Detecting Changes of

Milk Quality in Dairy

Cattle to Predict a
Subclinical Mastitis
Inflammation
Lauren Harper
Marine Academy of Technology and
Environmental Science
Manahawkin, NJ
January 4, 2016
Advisors: Dr. John Wnek and Mr. Jason Kelsey

Table of Contents
Abstract...page 1
Introduction.page 2
Materials..page 5
Methodology...page 5
Data.....page 8
Results...page 14
Discussion.page 15
Conclusion.....page 16
Future Research.page 17
References.page 18
Acknowledgements...............................................................................................................page 20
Abstract
Mastitis is considered to be the costliest malady in the entire dairy industry. Costs are attributed to
purchasing antibiotics, throwing away milk while the animal is on the antibiotics, and the
permanent reduction of the quantity and quality of milk that the animal will produce following the
mastitis inflammation. Mastitis is defined by the inflammation of mammary tissue as a result of a
bacterial or viral infection, which makes mastitis hard to treat and (in minute cases) to detect. The
current most accepted and cost effective method of diagnosing mastitis is the California Mastitis
test; however, this test is qualitative and only detects severe mastitis cases at which point the
inflammation has already cost the farmer thousands of dollars, over the rest of the cows life. The
goal of this experiment was to determine which milk quality parameters most accurately diagnose
a case of subclinical mastitis. In stage one of this experiment, a mastitis infection was simulated
through the use of unpasteurized milk, E. coli, and an incubator. Multiple milk quality parameters
were measured at regular intervals over the course of a 24-hour period, and the collected values
were plotted to display the trends during an infection. Stage two of the experiment involved
sampling bovine milk prior to milking and testing the same parameters as in stage one. In the final
stage of this experiment, the data was analyzed to determine which milk quality parameter best
predicted a mastitis infection, based on the known infected samples collected from the cows (as
diagnosed by the farmer). The results suggest that conductivity, pH, and teat temperature produce

Page 1 of 21

the most accurate diagnosis of subclinical mastitis. Daily samples and recordings of these
measurements can save the farmer up to $129.37 dollars per cow per year.
Introduction
Mastitis is often considered the costliest 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 indications of an infection such as swelling and milk abnormalities) (Questions
about Milk, 2013). Clinical mastitis can occur in three primary forms: acute: a sudden flare up
in an otherwise healthy cow, per-acute: quick onset of severe inflammation and pain causing the
cow to become terribly ill within a short period of time, and sub-acute: minor displays of
symptoms such as clotting in milk and slight inflammation (Down, Green, and Hudson 2013).
Subclinical mastitis is considered the costliest form of mastitis because it is often reoccurring,
undetected, highly contagious, and reduces overall milk quality and yields (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). Milk from a cow with subclinical mastitis will vary
in composition from the cow's healthy average, which lowers the quality of her milk at market.
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 an inflammation of the mammary cells within the cows udder due to a release
of leukocytes because of bacteria invading the teat canal (Jones 2009). Leukocytes are often
referred to as somatic cells or white blood cells. High levels of somatic cells are unfavorable to
large distributing companies as it can reduce the grade of their milk product (Ruegg 2002).
Farmers that supply milk with high somatic cell counts are often fined furthering the
deteriorating economic effect of mastitis. High concentrations of somatic cells result in reduced
milk production and an altered milk composition (Jones 2009). There are multiple parameters
that can be used independently to test for mastitis. The current most cost effective method of
testing for mastitis is using the California Mastitis Test. The California Mastitis Test is a
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qualitative measurement of somatic cells. The CMT kit uses a four chambered paddle where strip
samples of milk are placed, and then measured to a standardized amount. Strip samples are one
pull of the teat obtained prior to milking, which is a method used in most dairy farms because the
milk within the teat typically holds the highest concentration of bacteria. After adding the strip
samples to the paddle, a detergent and a bromcresol indicator solution is placed into each
chamber in standardized amounts (Ruegg 2002). The solution disturbs the cell membranes of any
cells present in the milk sample, allowing the DNA in the sample to react with the test reagent
forming a gel (White et al. 2005). The reaction can take place within 15 seconds and simply be
the collection of a slightly purple area that quickly disappears. The problem with the CMT is that
the somatic cell count given by the test has a wide range given as a qualitative parameter (Table
1) (Ruegg 2002).
Table 1: The somatic cell count ranges given for each level of visible reaction for the California
Mastitis Test.

