Total And Viable Counts Of Microorganisms Biology Essay
The term micro-organism besides called bugs refer to little bantam individual celled beings normally non seeable to the bare oculus. Microorganisms require magnification for proper visual image and declaration of their construction. These involve usage of microscopes or amplifying lens with appropriate degree of magnification ( Tortora, Funke and Case 1995 ) . Microorganisms are found in all living things all over the universe i.e.
workss and animate beings. They exist in a assortment of home grounds. They can populate in air, on land or in H2O both fresh and salty. The three chief categories of micro-organisms are bacteriums, Fungis and viruses.
Different micro-organisms have different effects. Some are harmful while others are good. Pathogenic and spoilage bugs do diseases and nutrient spoilage severally therefore referred to as harmful micro-organisms. We besides have others that are needed by populating things to last and they are termed as good bugs ( Tortora, Funke and Case 1995 ) .
Microbes are classified either by puting them in procaryote or eukaryote group or by sorting them harmonizing to the temperature in their surrounding environment where they are classified as thermophilic bugs ( thermophiles ) , mesophilic bugs ( mesophiles ) or psychrophilic bugs ( psychrophiles ) . Thermophilic bugs are those bugs that grow good in high temperatures above the human organic structure temperature. Mesophilic bugs are those that grow good in temperatures equal to that of human organic structure while psychrophililic bugs are those that grow good in low temperatures below that of human organic structure ( Jaggi 1985 ) .Like any other life being, microbes reproduce for sweetening of their species. They reproduce by agencies of either sexual or nonsexual reproduction.
Sexual reproduction occurs by copulating of a male and a female while nonsexual reproduction is by cell division either by mitosis, miosis or binary fission.
3. Count of micro-organisms.
Microorganisms can be counted. They need to be detected first before being counted. There are several techniques involved when numbering bugs.
One or more of the following techniques may be used when numbering bugs. Direct numbering techniques can be used. The oldest of these is microscopy which involves magnification of single cells to go seeable to the bare oculus. Direct numeration techniques do non trust on cell population growing. A more recent of the direct numeration techniques involves usage of immunofluorescence and epifluorescence versions of cell labeling used in concurrence with cytometry. In both engineerings, the trigger for a count is derived from individual cells ( Diaper et Al. 1992 ) .We besides have civilization techniques that rely on growing of bugs to a degree where they are seeable.
This is done under specific conditions of temperature, O, clip and foods among others.The last technique employed is that of newsman checks which asses the population of bugs through their metabolic activities. The population does non needfully hold to be turning. Examples of such techniques include conductance, colorimetric analysis, adenosine triphosphate ( ATP ) and turbidometry ( Bowden 1977 ) .Count of micro-organisms can be done by different methods. In most instances the sample to be counted is foremost diluted to avoid overpowering the whole numeration process. However in some instances, the sample may be excessively dilute to give the needed minimal count to be able to gauge the microbic population of that sample.
In such instances, concentration of the sample is carried out.Count can be done by usage of a slide and a screen faux pas. A bead of the diluted sample is put on the slide with a suited agent for proper visual image of the sample. It is so covered humor a screen faux pas and put under a microscope and observed at a suited magnification. The centre country can be dimensioned with engraved grids. The figure of bugs in the grids is multiplied by the dilution factor to acquire the figure in the original sample ( Black 1996 ) .The Petri dish count is where the sample is diluted to a point where the settlements will be statistically important to be counted but non so many to overgrow each other. This method takes clip for the single cells to turn into settlements.
The settlements counted are multiplied by the dilution factor to acquire the figure in the original sample. The consequences here are expressed in settlement organizing units per millilitre i.e. CFU/ML. The clip taken for the cells to turn into single settlements is called the incubation period.
Count of bug is of import as it enables us gauge the microbic population in a assortment of merchandises ( Breeuwer et al. 1994 ) .
4. Entire count.
Entire count is besides termed as standard home base count or settlement count. It gives the entire figure of bugs both feasible and non-viable. All cells are counted.
These include bacteriums, barms and molds. It is normally done by pour home base method. Entire count by and large requires employment of a microscope.
For case, when finding entire microbic count in H2O by pour home base, a known volume of H2O is assorted with liquefied yeast-malt infusion agar and given clip to solidify. This is done on several home bases. One set of home bases are incubated at 37 & A ; deg ; C for approximately 24 hours and the other set of home bases are incubated at 20-22 & A ; deg ; C for 3 yearss. You will happen that most bacteriums capable of growing in H2O do so good at 22 & A ; deg ; C than at higher temperatures. While the bugs that grow good at 37 & A ; deg ; C will non turn really good in H2O. This means that the two types of micro-organisms need to be counted otherwise since they differ in their growing form. In this instance, transporting out of entire count on H2O is good in several ways ( Paulse, Jackson and Khan 2007 ) . It helps to measure the efficiency of certain H2O intervention processes like curdling, flocculation and disinfection.
It besides gives an indicant of the degree of cleanliness of the H2O distribution system. It can besides be used it find the suitableness of H2O supply to houses where nutrient and drinks are prepared on big graduated table.Entire count is achieved either by usage of direct or indirect counts. One method of direct count is the usage of a haemocytometer. A haemocytometer is a specialised microscope slide of import in cell numeration. The cardinal portion of this slide has etched grids with exactly spaced lines to enable accurate numeration. In order to acquire an accurate count in this method, the cell figure should run between 40 to 70 cells in a one millimeter square.
