Biology 代写 生物论文代写
Milk can be defined as the lacteal secretion obtained from the mammary glands of mammals by the process called milking. In Kenya, there are many different sources of milk, from cows, goats, sheep and even camels. The most common source of milk, however, is from cows as they are the most common animals in the country. The amount of milk from these cows is enough to satisfy the needs and wants of the country’s population.
As of the year 2008 the estimated cattle population was estimated to be about 3.9 million.
In this experiment, there were 3 different types of cow milk used, Raw cow milk or Unpasteurized cow milk, HTST Pasteurized cow milk and UHT pasteurized cow milk.
Raw cow milk can be defined as the naturally unaltered milk. This milk is not heated above the animal’s highest body temperature which would usually be 38 degrees Celsius. Raw cow milk is therefore not pasteurized, homogenized or frozen, nor has it been altered with additives, chemicals, light or homogenization.
Pasteurization can be defined as the process which slows microbial growth in milk by heating the milk to a specific temperature for a definite length of time, and the cooling it immediately. The process was named after its creator, a French chemist and microbiologist, named Louis Pasteur. The first ever pasteurization test was conducted by Louis Pasteur and Claide Bernard on April 20th 1864. This process was originally created as a way of preventing wine and beer from souring.
This process doesn’t destroy or kill all pathogenic micro-organisms in the milk; it decreases the number of viable pathogens that are likely to cause illness among its consumers. The pasteurization of milk inactivates or kills the organisms that grow rapidly in the milk but does not destroy the organisms that grow slowly or produce spores. Pasteurization of milk typically requires temperatures below boiling since at very high temperatures casein micelles will irreversibly curdle.
Two out of the three types of cow milk I used during this experiment came from some form of pasteurization. These types of pasteurization are: high temperature, short-time pasteurization (HTST) and Ultra high temperature pasteurization (UHT).
Ultra-high temperature pasteurization involves exposing the milk to temperatures exceeding 135 degrees Celsius (275 degrees farenheight) for one or two seconds.
High temperature, short-time pasteurization involves the milk being exposed to heat or temperatures as high as 160 to 165 degrees farenheight (71 to 74 degrees Celsius) for 10 to 20 seconds.
The most effective pasteurization technique is the Ultra high temperature technique as it kills all the pathogens in the milk. Milk that has undergone UHT pasteurization has a longer shelf life and can be kept at room temperature as long as its container is closed.
In Kenya, UHT processing is not as popular as the milk that has undergone HTST pasteurization. This is because the UHT processed milk has a ‘burnt’ taste that does not appeal to the milk consumers in my country.
Biology 代写 生物论文代写
In Kenya, many of our people prefer raw cow milk over pasteurized milk. This is because it is 20 to 50 % cheaper than pasteurized milk, its supply involves fewer costs, people prefer its tastes and its high butter milk content and most importantly consumers feel that simply boiling the milk removes most health hazards. This type of mentality doesn’t occur with Kenyans in the ‘higher-income bracket’ who would much prefer pasteurized milk to raw milk due to its health benefits.
The making of yoghurt involves both chemical and biological processes. According to the code of Federal Regulations of the FDA (FDA,1996,c), yoghurt is defined as the food produced by culturing one or more optional dairy ingredients with a characterizing bacteria culture that contains the lactic acid bacteria, Lactobacillus bulgaricus and Streptococcus thermphilus.
Fermentation is the action of milk bacteria upon the milk sugar which converts it into lactic acid which gives the sour taste to the milk and causes the milk protein to curdle. After the lactic acid is produced, its presence in the milk makes it hard for bacteria to grow. After the souring/ fermentation, it is said that the milk would for sometime not undergo any further changes and this can be detected by noting the pH.
Yoghurt was accidentally invented thousands of years ago but has gained a lot of popularity around the world. This is because of the introduction of fruit and other flavourings put in the yoghurt and it being convenient as a ready made fast food and the image of yoghurt as a low fat healthy food.
“Milk normally has a pH of about 6.5-6.7 and at this pH value the casein is without protons and has a negative charge which makes the casein micelles relatively soluble as they repel each other. In an acidic environment, the milk coagulates because the casein has its Isoelectric point at about a pH of 4.6. At this pH value it has an equal number of positive charges and negative charges. The positive part of each micelle is attracted by the negative part of the other causing the formation of ionic bonds among the micelles which work against the dipole-dipole bonds with water so that the protein precipitates in the form of deminerilized casein and in the solution remains soluble calcium salts. So the increased acidity due to the conversion of lactose to lactic acid causes milk proteins (casein) to tangle into solid masses.”
Equation for lactic acid formation from lactose:
C12H22O11 + H2O 4C3H6O3
Structural formula of lactic acid:
Lactic acid is a weak acid because it only partially dissociates. When this dissociation occurs, an ion lactate and H+ ions are formed. The reaction is reversible.