Other regular somatic cell count analyses are generally geared toward bulk tank somatic
cell counts (The Value and Use). The bulk tank is an insulated container that receives and
stores the milk from every cow prior to it being shipped to be pasteurized. However, this type of
sampling does not allow the farmer to tell which cows are infected. This type of sampling is used
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for the milk product buyers and distributors to make sure that the milk is within their quality
ranges. However, by only using the average milk quality of the bulk tank to test for mastitis, the
disease can spread throughout the farm unnoticed until every cows milk output decreases in
quantity and quality (The Value and Use). Early diagnosis is also key because it reduces the
permanent damage that prolonged inflammation causes to the mammary cells. Therefore, the
best way to diagnose mastitis is early and individually; however, it must be in a quick and
efficient way so that the farmers will not lose milking time from the device.
Previous research shows 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 also 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) then a final number and range of somatic cells will be given
(Broadhagan 2012). The iPhone tool is quick and convenient; however, it is not feasible because
the chambers needed for the test are not reusable. Mastitis needs to be monitored on every farm
because as of 2009 mastitis caused the average United States Dairy Farm to lose 171 dollars per
cow annually (Jones 2009).
The parameters selected to measure for this research project were: teat temperature, milk
temperature, conductivity, turbidity, pH, phosphates, CMT, microorganisms, and milk
composition. Teat temperature indicates a possible inflammation, this is because the
inflammatory response opens blood vessels near the infection which causes redness and an
increase in temperature to respond to the possible bacterial infection. Milk temperature is used
similarly, however for this research, more than a strip sample was used to assure a sufficient
amount of milk to analyze all parameters; therefore, this measurement was more indicative of the
entire udder quarter instead of just the teat. Turbidity was measured to detect for material
clumping, cows with mastitis will have milk that clumps more often and settles out, making the
Page 4 of 21

rest of the milk clearer. The pH of the milk was measured because "Elevated milk SCC is
associated with altered protein quality, change in fatty acid composition, lactose, ion and mineral
concentration, increased enzymatic activity, and a higher pH of raw milk" (Ogola, Shitandi, and
Nanua 2007). The parameter of phosphates was measured because phosphates are known to
decrease in clinical mastitis cases (Raji, Ezzatpanah, and Givianrad 2010). The California
Mastitis Test will also be used to compare other collected values to. The microorganisms test
will assist in determining the presence and abundance of microorganisms in the milk sample,
which may affect the results otherwise. Finally milk composition will be tested to determine
compare values to considering that milk composition is known to alter given an inflammation
such as mastitis.
By reducing presence of mastitis in the dairy industry, the cost to the farmer to produce
milk will be reduced; in turn, the cost of milk to the consumer can be reduced. Therefore, the
goal of this research project was to use various water quality parameters and test milk samples
with them to identify the parameters with the most accurate identification of a cow with mastitis
and without. After data collection and analysis, the objective of this project is to determine the
most time and cost effective method of detecting subclinical mastitis in dairy cattle.
Materials
Field Journal

