Source: UNIV OF WISCONSIN submitted to NRP
EFFECTS OF INCREASING RESIDUAL MILK AND MILKING FREQUENCY ON LACTATION PHYSIOLOGY, MILK YIELD, MILKING TIE, AND TEAT TISSUE STRESS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1014216
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Oct 1, 2017
Project End Date
Aug 31, 2019
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
UNIV OF WISCONSIN
21 N PARK ST STE 6401
MADISON,WI 53715-1218
Performing Department
DAIRY SCIENCE
Non Technical Summary
Current teatcup removal strategies applied to both conventional and automatic milking installations have been derived from studies in the 1990's performed in conventional parlors. AMS installations have the ability to determine milk flow at the quarter level and can detach during milking also at the quarter level. The economic return on an AMS installation is partly driven by the number of cows milked per hour. Given that the capital cost of a single AMS unit is high, any strategies that improve the number of cows that can be milked through a single unit without compromising milk yield can be highly useful in facilitating the use of this technology. Additionally, the allowable milking frequency applied to cows in an AMS installation is derived from milking frequency studies conducted in conventional farms and parlors. Further work is required to clarify the optimal milking frequency settings for AMS parlors.The experimental design for this proposal affords the opportunity to further explore the physiological mechanisms responsible for reduced milk yield where cisternal milk left behind at the end of milking is increased resulting in an undermilked cow and the potential interaction with milking frequency. Previous experimental work conducted in this area has only focused on reducing milking interval and not via increased cisternal milk.The quarter milker device proposed for this research will mimic the role of an AMS installation so that research findings can then be applied to commercial AMS devices. Furthermore, milk harvesting strategies resulting in higher residual milk yields applied in conventional parlors could be derived from this research.
Animal Health Component
50%
Research Effort Categories
Basic
50%
Applied
50%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
30534101020100%
Knowledge Area
305 - Animal Physiological Processes;

Subject Of Investigation
3410 - Dairy cattle, live animal;

