User:Ghenzmitz

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THIS IS A SUMMARY OF TOPICS IN MICROLAB

Microbiology – the study of little organisms that cannot be seen by our naked eye Micro – “little or minute” Biology – “study of life” Microorganisms – they could be pathogenic (cause disease) or ubiquitous (can be found anywhere) in nature

Microscope – special instrument of precision used to magnify organisms that cannot be seen by our naked eye.

2 kinds of Microscope a. simple – uses one lens, little magnifying lens and can magnify 3-20x the actual size b. compound – uses 2 or more lenses, can magnify 40-100x the actual size

Mechanical Parts of Microscope 1. Draw tube – Holds the eyepiece 2. Body tube – Holds the principal lenses 3. Substage – holds the condenser and iris diaphragm 4. Stage – where the slide with specimen is being placed 5. pillar – connects upper part to the base 6. arm – used for holding 7. Base – supports the whole microscope 8. Revolving nose piece – use to adjust principal lenses as desired 9. adjustable mirror – collects and reflects light up in to the microscope 10. inclination joint – allow upper part to be tilted back to any degree desired 11. iris diaphragm – controls the amount of light that enters the condenser 12. stage holder /stage clip – holds the slide 13. slide adjustment knob – moves the slide sideward and forward and backward 14. Coarse adjustment knob – used while focusing under low power objective. 15. fine adjustment knob – used while focusing under HPO and OIO

Optical Parts of the Microscope 1. Principal Lenses i. Low power objective (LPO) – yellow ring in color, with 10x magnification \; used when we are focusing protozoa, parasites or larger microorganisms ii. High Power Objective (HPO) – color of ring is green. With 40x magnification – used when examining bacteria or organisms suspended in fluid or water iii. Oil Immersion Objective (OIO) – color of the ring is white – with 100x magnification – used for sputum specimen or stained organism – it is called OIO because the objective is immersed to the oil on the slide – use olive oil or cedar oil 2. Eye piece – magnifies the image formed by the objective 3. Condenser – concentrates the amount of light on the specimen

How to manipulate Microscope: 1. Place the microscope on a flat surface and the mirror points towards the source of light and take a comfotabgle position at the instrument. 2. with the object on the stage, swing into place the objective you wish to use and while watching from the side, put the objective just under position it will have when in focus. 3. look into the microscope. Keep both eyes open(to avoid eye straining) . secure proper amount of light by manipulating the mirror, condenser and diaphragm 4. if the field is not uniformly bright, adjust the mirror, make certain that the diaphragm is fully opened. 5. to focus the object, while looking into the ocular, move the tube downward with the coarse adjustment until it is short distance away from the slide mounted. Then look into the ocular and reverse the motion with the coarse adjustment until the object is visible. Do so slowly. 6. then swing the HPO into position. It will be in focus because most microscopes are parfocal (focal distance of the lens having the same magnification as the objective). To bring it to focus, use the fine adjustment turning slowly. 7. now raise the tube with coarse adjustment and swing the OIO into position. Place a small drop of cedar oil on the portion of the slide to examine, avoiding an excess of oil. a. With the coarse adjustment knob, bring the tube down until it the OIO strikes the oil, turning a little further until it is completely battered in but not touching the slide of the object b. Then, looking into the ocular and by means of find adjustment, focus by drawing the tube away from the slide. Before focusing, determine in which direction the fine adjustment must be turned so as to move the objective away from the slide. 8. to focus the OIO lens, the fine adjustment must be moved very slowly because of the rapidity in which the object will go out of focus owing to the short looking distance. a. If the high source is not properly centered, the object on the slide will appear to be displaced in a horizontal direction when focusing. b. This condition can be remedied by adjusting the mirror until there is no more apparent horizontal movement in the raising and lowering of the objective. c. With the OIO lens, one may find that the entire field is not in focus, so that for instance, with a slight turn of the fine adjustment the periphery of the field may be clear and then with another adjustment the center is in focus. The look of uniform focus is due to Spherical Aberration – a loss of definition in the image produced by the spherically curved mirror lens.

