cases were assigned to either the blanket therapy or pathogen-based therapy group. Cows in the blanket therapy group received immediate on-label intramammary treatment with ceftiofur hydrochloride for 5 d. Upon receipt of 24 h culture results, cows in the pathogen-based group followed a protocol automatically assigned via Dairy Comp 305 (Valley Agricultural Software, Tulare, CA): Staphylococcus spp., Streptococcus spp., or Enterococcus spp. were administered on-label intramammary treatment with cephapirin sodium for 1 d. Others, including cows with no-growth or gram-negative results, received no treatment. A total of 725 cases of clinical mastitis were observed; 114 cows were not enrolled due to severity. An additional 122 cases did not meet inclusion criteria. Distribution of treatments for the 489 qualifying events was equal between groups (pathogen-based, n = 246; blanket, n = 243). The proportions of cases assigned to the blanket and pathogen-based groups that received intramammary therapy were 100 and 32%, respectively. No significant differences existed between blanket therapy and pathogen-based therapy in days to clinical cure; means were 4.8 and 4.5 d, respectively. The difference in post-event milk production between groups was not statistically significant (blanket therapy = 34.7 kg; pathogen-based = 35.4 kg). No differences were observed in test-day linear scores between groups; least squares means of linear scores was 4.3 for pathogen-based cows and 4.2 for blanket therapy cows. Odds of survival 30 d postenrollment was similar between groups (odds ratio of pathogen-based = 1.6; 95% confidence interval: 0.7-3.7) as was odds of survival to 60 d (odds ratio = 1.4; 95% confidence interval: 0.7-2.6). The one significant difference found for the effect of treatment was in hospital days; pathogen-based cows experienced, on average, 3 fewer days than blanket therapy cows. A majority (68.5%) of moderate and mild clinical cases would not have been treated if all cows on this trial were enrolled in a pathogen-based protocol. The use of a strategic treatment protocol based on 24-h postmastitis pathogen results has potential to efficiently reduce antimicrobial use.

An examination of hyperconjugative and electrostatic effects in the hydride reductions of 2-substituted-4-tert-butylcyclohexanones.

To better understand electronic effects on the diastereoselectivity of nucleophilic additions to the carbonyl group, a series of 2-X-4-tert-butylcyclohexanones (X = H, CH(3), OCH(3), F, Cl, Br) were reacted with LiAlH(4). Reduction of ketones with equatorial substituents yields increasing amounts of axial alcohol in the series for X [H < CH(3) < Br < Cl < F << OCH(3)]. These data cannot be explained by steric or chelation effects or by the theories of Felkin-Anh or Cieplak. Instead, an electrostatic argument is introduced: due to repulsion between the nucleophile and the X group, axial approach becomes energetically less favorable with an increase in the component of the dipole moment anti to the hydride approach trajectory. The ab initio calculated diastereoselectivities were close to the experimental values but did not reproduce the relative selectivity ordering among substituents. For reduction of ketones with axial substituents, increasing amounts of axial alcohol are seen in the series for X [Cl < Br < CH(3) < OCH(3) < H < F]. After some minor adjustments are made, this ordering is consistent with both the electrostatic model and Felkin-Anh theory. Cieplak theory cannot account for these data regardless of adjustments. Ab initio calculated diastereoselectivities were reasonably accurate for the nonpolar substituents but were poor for the polar substituents.