Specimen Cups

Colorimeter

Phosphate Test Kit

Thermometer

Laser Temperature Gun

YSI-85

pH probe

CMT Test Kit

Methyl-Blue

Incubator

Vacuum Filtration System

Balance

Hot Plates

Bulk Tank Sample

Needles

Blood Test Tubes

Centrifuge

E. Coli

Lap Top

Clean Room

Petri Dishes

Micropipettes

Refrigerator

Distilled Water

Waste Container

Excel

Graduated Cylinder

Methodology
Study Site:
Page 5 of 21

The primary study site was my own home; all of the milk samples were tested there. In
addition, some of the samples were tested at my school (the Marine Academy of Technology and
Environmental Science). The school was used for the centrifuge, to grow the E. Coli, and to
weigh the milk components in the analytical balance. The milk was obtained from Brynmore
Dairy Farm located in New Egypt, NJ. Brynmore Dairy Farm was selected because it was one of
the few farms that were willing to allow me to sample from their cows, and for convenience
based on distance from my home.
Procedure:
1. Presented research proposal to school for approval, and submitted required forms:
general, working with bacteria, and working with animals.
2. Found local dairies that are willing to participate in this research project by allowing
samples to be obtained from their cows with compensation. Located at least three dairies
within 30 minutes of the testing location. Two of the Dairy Farms were willing to
participate; however, one of the farms would only allow for bulk tank samples at 6:00am.
Therefore, a single Dairy Farm was used: Brynmore Dairy Farm.
3. Contacted the local large animal veterinarian and asked for his ability to mentor this
project and assist where needed such as the official diagnosis of mastitis in the animals.
The veterinarian felt uncomfortable working with bovine, but referred me to multiple
other veterinarians each of which were unable or unwilling to assist with this research
project. Therefore, the diagnosis given by the head farmer at Brynmore Dairy Farm was
accepted as true; however, it should be noted that this could lead to error.
4. Researched probes that can be used in the model machine that could record data into a
computer program for each cow with strong accuracy. Researched the cost of these
probes to be included in the design of a machine that will be able to detect early signs of
mastitis accurately through the combination of all probes used. Researched what is
needed to maintain the probes in apex performance. Created a model machine for future
research, costs were recorded to be used to analyze the financial benefit of testing for
mastitis regularly.
Page 6 of 21

5. Obtained fresh whole milk samples from healthy cows, samples of milk from cows that
are expected to have mastitis according to the head farmer at Brynmore Farm, and
samples from cows that may have early signs of mastitis such as a fever. Samples were
also collected from a cow that was going dry (slowly producing less milk right before
giving birth again).
6. The samples were collected at Brynmore Dairy Farm (approximately 3 oz. per teat of
each cow sampled or 12 oz. per cow sampled). First the cows teat temperature was
taken, then the teats were washed and stimulated with a warm wet cloth. Following the
stimulation, I collected the samples in separate specimen cups, and recorded the
temperature of the milk; however, my sampling occurred in January and the surrounding
temperature might have caused error.
7. The samples were brought back to my home, and I began testing the various parameters
a. Conductivity was measured using the YSI 85 the probe was either placed directly
into the sample, or a dilution was made first if the conductivity was over range,
which is another source of error. When the solution was diluted, the value red off
of the meter was multiplied by the reciprocal of the factor of dilution plus one.
b. Turbidity was measured as a dilution which included 1% of the actual milk
sample placed into a vial and selected as a test on the colorimeter provided by my
school.
c. pH was measured using a pH probe placed directly into the milk sample.
d. phosphates were measured following a dilution using the LaMotte phosphate low
range indicator kit, and placing the sample into the colorimeter.
e. The California Mastitis Test was performed by placing equal portions of milk and
CMT solution and mixing while watching for a coagulation which indicated
various ranges of somatic cell counts.
f. Microorganisms were measured by placing the milk and 8 drops of methyl-blue
into an incubator for 30 minutes while observing the changes every 5 minutes.

Page 7 of 21

g. Finally, milk composition was measured by following lab instructions for Milk
Analysis found online at http://course1.winona.edu/jfranz/lab/milklab.html
i.

First the (protein and fat) and (lactose and phosphates) are separated then
each group was separated respectively and the total percentage of each
was recorded based on mass.

8. A control mastitis sample was also created through the incubation of milk sampled from
the bulk tank and infused with the E. coli bacterium. Initial measurements were made,
then the sample was left to incubate for approximately 8 hours. Then regular samples
were taken every hour. Each sample taken was then tested using all of the parameters
mentioned in Methodology point 7. During each sample, white blood cells were added to
the control to simulate the change that white blood cells induce in an actual mastitis
inflammation. All lab safety parameters were followed.
9. Created graphs of the results of the milk analyses in step 7 and 8 for each parameter
separately. Used T-tests to determine if significant differences between the milk supplied
by healthy cows and those which were diagnosed to have an inflammation. Plotted values
together on a graph and created 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.
10. All of the parameters were given scores based on the accuracy of diagnosing a cows
quarter of having mastitis. Each parameter was also given a price which were recorded
previously. Then the most accurate and cost effective parameters were selected as the
best methods for regularly detecting mastitis.
Statistical Analysis:
Correlations, Regressions, and t-tests were used to statistically analyze the data. Graphs
were also created with every parameter measured compared to the likelihood that the cow had
mastitis based on the diagnosis given by the farmer.