Ways of Taking care of Microscope 1. when using the microscope put in flat surface and in comfortable position 2. handle the microscope with care by holding the arm and support the base 3. cover the microscope with plastic when not in use 4. place the microscope points towards the light 5. Know the different parts of the microscope and its use in order to save time.

Sizes of Bacteria a. 0.2-2 um(micron) in diameter b. 2-8 um in length

Shapes of Bateria A. Spherical or Cocci a. Diplococci – which remain in pairs b. Streptococci – cling together like chains c. Tetrads – divided in 2 planes or grouped in 4 d. Sarcinae – divided in 4 planes or grouped in 8 e. Staphylococci – divided in multiple plane or grape-like structure B. Rod-shape or Bacillus: a. Single bacilli b. Diplobacilli c. Streptobacilli d. Coccobacillus C. Spiral – shaped like a coile wire string a. Vibrios (curve rods) b. Spirilla (helical in shape, fairly rigid bodies c. Spirochetes (flyable, several complete, spiral turns)

How to focus Organisms under OIO a. place the microscope in flat surface and mirror towards the light b. then 1st use LPO then adjust using the coarse adjustment knob c. move the revolving nosepiece to focus in HPO then adjust the fine adjustment knob d. move the revolving nosepiece to focus in OIO, swing the HPO and place a drop of cedar oil to the slide e. focus to OIO then adjust using the fine adjustment knob f. be sure that the OIO touches the cedar oil to focus g. if it is dark, adjust the movable mirror

Gram positive: 1. streptococcus pneumoniae (spherical shape) 2. Staphylococcus Aureus (spherical) 3. Mycobacterium tuberculosis (rod shape) 4. Bacillus Subtilis (rod shape)

Gram Negative – 1. Vibrio Cholerae (curved) 2. Shiglella Dysenteriae (rod) 3. Neisseriae Gonorrhea (spherical) 4. Proteus Vulgaris (rod shaped) 5. Salmonella Typhosa (rod shape)

STAINING 4 purposes: 1. to make it visible under the microscope 2. reveals the chemical nature of the organism 3. shows the shapes, structure, size 4. inhibits the growth and development of the bacteria or organism so that other organism growing simultaneously may be studied.

4 types of Staining 1. simple staining./ direct staining – uses only one stain and direct color of organism 2. differential staining – uses 2 stains of organisms; (used to different one organism to the other) • initial stain • counter stain examples: • gram staining • acid fast staining 3. Special/Selective Staining – reveals the special structure of organism 4. Indirect Staining – cannot stain the organism only the background is being stained

Steps in Acid Fast Staining 1. smearing – process of putting the specimen on the slide and spread it into thin field by the use of applicator stick 2. fixing/flaming – passing the slide over the flame 3x a. so that the organism will stick to the slide b. to kill the organism c. to prevent the organism from spreading 3. flood the reagent Carbol Fuchsin (red) – initial sign 4. Wash with water 5. Decolorize with Acid Alcohol - decolorizer 6. Wash with water 7. flood with methylene blue (blue) – counterstain 8. air dry and examine

Principle of Acid Fast Staining 1. color red resist the decolorization 2. to show the chemical nature of organism 3. to see the morphological orrangement, structure, size and shape of organism

Reagent used in Acid Fast Staining 1. carbol fuchsin 2. acid alcohol 3. methylene blue

Steps of Gram Staining 1. Smearing 2. Flaming.fixing 3. gentian/crystal violet (initial stain) 4. 1 drop 5% sodium carbonate – to prevent overstaining(neutralizer) 5. wash with water 6. flood with gram iodine – (mordant)- renders the cell wall of the organism impermeable 7. wash with water 8. decolorize with ethanol – other are acetone, alcohol, ethyl alcohol 9. wash with water 10. counter stain with safranin(red) counter stain

Principle of Gram Staining a. gram (+) = purple b. gram (-) = red c. resist decolorization, retain the initial stain

Reagents used in gram staining 1. gentian/rystal violet 2. 5% sodium carbonate 3. gram iodine 4. ethanol 5. safranin

