Group's members

Before exploring our blog, let's introduce ourselves first........


Hi, I am Teoh Leong Sin. XD

Hi, I am Ooi Sin Ying.(",)

Wow, is my turn, I am Tan Hwee Feng. (~^^~)

Welcome to our blog, I am Tang Su Xian. (^6^)

Thursday, 19 April 2012

Lab 4 report written by TANG SU XIAN

Name: Tang Su Xian
Matric's card no.: 111431




Lab 4 : Sources of contamination and infection
Introduction
Contamination is the presence of a minor and unwanted constituent (contaminant) in material, physical body, natural environment, at workplace and so on. In biological sciences, accidental introduction of “foreign” material (contamination) can seriously distort the results of experiments where small samples are used. In cases where the contaminant is a living microorganism, it can often multiply and take over the experiment, especially cultures and render them useless.

An infection is the invasion of body tissues by disease-causing microorganisms, their multiplication and the reaction of body tissues to these microorganisms and the toxins that they produce.  Infections are caused by microorganisms such as viruses, prions, bacteria, and viroids, though larger organisms like macroparasites and fungi can also infect.

  Airborne microorganisms are usually carried on dust particles, although some (fungal spores, for example) may be carried directly by air currents. It is important for microbiologists to be aware of the potential for contamination by airborne microorganisms. Carefully observation of simple precautions dramatically reduces the risk of contamination of the cultures.

  Every human is colonized by billions of microorganisms. These microorganisms, some of which are vital to our wellbeing, constitute our resident or normal microflora. Resident microorganisms are nourished by the chemicals and moisture excreted by human body. In moist areas such as the armpit, there may be one million bacteria per centimeter cube; on the drier skin of the forearm, there may be 10000 bacteria per centimeter cube.

  Resident microorganisms are either nonpathogenic or are prevented from infecting the body by an array of mechanical and chemical defences. Some resident microbes are, hower , opportunistic pathogens which may cause infection if the body’s defences are breached. For example, if the skin is broken.

  Transient microorganisms are picked up from our environment. For instance,from faecal contact or from soil and usually fail to become permanent skin residents. One of the most important reasons for failure to gain permanence is that the established residents are better able to compete for nutrients. Since transients generally originate in the other environments, they are poorly adapted to conditions on the skin and usually disappear within 24 hours of arrival.

Objective
To determine the microorganisms in the air and from healthy humans.

Results:

1.) Colonies on ear

2.) Colonies in air
3.) Colonies from violent coughing





4.) Colonies in normal breathing


5.) Colonies on hands


Discussions:
1.) In this experiment, there are some precaution steps to be taken:
 #Spray some Lysol solution on the table and rub it with a sponge before the experiment is conducted.
#The sterile Petri dishes must be opened near the Bunsen burner to avoid contamination.
# The Petri dishes containing culture medium agar and the hand-washed water should be rotated lightly to make sure that the contents mixed together.

2.)  Atmosphere contains all major groups of microbes ranging from algae to the viruses. In addition to gases, dust particles and water vapour, air also contains microorganisms. There are vegetative cells and spores of bacteria, fungi and algae, viruses and protozoan cysts. Since air is often exposed to sunlight, it has a higher temperature and less moisture. So, most of these microbial forms will die.

3.) Although bacteria consist of only one single cell, they're an amazingly complex group of living creatures. They can live in extreme temperatures, ranging from above the boiling point to low temperatures that would freeze human blood. Bacteria "eat" everything from sugar and starch to sunlight, sulfur and iron. There's even a species of bacteria" Deinococcus radiodurans" that can withstand blasts of radiation 1,000 times greater than would kill a human being. The entire ecosystem depends heavily upon the activity of bacteria. In their various forms, they're crucial for recycling organic matter such as leaves that fall off trees. Others digest some food within the human body that would otherwise be difficult to break down. Most bacteria fall into one of three groups, namely aerobic bacteria, anaerobic bacteria and facultative anaerobes. The most common bacteria in the air are E.coli , S.marcesens, S.aureus.

