OBJECTIVES

1. To conduct serial dilutions and plating (pour plates and spread plates) to establish viable
bacterial cell count.
2. To perform an aseptic and free contamination experiment of microbial load by using
sterilization and aseptic technique when handling the lab supplies, media, reagents and
cultures.
3. To understand plate count technique in order to determine the Colony Forming Units
(CFUs) in the dilution made.

QUESTIONS
1. Discuss the importance in determining the microbial loading in a food sample.
Microbial microorganism population in food can depends on the level of sanitation used at
all phases, processing and preservation methods used to kill and prevent microorganisms growth
and the degree of abuse in which can leads to microbial growth for example temperature abuse in
preparation of food which can lead to food spoilage which happened due to presence of
microorganisms. Microorganism that present in food leads to contamination in which mostly due
to poor sanitation during preparation handling and processing process.
The importance of determining microbial loading in food sample which is to conduct
microbiological examination towards foods and food ingredients is to help in assessing the food
safety to consumers, to study about their stability and shelf life under normal storage conditions
and the level of sanitation used during handling. Furthermore, microbial load can be important in
determining whether food ingredient or food that is prepared meet acceptable standards, guidelines
and specifications.

2. You are a process engineer and have developed a bioreactor for microbiological
production of bioplastic by using bacterial cells, Pseudomonas sp. The determination of
microbial growth gives detailed insight to monitor and optimize the bioreactor so that the
bioreactor produces the maximum amount of bioplastics. Explain how you can determine
the growth of Pseudomonas sp. in your bioreactor for a period of 48 hours.
Pseudomonas are motile, rod shaped and a gram negative aerobe bacteria. They are found almost
everywhere in water, plant, animals and in soil where in most cases it is not pathogenic and can be
put into benefit.
Pseudomonas can be tested by using several microbial tests. Pseudomonas gives negative Voges
Proskauer, indole and methyl red tests, but a positive catalase test. While some species show a
negative reaction in the oxidase test, most species, including P. fluorescens, give a positive result.
3. Certain Pseudomonas species may also produce additional pigments, such as pyocyanin (blue
pigment, a siderophore) by P. aeruginosa4, quinolobactin (yellow, dark green in presence of iron,
a siderophore) by P. fluorescens5, a reddish pigment called pyorubrin and pyomelanin (brown
pigment). On blood agar a hemolytic reaction can be observed. (Sigma Aldrich, 2017)