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THIS DOCUMENT SHALL NOT BESUBJECTED TO ANY MEANS OFREPRODUCTION, ELECTRONIC,MECHANICAL, INCLUDING PHOTOCOPYING,RECORDING, AND ANY FORM OFINFORMATION STORAGE DATABASEWITHOUT PERMISSION FROM THE AUTHORTHIS MATERIAL IS ALLOTTED FORPLM BS BIOLOGY STUDENTS USE ONLY.PLM-BS. BIOLOGY BATCH 2023 | MICROBIOLO...

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THIS DOCUMENT SHALL NOT BE SUBJECTED TO ANY MEANS OF REPRODUCTION, ELECTRONIC, MECHANICAL, INCLUDING PHOTOCOPYING, RECORDING, AND ANY FORM OF INFORMATION STORAGE DATABASE WITHOUT PERMISSION FROM THE AUTHOR THIS MATERIAL IS ALLOTTED FOR PLM BS BIOLOGY STUDENTS USE ONLY.

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Lecture 1

Lecture 2

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CONTRIBUTORS Notes Patrizia Starr A. Morados Roger John F. Peñas Leslie E. Traqueña Stephen Kyle C. Arcan Angelica Cacho Ericka Renee H. Gonzales Patrizia Starr A. Morados Roger John F. Peñas Leslie E. Traqueña

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Trans creator Stephen Kyle C. Arcan Jazzmine Rose M. Mones Leslie E. Traqueña Jazzmine Rose M. Mones Leslie E. Traqueña

Lecture 3 Lecture 4 Lecture 5

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PLM-BS. BIOLOGY BATCH 2023 | MICROBIOLOGY 22 1

HISTORY OF MICROBIOLOGY Modul 1 Lecture 1 b Prof. Emmanuel Divinagracia

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Outline I. Introduction to Microbiology II. Scientific Method III. Theories a. Theory A. Spontaneous Generation B. Germ Theory IV. Timeline V. Taxonomy

page 3 page 3 page 4 page 4,5

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I. INTRODUCTION TO MICROBIOLOGY Microbiology is a branch of biology that studies microorganisms that are too small to be seen without magnification, and their effects on humans, animals, plants, and other organisms.

Types of Microorganisms:

Results must be published and repeated by other investigators.

If evidence of a theory is so compelling that the next level of confidence is reached, it then becomes a Law or principl . If hypothesis is supported by a growing body of evidence and survives rigorous scrutiny, it moves to the next level of confidence by becoming theory.

Using the Scientific Method to Investigate Bacterial Endospores Observation / Information Gathering (1) Dr. Jenner observe the cows had a form of pox like smallpox. Jenner also noted that milkmaids acquired cowpox only in the hand and they appeared to be immune to smallpox.

Formation of Jenner’s hypothesis (2 Jenner deduced that the cowpox was closely related to smallpox and could possibly be used on patients to provide protection like that of the milkmaids he had seen.

• Bacteria Testing the hypothesis experiment I (3) Jenner took scrapings from cowpox blisters on the hand of a milkmaid and inoculated them into a boy who had not had smallpox. He developed minor symptoms but remained healthy.

• Viruses • Fungi • Protozoa • Helminths (Worms) • Algae II. SCIENTIFIC METHOD

Testing the hypothesis experiment II (4) After a few weeks, the child was exposed twice to the pus from an active smallpox lesion. He did not acquire smallpox and appeared to that immune protection.

Francis Bacon (1561 – 1626) Started scientific method • Approach to explain certain natural phenomena Form a hypothesis – a tentative explanation that can be supported A lengthy process of experimentation analysis and testing

Reproducibility of result (5) Jenner went on to inoculate 23 other test subjects with cowpox. For the first time, he used lesion from one hild to inoculate another. All subjects remained protected from smallpox. Publishing of results; other medical testing (6) Jenner wrote a paper detailing his experiment. He called his technique vaccinatio from the Latin vacca for cow.

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Other local English physicians began to vaccinate patients with some success.

o o

Vaccination theory become widespread (7)

through the time the broth became cloudy organic matter contains vital forces that could give rise to life.