CH3CH(OH)CO2H CH3CH(OH)CO2 + H+ (Ka=1.38×10-4)
Lactose is a sugar molecule called a disaccharide. It is composed of two molecules: one glucose molecule and one galactose molecule joined together to make one molecule of lactose.
Since lactic acid is weakly acidic, the rate of fermentation reaction can be monitored by measuring pH. The pH probe helps us to monitor the pH by measuring pH as the activity of hydrogen ions surrounding the thin-walled glass bulb at its tip. The probe produces small voltages (about 0.06 volt per pH unit) that is measured and displayed as pH units by the meter.
In this experiment I compared the rate of fermentation of 3 different types of milk with and without a bacterial starter culture. I expect to find that the milk samples with the starter culture will ferment much faster than the milk samples without the starter culture, this is because I would find that the enzymes of the bacteria would speed up the reaction. Also I would expect to find that the Unpasteurized milk will ferment much quicker than the rest of the milk while the UHT milk will ferment much slower because the UHT milk has gone through such a high temperature processing that virtually all the bacteria in the milk would have been killed.
MATERIALS AND METHODS
VARIABLES
Independent Variable: -milk type
-presence or absence of culture
Dependent Variable: -pH change with time
Controlled Variable: -milk volume
-culture mass
-temperature of milk samples
Biology 代写 生物论文代写
MATERIALS USED
100mL of Long Life UHT cow milk (whole milk)
100mL of Pasteurized cow milk (whole milk)
100mL of Unpasteurized cow milk (raw milk)
18 test tubes
Three 10mL measuring cylinders (+/- 0.05mL)
Water bath
Weighing scale (+/- 0.005g)
Vernier pH probe
6 containers
1 thermometers
Clock
Naming tags
Test tube racks
Thermophilic starter culture containing Lactobacillus bulgaricus and Streptococcus thermphilus
SOURCE OF MATERIALS USED
All the milk, I personally bought with the exception of the raw cow milk that was given to me by a farmer in the rural Kiambu area of Central Kenya. The raw milk had not gone through any further processing after the cow was milked and was packed in a sterilized white plastic bottle.
The Thermophilic starter culture was acquired courtesy of and with permission from the manager of the Palmhouse Dairies in Kiambu. The manager also took time to carefully explain to me the precautions, storage and handling of the Thermophilic starter culture.
The rest of the equipment was provided to me by the Braeburn College laboratory with the help of one lab technician.
D. PROCEDURE UNDERTAKEN
All equipment needed was first sterilized before the experiment was conducted in hot boiling water for 15minutes.
pH of the milk samples was taken before the experiment began. The pH probe after each pH recording was rinsed with distilled water and wiped with tissue
18 test tubes were placed in racks in the water bath of constant temperature. 9 test tubes were placed in each rack so as to differentiate the ones that had the Thermophilic starter culture from the ones that didn’t have. They were then correctly labeled( using milk type and trial)
Three 10mL measuring cylinders were used to measure the volume of each milk type. I used 10mL of milk for the experiment for the different types of milks and for each trial. Different measuring cylinders were used so as to avoid any mixing between the different milk types as these could affect the results I get.
0.11g of the Thermophilic starter culture was weighed on a weighing scale. A white sheet of paper was placed on the weighing scale to avoid the culture form sticking on the surface of the weighing scale. This also made it easier in transporting the culture and putting it in the 9 milk samples that required it.
The culture was mixed thoroughly with the milk using a sterilized wooden splint till all the culture was dissolved.
pH was taken using a pH probe, for the first hour, taken after every 20minutes then after that, after every hour for a total of 7hours.
NOTE:
The pH of the milk was determined by inserting the Vernier pH probe directly into the milk before the Thermophilic culture was added into the milk and also during the fermentation process of all the 18 samples. pH of the milk without the Thermophilic starter culture was taken beforehand so as to avoid contamination with the ones that contained the Thermophilic starter culture. Between pH measurements of each and every sample, the pH electrode was rinsed with distilled water and wiped with tissue after each measurement was taken because the glass bulb in the pH probe is sensitive to heat so I ruled out rinsing it in boiling water.
For the first hour of the experiment, pH was taken after 20minutes. This was done so as to detect how fast the fermentation process was taking place for each and every sample. After the first hour, pH measurements were taken after every hour for the next 6 hours and then finally the pH reading was taken after 24 hours were finalized.
During pH readings, the milk samples were in the water bath at a constant temperature.
Biology 代写 生物论文代写
RESULTS:
Samples with starter culture
During the first hour of the experiment all the milk sample pH’s dropped significantly with the Unpasteurized milk having the highest drop in pH of 2.25 while the Pasteurized milk came in 2nd with a pH drop of 2.18 and the UHT milk came in 3rd with a pH drop of 2.16. So far, this is what I would expect considering the heat treatment (or lack of) that these milk samples have gone through.