Data
Page 8 of 21

Microorganisms as Controls progressed

Value scored for result

3
2.5
2
1.5
R = 0.7394

1
0.5
0
0

200

400

600

800

1000

1200

1400

1600

1800

Length of Incubation (minutes)

Figure 1: The three colors displayed symbolize three different control samples. The scale for
presence of microorganisms ranges from 0-3, but values recorded ranged from 1-3. (n = 22;
Regression p-val <0.001).

Conductivity (microSiemens/cm^3)

Conductivity as Control Progressed

14000
13000
12000
11000
10000
9000
8000
7000
6000
5000
4000

R = 0.9434

200

400

600

800

1000

1200

1400

1600

1800

Length of Incubation (minutes)

Figure 2: The blue line signifies the initial (bulk tank) conductivity, the black line represents the
highest accepted bulk tank value. Values collected range from 6000-13320 microSiemens (n =
22; Regression p-val <0.001).
Page 9 of 21

pH

pH of Milk as Controls Progress

7.5
7
6.5
6
5.5
5
4.5
4
3.5
3

R = 0.9228
0

200

400

600

800

1000

1200

1400

1600

1800

Length of Incubation (minutes)

Figure 3: The blue line signifies the initial (bulk tank) pH, the black line represents the accepted
average milk pH. Values collected range from 6.8-3.81 (n = 22; Regression p-val <0.001).

Turbidity as Control Progressed

Turbidity (FTUs or NTUs)

50000
40000

R = 0.6399

30000
20000
10000
0
0

200

400

600

800

1000

1200

1400

1600

1800

Length of Incubation (minutes)

Figure 4: The blue line signifies the initial (bulk tank) turbidity. Values collected range from
8700-43200 NTU (n = 22; Regression p-val <0.001).

Page 10 of 21

Phosphates as Controls Progressed

Phosphates (ppm)

6000
5000
4000
R = 0.3382

3000
2000
1000
0
0

200

400

600

800

1000

1200

1400

1600

1800

Length of Incubation (minutes)

Figure 5: The blue line signifies the initial (bulk tank) phosphates. Values collected range from
1240-5320 ppm (n = 22; Regression p-val < 0.01).

Milk Temperature Reflecting Mastitis

Milk Temperature (C)

41
39
37
35
33
31
29

R = 0.4441

27
25
0

10

15

20

25

30

35

40

Likelihood Cow has Mastitis or Displays Similar Characteristics

Figure 6: Green symbolizes the samples from cows diagnosed as healthy, yellow is a cow that is
going dry, and red is a sample from a cow diagnosed with mastitis (by the farmer). The space in
between the black lines and including the black lines is the accepted range of a healthy cows
body temperature. Values collected range from 32.25-37 (C) (n = 34).
Page 11 of 21

Teat Temperature Reflecting Mastitis

Teat Temperature (C)

45
40
35
30
25
20
R = 0.7601

15
10
0

10

15

20

25

30

35

40

Likelihood Cow has Mastitis or Displays Similar Characteristics

Figure 7: The space in between the black lines and including the black lines is the accepted
range of a healthy cows body temperature. Values collected range from 22.2-37.1 (C) (n = 34;
Regression p-val < 0.001).

Milk pH Reflecting Mastitis

7.3
7.2

pH of Milk

7.1
7
6.9
6.8
R = 0.6948

6.7
6.6
6.5
0

10

15

20

25

30

35

40

Likelihood the Cow has Mastitis

Figure 8: The blue line signifies average (bulk tank) pH of milk. Values collected range from
6.58-7.17 (n = 34; Regression p-val <0.001).
Page 12 of 21

Conductivity (microSiemens/cm^3)

Milk Conductivity Reflecting Mastitis

12000
10000
8000
6000
4000
2000

R = 0.2217

0
0

10

15

20

25

30

35

40

Likelihood the Cow has Mastitis

Figure 9: The blue line signifies the average (bulk tank) conductivity of milk. Values collected
range from 3137-9560 microSiemens (n = 34; Regression p-val <0.05).

