20142015

The Detection of Early

Signs of Subclinical
Mastitis in Dairy Cattle
Through the Creation of a Computer Based
Program to Predict a Mastitis Outbreak

Lauren Harper
Marine Academy of Technology and Environmental Science (MATES)
2014-2015

Introduction/Background Research: Mastitis is often considered the most costly disease in

raising dairy cows (Ruegg 2002). Mastitis can occur in two forms clinical mastitis (in which
there are visible signs of mastitis in the milk and udder) and subclinical mastitis (the
inflammation of the mammary gland does not cause visible signs such as swelling and milk
abnormalities) (Questions about Milk, 2013). Subclinical mastitis is often seen as the most
economically important form of mastitis because it is often reoccurring, undetected, reduces
overall milk yields, and it is highly contagious to surrounding cows (Ruegg 2002). The cows
with subclinical infections will produce milk that visually appears normal; however the cow will
produce less milk and lower quality milk due to persistent and chronic infection that damages
milk secreting cells (Ruegg 2002). As of 2009 mastitis caused the average United States Dairy
Farm to lose 171 dollars per cow annually (Jones 2009). Mastitis also causes the dairy industry
to lose 1.7-2 billion dollars annually or 11 percent of total U.S. milk production (Jones 2009).
Mastitis is the inflammation of mammary cells within the cows udder due to a release of
leukocytes because of an invasion by bacteria of the teat canal (Jones 2009). Leukocytes are
often referred to as somatic cells or white blood cells. High concentrations of leukocytes result in
reduced milk production and an altered milk composition (Jones 2009). High levels of somatic
cells are unfavorable to large distributing companies as it can reduce the grade of their milk
product (Ruegg 2002). There are multiple different parameters that can be used independently to
test for mastitis or common parameters related to identifying mastitis. The best current method of
testing for mastitis is using the California Mastitis Test. The California Mastitis Test is a
qualitative measurement of somatic cells. The CMT kit uses a four chambered holder where strip
samples of milk are placed, and then measured to a standardized amount. Then detergent and a
bromcresol indicator solution is placed into each chamber in standardized amounts (Ruegg

2002). The reaction however can take place within 15seconds and simply be the collection of a
slightly purple area that quickly disappears. The problem with the CMT as well is that the
somatic cell count given by the test has a wide range given for a qualitative parameter (See Table
1) (Ruegg 2002).
Other somatic cell counts are generally geared toward Bulk Tank Somatic Cell Counts
(The Value and Use). This type of sampling however does not allow pinpointing which cows
are infected. This type of sampling is used for the product buyers and distributors to make sure
the milk is within their quality ranges. However, by only using this method of mastitis testing,
the disease can spread throughout the farm and cause every cows milk output to decrease and be
of a lower standard (The Value and Use). Therefore, the best way to diagnose mastitis early is
individually, however it must be in a quick and efficient way so that the farmers will not lose
milking time from the devise.
Previous research projects have shown that conductivity can detect clinical and
subclinical mastitis (Ilie, Tudor, and Gali 2010). The study by Ilie, Tudor, and Gali found that
conductivity correctly detected 81.4% of clinical cases of mastitis and 44.0% of subclinical cases
(2010). The conductivity detection method also only incorrectly determined healthy cows as
infected 26.5% of the time (Ilie, Tudor, and Gali 2010). There is a new method of measuring
somatic cell count through the use of an iPhone (Dairy Quality SCC 2014). The device takes a
milk sample into a chamber and then the iPhone reads the sample after a chemical reaction
occurs (much like a color concentration analysis would occur) then a final number and range will
be given of SSC (Broadhagan 2012). This is a quick method; however it will not be possible, or
feasible for a farmer to test 4 cow teats, 2 times a day, with up to a few hundred head of cows.

