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What is the purpose of pH adjustment and autoclaving during medium preparation?

pH adjustment
- To neutralize chemical reaction in the solution which affects the physical appearance of culture media and the ability to grow microorganisms.
- pH changes in the cell culture environment can cause organelles malfunction which alter cellular process like metabolism, cell growth, and membrane potential.
-to provide optimal growing environment for the cell or microorganism culture to produce the target product at optimum level.
- Various pH indicator is used to defferientiation the microbes in the culture.

Autoclaving
- Used to decontamination certain biological waste and sterilize media, instruments and lab ware by killing unwanted microorganisms and spores through high temperature and pressure.
- As a final sterilization for culture media.
- To prevent any unwanted microorganism to compete the nutrients with the target cells or microorganisms

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What are the purposes of glucose and peptone in microbiological media

The purposes of glucose in microbiological media are :

-Glucose is a main fuel source to supply energy within microbiological media.

-Energy that contains in glucose is able to synthesize ATP both in the interconnected and independent ways.

-Glucose is able to act as a reducing power that is required in maintaining the pool of NADPH for microbiological culture.

-Glucose provides the reducing power needed to neutralize oxidative species which is oxidative stress that forms in vivo and in vitro.

-Metabolized principally by glycolysis to form pyruvate which may convert to lactate or acetolactate and enter the citric acid cycle to form CO2.

The purposes of peptone in microbiological media are :

-Peptone is a soluble protein that acts to provide primary organic nitrogen source in the media.

-Peptone can also be used as the source of carbohydrates.

-Provide amino acid for the microbes in the media.

-Peptone provides water-soluble minerals and micronutrients for microbiological culture.

-Peptone is a complex mixture that provides crucial nutritional support for the growth and proliferation of microbiological cells in boosting the fermentation process.

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What is the purpose of isolation of microorganisms from their natural environment?

The purpose is to separate one type of microorganisms from other microorganisms to obtain pure cultures. With this, roles of the type of microorganisms can be identified. This may be performed by growing it on solid medium, where microbial cells will form a colony of cells that will remain in place.
Besides that, it also to separate one type of microbe with other microbe derived from a mixture of various microbes. This is because bacteria live as a community with other microbial species, rather than living alone. Hence, the best way to identify and characterize the desirable species of bacteria, as well as to minimize the risk of contamination, is to isolate them from their natural environment.

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What are microbiological test and their importance?

Microbiological test is the use of biological, biochemical, molecular or chemical methods for the detection, identification or enumeration of microorganisms, as well as the characteristics of these microorganisms in the sample. It is often used in food or drink products, environmental or clinical sample to analyse any harmful pathogens, allergens, contaminants, and bacteria such as fungi, mould, E.coli and more. Microbiological testing is very much applied in pharmaceutical centres, healthcare, textiles and food safety. While many microbes are harmless to humans, others can be dangerous. They have the potential to contaminate food, introduce toxins, cause disease and cause a slew of other issues. Therefore, the significance of microbiological testing is in identifying these contaminants quickly and treating them before they cause irreversible damage to the human body. Microbiological test also enable the  quick identify the identity of microorganism and then recognised any contaminants or harm they bring so that the treat can be done before they do irreversible damage. It is important to identify any potentially harmful  contaminants in the sample and this testing is done to make sure that researches are performed in a controlled environment. Microbiological testing are essential as microbes can cause serious problems; food spoilage, inducing toxins, and may cause diseases too. With proper microbiological testing, we can further detect,  analyse any unknown microbes and further treating the contaminants before any irreversible damage. Besides, microbiological testing is also important in verifying environmental and equipment sanitation practices, monitor the water quality, and screening raw products prior to shipment. Such examples of common microbiological testing are culture media, immunoassay, Polymerase Chain Reaction (PCR) and more. Not only in food safety, microbiological test is also important in pharmaceutical and healthcare sectors. Since pharmaceutical industry is responsible for the development and testing of medication, microbiological testing must be vigorous and through to maintain high quality standards and virtually free of germs to ensure the medication are sterile. Microbiological testing is important to avoid the spreading of harmful germs in healthcare industry too by establishment of test regularly.