3.)  Fungi are neither animals nor plants - in fact, they command one of the "Five Kingdoms of Life" all to themselves. There are over 100,000 species of fungi, but only about 400 cause diseases relevant to man, animals, or plants. Essentially, Fungi are plant-like organisms that lack chlorophyll. Without chlorophyll, which transforms light into energy, fungi must absorb food from others. Since they don't use light to make food, fungi can live in damp and dark places. Many strains of fungi are extremely useful. The job of Fungi is to "eat" things when they are dead - a kind of natural recycling system. Fungi find nutrients within our rubbish -they eat the waste and turn it into soil. There are also "good" fungi such as the mushrooms we eat, and the yeast we use to make both bread and beer. Bad fungus is just good fungus trying to do its job far too early - in other words, trying to eat before their food is actually dead. This includes the nutrients from the tissue of human beings (as in tinea or ringworm) and inside the lung and nasal passages (which causes diseases such as sinusitis).

4.) Mould is the common name for fungi that grow in a filamentous (thread-like) fashion and reproduce by means of spores; all moulds are fungi, but not all fungi are considered "moulds". There are over 20,000 known species of moulds. The filaments of mould are known as hyphae. At the tips are rounded, blob-like forms called conidia or spores. These round fungal forms are relatively metabolically inactive - all they're doing is trying to find a nice, warm, moist place to live and multiply. Moulds reproduce by making spores. Spores are microscopic; they vary in shape and size (2-100 micrometers - a micrometer is one millionth of a metre). The spores allow the organism to survive during periods where the conditions aren't ideal for growth (for example, some species don't like dry, cool environments). Spores travel in several ways in search of a place to live - passively (by wind, for example), when disturbed (by a person), or when actively discharged by the mould (usually under moist conditions or high humidity). Mould spores waft through the indoor and outdoor air continually. When mould spores land on a damp spot indoors, they begin eating whatever they are growing on. Moulds can grow on virtually any organic substance, such as oil, dust, compact discs (CD's), skin cells and dirt providing moisture and oxygen are present.


 5.)Microcoocus is a Gram-positive, aerobic bacterium which is a member of the Micrococcaceae family. Micrococcus cells can be observed under the microscope as spherical cells forming pairs or clusters. If cultured in broth or on nutrient agar, the colonies may be red or yellow when observed unstained. Although these bacteria are a common human skin contaminant, they are relatively harmless to humans because they maintain a saprophytic lifestyle. They can also be found in freshwater environments or in soil. Three common species of Micrococcus are M. luteus, M.roseus, and M. varians.

6.) Diseases spread through indirect contact with respiratory secretions. Microorganisms that may be transmitted through this route include influenza, Streptococcus, respiratory syncytial virus (RSV) and the common cold. Because these diseases may be spread indirectly by hands contaminated by respiratory discharges of infected people, illness may be avoided by washing hands after coughing or sneezing and after shaking hands with an individual who has been coughing and sneezing.  Diseases may also be spread when hands are contaminated with urine, saliva or other moist body substances. Microorganisms, which may be transmitted by one or more body substances, include cytomegalovirus, typhoid, staphylococcal organisms, and Epstein-Barr virus. These germs may be transmitted from person to person or indirectly by contamination of food or inanimate objects such as toys.

7.) Bordetella organisms are small, Gram-negative coccobacilli which are strict aerobes. The three species of this genus vary in motility and certain biochemical characteristics. The most important species in this genus is B. pertussis, the organism which causes whooping cough. This highly contagious bacterium makes its way into 53 respiratory tract via inhalation and subsequently binds to and destroys the ciliated epithelial cells of the trachea and bronchi. It does this through the use of several toxins: Pertussis toxin, Tracheal cytotoxin, Hemoagglutinin.


Conclusion

In conclusion, there are various microorganisms in the air and human's bodies. From this experiment,  i found that there are various microorganisms in air. From the morphology of the colonies, i concluded that the culture might contaminated with the fungus and bacteria as they exposure to the air. 

References




Monday, 16 April 2012

LAB 4 : SOURCES OF CONTAMINATION AND INFECTION

Procedure :

Step 1 : Sterilizer work bench with 70% ethanol.