For each test, a new chamber with chemical reactions is needed; this whole process adds time
and money.
Hypothesis: Through the use of various indicators of mastitis a bell curve will be constructed
that is able to predict a mastitis infection in dairy cattle.
Materials/Methods:
1. Present research proposal to school for approval, and submit required forms general, bacteria,
and working with animals.
2. Find local dairies that are willing to participate in this research project by allowing samples
to be obtained from their cows with compensation. Locate at least three dairies within 30
minutes of the testing location.
3. Contact the local large animal veterinarian and ask for his ability to mentor this project and
assist where needed such as the official diagnosis of mastitis in the animals.
4. Research probes that can be used in the model machine that could record data into a
computer program for each cow with strong accuracy. Research the cost of these probes to be
included in the design of the machine that will be able to detect early signs of mastitis
accurately through the combination of all probes used. Research what is needed to maintain
the probes in apex performance.
5. Obtain healthy fresh whole milk samples (12; 4/farm), samples of milk from cows that are
diagnosed with mastitis from the veterinarian (12), and samples from cows that may have
early signs of mastitis such as a fever (36). Collect from the same cows (4 healthy, 4 that
show signs of mastitis, and possibly 4 that have mastitis) 4- 6 different times approximately a
week apart depending on time restrictions. Compare all of the parameters (temperature, pH,
conductivity, turbidity, colorimeter directly after a California Mastitis test is performed) of
all milk samples then further milk analysis will be performed to find the best and most

accurate method to detect for subclinical mastitis. Some of the milk analyses include the
detection of microorganisms in milk, the separation of the components of milk (casein,
lactose, and milk fat) and their concentrations in the milk. Finally, leucocytes or the white
blood cells that cows release into the milk when there is a sign of infection will be counted.
a. A control mastitis sample can also be created through the incubation of healthy
milk infused with the E. coli bacterium and incubated to be removed at standard
time increments (possibly infused then with leukocytes as a cow would release)
and tested for all parameters then replaced for further incubating to create a graph
of simulated mastitis progression. All safety parameters will be followed.
6. Create graphs of the results of the milk analyses in step 5 for each parameter separately.
Determine if there are any significant differences between the healthy milk samples and the
mastitis milk samples. Plot those values then together on a graph and create a line of best fit
where all area in between the healthy and unhealthy milk samples will be referred to as the
samples which could be signs of subclinical mastitis.
a. Then all of the parameters can be given scores based on the likely hood of the
cows quarter having mastitis. These scores can then be averaged from all
parameters to create a graph that will state the likelihood that the cow has
subclinical mastitis and will alert the farmer to check the cow or call a
veterinarian when appropriate for medication or proper treatment.
7. Make a computer model of the machine that is the most accurate and cost effective. Create a
model of the machine which can include other attachments such as an infrared camera that
can scan a field of cattle for elevated temperatures. Include prices of all parts and total cost
for the farmer, and the number of correct diagnoses needed for the farmer to break even.
8. Create a computer program that will take inputs of data points to create a cows trend and
locate significant changes and when the milk is between healthy and mastitic regions.

9. Test milk samples that are known to be from cows with mastitis to see if the program
accurately diagnosis them (36). Booties will be worn to protect farms from cross
contamination.
Budget:
Item
3 oz Plastic Vials

Igloo Playmate cooler

Thermocouple
Milk Sample Reimbursement

Possible Veterinarian
Reimbursement
Go Wireless Temp With
Vernier
Go Wireless pH with Vernier
Conductivity probe
Turbidity Sensor
Colorimeter
Methylene blue
Incubator
HCl
Calcium Carbonate
Ethanol
Activated Carbon

Blood Agar Plate 10pk

Total:

Cost & Vendor

$33.60 + tax and shipping and
handling
Uline model number S-19523
24.99 Currently own
28.00 School owned
(96cups +2cups(mistakes))/16
= 6.125 gallons * 3.77=
$23.09
$50/farm visit * 3 farms =
$150
$75.00 School owned
$99.00 School owned
$95.00 School owned
$112.00 School owned
$115.00 School owned
$3.75
The Science Company
$305.95
School owned
$17.00
School owned
$3.99
Marineland Premium
activated carbon 8 oz.
$27.95
Online Science Mall
Out of Pocket:268.78
School Owned Utilities:
644.95

Purpose
To collect samples and
transport after collection
To transport samples
Monitor temperature inside
the cooler
Sampling