Over the next 100 years vaccination was brought to the rest of the world through local programs. Scientist used Jenner’s methods to developed vaccines for other pathogens. The theory of artificial immunity became well established. Smallpox is eradicated from the world. (8) A massive vaccination campaign aimed to reduce cases and to stamp out the disease completely. Billions of doses given over a decade reduce smallpox to zero. The last cases occurred in 1977 and 1979 the disease was declared eradicated. III. THEORIES Spontaneous Generation – an early belief that some forms of life could arise from vital forces present in nonliving or decomposing matter (flies from manure, etc.) Aristotle 384 - 322 B.C o o

Figure 1.2 Needham’s Set up

Lazzaro Spallanzani (1729 – 1799) o o o o

Against to spontaneous generation he improved the experiment of Needham he boiled the broth longe Air carried germs on cultured medium

First person to develop the idea of spontaneous generation Simpler invertebrates could arise from spontaneous generation

Francesco Redi (1626 – 1697) o o o

Against to spontaneous generatio Experiment on decaying mea No maggots/flies produce if the meat is covered

Figure 1.3 Spallanzani’s Set up

Theodore Schwann (1810 – 1882) Figure 1.1 Redi’s Set up

o o o o

John Needham (1713 – 1781 o o

Against to spontaneous generation He allowed the air to enter the flask that has sterile nutrient solution Heated the incoming gas No growth observed in boiled broth media

support spontaneous generation. he boiled the broth and poured into covered flask.

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Figure 1.4 Schwann’s Set up Figure 1.5 Pasteur’s experiment with the swan-necked flask George Friedrich Schroder (1810 – 1885) and Theodor von Dusch (1824 – 1890) o o o

Allowed air to enter flask. Flask is heater and sterilized. No growth in the medium kahit hindi heated ung air.

The Germ Theory of Disease Many diseases are caused by the growth of microbes in the body and not by sins, bad character, or poverty, etc Two major contributors: o Louis Pasteur (Anthrax, Fowl cholera, rabies) o Robert Koc (Anthrax, cholera, Tb)

Louis Pasteur (1822 – 1895) o

o

he debunked the spontaneous generation and showed that fermentation, spoiling, and contamination of organic substances was due to the presence of environmental microorganisms. He was challenged to a debate that offers a large prize money to whoever that can refute the spontaneous generation. He won and garnered recognition.

o

Proved the germ theory of disease and launched the modern microbiology.

o

In his debate with Archime de Pouchet, he elegantly showed how sterilizing, chemical sterilization and filtration of air and water could maintain organic materials in sterile conditions without microbial growth. This became the Pasteurization method. His fame for his rabies research led the French Government in 1888 to build the Pasteur Institute located in Paris. It is originally established as a clinical center for treatment for rabies and other infectious diseases.

IV. TIMELINE Girolamo Fracastoro o o o

“de Contagione” (1546) – treatise on the germ theory He surmised that tiny, free living organism, which he referred to as “seeds of disease” existed in nature. He postulated that these disease – causing organism could be transmitted from person to person directly or via for mite intermediaries, thereby spreading contagion.

Antoine van Leeuwenhoek (1632-1723) o o o o o

Dutch linen merchant Invention of microscope (*1677) made him famous. First to observe living microbes Single lens magnified up to 300X. The founding father of microbiology

Leeuwenhoek’s Work: He used home-made microscopes (single lens magnified up to 300X) to discover the invisible world of micro-organisms.

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Through his microscopic observations of organisms such as bacteria and protozoa, Antoine van Leeuwenhoek effectively began the discipline of microbiology. His studies of insects, mollusks, and fish showed that these animals did not begin their life cycle with spontaneous generation, from nonliving matter. The development of microscope confirming Fracastoro’s hypothesis. He first observed bacteria in the year 1676 and called them 'animalcules' (from Latin animalculum' meaning tiny animal). Most of the animalcules are now referred to as unicellular organisms, although he observed multicellular organisms in pond water.

John Snow (1813 – 1858) o o o

o

Founding father of epidemiology He speculated that cholera was a waterborne or foodborne, intestinal illness (1849) He observed the highest incidence of disease was at the site of pumping water station. The water intake was drawn from Thames River. He deduced that the infection was transmitted by contaminated water.

Agostino Bassi (1773 – 1856) o o o

Showed that microorganism could cause disease Disease cause by fungal infectio Disease could be due to microbial infectio .

Reverend Miles Joseph Berkeley Marcuz Plenciz o

Proposed germ theory of diseases (1762)

Jacob Henle o

o o o

Advanced the germ theory concept (1840

Discovery of Spores and Sterilization

John Tyndall (1820 – 1893) and Ferdinand Cohn (1828 – 1898) o each demonstrated the presence of heat resistant forms of some microbes. o Cohn determined these to be heat resistant bacterial endospore o John Tyndall deals in the final blow of spontaneous generation; demonstrated the dust carried germs.

Potato blight cause by water mold (*1852) He noted the unmistakable presence of microscopic mold elements in diseased plants. Potato blight led to the death of one million Irish and result in the mass emigration of approximately two million of their countrymen.

