.

Very simply:
The gram positive bact. Bind to the stain and never let go. And the gram negative bact. Stick to the stain but let go of it when exposed to the buffer. :)

 cocci bacilli rod round

Gram -ve

Cocii

Gram -ve

Bacilli

The Basic idea of the section is to detect and identify the fungal infection in the patient we suspect to be infected.

Sowhen a patient comes to the doctor with presumptive symptoms the doctor makes a clinical diagnosis. But that is not enough, so he orders some laboratory tests which may include: Histopathological examination. Culture media. Serological examination. Experimental animals.

To make sure there is a fungal infection and identify the type of infection we need to: Get a sample. Examine it under the microscope and by the naked eye( micro & macroscopically). Grow a culture media. And finally identify it.

Sampling:
The samples are collected in sterilized containers (petri dishes tubesetc.). Inside these containers there is sabourouds brotha sugar solution which is full of nutrients to keep the fungus alive till examination.

Site of infection Skin, nails, hair

Sample

Tool

Mucous membrane

Bronchopulmonary Systemic mycosis

Urinary tract

Cerebrospinal

Pleural & peritoneall

Lumbar puncture

Needle Aspiration

Examination
The sample is then placed in a 10-30% KOH solution which helps in dissolving the keratin from the sample for a better examination ( de ma3looma exclusive ;) :P ). Examine widely by low power to find the infected area then use high power. spores . Mycelium

Fungus

Identified by

picture

Mycelium through (filamentous identification of fungi). the arthrospores in chains or single.

Yeast-like
Candida

Budding yeast cells and filaments.

Culture
To increase the mass of the fungus enough to observe it we put it in a medium that is suitable for growth & for that culture media we usually use Sobaurauds Dextrose Agar SDA. Its composition is:

Dextrose Peptone Agar H2O

20 g 10 g 20 g 1L

And It has a slightly acidic pH of 5.4 .

 The culture grown in tubes is slopes to allow us to see the medium recto and verso.

The sample might contain bacteria or unwanted fungi that contaminate the medium and grow on the expense of the fungi. So we use anti-biotics as chloramphenicol , Cycloheximide (for dermatophytes )

 The stain Characteristics & morphology of the sample Histochemical composition.

Color. Texture(wooly , spongy, powdery etc.). Type ( yeast-like or filamentous). The pigment and its distribution.

 1. 2. 3. 4. H & E. Periodic acid-Schiff stain (PAS). Gomori methenamine Silver stain (GMS). Mayers mucicarmine stain.

Not very specific because some fungi share the antigens, so it helps in reaching a presumptive diagnosis. Examples: 1. Double Diffusion method. 2. Counter-immuno electrophoresis. 3. Latex agglutination. 4. Indirect Immunofluorescence. 5. Indirect haemagglutination. 6. ELISA.

Slides
Stain: Gram stain+. Color: Violet. Round, oval looking spores. Oil emersion lens. Yeast-like fungi.

Candida

 High power. Rough thick wall. 6-8 per macroconidia.

Macro conidia of microsporum canis

 Low power. Black, large, round vesicle. Biseriated phyalides.

 High power. Greyish blue, club shaped vesicles.

Uniseriated phyalides.
The spores occupy two thirds of the vesicles. Conidia are spores that separate from the vesicles to aid in asexual reproduction.

Petri dishes:
Mycelium ( filamentous fungus). Medium:SDA Color: Black.

Verso (lower)

Recto (upper)

Medium: SDA. Yeast-like fungus. Budding and filaments. Color: Creamy-white

Verso (lower)

Recto (upper)

 Stain: SDA. Mycelium ( filamentous). Color: Green.

Section3 Protozoa & Helminthes

Gametocyte of P. vivax Microspora

Sarco-mastigophora

Cyst of balantidium coli

Sarco Dina Entamoeba Enamoeba Histolytica

Mastigophora Leishmania

Cyst of balantidium coli

Size: 60-70
Shape: Ovoid with a distinct cell wall. Nuclei: 2 large shaped nuclei(macronucleus & a smaller micronucleus in its cavity.

Macronucleus

Sarco-mastigophora
Sarco Dina Entamoeba Histolytica

Size:6-18.

Shape: Rounded.
Nuclei:4 vesicular nuclei with vesicular dots representing the karyosomes. Chromatoid bodies: Refractile rods with rounded tips, black in color.

Sarco-mastigophora Mastigophora Leishmania Shape: Spindle.

Size: 15-20.
Nucleus: Central.

Kinetoplast: Anteriorly situated.

Flagellum: Long, free, originating from the basal granule.

