LAB 4: SOUCES OF CONTAMINATION AND INFECTION

SCHOOL OF TECHNOLOGY INDUSTRY

UNIVERSITY SCIENCES MALAYSIA

DEGREE IN BIOPROCESS

IBG 102

BIOLOGY FOR TECHNOLOGIST

Name	1.     LOH SHI WEI (137602) 2.     LAI CHONG SING (137592) 3.     SITI NORASYIKIN BINTI SALMI (137673) 4. SITI NUR SUHAILI AFIQAH BINTI SARIMAN (137674) 5.     NUR LIYANA ATHILAH BINTI MOHD AFFANDI (137636)
Title	LAB 4: SOUCES OF CONTAMINATION AND INFECTION
Date of practical	03/10/17
Date of Report Submitted	10/10/17
Lecturer	DR. TYE

LAB 4: SOUCES OF CONTAMINATION AND INFECTION

INTRODUCTION

Airborne microorganisms are usually carried on dust particles. The sources of contamination which can infect the human’s health could be come from food, water, chemical substances, building area and etc. Mostly, bacteria, viruses, yeasts, fungi, and protozoa are the microbiological pathogen which could cause contamination and thus infections. If the victim is infected from the disease, he could spread the virus through the air or skin contact to other people.

Every human is colonized by billions types of microorganisms. There are two types of microorganisms found in human body which are resident microorganisms and transient microorganisms.

Resident microorganisms are the microorganisms which stay permanent to our skin. They are not removed by routine hand-wash. They are normally colonised on the surface of the skin, mucous membranes, digestive tract, upper respiratory system and distal portion of the urogenital system throughout life. They do not cause any harm to the human’s health but can protect the human from the disease. Example of resident bacteria are Staphylococcus epidermidis which residue in the skin and lactobacillus which residue in the intestines

Transient microorganisms are the microorganisms which can pick up from the environment. These include bacteria, fungi and viruses, which reach the hands, for example, by direct skin-to-skin contact or indirectly via objects.  Basically, they do not produce disease and establish themselves permanently on the surface as they are weak to compete for nutrients compare to resident microorganisms. However, if the resident microorganisms are disturbed, transient microorganisms may colonize, proliferate and cause disease.

Basically both types of microorganisms do not cause harm to our body. They become opportunistic pathogens only when immune suppression of the host, reduction in microbial antagonism, and introduction of normal microbiota into an unusual site in the body.

OBJECTIVE

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

MATERIALS AND REAGENTS

Molten nutrient agar

Sterile water

Sterile petri dishes

Sterile clinical swab

Pipette and tips

PROCEDURE

Air:

1.      The molten agar was poured into sterile petri dish and cool.

2.      The lid of petri dish was removed and leave it resting, faced down on the side of the plate. The plates was exposed for 5 minutes.

3.      The lid of plate was replaced and incubated at 37ᵒC for 48hours.

Hands:

1.      Sterile water was used to wash hand. No soap was used.

2.      Automatic pipette was used to transfer 1ml of wash water to the petri dish.

3.      Molten nutrient agar was added to petri dish.

4.      The lid of the petri dish was replaced and the petri dish was gently rotated until water and agar was thoroughly mixed. The agar was not allowed to contact with the lid of petri dish.

5.      The petri dish was inverted after the agar has set and incubated at 37ᵒC for 48hours.

Ear:

1.      The molten agar was poured into sterile petri dish and cool.

2.      A sterile swab was used to rub into ear of the subject with extreme care.

3.      The swab was used to inoculate the labelled plate. Streak method was used.

4.      Petri dish was incubated at 37ᵒC for 48hours.

Normal breathing:

1.      The molten agar was poured into sterile petri dish and was cooled.

2.      The lid was removed and the plate was hold about 15cm from mouth. Directly breathe normally onto the plate for 1 minute. The lid was replaced.

3.      Petri dish was incubated at 37ᵒC for 48hours.

Violent cough:

1.      The molten agar was poured into sterile petri dish and was cooled.

2.      The lid was removed and the plate was hold about 15cm from mouth. Cough violently onto the plate. The lid was replaced.

3.      Petri dish was incubated at 37ᵒC for 48hours.

RESULT

TEST	SELF-MADE MEDIA	COMMERCIAL MEDIA
Air
Hands
Ears
Normal breathing
Violent coughing

Morphology of Bacteria Colonies	Air	hands	Ears	Normal breathing	Violent coughing
Elevation	Flat, pulvinate, umbonate	Raised, flat, umbonate	Flat, raised	Flat, umbonate	Flat, pulvinase
Form	Circular, filamentous	Circular, rhizoid, irregular	Circular, filamentous, irregular	Circular	Circular
Surface	Shiny and smooth,	Shiny and smooth,	Dull and wrinkled	Shiny and smooth	Shiny and smooth
Texture	Dry	Moist	Moist	Dry	Dry
Colour	Buff	Buff	Buff, white	Buff	Buff
Opacity	Opaque	Opaque, cloudy	Opaque, cloudy	Opaque, cloudy	Clear
Margin	Entire, filiform,	Entire Lobate, filiform	undulate	Filiform	Filiform