Step 2 : Lighten the bunsen burner



















All events is done near the bunsen burner.

AIR , EAR , NORMAL BREATHING , VIOLENT COUGHING

Step 1 : Take the molten agar and pour in petri dish and cool.



Step 2 :

AIR - Remove lid, leave it for 5 minutes and replace the lid.


EAR - Rub a sterile swab moistened with sterile isotonic solution into ear and use the swab to
           inoculate the labeled plate.
  
NORMAL BREATHING -  Remove the lid, hold about 15cm from mouth and breathe normally and
                                              directly onto the plate for1 minute.




















VIOLENT COUGHING - Remove the lid, hold the plate about 15 cm from mouth and cough
                                           violently onto the agar.

Step 4 : Replace the lid and incubate at 37ºC for 48 hour.


HANDS :

Step 1 : Wash hand with distilled water, take 1ml of wash water to the petri dish.

Step 2 : Pour the molten agar into petri dish.


Step 3 : Replace the lids of petri dish and rotatethe dish until the wash water is thoroughly mixed
             with the molten agar.

Step 4 : After the agar has set, invert the dish and incubate at 37ºC for 48 hours.

Let share our  LAB 4 REPORT ...^^

Tuesday, 10 April 2012

Lab 3 report prepared by Teoh Leong Sin


Name : Teoh Leong Sin
Matric No : 111433

Introduction

Culture media are available commercially as powders; the require only addition of water. Nutrient medium is a general preparation for culturing microorganisms which are not nutritionally fastidious.The broth contains:
3.0 g/L "Lab-lemco" powder (a beef extract)
2.0 g/L yeast extract
5.0 g/L peptone (a nitrogen source)
5.0 g/L sodium chloride
15.0 g/L agar powder
The agar has the same composition, except that it contains 15g/L agar. The final pH of both media is 7.4.
     Autoclaving is a process that use moist heat and pressure so that all parts of the material to be sterilized reach 121 degree C for 15 minutes. An autoclave is, in essence, a large pressure cooker; a chamber which may be sealed off against surrounding air. Materials for sterilization are placed in the chamber, the door is sealed, and pressurized steam is forced into the chamber. The incoming steam displaces cooler air through an exhaust valve; this valve closes when the cell cooler air has been vented.
     Steam is continually forced into the chamber until the pressure reaches 103kPa above atmospheric pressure; at sea level, this pushes the temperature in the chamber to 121 degree C. The high pressure prevent solutions from boiling over at this temperature. Larger volumes require longer than 15 minutes to heat up to 121 degree C throughout. After sterilization, the steam pressure is slowly decreased to atmospheric pressure. The sterilized objects can then be removed.


Objective

To prepare sterile nutrient agar for culturing microorganisms.


Discussion
1) Precaution steps of preparing culture medium
-weigh appropriate amount of broth and then mix it into a beaker. After that, add 200mL of distilled water into the beaker 
-heat up the solution and stir it by using a rod to let the culture medium completely dissolve
-After boil,pour the culture medium into Scott bottle and loosely recap the bottle.
-Sterilize the culture medium by using autoclaving method
- After sterilization, the steam pressure is slowly decreased to atmospheric pressure. Scott bottles is removed from the autoclave machine and cooled it in laminar air flow cabinet




2) Autoclaving method
-a most effective and most efficient method of sterilization.
-culture medium steamed under high pressure at a standard temperature of 121 degree celcius for 15 minutes.
-time to autoclaving depend on the volumes of culture medium.
-when proper condition and times are employed, there are no living organisms will survive.

3) Laminar flow
-physical containment devices that act as primary barrier to protect from contamination and infection 
-horizontal-flow clean bench and biological safety cabinet
-Horizontal laminar flow use a high effiency particulate air (HEPA) filter and blowers that generate a non-mixing stream of air 
-Vertical laminar flow(biological safety cabinet)  are used for work with low-risk substances and materials, when protection of working material from environment is required or work with item requires a sterile working zone. Cabinets are used for equipment of local workplaces in medical, pharmaceutical and other institutes with high requirements for air cleanliness in the working zone.