Gametocyte of P. vivax
Shape: Rounded enlarged mass of compact cytoplasm with a single nucleus which may be compact & peripheral or diffuse and central. Size: Large filling the enlarged R.B.C. Infected R.B.C is enlarged, can be irregular with schuffners dots.

Taenia Saginata

Ascaris Lumbricoids

Fasciola Hepatica

Schistosoma Mansoni

Mature segment of Taenia Saginata Shape: Square. Common genital pore that opens in the lateral margin.

Testis: scattered follicles in the lateral fields,

Ovary: Bilobed in the posterior zone.

Vitelline glands appear as triangular mass behind the ovary.

Uterus: Blind elongated tube in mid-plane.

Genital pore

Uterus

Ovary Testis

Vitelline gland

Ascaris Lumbricoids
Shape: cylindrical. Color: Ivory white. Male: 15-20 cm. in length with curved tail. Female(wakleen 7a2ena fe kol 7eta :P 72o2 el mar2a ba2a) 2040 cm. with a blunt tail.

Schistosoma Mansoni
Size: 8mm.1mm. Suckers: Two oral and a large ventral one. Testes: 6-9 small testes, posteriolateral to the ventral sucker.

Fasciola Hepatica
Size: 2-3cm. Shape: Flat, leaf-like with converging sides. Prominent cephalic cone & shoulders. Suckers: Oral ventral suckers. Intestinal caeca: Blind, The lateral branches are more branched than the medial ones. Ovary & Testis: highly branched. Uterus: anterior to ventral suckers, packed with brown eggs. Vitelline gland: small follicle at the lateral end.

http://www.mediafire .com/?kfpi09j2o7824 b8

SECTION 4 ENTOMOLOGY

Class
Crustacea Order Arachnida Order
Scorpionida Acarina Bugs Sub-order Fleas

Insecta Order
Lice

Cyclopes

Ticks
Diaptomas

Mites

Diptera
Suborder

Flies

Mosquito

Ticks
*Male Hard Tick Unsegmented, oval in shape. Anteriorly protruded mouth. The dorsal surface is entirely covered with scutum. four pairs of legs.

*Female Hard Tick Unsegmented, oval in shape. Anteriorly protruded mouth. The anterior third of the dorsal surface is covered with scutum. four pairs of legs.

*Adult Soft Ticks Unsgmented.

Not covered with scutum.

The mouth part originates ventrally. Four pairs of legs. The body has rounded borders. ):
http://www.mediafire.com/?6uw8x7vbz5u 2bl7

*Sarcoptes Scabiei
Size:300 Rounded or oval in shape Unsegmented Four pairs of legs Tegument "outer body covering"Is covered with hair or bristles

*Cyclopes Size: 1-2 mm. Shape: pyriform Cephalothorax is broad, two pairs of long & short antenae Five pairs of legs 4 Segments, the last segment is biforked

 Length: 12cm.

*Scorpion

Color: yellowish brown. Body: cephalothorax & abdomen Cephalothorax: 4 pairs of legs. Abdomen:12 segments, the first seven are broad, the rest is narrow. The last segment carries the telson (the last division of the body of a crustacean) which carries the stinger.

 Head: narrow, carrying two compound eyes. Protruded mouth used to suck blood. 3 pairs of legs Large abdomen.

 Size: 2-3mm. Shape: compressed dorsoventrally. Head: a pair of short antennae & a pair of simple eyes. Thorax: Fused, non-segmented and bares three pairs of legs. No wings. 8 Abdominal segments.

the last segment is shaped like an inverted V.

The last abdominal segment has a triangular penis.

Egg pediculus humanus (nit)

Attached to the hair shaft. The is with an ornamented anterior led.

Phthirus Pubis

Shape: Crab-like, compressed dorso-ventrally. Thorax: Fused, Non-segmented.

Three pairs of legs and NO wings.

Segmented antenae.

Abdomen: last four segments carry finger-like projections, the last one is indented.

1-Combed: Ctenocephalides Canis

Compressed laterally. Head: with a pair of simple eyes. Combs: One on the head. The other one is on the posterior margin of the prothorax( first segment). Three legs, the posterior one is the most powerful( for jumping ). Posterior end is pointed with coiled penis.

2-Non-Combed:
Xenopsylla Cheopis"red flea"
White in color. Compressed laterally. Head: with pair of simple eyes. Thorax: No wings, Three pairs of legs. Presence of mesopleural suture, which is a thick rod-like structure, over the attachment of the second leg. Posterior end is rounded with coiled penis.

No Comb

Suture

Pulex irritance"human flea"

Same with no suture White in color.