Pyogenic – neisseriae gonorrhea = pus forming organism (ex. All cocci grouped) Tetanus – means lock jaw

• all cocci are (+) except Neisseria Group (Neisseriae Gonorrhea)

• all bacilli are (-) except:

• Mycobacterium group o M. tuberculosis o M. leprae • Coryne Bacterium o Diphtheriae • Spore bearer o Aerobic  Bacillus anthracis  Bacillus subtilis o Anaerobic  Clostridium tetani  Clostridium botulinum  Clostridium novyi  Clostridium septicum  Clostridium perfringens

Blood - red fluid - sticky red fluid filling and circulating through the heart and blood vessels

Element of Blood (components) a. red blood cell – erythrocytes a. removes CO2 and responsible in acid base balance and viscosity and specific gravity b. White Blood Cells – Leukocytes a. Soldiers in the body c. Platelets – thrombocytes a. Responsible in blood clotting

Blood Types:- discovered by Karl Landsteiner and Co-workers (1900-1902) “A” “B” “AB” “O”

(Ag) Antigen – substance foreign to the body - used as stimulator - the number of CHON introduced in the body that stimulates the formation of antibodies - a substance that causes the formation of antibodies (ex. Toxin, allergens, agglutinogen, precipitinogen ) - Agglutinogen – present in RBC

(Ab) – Antibodies – defense to prevent illness

    -serum CHON (immunoglobulin) produce by lymphocytes and plasma cells in the response to antigenic stimulation
   - ex. Antitoxin, antiallergenic, agglutinin (present in plasma and serum), precipitin

Plasma – fluid portion of the circulating blood Serum – fluid portion of the coagulated blood

Agglutination Reaction – occurs when Agglutinogen of Type A meets the Type A Agglutinin >Agglutinogen of a RBC should not meet the agglutinin of the plasma type of blood

Crossmatching – a test for compatibility of the donors blood with the recipient

Types of Crossmatching 1. Major Crossmatching – donors RBC and the pts. Serum i. Most important because it is the donors RBC tested for compatibility of the pts. Serum 2. Minor Crossmatching – donors serum and pts cells

1st drop of Blood (should be wiped with dry cotton balls) – contains tissue juices and contaminated with extraneous materials that clings to the skin of the finger and it might not be the true representative of your blood

Antiserum A – color blue Antiserum B – color yellow

Donors Blood Group Agglutinogen in Donor RBC Agglutinin in Pts. Plasma Patients Blood Group “A” A anti b A “A” A None AB “B” B anti a B “B” B None AB “O” None anti a B None anti b A “AB” A B none A B “O” None anti a anti b B A O

“O” – universal donor because there is no agglutinogen present that would react to the patients agglutinin therefore no agglutination reaction

“AB” – universal recipient because there is no agglutinin in the plasma that would react to the donor’s agglutinogen, therefore no agglutination reaction

Increased RBC – polycythemia Decreased RBC – anemia

Plasma – fluid portion of the circulating blood Serum – fluid portion of coagulated blood

Blood -cells -plasma -serum

Normal level of RBC a. male – 4.6 – 6.2x1012/L b. female – 4.2 – 5.9x1012/L

RBC – carries O2 into the tissue and removes CO2 in the blood

WBC – protect body from infection or against pathogenic organism by ingesting the bacteria called Phagocytosis - attack the bacteria through microphages - cooperate in promoting tissue repair and regeneration

Blood typing – is a test to classify blood by determining the presence and absence of antigen in RBC and antibodies in the plasma

• the same agglutinogen and agglutinin should not meet because agglutination will occur

“O” – no agglutinogen in RBC “AB” – no agglutinin in plasma

Rh factor – by Karl Landsteiner and Weiner - has no naturally agglutinin in pt. serum and plasma - when the mother is Rh(-) and the baby or fetus is Rh (+) give RhIG - RhIG – given o Within 72 hrs. after delivery o 7-9 montths if mother is pregnant or 7-9 months preterm o In late pregnance RhIG will not cross in the placenta and destroy RBC