 Conclusion
The best way of sterilization is autoclaving, it can completely inactivate all microorganism  in the medium. 



References
http://en.wikipedia.org/wiki/Autoclave 
http://www.ehow.com/how_4913374_use-autoclave.html
http://en.wikipedia.org/wiki/Laminar_flow_cabinet
http://www.terrauniversal.com/laminar-flow-hoods/vertical-laminar-flow-hoods.php











Lab 3 Report written by Tan Hwee Feng

Name : TAN HWEE FENG
Matric No : 111427

Introduction

Culture media are available commercially as powders; the require only addition of water. Nutrient medium is a general preparation for culturing microorganisms which are not nutritionally fastidious.The broth contains:
3.0 g/L "Lab-lemco" powder (a beef extract)
2.0 g/L yeast extract
5.0 g/L peptone (a nitrogen source)
5.0 g/L sodium chloride
15.0 g/L agar powder
The agar has the same composition, except that it contains 15g/L agar. The final pH of both media is 7.4.
     Autoclaving is a process that use moist heat and pressure so that all parts of the material to be sterilized reach 121 degree C for 15 minutes. An autoclave is, in essence, a large pressure cooker; a chamber which may be sealed off against surrounding air. Materials for sterilization are placed in the chamber, the door is sealed, and pressurized steam is forced into the chamber. The incoming steam displaces cooler air through an exhaust valve; this valve closes when the cell cooler air has been vented.
     Steam is continually forced into the chamber until the pressure reaches 103kPa above atmospheric pressure; at sea level, this pushes the temperature in the chamber to 121 degree C. The high pressure prevent solutions from boiling over at this temperature. Larger volumes require longer than 15 minutes to heat up to 121 degree C throughout. After sterilization, the steam pressure is slowly decreased to atmospheric pressure. The sterilized objects can then be removed.

Objective

To prepare sterile nutrient agar for culturing microorganisms.


Discussions

1) We need to weigh and mix all the ingredients into the beaker to prepare culture medium. After  mixing the ingredients, we need to add 200 mL distilled water into the beaker which has been  measured using measuring cylinder.

2) After preparing culture medium solution, we heat up culture medium until it boils. While heating, we use rod to stir the media so that the all powder form culture medium dissolved in water. Next, we pour whole culture medium into Scott bottle and loosely recap the bottle.

3) Culture medium in Scott bottle need to be autoclaved. Autoclaving is most effective and most efficient means of heat sterilization.Culture medium should be sterilized reached 121 degree C for 15 minutes to ensure there are no microorganisms contamination.

4) The autoclave operates by using steam under high pressure as sterilizing agent. High pressure enable steam to reach high temperature which can increase heat content and killing power.

5) After autoclaving, Scott bottles were removed from autoclave machine. Sterile media is then put into vertical laminar flow cabinet ( biological safety cabinet) for the following process.

6) Vertical laminar flow cabinet provide a clean, safe environment for sterile media. It use a high-efficiency particulate air (HEPA) filter and blowers that generate a non-mixing stream of air.


Conclusion

We are able to prepare culture medium with correct quantities of ingredients. Preparing culture media need to be go through some process such as weighing the ingredients, mixing ingredients with distilled water, heating media until it boils and autoclaving. Finally, we have prepared culture medium successfully.

References

http://www.invitrogen.com/site/us/en/home/References/gibco-cell-culture-basics/cell-culture-equipment/laminar-flow-hood.html
http://www.aircleansystems.com/process_vertical.htm
http://www.cabri.org/guidelines/micro-organisms/M203Ap1.html
http://www.cib.csic.es/en/servicio.php?iddepartamento=23





