Mikrobiologi Fak Peternakan Disiapkan oleh Prof Dr.drh.Pratiwi Ts, MS 7/5/2018 mikro PTS - pet - 2 2014

Ragam mikrobiologi Sub mikroscopis 7/5/2018 mikro PTS - pet - 2 2014

Bacteria The smallest cells Visible only with the aid of a microscope The smallest bacteria: Chlamydia and Rickettsia-0.1-0.2 micrometer Larger bacteria: many microns in length 7/5/2018 mikro PTS - pet - 2 2014

I. The same microorganism must be present in every case of the disease Koch’s Postulates: Set of guidelines which provides a definite correlation between the pathogen and the associated disease it cause in it’s host I. The same microorganism must be present in every case of the disease II. The pathogen must be isolated and retrieved from the diseased host, grown in pure culture and identified III. The same disease must be produced when the pathogen from pure culture is introduced into a healthy host IV. The pathogen must be recovered from the inoculated animal and must be shown to be the original organism (?) 7/5/2018 mikro PTS - pet - 2 2014

7/5/2018 mikro PTS - pet - 2 2014

Introduction: Microorganisms – several classes of living beings Based on the organization of their cellular structures, all living cells can be divided into two groups: eukaryotic and prokaryotic Eukaryotic cell types - Animals, plants, fungi, protozoans, and algae Prokaryotic cell types - bacteria & blue green algae 07.09.08 Dr Ekta Chourasia

SIZE OF BACTERIA Bacteria of medical importance Unit for measurement : Micron or micrometer,μm: 1μm=10-3mm Size: Varies with kinds of bacteria, and also related to their age and external environment. Bacteria of medical importance 0.2 – 1.5 µm in diameter 3 – 5 µm in length 7/5/2018 mikro PTS - pet - 2 2014

Ukuran dan morphologi 7/5/2018 mikro PTS - pet - 2 2014

Methods Simple staining; Microscope Light Microscope Electron Microscope Darkfield Microscope Phase Contrast Microscope Fluorescence Microscope Cofocal Microscope） Staining Methods Simple staining; Differential staining ( Gram stain, Acid-fast stain), Special staining( Negative stain, Spore stain, Flagella stain) 7/5/2018 mikro PTS - pet - 2 2014

Microscopic measurement Cocci: sphere, 1μm Bacilli: rods , 0.5-1 μm in width -3 μm in length Spiral bacteria: 1~3 μm in length and 0.3-0.6 μm in width 7/5/2018 mikro PTS - pet - 2 2014

Structure of Bacteria Particular structures capsule flagella pili spore 7/5/2018 mikro PTS - pet - 2 2014

Essential structures cell wall cell membrane Cytoplasm nuclear material 7/5/2018 mikro PTS - pet - 2 2014

Shape of Bacteria Cocci – spherical/ oval shaped major groups Bacilli – rod shaped Vibrios – comma shaped Spirilla – rigid spiral forms Spirochetes – flexible spiral forms Actinomycetes – branching filamentous bacteria Mycoplasmas – lack cell wall 7/5/2018 mikro PTS - pet - 2 2014

Arrangement of bacteria: Cocci Coccus Cocci in pair – Diplococcus Tetrad – groups of four Cocci in chain - Streptococci Sarcina – groups of eight Cocci in cluster - Staphylococci 07.09.08 Dr Ekta Chourasia

Bacterial shape Sperical (coccus) Rod-shaped Spiral Spiral or spirillum Helix or spirochete 7/5/2018 mikro PTS - pet - 2 2014

Bacterial arrangement Diplococcus Chains: streptococcus Clusters: staphylococcus Packets of eight:sarcina 7/5/2018 mikro PTS - pet - 2 2014

Other shapes of bacteria Comma shaped Spirilla Spirochetes 07.09.08 Dr Ekta Chourasia

Arrangement of bacteria: Bacilli 07.09.08 Dr Ekta Chourasia

Shape Spherical: coccus (Staphyloccus) Rod-shaped: bacillus (Escherichia) Snakelike: spirillum (Treponema) Branched filamentous (Nocardia and Actinomyces) ( Clusters: diplococcus (Neisseria) chains (Streptococcus) grapelike (Staphylococcus) ) 7/5/2018 mikro PTS - pet - 2 2014

Anatomy of A Bacterial Cell Outer layer – two components: Rigid cell wall Cytoplasmic (Cell/ Plasma) membrane – present beneath cell wall Cytoplasm – cytoplasmic inclusions, ribosomes, mesosomes and nucleus Additional structures – plasmid, slime layer, capsule, flagella, fimbriae (pili), spores 07.09.08 Dr Ekta Chourasia

Staining Simple Differential: Gram and Acid-fast stain Gram-stain : cell wall Acid fast stain : Mycobacterium Negative stain : Indian ink (capsule) Special staining 7/5/2018 mikro PTS - pet - 2 2014

