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    Assignment 4 Micro

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    Awais Yousaf
    This document contains an assignment submitted by Hamza Rehman to Rabia Masood for Botany course at Hazara University. It includes 3 questions asking to differentiate lyophilization and cryopreservation, describe steps of periodic transfer in microbial preservation, and define agar slant culture, viable bacterial cell and nutrient broth. The response provides detailed explanations for each question answering the key differences between lyophilization and cryopreservation, outlining the steps of periodic transfer as refrigeration, paraffin method and defining agar slant culture as agar mixed with nutrients and placed in test tubes at an angle, a viable bacterial cell as a living cell, and nutrient broth as a general purpose medium for cultivating microorganisms.

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    Assignment 4 Micro

    Uploaded by

    Awais Yousaf
    33 views11 pages
    This document contains an assignment submitted by Hamza Rehman to Rabia Masood for Botany course at Hazara University. It includes 3 questions asking to differentiate lyophilization and cryopreservation, describe steps of periodic transfer in microbial preservation, and define agar slant culture, viable bacterial cell and nutrient broth. The response provides detailed explanations for each question answering the key differences between lyophilization and cryopreservation, outlining the steps of periodic transfer as refrigeration, paraffin method and defining agar slant culture as agar mixed with nutrients and placed in test tubes at an angle, a viable bacterial cell as a living cell, and nutrient broth as a general purpose medium for cultivating microorganisms.

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    This document contains an assignment submitted by Hamza Rehman to Rabia Masood for Botany course at Hazara University. It includes 3 questions asking to differentiate lyophilization and cryopreservation, describe steps of periodic transfer in microbial preservation, and define agar slant culture, viable bacterial cell and nutrient broth. The response provides detailed explanations for each question answering the key differences between lyophilization and cryopreservation, outlining the steps of periodic transfer as refrigeration, paraffin method and defining agar slant culture as agar mixed with nutrients and placed in test tubes at an angle, a viable bacterial cell as a living cell, and nutrient broth as a general purpose medium for cultivating microorganisms.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    33 views11 pages

    Assignment 4 Micro

    Uploaded by

    Awais Yousaf
    This document contains an assignment submitted by Hamza Rehman to Rabia Masood for Botany course at Hazara University. It includes 3 questions asking to differentiate lyophilization and cryopreservation, describe steps of periodic transfer in microbial preservation, and define agar slant culture, viable bacterial cell and nutrient broth. The response provides detailed explanations for each question answering the key differences between lyophilization and cryopreservation, outlining the steps of periodic transfer as refrigeration, paraffin method and defining agar slant culture as agar mixed with nutrients and placed in test tubes at an angle, a viable bacterial cell as a living cell, and nutrient broth as a general purpose medium for cultivating microorganisms.

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    DEPARTMENT OF BOTANY,HAZARA

    UNIVERSITY,MANSEHRA
    ASSIGNMENT
    Assignment title: Ass [4] : 3 Questions

    Student Name: Hamza Rehman

    Roll.No : 51196

    Course Name: BS

    Department : Botany

    Semester: 4th

    Teacher Name: Rabia Masood


    From: Balakot

    Submission: 07 july 2020

    Remarks:
    Q.No 1: Differentiate between lyophilisation and
    cryopreservation?
    Cryopreservation:
    The process of cooling and storing cells, tissues, or organs at very low
    temperatures to maintain their viability. For example, the technology of cooling
    and storing cells at a temperature below the freezing point ('196' C) permits high
    rates of survivability of the cells upon thawing.

    Lyophilization
    Lyophilization and freeze drying are synonymous. Lyophilization is a water
    removal process typically used to preserve perishable materials, to extend shelf
    life or make the material more convenient for transport. Lyophilization works by
    freezing the material, then reducing the pressure and adding heat to allow the
    frozen water in the material to sublimate.

    Lyophilization’s 3 Primary Stages


    Lyophilization occurs in three phases, with the first and most critical being the
    freezing phase. Proper lyophilization can reduce drying times by 30%.
    Freezing,primary,secondary
    Cryopreservation

    Cryopreservation (i.e., freezing in liquid nitrogen at-196°C)


    helps survival of pure cultures for long storage times.
    In this method, the microorganisms of culture are rapidly
    frozen in liquid nitrogen at -196°C in the presence of
    stabilizing agents such as glycerol, that prevent the formation
    of ice crystals and promote cell survival.