Sunday, 8 April 2012

Lab Report 3 by Ooi Sin Ying

Name           : Ooi Sin Ying
No. Matrice : 111410

Introduction:
Culture media are available commercially as powders; the require only addition of water. Nutrient medium is a general preparation for culturing microorganisms which are not nutritionally fastidious.
The broth contains:
 - 3.0 g/L "Lab-lemco" powder (a beef extract)
 - 2.0 g/L yeast extract
 - 5.0 g/L peptone (a nitrogen source)
 - 5.0 g/L sodium chloride
 - 15.0 g/L agar powder
The agar has the same composition, except that it contains 15g/L agar.
The final pH of both media is 7.4.
Autoclaving is a process that use moist heat and pressure so that all parts of the material to be sterilized reach 121ºC for 15 minutes. An autoclave is, in essence, a large pressure cooker; a chamber which may be sealed off against surrounding air. Materials for sterilization are placed in the chamber, the door is sealed, and pressurized steam is forced into the chamber. The incoming steam displaces cooler air through an exhaust valve; this valve closes when the cell cooler air has been vented.
Steam is continually forced into the chamber until the pressure reaches 103kPa above atmospheric pressure; at sea level, this pushes the temperature in the chamber to 121ºC. The high pressure prevent solutions from boiling over at this temperature. Larger volumes require longer than 15 minutes to heat up to 121ºC throughout. After sterilization, the steam pressure is slowly decreased to atmospheric pressure. The sterilized objects can then be removed.

Objective :
To prepare sterile nutrient agar for culturing microorganism

Dicussion :
1. Preparation of nutrient medium.
 - Weigh appropriate amount of broth (with agar) powder into Scott bottles and dissolve with 200mL
   of distilled water. Mix well.
 - Loosely recap the bottles and set aside for sterilization.
 - Sterilized all media at 121ºC for 15 minutes.
 - After autoclaving, remove the media. Allow the broth preparation to cool then tighten the cap of
   each bottle.

2. Principle od autoclave
 - most effective and most efficient means of sterilization.
 - Higher temperature ensure more rapid killing.
 - The standard temperature and pressure of autoclave is 121ºC and 15psi.
 - Autoclave operates by using steam under pressure as a sterilizing agent.
 - High pressures enable steam to reach high temperature which increase heat content and killing
    power.
 - Ideal for sterilizing biohazardous waste, surginal dressing, glassware, microbiologic media, liquids.
 - Plastics and certain medical instrument such as fibre-optic endoscopes cannot withstand
    autoclaving.
 - Materials with bioactive compounds cannot be autoclaved.

3.Principle of laminar flow
 - Physical containment devices that acts as primary barriers to protect  from worker-generated or
   environmental sources of contamination and protect the laboratory worker and laboratory
   environment from expose to infectious or other hazardous materials that are present within the
   hood.
 - Two types of laminar flow :
   (a) Horinzontal laminar flow
   (b) Biological safety cabinet
 - Both use a high-efficiency particulate air (HEPA) filter and blowers that generate a non-mixing
   stream of air.
 - Horizontal laminar flow provide a near-sterile environment for clean handling of non hazardous
    material such as sterile media or equipments.
 - Because of the flow of air within the hood directs back to the user and the room, horizontal flow
   hoods are never to be used with infectious agents or toxic chemicals.
 - Flame sterilization should be minimized in laminar flow as the turbulence generated by the flame
   can disturb the sterile air stream.
 - Turn on the blower and UV light without work for about 60 mins before use and after used.


Conclusion :
The nutrient medium is prepared sucessfully.
Autoclaving is a most effective and most efficient means of sterilization.
Laminar cabinet can provide a contamination free condition during experiment.

Reference :
1.http://en.wikipedia.org/wiki/Autoclave
2.http://wiki.answers.com/Q/The_principle_involved_in_the_operation_of_autoclave
3.http://www.globalrph.com/aseptic.htm
4.http://wiki.answers.com/Q/What_is_the_principle_of_Laminar_air_flow

Lab 3 report written by Tang Su Xian

Name: Tang Su Xian
Matric card no.: 111431

Lab 3: Preparation and sterilization of culture media

Introduction:
Culture media are available commercially as powders; they require only the addition of water. Nutrient medium is a general purpose preparation for culturing microorganisms which are not nutritionally fastidious. The broth contains:
3.0g/L “Lab – lemco” power ( a beef extract )
2.0g/L yeast extract
2.0g/L peptone( a nitrogen source)
5.0g/L sodium chloride
15.0g/L agar powder
  
  The ready-made nutrient agar contains 15 g/L agar and the same contents as the manually-made nutrient medium. The final pH of both media has to be 7.4.
    