Diagnosis of mastitis and

how far along the mastitis is.
Temperature of fresh milk
samples
pH of milk samples
Conductivity of milk samples
Turbidity of milk samples
Quantitative measurement
from milk samples after CMT
Detecting micro-organisms in
milk
Detecting micro-organisms in
milk
Separating components of
milk
Separating components of
milk
Growing E. coli bacteria in
Milk

Literature Cited:
Broadhagan, A. (2012, September 18). New iPhone App Can Test Somatic Cell Count.
Farmscom RSS. Retrieved June 23, 2014, from http://www.farms.com/ag-industrynews/new-iphone-app-can-test-somatic-cell-count-327.aspx
Chanie, M., & Tadesse, A. (2012). Study on the Occurrence of Bovine Mastitis in
Addis Ababa Dairy Farms and Associated Risk Factors. <i>Academia.edu</i>. Retrieved
June 23, 2014, from
http://www.academia.edu/4213326/Study_on_the_Occurrence_of_Bovine_Mastitis_in_A
ddis_Ababa_Dairy_Farms_and_Associated_Risk_Factors
Dairy Quality SCC Testing. (2014). Dairy Quality SCC Testing. Retrieved June 23,
2014, from http://www.dairyconsultinggroup.com/Milk-Guardian.html
Harmon, R. (2001). Somatic Cell Counts: A Primer. . Retrieved June 23, 2014, from
http://www.nmconline.org/articles/sccprimer.pdf
Ilie, L., Tudor, L., & GALI, A. (2010). The Electrical Conductivity Of Cattle Milk And The
Possibility Of Mastitis Diagnosis In Romania. . Retrieved June 23, 2014, from
http://www.usab-tm.ro/vol10bMV/38_vol10b.pdf
Ingalls, W. (2001, July 16). Testing for pre-clinical mastitis. Retrieved December 6, 2014, from
http://www.milkproduction.com/Library/Scientific-articles/Animal-health/Testing-for-preclinical-mastitis/

Jones, G. (2009, May 1). Understanding the basics of mastitis. Retrieved from
http://pubs.ext.vt.edu/404/404-233/404-233.html
O'Mahony, F. (1988, March 1). Milk Chemistry an Introduction. Retrieved December 6, 2014,
From: http://www.ilri.org/InfoServ/Webpub/fulldocs/ilca_manual4/Milkchemistry.htm
Pathogens. (2014). Pathogens. Retrieved June 23, 2014, from
http://www.dairyco.org.uk/technical-information/animal-healtwelfare/mastitis/pathogensthe-cause-of-mastitis/#.U6Ymx_ldWt0
Questions about milk quality: What is the difference between clinical and subclinical mastitis.

(2013, May 14). Retrieved from

http://www.progressivedairy.com/index.php?option=com_content&view=article&id=105
91:questions-about-milk-quality-what-is-the-difference-between-clinical-and-subclinicalmastitis&catid=143:herd-health&Itemid=210
Ruegg, P., & Reinemann, D. (2002). Milk Quality and Mastitis Tests. . Retrieved June 23, 2014,
from http://milkquality.wisc.edu/wp-content/uploads/2011/09/milk-quality-and-mastitisdiagnostic-tests.pdf
Somatic Cell Count. (2014). Somatic Cell Count. Retrieved June 23, 2014, from
http://www.dairyco.org.uk/technical-information/animal-healthwelfare/mastitis/symptoms-of-mastitis/somatic-cell-count-milk-qualityindicator/#.U6YnffldWt0
The Value and Use of Dairy Herd Improvement Somatic Cell Count. (n.d.). The Value and Use
of Dairy Herd Improvement Somatic Cell Count. Retrieved June 23, 2014, from
https://nmconline.org/dhiscc.htm
Urech, E., Puhan, Z., & Schllibaum, M. (1999, November). Changes in milk protein fraction as
affected by subclinical mastitis. Retrieved December 6, 2014, from
http://www.ncbi.nlm.nih.gov/pubmed/10575607

Figures and Tables:

Table 1: The somatic cell count ranges given for each vague level of visible reaction.