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Synthetic medium is a defined medium where all the chemicals involved are known. This medium act for the most consistent cell culture environment as well as the purest, and is free of animal-derived component.

The differences between natural medium and synthetic medium are as follows:

Natural Medium:
-Composed of natural biological components which are chemically undefined.
-Media type : biological fluids, tissue extracts, clots

Synthetic Medium:
-Prepared artificially with the composition of chemically known organic and inorganic components
-Media type : balanced salt solutions, basal media, complex media

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The effectiveness of the media fed to the microorganisms can be assessed by examining the growth of the microbes. The media is considered effective if the growth and size population of the microorganisms are high. This can be done by using direct and indirect methods. Dry cell weight measurement, total cell number and viable cell number are the examples of direct methods that can be utilised to measure the growth of the cell. When these factors available in large amount, this mean the microorganisms are growing actively and the media formulation are suitable. As for indirect methods, rate of substrate consumption or product formation and viscosity can be used to monitor the growth of microorganisms in the media. Increase in substrate consumption, product formation and viscosity of the media indicates a positive grow of the microorganisms. On the other hand, low substrate consumption, product formation and viscosity of the media means the growth rate is low and the media is considered unsuitable or less effective.

Apparently, these methods do have some advantages and disadvantages. For example, the total cell number is a rapid method and requires low time consumption, however it is difficult to distinguish the differences betweed dead and living cells due to the extremely tendious condition, which lead to inaccurate result. Therefore it is important to choose the best method to handle depends on the media and microorganisms used.

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What is medium formulation and its importance?

A critical stage in the design of effective laboratory experiments, pilot-scale development, and manufacturing processes is medium formulation. The elements of a medium provide essential nutrients and water that can satisfy the elemental requirements for cell biomass and metabolite synthesis. The medium, together with optimal temperatures, light levels, and favourable environmental circumstances, creates the ideal environment for microbial development.While manipulating a medium for the goal of a wide spectrum of production, there are a few things to keep in mind.

• It should be able to produce the maximum amount of product.

• It should produce the least amount of unwanted product possible.

• Affordable

• Aeration, medium formation, sterilisation, agitation, extraction, purification, and waste treatment should all be as minimum as possible.

• It must be able to produce the highest concentration of biomass and be available all year.

• The media must meet all of the organism's nutritional requirements as well as the process's objectives.

Developing an equation based on stoichiometry to balance growth and product generation is the first step in designing the medium formulation. This equation, which may be expressed quantitatively, aids in determining the most cost-effective media design by estimating the minimum quantity of nutrients required to generate a specific amount of biomass. To get the most out of the fermentation process, the medium composition needs to be well-suited to the fermentation process. By identifying the substrate concentration required to create the requisite product yield, the medium formulation is essential for determining the specific amount of biomass required to produce a particular amount of product. In addition, the formulation of specialised media is critical for obtaining products of consistent quality while minimizing substrate material and transportation costs. The well-formulated medium can provide the appropriate amounts of macronutrients, micronutrients, trace elements, dissolved oxygen, vitamins, enzymes, other dissolved gases, and inhibitors to meet the fermentation process' requirements. These components can achieve the elemental requirements for metabolite and biomass formation as well as suitable energy provision for microbial fermentation by determining the medium formulation.

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Media optimisation is an analytical process where multiple parameters screening for each type of media component are manipulated to observe the concentration and yield of a particular fermentation product. Yield of the product would increase when the bacteria are set to grow in an optimal condition suitable for their growth and stability. Media optimisation is indeed important to ensure stable production of products. Other than that, it can be used to tune the transition between exponential growth and stationary phase, which is commonly used to separate the growth and production phases in microbial bioprocesses.

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Media is food to microorganisms. It contains high amounts of nutrients and optimum conditions required by microorganisms to grow and produce final products. It is high in Carbon and Nitrogen source. Its minor components include inorganic salts, vitamins, growth factors, anti-foaming agents, buffers, dissolved oxygen, other dissolved gases, growth inhibitors and enzymes.