  Autoclaving is a process that use moist heat and pressure so thst all parts of the material to be sterilized reach 121 ̊C for 15 minutes. An autoclave is, in essence, a large pressure cooker; a chamber which may be sealed off against surrounding air. Materials for sterilization are placed in the chamber, the door is sealed, and pressurixed steam is forced into the chamber. The incoming steam displaces cooler air through an exhaust valve; this valve closes when the cell cooler air has been vented.
  
   Steam is continually forced into the chamber until the pressure reaches 103 kPa above atmospheric pressure; at sea level, this pushes the temperature in the chamber to 121 ̊C. The high pressure prevents solutions from boiling over at this temperature. Larger volumes require longer than 15 minutes to heat up to 121 ̊C throughout. After sterilization, the steam pressure is slowly decreased to atmospheric pressure. The sterilixed objects can then be removed.

Objective:
To prepare sterile nutrient agar for culturing microorganisms

Discussions:
1.)    There are several precaution steps to be taken in this experiment, which include:
a.)    The pan and the balance must be cleaned before weight the culture medium powders. This is to ensure that the culture medium powders are not containminated by other things and can get more accurate amount of culture medium powders.
b.)    Besides, distilled water should be used to mix with the culture medium powders instead of using tap water. This is because the tap water might contain high amount of minerals.
c.)     The culture medium should be stirred during the heating process of culture medium until the medium boils and the colour of medium changes from murky to clear. This is to ensure that all the agar powder has dissolved in the culture medium.
d.)    The caps of two Scott bottles that filled with distilled water and culture medium that have been heated should be recaped loosely before putting into the autoclave machine. This is to prevent the Scott bottles from broke during autoclaving process.
2.)    Sterilization of culture medium
a.)    Although sterilization of culture media is best carried out in a steam autoclave at temperatures between 121-134°C, it has to be recognised that damage is caused to the medium by the heating process.
b.)    It is important, therefore, to optimise the heating process so that a medium is sterile after heating but minimal damage is caused to the ingredients of the medium. As a general rule it is accepted that short-duration, high-temperature processes are more lethal to organisms and less chemically damaging than are longer, lower temperature processes e.g. 3 minutes at 134°C is preferable to 20 minutes at 115°C..  
3.)    When removed the Scott bottles from the autoclave machine,they should be allowed to cool down in a laminar air flow cabinet. Alternatively Scott bottles may be sterilized in a jar which is covered by a piece of felt which effectively protects the Scott bottles from infection by air- borne microorganisms. Caps of Scott bottles are screwed down tightly after the culture medium have been cooled to ambient temperature.
4.)    Laminar air flow is based on the flow of air current to create uniform velocity, along parallel lines, which helps in transforming microbial culture in aseptic conditions. When fresh air is passed in the laminar air flow it replaces the comtamintaed air inside and keeps it contamination free. 

Conclusion:
I have learned the way to prepare and sterilize the culture medium. During the preparation of culture medium, I found that there are a lot of precaution steps are needed to be taken in order to prepare a sterilized culture medium successfully.

References:
http://www.secondaryteachersstore.net/all_about_agar.pdf
http://en.wikipedia.org/wiki/Laminar_flow_cabinet
http://en.wikipedia.org/wiki/Autoclave

LAB 3 : PREPARATION AND STERILIZATION OF CULTURE MEDIA

Procedure :

Step 1 : Weight appropriate amount of broth (with agar )power into beaker and dissolve with 200mL
             of distilled water. Mix well.


Step 2 : Stir the medium while heating until it boils.



















Step 3 : Transfer the medium into scott bottles















Step 4 : Loosely recap the bottles and put into autoclave for sterilization.



Let have a look at our group members' lab 3 report...^^