    Lyophilisation (Freeze-Drying)

    In this method, the culture is rapidly frozen at a very


    low temperature (around -70°C) and then dehydrated
    by vacuum.
    Under these conditions, the microbial cells are
    dehydrated and their metabolic activities are stopped;
    as a result, the microbes go into dormant state and
    retain viability for years.
    Lyophilized or freeze-dried pure cultures and then
    sealed and stored in the dark at 4°C in refrigerators.
    Freeze- drying method is the most frequently used
    technique by culture collection centres.
    Q.NO:2 What are different steps of periodic transfer
    in microbial preservation?

    Periodic Transfer to Fresh Media


    Strains can be maintained by periodically preparing a fresh culture from the
    previous stock culture. The culture medium, the storage temperature, and the
    time interval at which the transfers are made vary with the species and must be
    ascertained beforehand. The temperature and the type of medium chosen should
    support a slow rather than a rapid rate of growth so that the time interval
    between transfers can be as long as possible. Many of the more common
    heterotrophs remain viable for several weeks or months on a medium like
    Nutrient Agar. The transfer method has the disadvantage of failing to prevent
    changes in the characteristics of a strain due to the development of variants and
    mutants.
    Refrigeration
    Pure cultures can be successfully stored at 0-4°C either in refrigerators or in cold-
    rooms. This method is applied for short duration (2-3 weeks for bacteria and 3-4
    months for fungi) because the metabolic activities of the microorganisms are
    greatly slowed down but not stopped. Thus their growth continue slowly,
    nutrients are utilized and waste products released in medium. This results in,
    finally, the death of the microbes after sometime.
     
    Paraffin Method/ preservation by overlaying cultures with mineral oil

    This is a simple and most economical method of maintaining pure


    cultures of bacteria and fungi. In this method, sterile liquid paraffin is
    poured over the slant (slope) of culture and stored upright at room
    temperature. The layer of paraffin ensures anaerobic conditions and
    prevents dehydration of the medium. This condition helps
    microorganisms or pure culture to remain in a dormant state and,
    therefore, the culture can be preserved form months to years (varies
    with species).
    The advantage of this method is that we can remove some of the
    growth under the oil with a transfer needle, inoculate a fresh
    medium, and still preserve the original culture. The simplicity of the
    method makes it attractive, but changes in the characteristics of a
    strain can still occur.
    Q.No:3 Define these terms with examples:

    1 Agar slant culture


    2 Viable bacterial cell
    3 Nutrient broth

    Agar slant culture


    Agar, a gelatin-like substance extracted from red algae, is commonly used to
    culture microorganisms. Various nutrients are added to agar to enhance the
    growth of bacteria in either shallow plates or test tubes. When agar media is
    placed in test tubes it is in liquid form. The test tubes are placed on an angle to
    cool and congeal, creating a slanted surface, or an agar slant.
    Agar
    Agar is a substance extracted from the cell walls of red algae. While it is
    commonly used to grow bacteriological cultures, the agar must be impregnated
    with nutrients such as beef extract and peptone in order to support life. Various
    substances mixed into the agar promote the growth of very specific types of
    bacteria. For instance, mannitol salt agar, which contains sodium chloride, favors
    the growth of only staphylococcus bacteria and can differentiate between
    pathogenic and non-pathogenic strains.
    Slanting
    Agar slants are created by bringing agar to the boiling point and pouring it into a
    test tube. Before the agar cools and solidifies, the test tube is set on its side. Once
    the agar is cooled, the test tube can be stored upright, and the agar inside has a
    slanted appearance.
    Advantages
    Slanting the surface of the agar gives the bacteria a greater surface area on which
    to grow in a test tube. Furthermore, slants are created in test tubes that can be
    capped, which minimizes water loss. This is important because of the high
    moisture content of agar media.
    Uses
    Agar slants can be used to culture bacterial cells for identification. Attempting
    to identify bacteria from a large sample is difficult because bacteria are small and
    can be difficult to find. However, when placed on a nutrient agar slant, bacterial
    cells will dividuced enough cells to examine microscopically.
    Viable Bacterial Cell

    Nutrient Broth
    Nutrient Broth is a general purpose medium used for cultivating a broad
    variety of fastidious and non-fastidious microorganisms with non-exacting
    nutritional requirements. Peptone and yeast extract provide nitrogenous
    compounds, vitamin B complex, amino acids and other essential growth nutrients.

    End

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