If this demand is non met, necessary accommodations by either dilution or concentration are done as necessary ( Rapposch, Zangerl and Ginzinger 2000 ) .In indirect counts, one method is by usage of a tintometer. As the micro-organisms turn with clip, they make the agar more and more turbid. This turbidness can be measured by usage of a tintometer where optical denseness is measured. The greater the optical denseness the greater the figure of bugs ( Breeuwer et al.
1995 ) .There is besides a step of dry weight. This method involves centrifugation followed by weighing to acquire the dry weight. The restriction of this method is that cells are destroyedThe other indirect count method involves the usage of a colter counter.
A colter counter is a investigation which measures fluctuation in conduction of a solution as a bacterium passes through a narrow spread ( Daley 1979 ) .The advantage of direct and indirect counts is that the procedure can be automated but the disadvantage is that they can non distinguish dead cells from populating 1s.
5. Feasible count.
Feasible cont involves numbering of settlements produced by merely feasible cells under favourable growing conditions. This can be accomplished by techniques like pour plating, spread plating and most likely figure with an premise that each and every feasible cell gives rise to a pure settlement.
( Black 1996 ) .In pour plating, the liquid media and the diluted sample are poured together in Petri dishes while still in liquid signifier and left to solidify. After solidifying, the Petri dishes are incubated at appropriate temperature for the needed period of clip during which the growing is realized.
The home bases are so removed and distinguishable settlements counted and expressed in settlement organizing units per milliliter.In spread plating, the media is prepared individually and poured in Petri dishes while still in liquid signifier. It is so left to solidify. After solidifying, a little known volume from the diluted sample is put to each Petri dish and with the aid of a sterilised spreading rod, the sample is equally spread over the media.
The home bases are so incubated at an appropriate temperature for a given period during which growing is realized. The home bases are so taken for numeration of settlements utilizing amplifying lens ( Black 1996 ) .Colonies grown in Petri dishes by assorted methods excepting streaking method may be used to gauge the count of feasible bugs since home base counts assume that every settlement is founded by a individual cell and that the cell must hold been alive to turn and organize that settlement.Problems with home base counts are several.
They require a long clip of incubation to be able to visualise the settlements. Clumping of cells can take to undercounting of feasible cells. Cases of excessively many or excessively few settlements on a home base to accurately gauge feasible count are common. Consecutive dilution is frequently required to forestall instances of overcrowding of cells.
Too few cells require concentration by either centrifugation or filtration. In a instance where excessively few settlements are present so the original civilization must be concentrated before finding the home base count. Filtration is a method used to concentrate micro-organisms by screening bugs out of the medium. Centrifugation is besides a separation method based on the denseness. This helps divide the bug from the medium since both have different densenesss ( Pettipher, Mansell, McKinnon and Cousins 1980 ) .
. In consecutive dilution, increases are made in 1000, 100, or 10. The figure of dilutions to be done depends on the concentration of the original solution and the needed concentration. The volume of the solution needed is besides critical. If little measures of solutions are needed so greater Numberss of dilutions are necessary. Consecutive dilution allows little aliquots to be diluted alternatively of unneeded large volumes of stuffs.
When transporting out consecutive dilution, a little sum of original sample is removed to another container and its volume adjusted to original volume utilizing a suited buffer or distilled H2O e.g. if 1ml of the original solution is taken and 10µL removed and put in 990µL of media or H2O so we will hold made a 1:100 dilution. If the original solution contained 5 ten 106 cells/mL so we now have a concentration of 5 tens 104cells/mL because we have divided the concentration by 100 ( Pettipher, Mansell, McKinnon and Cousins 1980 ) .Another method of finding approximative feasible count is by usage of most likely figure. This method involves thining the growing civilizations and so turning the dilution civilizations in broth tubings. This method is utile where it is good to utilize broth other than solid media particularly for extremely motile beings which are hapless in organizing settlements.
The most likely figure method is largely used when the being in inquiry is non able to turn on solid agar or in state of affairss where the micro-organisms are excessively few to give dependable step of population size by the standard home base count method ( Black 1996 ) .The advantage of feasible count is that the method can be made really sensitive and that one can be able to number subsets of population. However the disadvantage is that sometimes the settlement organizing units may undervalue the figure of cells because of clip-clop or ironss of cells.
It is besides clip devouring as the counts require at least few hours or overnight for incubation ( Bowden 1977 ) .Feasible count of bugs is largely applicable in nutrient processing industries largely dairy and meat processing workss where microbiology is most applicable. It helps in appraisal of shelf life of processed nutrient merchandises every bit good as rating of healthful conditions under which the merchandises were manufactured. The efficiency of certain intervention procedures like pasteurisation, sterilisation and cold storage done during production is besides evaluated by feasible microbic count.
Entire and feasible counts of micro-organisms are of import patterns in microbiology applicable in Fieldss of medical specialty, nutrient among others. The chief difference between the two is that entire count determines the count of all cells both dead and alive while feasible count estimation the figure of feasible or unrecorded cells merely capable of turning into distinguishable settlements.