However when the experiment goes on steadily with a decrease in pH, I noticed that some fluctuations in pH occurred between the 4th and 7th hour. No fluctuation in pH was seen in the UHT milk.
In the first 7 hours of the experiment the highest drop in pH came from the Unpasteurized milk with a pH drop of 2.59, then the UHT milk came in 2nd with a pH drop of 2.53 while the Pasteurized milk came in 3rd with a pH drop of 2.50.
In 24hours all the pH’s seemed to settle(as there was no evident change in pH) and the highest drop in pH came from the Unpasteurized milk with a pH drop of 3.03 settling at a pH of 4.12, the Pasteurized milk coming in 2nd with a pH drop of 2.77 settling at a pH of 4.23 and the UHT milk coming in 3rd with a pH drop of 2.66 settling at a pH of 4.27.
Samples without starter culture
During the first hour of the experiment, the pH does drop in the first 20 minutes but the drop in pH in the full hour is minimal. In the case of the Unpasteurized milk, the pH in the first 20 minutes does drop by a pH of 0.35 but there is no change till after the 1st hour. In the case of the Pasteurized and UHT milk there are some fluctuations in the pH of the milk in the 1st hour. In the 3 hours during the experiment, fluctuations are seen to occur in all the milk samples and after that the pH of the milk samples decrease as per normal.
In the first 7 hours the highest drop in pH came from both the Unpasteurized and the UHT milk with a pH drop of 1.37, the Pasteurized milk had a pH drop of 1.09.
After the 24hours, the highest drop in pH came from both the Unpasteurized and the UHT milk with a pH drop of 2.74 while the Pasteurized milk had a pH drop of 2.72.
DISCUSSION:
The addition of a starter culture, in theory, is meant to increase the rate at which the different types of milk samples ferment, because the enzymes in the bacteria of the starter culture are biological catalysts that are made up of proteins and are used to speed up chemical reactions. This in practice is true, using the results obtained during the experiment. The Thermophilic starter culture increases the rate at which the lactose in the milk is turned into lactic acid therefore decreasing the time it would take the different milk samples to ferment naturally.
To make yoghurt a Thermophilic starter culture is used that normally has a combination of both Lactobacillus bulgaricus and Streptococcus thermphilus. The growth of these two bacteria is independent but when combined and used together, the rate of production of lactic acid from the lactose in the milk is much higher than when they are used individually. The growth of Streptococcus thermphilus occurs much faster and produces both acid and carbon dioxide. The formate and carbon dioxide produced stimulates the growth of the Lactobacillus bulgaricus. The proteolytic activity of the Lactobacillus bulgaricus produces stimulatory peptides and amino acids used by the Streptococcus thermphilus. The lactic acid produced during milk fermentation causes proteins to coagulate leading to curds and a thickening of the milk texture. A further increase in the lactic acid in the milk causes a further drop in pH. The initial drop in pH is caused by the Streptococcus thermphilus to pH values of around 5.0. Using the results obtained this is true as all the milk samples had a drop in pH of values around 5.0. The bacterial growth of Lactobacillus bulgaricus is responsible for the further decrease in pH to values around 4.0. This information is consistent with the results I got from the experiment although there was no way I would have been able to know when each of the bacteria was growing.
The fermentation process of the milk samples without the Thermophilic starter culture occurred more slowly. Natural souring depends on the heat treatment or lack thereof of the milk. In theory it is said that milk contains bacteria and depending on the heat treatment of the milk, the souring/fermentation process will take place at different rates. For instance, the Unpasteurized milk hasn’t gone through any kind of heat treatment this would therefore increase the rate at which it sours/ferments as it contains more bacteria in it that will be used to convert more lactose into lactic acid. The HTST Pasteurized milk has undergone heat treatment but this heat treatment kills the bacteria that cause disease and not all the bacteria in the milk will be removed. This means that the milk will sour but at a slower rate than the Unpasteurized milk. In the case of the UHT milk, the heat treatment this milk undergone virtually kills all the bacteria in the milk so I wouldn’t expect it to sour, but it does. This is because as long as the UHT milk is kept sealed, it will not spoil but when it is left in open air it will spoil (a condition the manufacturers’ state on packages). This is because bacteria enter from the air and also through contamination by the probe. The bacteria from the air (and/from the probe) would cause a much slower souring process than in the Pasteurized milk. This means that I would expect the UHT milk to be the slowest fermentation/souring process in the experiment.
Fluctuations were recorded in the Unpasteurized and Pasteurized milk samples that contained the Thermophilic starter culture and in all the milk samples that didn’t have the Thermophilic starter culture. The fluctuations in these milk samples could be due to a possible increase and decrease in microbial activity. Fluctuations in pH could mean that the H+ ions are being used up in some other biochemical processes or just maybe that the probe was faulty!