1884: Christian Gram: First publication for the Gram stain method) Editor's note: I would like to testify that I have found the Gram method to be one of the best and for many cases the best method which I have ever used for staining Schizomycetes. Gram + Gram - Cell wall Cell (inner) membrane Outer membrane Ribosomes Granule Nucleoid Cell membrane Capsule Flagellum Pili Gram, C. 1884. Ueber die isolirte Farbung der Schizomyceten in SchnittÄund Trockenpraparaten. Fortschritte der Medicin, Vol. 2, pages 185-189. 7/5/2018 mikro PTS - pet - 2 2014

Figure: 04-28 Caption: Cell walls of Bacteria. (a,b) Schematic diagrams of gram-positive and gram-negative cell walls. 7/5/2018 mikro PTS - pet - 2 2014

Differential staining ( Gram stain, Acid-fast stain), Staining Methods Simple staining; Differential staining ( Gram stain, Acid-fast stain), Special staining( Negative stain, Spore stain, Flagella stain) 7/5/2018 mikro PTS - pet - 2 2014

Gram stain Two major classes of bacteria are distinguished: Gram-positive and Gram-negative bacteria Except: Mycobacteria (waxy outer shell , distinguished by acid fast stain) Mycoplasmas(no peptidoglycan) 7/5/2018 mikro PTS - pet - 2 2014

Figure: 04-04a-01 Caption: The Gram stain. (a) Steps in the Gram stain procedure. 7/5/2018 mikro PTS - pet - 2 2014

Figure: 04-04a-02 Caption: The Gram stain. (a) Steps in the Gram stain procedure. 7/5/2018 mikro PTS - pet - 2 2014

Bacterial growth curve Lag phase Exponential phase Stationary phase Death phase 7/5/2018 mikro PTS - pet - 2 2014

Figure: 06-01 Caption: The general process of binary fission in a rod-shaped prokaryote. For simplicity, the nucleoid is depicted as a single circle in green. 7/5/2018 mikro PTS - pet - 2 2014

Figure: 06-08 Caption: Typical growth curve for a bacterial population. See Sections 6.5 and 6.6 for a description of the counting methods employed. 7/5/2018 mikro PTS - pet - 2 2014

Bacterial growth Bacterial replication Two equivalent daughter cells Binary fission (Escherichia coli: 20 minutes Mycobacterium much slower: 12-24h) 7/5/2018 mikro PTS - pet - 2 2014

35 kinds of infectious diseases class one: plague, cholera class two: viral hepatitis, bacillary dysentery and intestinal amebiasis, typhoid fever and paratyphoid fever, AIDS, gonorrhea , avariosis, poliomyelitis, measles, pertussis, diphtheria, epidemic cerebrospinal meningitis, scarler fever, epidemic hemorrhagic fever, rabies, leptospirosis, brucellosis, anthrax, typhus, kala-azar, epidemic encephalitis B, malaria, dengue fever, lung tuberculosis, fetal tetanus class three:schistosomiasis , filariasis, hydatid disease, leprosy , influenza, mumps, rubella, infectious dysentery, acute hemorrhagic conjunctivitis 7/5/2018 mikro PTS - pet - 2 2014

Spores Resistant to harsh conditions (loss of nutritional requirement, dessication, intense heat, radiation and attack by most enzymes and chemical agents) 6-8 hours 7/5/2018 mikro PTS - pet - 2 2014

Kinds of spores Endospore: Bacillus subtilis Terminal endospore: Clostridium tetani ‘drumstick’ Subterminal: Clostridium botulinum 7/5/2018 mikro PTS - pet - 2 2014

Endospores (spores) Identification of Bacteria Pathogenesis Resistance 7/5/2018 mikro PTS - pet - 2 2014

Endospores (spores) Dormant cell Resistant to adverse conditions - high temperatures - organic solvents Produced when starved Contain calcium dipicolinate DPA, Dipicolinic acid Bacillus and Clostridium 7/5/2018 mikro PTS - pet - 2 2014

Figure: 04-62 Caption: Formation of the endospore. Phase contrast photomicrographs are of cells of Clostridium pascui. 7/5/2018 mikro PTS - pet - 2 2014

Shape & position of bacterial spore Oval central Spherical central Non bulging Oval sub terminal Oval sub terminal Bulging Oval terminal Spherical terminal Free spore 07.09.08 Dr Ekta Chourasia

Metabolism Catabolism: substrate breakdown and conversation into usable energy Anabolism: synthesis of cellular constituents (cell wall, proteins,fatty acids, nucleic acids 7/5/2018 mikro PTS - pet - 2 2014

Bacterial growth Bacterial replication Two equivalent daughter cells Binary fission (Escherichia coli: 20 minutes Mycobacterium much slower: 12-24h) 7/5/2018 mikro PTS - pet - 2 2014

Figure: 01-04 Caption: The machine and coding functions of the cell. In order for a cell to reproduce itself there must be an adequate supply of energy and precursors for the synthesis of new macromolecules, the genetic instructions must be replicated such that upon division each cell receives a copy, and genes must be expressed (the processes of transcription and translation) to form the proper amounts of necessary proteins and other macromolecules that will make up the new cell. 7/5/2018 mikro PTS - pet - 2 2014

7/5/2018 mikro PTS - pet - 2 2014