Phosphates Reflecting Mastitis

Phosphates (ppm)

3000
R = 0.7279

2500
2000
1500
1000
500
0
0

10

15

20

25

30

35

40

Likelihood the Cow has Mastitis

Figure 10: The blue line signifies the initial (bulk tank) phosphate value. Values collected range
from 889.6-2444.8 ppm (n = 34; Regression p-val <0.001).

Page 13 of 21

Reaction Caused by California

Mastitis Test

California Mastitis Test Accuracy

1.6
1.4
1.2
1
0.8
0.6
0.4
0.2
0
-0.2 0
-0.4

R = 0.3976
5

10

15

20

25

30

35

40

Likelihood the Cow has Mastitis

Figure 11: Observed data was given a value based on the prominence of the chemical reaction
from 0-1.5. Values collected range from 0-1.5 (n = 34; Regression p-val <0.05).
Results
Conductivity was found to correlate best with the diagnosis of mastitis both in the control
sample and in the data collected from the cows; however, the correlation was much stronger in
the control than in the actual samples. pH also showed a strong correlation to diagnosing mastitis
in both the control and actual samples; the pH of milk from a cow suspected to have mastitis or a
cow that is going dry was significantly higher than clean cows. Phosphates showed a strong
correlation specifically in the actual samples. Cows that were going dry were found to have
significantly higher milk temperature and turbidity. Cows suspected to have mastitis had
significantly lower teat temperatures than a healthy cow. Cows suspected to have mastitis had
significantly lower turbidity values. Cows that were going dry or suspected to have mastitis
showed significantly lower phosphate values. The correct diagnosis of mastitis in a dairy cow
through the CMT was insignificant. When analyzing the milk components, it was found that the
Page 14 of 21

milk component (Ca3(PO4)2) is significantly reduced in cows with mastitis. Finally, no methylblue microorganism test turned entirely white.
Discussion
There were two phases of this project, creating control trend lines from E. coli laden
samples and sampling lactating bovine of which 2/9 of the samples were from cows suspected to
have mastitis and 1/9 were going dry and thus have higher somatic cells, signs of mastitis, and
are the most likely subjects to have mastitis. Conductivity readings from the control sample most
closely correlated to the progression of E. Coli growth and thus the simulated mastitis. Control
sample 350 A (green) most closely correlated with an R^2 value of 0.99. High somatic cell
counts from the milk, and therefore mastitis, results in the ionization of many typically unionized
milk components (Ogola, Shitandi, and Nanua 2007). The pH of the control did not reflect the
data collected for cows with mastitis and clean cows, which is likely due to the fermentation
process occurring with the lactic acid in the milk (Fankhauser 2010). The milk actually becomes
more basic as mastitis develops. It becomes more basic because the inflammation causes reduced
secretion of milk and higher permeability of the mammary epithelial cells, which can lead to
transfer of components of the blood into the milk such as citrates, bicarbonates, and Na and Cl
ions (Ogola et. al 2007). Therefore, the trend line developed from the sampled data and not the
control is more accurate. Unlike the hypothesis predicted, the milk becomes less turbid as
mastitis progresses, this is because the milk curdles within the udder and is released in chunks of
the protein casein and milk fat surrounded by a liquid composed primarily of water, calcium,
vitamins, and sugars such as lactate and glucose. Therefore, the curdles within the sample
quickly settle to the bottom removing their original turbidity from the sensor within the
colorimeter. The cow within the samples that was going dry had really high turbidity readings
because the udder is cleansing itself and reducing the quantity; thus not releasing as much water
into the milk. The milk temperature of the cow steadily increased with likelihood of mastitis
because the milk temperature accurately represents internal body temperature. The temperature
of the milk of the cow that was going dry was significantly higher than all other data points
Page 15 of 21