Using the H+ concentration graphs, we note that there is barely any increase in H+ concentration in the milk samples without the culture. This is because the rate of fermentation is affected by the concentration of reactants and bacteria number (and hence enzyme concentration). A sharp rise occurs after 7 hours because bacteria got in and reproduced causing concentration to increase and speed up the reaction.
Biology 代写 生物论文代写
CONCLUSION:
When I began the experiment I assumed that if I started measuring the pH changes of the milk WITHOUT the Thermophilic starter culture then measure the pH of the milk WITH the Thermophilic starter culture and use the time interval between the next reading as a scope that would be able to eliminate the chance of any bacteria/ Thermophilic starter culture to be caught in the probe as I would make sure that I sterilized the probe with distilled water each time after I took pH values of the milk and note the expected pH of the distilled water (when I placed the pH probe in the distilled water).
The inconsistency of the results obtained during the experiment and the assumptions made before came from the rate at which the individual milk types ferment. I had previously assumed that the rate of fermentation of each individual milk type would be fastest in the Unpasteurized milk then the Pasteurized milk and then followed by the UHT milk due to their different (or lack thereof) heat treatments they had undergone through. My experimental results contradicted the theory behind it leaving my experiment with a number of flaws that could have caused this contradiction.
From research, when UHT milk is left in open air it will spoil and this is actually a condition the UHT milk processors state on their packaging and as long as the UHT milk is sealed it will not spoil. However this probably doesn’t explain why it fermented as fast as it did. The only possible explanation for this would have been that there was contamination between the samples caused by the use of the pH probe, ineffective sterilization or very rapid entry of bacteria from the air. I could distinguish between these possibilities by repeating with a more effective sterilization technique.
I will now look at the possible weakness and improvements that could be used to improve this experiment.
WEAKNESSES AND LIMITATIONS
IMPROVEMENTS
Distilled water was used to clean the pH during the experiment and this could have proved inefficient this is because the bacteria/culture that was on the pH probe wouldn’t have been killed in between pH measurements.
Use of alcohol/ ethanol as a form of sterilization. One would need to use 70% ethanol mixed in 30% water because the water helps to penetrate the bacteria cell wall around the pH probe. If one uses 100% ethanol, the bacterial cell wall could resist the effect of the ethanol.
Use of 1 pH probe during the experiment due to the fact that the laboratory only had 1 that I could use at the time. This made taking pH readings and measurements difficult as I had to keep sterilizing the probe all the time. This also increased the chances of contamination in the milk.
The use of at least 6 pH probes so as to find the pH changes for the 3 milk samples with the culture and the 3 milk samples without the culture. This would avoid contamination of the culture to the milk samples without the culture and also cross contamination between the different types of milk. These pH probes would need to be calibrated using the same standard solution so as to increase result accuracy. Or running the trials successively which would greatly reduce the chances of contamination
Lactose content wasn’t stated on the different types of milk and this proved to be a problem because lactose content does affect the rate at which milk ferments. Higher lactose content in one of the milks results in the increase in the rate of fermentation and vice versa and this could have had a clear effect on my results.
Use milk types that have an indication of the lactose content in them. Also if the lactose content isn’t specified I could measure the lactose content by the use of a polarimetry.
The different milk types were taken from 3 different dairy plants. This could affect my results in the experiment as these milk types would have gone through different methods of heat treatments some more extreme than others and also using different types of cows to produce these milks
I could have obtained the different milk types from the same dairy plant making sure that the same cow was used to obtain the milk before any kind of heat treatment occurred. Also I could have done the heat treatment myself and be as accurate as possible to heat to the accepted temperatures. The only limitation with that is that the UHT milk goes through such a high heat treatment that it would be difficult to heat the milk that high temperature.
Immediately the culture was added it was difficult to take the pH measurements and it meant that some of the milk samples had a ‘head start’ in the fermentation process.
This would have been improved with the use of the 6 pH probes I mentioned earlier.
There were some successes during the experiment such as how I managed to keep the temperature of all the milk samples the same using a water bath. Also the general prediction that milk would ferment fastest with the addition of the culture was supported
However some questions remain unanswered even after the amount of research I did, eg, was there contamination? Where did it come from and how did it affect my results. I couldn’t find a good enough explanation for the fluctuation of pH during the experiment. During my research the process of putrefaction came up as a cause for the fluctuations but I didn’t understand how the decay process could be involved in the fermentation processes as I would assume that it would only occur after the fermentation process is complete. This is one of the phenomena’s that I didn’t understand and I’m hoping that with this Extended Essay I could inspire someone to explore the possible answers for it.
Biology 代写 生物论文代写