Field Of Science
1020 - Physiology;
Goals / Objectives
The overall objective of this research is to understand the influence of residual milk/udder fill, milking frequency and their potential interactions on milk yield and teat health. The outcomes of this research will be improved understanding of the physiological mechanisms controlling milk yield which will result in new strategies for managing milking.Specific Aim 1. Determine the effect of udder fill on milking time and milk yield.Specific Aim 2. Determine the main and interactive effects of udder fill and milking frequency on milking time, and milk yield.Specific Aim 3. Determine the underlying physiological and cellular mechanisms that account for milk yield suppression in response to changes in milking frequency and degree of udder evacuation.Specific Aim 4. Determine changes in teat end hyperkeratosis, teat barrel congestion, teat end congestion, and teat canal keratin levels, associated with different milking management strategies.
Project Methods
This project is a continuation of Hatch project 1006518-Experiment 1: Twelve healthy, multiparous cows will be enrolled in this experiment, which will be conducted at the UW Dairy Cattle Center. Assignment of the treatment and control half-udders shall be on an alternating basis as cows are enrolled. This will result in an N=12 treatment (20% of residual milk left behind) and N=12 control (0% residual milk left behind) half-udders. Any quarter treatments for mastitis shall remove the cow and all quarters from the study as will treatments for systemic illness. The take-off/residual milk settings for the treatment quarters shall be set based on previous individual milk yield information (specific aim 1). Cows will be enrolled on day 5 in milk after calving and will remain on study until day 47 in milk (resulting in a 42 d experiment) and milked 2x daily for the duration of the experiment. Hence, it will take about one month to enroll the required cows based on typical monthly calving records for the UW herd. The take-off settings/residual milk for each treatment half-udder shall then be adjusted on a weekly basis to take into account changes in overall milk production as days in milk increases. It is anticipated that the treatment half-udder will result in a 10% decrease in daily milk yield relative to the control half throughout the 42-day experimental period for each enrolled cow. Throughout the trial period enrolled cows shall be milked with a quarter milking device and half-udder milk yield will be recorded.Experiment 2: Individual cow enrollment into Experiment 2 will be as described for Experiment 1 (day 5 in milk through day 47 in milk, for a total of 42 days) although the number of cows enrolled will be higher than for Experiment 1. Experiment 2 will commence after the completion of Experiment 1. The protocol devised in Experiment 1 for residual milk settings will determine residual milk settings for treatment half udders. Again, throughout the trial period enrolled cows shall be milked with a quarter milking device. Healthy, multiparous cows (balanced for parity) will be enrolled and assigned randomly to a 2X milking group or a 3X milking group with equal numbers recruited into each group. Treatment and control half udders shall be assigned to each cow irrespective of milking frequency. The study design is a 2x2 factorial design (specific aim 2). Cows will be enrolled on day 5 in milk through day 47 in milk (a total of 42 days on experiment). Twenty-two multiparous cows (balanced for parity) will be enrolled on the study, and randomly assigned to 2x daily milking (N=11) or 3x daily milking (N=11). Additionally, half-udders on each cow will be assigned to a 20% increase in residual milk (treatment; N=22) or will be completely evacuated of milk (control; N=22). Half-udder milk yield will be recorded daily as described in experiment 1.Experiment 3: The purpose of this experiment is to examine the physiological processes underlying increasing residual milk, as well as the interaction between increasing residual milk and milking frequency. To this end in experiment 1, quarter milk samples will be taken weekly at the first milking of the specified days and will be analyzed for overall milk composition (fat, lactose, protein; AgSource) and somatic cell count (Milk Quality Laboratory). Additionally, we will measure milk sodium and potassium content (by Atomic Absorption) to assess alveolar cellpermeability in response to alterations associated with undermilking (14). An increase in sodium/potassium ratio in the milk isan indicator of mammary epithelial cell permeability (14). Blood samples (coccygeal vein) will be taken weekly immediately aftermilk sampling. Blood samples will be assessed for growth hormone (radioimmunoassay) and prolactin (radioimmunoassay) inorder to assess the endocrine status of the cow at the time of milking to ensure that animals are responding to treatmentsimplemented similarly.In order to determine the effects of residual milk and the interactions with milking frequency, we will collect samples duringexperiment 2. On days 0, 7, 14, 21, 28, 35 and 42 after enrollment, blood samples will be collected from the coccygeal veinand milk samples will be collected from each half-udder during the first milking of the specified day. Blood samples will beanalyzed for prolactin (RIA), growth hormone (RIA) and serotonin (ELISA). Milk samples will be analyzed for overall milkcomposition (fat, lactose, protein; AgSource), somatic cell count (Milk Quality Laboratory) as well as sodium and potassiumcontent (Atomic Absorption) to assess alveolar cell permeability in response to alterations associated with undermilking (14).Six cows from each of the 2X and 3X groups will biopsied (half-udder) on day 0 and day 42, resulting in a N=6 per treatmentdescribed in Figure 1. Mammary tissue for histological staining will be placed in 4% paraformaldehyle for 24 hr at 4°C and thenmoved to 70% ethanol until taken to pathology at RARC. Paraffin embedded tissues will be sectioned and stained withhematolxylin and eosin to assess alveolar structure, integrity, size and number in response to undermilking. Mammary tissuewill also be frozen immediately in liquid nitrogen for the purpose of mRNA extraction (TriReagent) and converted to cDNA usingiScript (Bio-Rad). cDNA samples will be sent to the UW Biotechnology center for Microarray Analysis, and data analysis will beconducted at the UW Biotechnology center. Genes that result in significant changes from the microarray analysis will then beanalyzed using real-time quantitative PCR (BioRad CFX 96) for confirmation of results. Mammary tissue will also be analyzedfor total serotonin content (Beckman Coulter; ELISA). Additionally quarter milk samples will be taken weekly at the first milkingof the specified days (days 0, 7, 14, 21, 28, 35 and 42) and will be analyzed for overall milk composition (fat, lactose, protein;AgSource) and somatic cell count (Milk Quality Laboratory). Additionally, we will measure milk sodium and potassium content(by Atomic Absorption) to assess alveolar cell permeability in response to alterations associated with undermilking (14). Anincrease in sodium/potassium ratio in the milk is an indicator of mammary epithelial cell permeability (14). Blood samples(coccygeal vein) will be taken weekly immediately after milk sampling. Blood samples will be assessed for growth hormone(radioimmunoassay) and prolactin (radioimmunoassay) in order to assess the endocrine status of the cow at the time of milkingto ensure that animals are responding to treatments implemented similarly.Experiment 4: The purpose of this experiment is to examine the effects of different milking strategies on teat health. To this end during experiment 1 and experiment 2, on days 0, 7, 14, 21, 28, 35 and 42, cows will undergo teat assessment (on all 4 teats) to determine teat hyperkeratosis, color and barrel scores according to the system as described by Neijenhuis et al (22). In addition to this scoring, teat canal keratin will be harvested utilizing the system described by Bright et al (21) to estimate keratin turnover and production in treatment and control quarter teat ends. These observations will then be analyzed to determine how treatment conditions might affect teat tissue stress during the milking process.