because it is a natural process as the cow goes dry, she cleanses the teats less often; therefore,
flushing the bacteria within her less frequently. So to prevent infection the udder is flooded with
somatic cells and the temperature is increased. The teat temperature of the cows were measured
in the center of the teat; however, for the first 23 data points, a thermocouple was used to collect
the temperature which required the wire to be pressed against the cow causing it discomfort.
Thus, those measurements were not used, and the teat temperature was re-measured using a laser
temperature-reading gun. Phosphates decreased as the likelihood of mastitis increased; however,
the control samples all displayed a drop in phosphates then an increase all below the average.
This could be attributed to the fact that the phosphates covalently bind to the protein casein,
which then forms ionic bonds to calcium ions forming a micelle, the casein and triglycerides
form the curdles observed; therefore, it is possible that some of the phosphates were captured in
the precipitate (Raji, Ezzatpanah, and Givianrad 2010). As the samples became more separated
and curdled they were stirred and the curdles were broken possibly re-releasing the trapped
phosphates. Finally, the most accepted method of diagnosing mastitis, the California Mastitis
Test, produced no significant correct data; thus, supporting my hypothesis that the California
Mastitis Test is not the best cow-side determinant of mastitis. The methyl-blue test produced no
samples that completely turned white, which is absurd considering the concentration of E. coli
within the control samples. However, it is likely that the E. Coli was primarily surrounding the
fat and protein curdles, which were white. Finally, finding the percentage of the main
components of the milk was removed from this study because of the time necessary to separate
each component that would be inefficient in a cow-side machine. However, there was one
significant difference between clean milk and milk from a cow with mastitis: the component of
Ca3(PO4)2 was significantly lowered in the later.
Conclusion
The data collected suggests that the best single method found in these experiments to
diagnose mastitis was pH according to its correlation with the simulated mastitis for the control
samples, and its correlation with suspected mastitis through samples collected. However, there is
error associated with the pH values and the best way to get around this error is to add additional
Page 16 of 21

tests that correlate well with the progression of mastitis. Therefore, the tests that would produce
the most accurate diagnoses in the most feasible manner are (in order) pH, turbidity,
conductivity, teat temperature, phosphates, mass of calcium within the milk (milk composition),
CMT, and milk temperature. Since 2 cows suffer through mastitis on average in every dairy farm
of 60 head, it would be beneficial for the farmer to monitor every one of his cattle every milking
to observe trends towards mastitis to detect, prevent, and treat early. A farmer of 50 head can
expect to make more than $13,226.45 per month from his cows milk; however, mastitis will cost
him $539.03 a month. A single purchase of equipment to test for the four most accurate and
cheapest tests can insure mastitis infections to diminish possibly saving the farmer $539.03 per
month or $6,468.36 a year of which he would only need to spend between $199.68 and $542.94
(depending on the quality of the equipment) and the cost of repairs. This is only for a small farm
of 50 head of cows. This technique will help small farmers hold together in the ever changing
market and help to prevent monopolies. In conclusion, regular monitoring should be considered
for financial benefit, and improved lifetime health of the dairy herd.
Future Research
If this project was replicated, there are a few changes that should be made. All of the
cows sampled from should be of the same breed (if possible) and in the same stage of the
lactation cycle. In addition, the control sample should be sampled every hour starting as soon as
the E. Coli is placed into the sample. The samples should also be sampled right away and not
placed into a cooler first (this requires taking one sample at a time).
In the near future, I will be further testing the best parameters to make a more accurate
trend line that indicates the possibility of a mastitis infection. Then, based on this trend line, I
would like to create a computer program to be able to record values from a milking and highlight
cows that the farmer should monitor for multiple milkings to determine if the cow is suffering
from a minute case of mastitis. Eventually, a claw could be made which tests for these
parameters and collects data from each cow at the same time.

Page 17 of 21

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Acknowledgements
I would like to thank my advisors Dr. John Wnek and Mr. Jason Kelsey for helping me
throughout the project. I would also like to thank Brynmore Dairy Farm for allowing me to
sample from their herd. Finally, I would like to thank my family and friends for assisting me in
this project; specifically, I would like to thank my parents and brothers for putting up with all of
the noises I made in the middle of the night while working on my project.

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