Compressed laterally.
Head: with pair of simple eyes. Thorax: No wings, Three pairs of legs. Presence of mesopleural suture, which is a thick rod-like structure, over the attachment of the second leg. Posterior end is rounded with coiled penis.

SECTION 7 DECONTAMINATION 1

 Decontamination 07 Betadine

 Chlorine Hydrogem peroxide Glutaraldehyde Cidex Lysol Different tissue cultures Infected and normal

Normal monolayer of tissue culture cells. Continuous sheet of normal cells.

Inoculated tissue culture cell line

showing cytopathic effect in the form of: Ballooning of the cells, and areas devoid of cells due to their lysis (days after inoculation).

Inoculated tissue culture cell line showing cytopathic effect in the form of:
a) plaques "areas devoidof cells due to lysis of the cells by virus b) Rounding of the cells "at the borders of the plaque ."

Fibroblast tissue culture cell line showing:

A central mass of infected cells with cytopathic effect. (hours after inoculation)

Inoculated tissue culture cell line showing:

Focal areas of infected cells (several hours after inoculation).

:)

Autoclave( the most efficient

method)

Note:

Hot Air Oven

short wave-length

Items for Y radiation & gaseous sterilization (all packed):

Filtering agent

fluid pointy :)

Monitoring the effectiveness of sterilization

Records presented by tools To measure physical conditionstemp.pressure.

For heat resistant bact. By culturing an appropriate media ( temp. and nutrients and time). Effective but takes a lot of time.

Tapes that carry chemicals arranged in lines that change color in the intended temp. and so, no mirobes.

 :) Ag-Ab reactions

Agglutination

Precipitation

Complement fixation

Ab labeling

Agglutination
The Ag Ab 6 .
. The Ab should be in the particulate form( not soluble) in a buffer

. Saline Abs specific for a certain group of bacteria Agglutination

 6 :).

Quantitative . 1\02 1\04, Ag Ag Ags End Point: It is the highest serum dilution that shows a positive result for the .Ag .) of the end point dilution( The titer: It is the reciprocal

 end point 1\061 1\061 061 Ags :) .

Passive Abs Ags Ags Ag

Activeinhibition inhibtion

 Latex . ): Abs or Ags

Staph aures Staph aures Coagulation

Scientifically speaking :D the Protein A leaves the Staph aures to bind to the Fc site on the IgG leaving the Fab site on the aures free Now, if we add a new bacteria it binds to the Fab site on the aures and causes coagulation

 erythroblastosis foetalis

1-Direct
Anti-human globulin .Ab for the Ab

:2-Indirect
Rh Abs Serum Rh+ cells Ags Anti-human globulin . . :P

percipitation
reaction in solution Ag Ab .

. . Ag & Ab hv 2 be in optimum conc

Slide percipitation

Tube periciptation

Agar (gel) diffusion tests

Double Immunodiffusion

Single(radial) immunodiffusion

Elek`s Test

Slide percipitation
RPR (rapid plasma reagin) is a screening test for syphilis. It looks for antibodies that are present in the blood of people who have the disease. The test is similar to the venereal disease research laboratory (VDRL) test.
Tube periciptation

As lancifield test for grouping Streptococci (ring test).

Double Immuno-difusion

Ab

Ag

Double Ab well Ag well Agar gel on a slide diffuse optimum .precipitation band Single (radial) Immunodifusion Perti dish wells well Ag Agarose gel .Ab Ag diffusion .

Ag

Ab in Agarose gel

 This technique is used for estimating the amount of different Ig classes in the serum using their anti-Ig.

Complement fixation
Abs Ags Complement system .Serum Complement guinea pigs .:P Ag . Complement

complement . Complement .Hemolysis

Ag Complement .Hemolysis R.B.Cs Complement

-ve( hemolysed)

+ve(non-hemolysed)

This test is used for diseases like brucellosis, whooping cough, gonorrhoea & typhus.

Ab labeling

For detection of

Direct Used in detection of rabies. We put the labeled Ab on the slide which has Ag.

Indirect We put non-labeled Ab. Add the labeled Ab Wash the cellsto remove excess Abs. Examine This method Is more economic( because we use a few labeled Abs only

We put serum with the wanted Ab. Then Ag. And finally Anti-human globulin.

Anti-human G. Ab Ag Ab Based on Immunological activity Ab Ag

For detection of Ag
(Double Ab sandwitch) We Put the Ab. Add the Ag. And finally add the Ab, but this time with a substrate that breaks when the reaction occurs and a color appears.

For detection of Ab
(Indirect ELISA) We put the Ag. Then the serum Ab(to be tested). And finally Anti-human globulin with a substrate that breaks when the reaction occurs and produces a color.

Colored

Not Colored

 , :)