TABLE 5.2: result of the morphology of bacterial growth on self-made media

Morphology of bacteria colonies	Air	Hands	Ears	Normal breathing	Violent coughing
Elevation	Flat, pulvinate, umboute	Raised, flat, umbonate	Flat, raised	Flat,	Flat
Form	Circular,  filamentous	Circular, rhizoid, irregular	Irregular,  circular	Circular	Circular, filamentous
Surface	Shiny and smooth, dull and wrinkled	Shiny and smooth, dull and rough	Dull and wrinkled	Shiny and smooth	Shiny and smooth
Texture	Dry	Moist	Moist	Dry	Dry
Colours	Buff	Buff and white	Buff	White	Buff
Opacity	Opaque	Clear	Opaque and cloudy	Opaque, cloudy	Clear
Margin	Entire, filiform, undulate	Entire, filiform	Undulate	Filiform	Entire,

TABLE 5.3: result of the morphology of bacterial growth on commercial media

DISCUSSION

The culture medium provides the nutrients for bacteria grow. Colonies with difference appearance is produced by different types of bacteria. Scientists use colony morphology to identify the type of bacteria growing on agar in the Petri dish. Each distinct colony represents an individual bacterial cell or group that has divided repeatedly. Being kept in one place, the resulting cells have accumulated to form a visible patch.

There are a few basic characteristic of colony morphology that are typically evaluated:

Elevation – This describes the side view of a colony. For example, flat, raised, umbonate, convex, pulvinate and crateriform

Form – The basic shape of the colony. For example, circular, filamentous, irregular and rhizoid

Size – The diameter of the colony. Tiny colonies are referred to as punctiform.

Surface – The surface of the colony appear. For example smooth, shiny, rough, wrinkled, dull or glistening.

Texture – The colony is dry, moist, mucoid, brittle or viscous.

Colour – Colony pigmentation. For example white, buff, red, purple, orange, yellow.

Opacity – For example, transparent, opaque, translucent, cloudy or iridescent.

Margin – The edge of a colony such as entire, undulate, lobate, curled or filifor

 (A)AIR

           Air mainly is transport medium for microorganisms. The most common genera of fungi in indoor air are Penicillium Aspergillus while the most common genera of bacteria found in indoor air are Staphylococci, Bacillus and Clostridium.

           Generally the colonies are distributed uniformly and similarly between both self-made agar and commercial agar. Most of the colonies in both types of agar appears to be flat or pulvinate, circular or filamentous, shiny and smooth, dry, buff, entire or filiform. There’s one colonies in commercial agar appears to be dull and wrinkled which is not found in self-made agar.

(B) HAND

This experiment shows that our hands contain lots of microorganisms, the main pathogenic bacteria which are more likely to be found on the hands are Staphylococcus, Corynebacteria, Streptococcus, E coli, Myobacteria, and Haemophilus.

Larger colonies found in self-made media than the colonies found in commercial media.

The colonies found in both media are raised, flat, umbonate, circular, rhizoid, irregular, shiny and smooth, moist, buff, entire, filiform. However, the colonies found in self-made media is opaque, cloudy but the colonies found in commercial agar is clear.

(C) EAR

           Since ears are exposed to the outside environment despite the best efforts of the ceruminous gland, the healthy outer ear still houses a variety of microbes. Staphylococcus epidermis, Turicellaotitidis, Alloiococousotitis, Pseudomonas aeruginosa, Corynebacterium, Staphylococcus aureus, and Streptococcus saprophyticum are some of the most common bacteria. The most common fungal microbe known to reside in the ear is Candida albicans. Microbes that are known to inhabit the middle ear are Streptococci, Haemophilus pneumoniae, Moraxella catarrhalis, and Mycobacterium.

           Large number of colonies are found in both self-made media and commercial media. The colonies found in both media are flat or raised, circular, irregular, dull and wrinkled, moist, opaque or cloudy and undulate. The colonies found in both media are similar but generally more and larger colonies are found in commercial media

(D) NORMAL BREATHING

The presence of colonies in the culture media shows that microorganisms can also be found in the mouth and nasal cavity. Compared to other culture media, there is relatively low number of colonies from the contamination of normal breathing. Examples of microorganisms areStreptococcus, Haemophilus, Micrococcus and Corynebacterium diphtheriae.

There’s only some colonies found in both self-made media and commercial media. The colonies isolated in both self-made media and commercial media are flat, circular, dry, white, shiny and smooth, opaque or cloudy and filiform.

(E) VIOLENT COUGHING

The observations are similar to that from the normal breathing due to same sources of contamination that is mouth and nasal cavity. However, coughing forces more air out of the mouth and therefore slightly more colonies are found in the culture media. Examples of microorganisms are Streptococcus pneumoniae, Streptococcus salivarius and Staphylococcus epidermidi

The colonies isolated in both self-made media and commercial media of violent coughing is similar to normal breathing, they are flat, circular, dry, shiny and smooth, and filiform. However, white, opaque or cloudy colonies are not found but clear is observed.

CONCLUSION
In conclusion, the objective of the experiment which is to determine the microorganisms in the air and from healthy humans is achieved as growth of microorganisms can be observed on the media. 

REFERENCES

http://microbeonline.com/colony-morphology-bacteria-describe-bacterial-colonies/

http://delrio.dcccd.edu/jreynolds/microbiology/2421/lab_manual/colony_morph.pdf

http://www.iknow2.net/what-kinds-of-bacteria-are-on-your-hands/

http://microbiologyonline.org/teachers/observing-microbes/observing-bacteria-in-a-petri-dish

https://www.scribd.com/doc/19079015/Air-Microbiology-2009