Progress 10/01/17 to 08/31/19

Outputs
Target Audience:The data from this project was presented at the American Dairy Science Association Meetings. This target audience consists of scientists focused on dairy. Specifically, it was in the lactation biology section, which focuses strongly on mechanisms to improve lactation outcomes in dairy cows. Furthermore, the results of this experiment were presented in extension type meetings to groups of farmers, nutritionists, dairy managers as well. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project provided research training to 3 different graduate students. It also allowed 6 undergraduates to participate in research associated with this project. How have the results been disseminated to communities of interest?We currently have 2 papers that are published, one in 2017 in the Journal of Dairy Science, and one in 2019 in the Journal of Animal Science. We also presented data at ADSA in 2017 and 2019. We are currently finishing up the sequencing analyses from the mammary biopsies and that will result in a presentation in 2020 at ADSA, and a 3rd research manuscript that should be published in 2020. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? The overall objective of this research is to understand the influence of residual milk/udder fill, milking frequency and their potential interactions on milk yield and teat health. Impacts are below: Aim 1: We determined that leaving 30% of milk in a respective quarter can result on a long-term negative impact on the milk yield of that quarter that was permanent for that particular lactation. Using our half udder model, we were able to demonstrate that this suppression in milk yield is regulated within a quarter, as the quarters that had no milk left behind did not have negative impacts on milk yield. We did not observe any specific effects on the time of milking. Aim 2: In our second experiment, we attempted to use increased milking frequency to recover the lost milk yield in a quarter seen by leaving 30% milk behind in that quarter. We compared a typical 2x daily milking to a 3x daily milking. As expected, we saw the milking 3x a day resulted in about 10% increase in milk yield, as has been previously reported. Surprisingly however, in quarters where we left 30% milk in the quarter, the 3x daily milking was unable to recover the lost milk yield to control levels. This suggests that if quarters are not completely milked out, that even increased milking frequency may not be able to correct this loss of milk yield. This may become problematic in robotic milkers, which are controlled at the quarter level. Future experiments would be directed and testing these hypotheses in a robotic system, as well as performing a "dose-response" experiment where we would test leaving behind, 10%, 20%, or 30% and making comparisons to determine thresholds on depressed milk production. It is also critical in the future to follow up if the suppression of milk yield seen in the cows where we left 30% milk behind, resulted in suppressed production in a subsequent lactation. Aim 3: We collected mammary gland biopsies from the quarters in our 2x vs. 3x lactation experiment to compare both 0% vs. 30% milk left behind to 2x vs. 3x milking. Those tissues are currently being statistically analyzed by our collaborator who performed all the sequencing in the summer of 2019. The data from this experiment will be presented at ADSA in 2020 and also result in a publication in 2020. Aim 4: We did not observe any significant changes at the level of the teat when leaving milk behind at a 30% rate.

Publications

  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Kuehnl, JM, Connelly, MK, Dzidic A, Lauber, M, Fricker, HP, Klister, M, Olstad, E, Balbach M, Timlin, E, Pszczolkowski, V, Crump, PM, Reinemann, DJ, Hernandez, LL. Journal of Animal Sci: 97 (6), 2424-2432. 2019 May 30. The Effects of Incomplete Milking and Increased Milking Frequency on Milk Production Rate and Milk composition.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2019 Citation: American Dairy Science Association
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Penry JF, Endres, EL, de Bruijn B, Kleinhans A, Crump PM, Reinemann, DJ, Hernandez, LL.Journal of Dairy Science: 100 (2), 1535-1540. Feb 2017. Effect of Incomplete Milking on Milk Production Rate and Composition With 2 Daily Milkings.


Progress 10/01/17 to 09/30/18

Outputs
Target Audience:During this year, we reported our results to both the community at the American Dairy Science Association, as well as in Journal format in the Journal of Dairy Science. We will be presenting new data also at the American Dairy Science Association in Cincinnati OH in June of 2019. We are also about to submit a second manuscript to the Journal of Dairy Science. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This grant allowed for the training of 2 graduate students (1 PhD and 1 masters student). How have the results been disseminated to communities of interest?We have reported our data to the scientific community by presenting at the Dairy Science Annual meeting, as well as through publication of the research. What do you plan to do during the next reporting period to accomplish the goals?During the spring of 2019, we will be analyzing mammary tissue biopsies through RNA sequencing to determine if there are any changes in transcriptional regulation in the mammary gland as a result of our treatments and that data will be published.

Impacts
What was accomplished under these goals? We determined that increased residual milk decreased milk yield, as well as after 10-12 days of treatment, resulted in a permanent decrease in milk yield. We determined that increased residual milk yield did not affect teat end hyperkeratosis, teat end barrel congestion, or teat canal ketatin levels. We determined that increased milking frequency resulted increased milk yield in quarters that had residual milk. We did not observe any significant effects on mastitis in dairy cows when residual milk was left behind, however we did see an increase in milk somatic cell count, although our cell counts were between 20,000 cells/ml-46,000 cells/ml, well below the threshold of what is optimal, which is suggested to be 100,000 cell/ml. We did see significant changes in milk fat (increased) and milk lactose (decreased) due to increased residual milk. During the Spring of 2019, we will be analyzing mammary tissue biopsies through RNA sequencing to determine if there are any changes in transcriptional regulation in the mammary gland as a result of our treatments and that data will also be published.

Publications

  • Type: Conference Papers and Presentations Status: Other Year Published: 2019 Citation: Increased milking frequency on milk production rate and milk composition. 2019. Abstract. American Dairy Science Association. Cincinnati, OH.
  • Type: Journal Articles Status: Other Year Published: 2019 Citation: Effects of increased residual milk and milking frequency on milk production. 2019. Journal of Dairy Science.