Parents Blood Group Possible Blood of Children Mother Father O O O O A O A O B O B A A A O A B A, B, AB O A AB A,B,AB B B B or O B AB A,B,AB AB AB A,B,AB O AB A,B,O

GHENDO QUICOY BSN II E Parasitology - study of parasites Parasites – organisms that live in the body of another for their survival - organism that regain living matter for nourishment

Host – one that harbours the parasite - animals where in or on which a parasite lives - a place of shelter; gives shelter

Classification of Parasites According to: 1. Habitat – dwelling place a. Ectoparasite – parasites that live outside the body of the host (e.g., louse/nits) 3 types of Human Lice 1. head louse – pediculosisi humanus capitis 2. body louse – pediculosis humanus 3. pubic or crab louse – with poor hygiene b. endoparasite – parasites that live inside the body of the host i. tapeworms ii. hookworms iii. pinworms 2. Effect of Parasite on Host a. Pathogenic – causes disease (tapeworms, ascaris; schistosomiasis – schistosoma) b. Non-pathogenic – do not cause disease (e.g., head louse )

2 GREAT DIVISIONS OF ANIMAL KINGDOM I. Protozoa – unicellular – single celled organisms

              >single cell organisms and lowest form of animals

1. amoebiasis – Entamoeba histolytica(causative agent) – intense dysentery or diarrhea with bloody, mucus filled stools waterly stools with blood 2. Trichomoniasis – Trichomonniasis vaginalis(causative agent) – widespread infection of the genitor-urinary tract; - infection is transmitted by sexual intercourse and is a venereal disease of generally unrecognized significance. 3. Trypanosomiasis – 1. American Trypanosomiasis – transferred to blood by small chagas Dse caused by Trypanosoma Cruzi; - fever, infection results from the contamination of the skin with insect feces not transferred by actual insect bite. 2. African Trypanosomiasis – Africal sleeping Dse – acute episodes of fever and inflammation of lymph nodes as the trypanosomes multiply on the bloodstream will multiply and invade the bloodstream.

Causes: a. trypanosoma gambiense] transmitted b. trypanosoma rhodiense ] by tse tse fly

4. Malaria/Malariasis Types: a. Tertian – most common type (sudden attack); paroxysm of chills and fever every 2 days (48 hrs); caused by: Plasmodium vivax b. Quartan – paroxysm of chills and fever every 72 hrs(3 days); caused by: Plasmodium malariae c. Malignant – no pattern ; >irregular paroxysm of chills and fever (caused by: Plasmodium falciparum) 5. Infection of Intestinal Flagellates a. Giardiasis – form of diarrhea  caused by: Giardia lamblia  found in the lumen of the upper small intestine s/s: persisting diarrhea, malabsorption and inflammatory changes

II. Metazoa – multicellular organisms 3 Phylla 1. Platyhelminthes (flatworms) 2 classes: a. Trematoda – flukes

                    - liver (eg. Clonorchis sinemais) fasciola hepatica
                    - blood (eg. Schistosoma japonicum , Schistosoma mansoni, 
                      Schistosoma haematoblum
                    - lungs (e.g. paragonimus westermani
                    - intestines (eg. Faciolopois buski

b. Cestoida – tapeworms (Cestoides)

                    - Tapeworms- 
              Common:
                *beef tapeworm – taenia saginata
                *pork tapeworm – Taenia Salium
                *Fish tapeworm – Diphyllobothrium latum
                *Dog tapeworm – dipylidium caninum
                *Rat tapeworm – hymenolepsis diminuta
                *Dwarf tapeworm – (whortest and smallest ) – hymenolepsis 
                                               nana

2. Nematoda (round worms) 1. Ascaris Lumbricoides – eel worm(bitok; largest known intestinal nematodes) 2. Enterobius vermicularis – pinworm; seatworm (scotch tape) 3. Trichiuris trichura – baboy 4. ancyclostoma duodenale – old worm 5. Necator americanus – new Warhol – Hood 3. Anthropoda (spider, mites, lice)