Heinrich de Bary o

Smut and rust fungal can cause disease (1853

Anton de Bary o

He proved that the etiology of potato blight was in fact a fungus (Phytophthora infectants) meaning plant destroye

Louis Pasteur (1822 – 1895) Sterility requires the elimination of all life forms including endospores and viruses. Ignaz Semmelweis o o

o

o

hand hygiene (1847 He recognized that the putrid odor associated with women dying of puerperal fever was similar to that emanating from corpse during autopsies by medical faculty and student He hypothesized that some form of “putrid matter” must be carried on the hands of physicians during their rounds between the autopsy and birthing tables and might be transmitted to pregnant women causing this highly lethal peripartum illnes Semmelweis found that washing hands using a dilute, chloride lime solution would remove the putrid odor

o o o o o

Showed microbes caused fermentation and spoilage. Disproved spontaneous generation of microscope. Proved the germ theory diseases. Developed pasteurization. He showed that heat sterilization, chemical sterilization, or filtration of air and water could maintain organic materials in sterile conditions indefinitely without any microbial growth.

Joseph Lister (1827 – 1912) o o o

Sterile technique (1867 He uses the sterile method to protect the wounds of trauma patient He investigated the use of chemical disinfectants as a method of preventing wound infection

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o

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The carbolic acid decreases the fetid odor, diluted solution of this chemical maintains the sterility of dressings, surgical instruments, and the hands of surgeons caring for injured patient Establishing principle of antisepsis

Charles Chamberland o o

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He identified anthrax bacilli in the blood of infected sheep and successfully transmitted the infection into healthy experiment animals. He employed the use of oil immersion microscope to study bacteria.

o

Developed new staining method for bacterial identification. Stains used for M. tuberculosis: Alkaline methylene blue in conjunction with a second stain (Bismarck brown)

o

Invented the procedure for the isolation of pure bacterial culture on solid media. /facilitated using aga as solidifying agent in flat Petri” dishes – Richard Petri.

Invented the autoclave – a water purification device that was later used in the discovery of viruse Developed Pasteurella vaccin

Alexandre Yersin o

Co-discoverer of plague bacillus

Jules Bordet o

discovered whooping cough bacillus and complement

Ilya Metchnikoff o

discovered the process of phagocytosis and provide the initial description of innate immunity

Albert Calmette o

discovered cobra antivenin and developed Bacillus – Calmette – Guerin – the first effective tuberculosis vaccine

o

The Koch Plate Technique – to obtain pure growth he insisted upon the use of single colony isolation Discovered microbial etiology of tuberculosis (1882 • Mycobacterium tuberculosi Established Koch’s postulate – a sequence of experimental steps that verified the germ theory.

Louis Pasteur (1822 – 1895) o o

o

o

o o

Attenuated vaccine (1879) Pasteur observed that after the serial passage the chicken cholera bacillus, now known as Pasteurella spp lost the capacity to cause lethality when injected to chickens. Chicken previously exposed to attenuated bacilli survived infection with virulent strains while the naïve chicken died rapidly. The serial passage of the bacteria at certain elevated temperature ranges and in the presence of oxygen resulted in organisms that could induce resistance to challenge using virulent forms of the same bacteria. Developed vaccine against anthrax (1881) Vaccine against rabies (1885) Robert Koch (1843 – 1910)

o o

Describe the life cycle of anthrax and the process of endospore formation (*1876 He began his investigations into the etiology of anthrax in sheep.

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o Tools for every criterion: ▪ 1. Microscopy, Staining = observe blood/tissue under the microscope ▪ 2. Laboratory Culture = Streak agar plate sample from either diseased or healthy animal. Next is inoculate healthy animal with cells of suspected pathogen. ▪ 3. Experimental animal o Remove blood or tissue sample and observe by microscopy. ▪ 4. Laboratory culture = pure culture (outcome -must be the same organism as before)

transforming principle or genetic material of Streptococcus pneumoniae was DNA not protein as previously postulated.

James Watson, Francis Crick, Rosalind Franklin and Maurice Wilkins o

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elucidation of DNA structure (1953)

First complete genomic sequence of bacteriophage (1977) First draft of human genome (2001)

__________________________________________________ Development of Aseptic Techniques The human body is a source of infection.

Max von Pettenkoffer o o

Sanitation theory Poor sanitation, unfavorable water, soil conditions and damp weather generated miasma poisons that subsequently caused illness.

• Dr. Ignaz Semmelweis – correlated infection with physicians coming directly from autopsy room to the maternity ward.

Rudolf Virchow o

• Joseph Lister – introduced aseptic techniques to reduce microbes in medical settings and prevent wound infections.

Founding father of cellular pathology

Paul Ehrich o

Co-discoverer of antibodies, antigens, chemotherapy for infectious diseases

• Dr. Oliver Wendell Holmes – observed that mothers of home births had fewer infections than those who gave birth in hospitals.

o Use of heat for sterilization and

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VI. TAXONOMY Richard Pfeiffer o o o

Discovered bacterial endotoxin. Discovered the phenomenon of bacteriolysis. Played a major role in the development of killed typhoid vaccine.

Emil von Behring o

Discover the serum therapy for diphtheria and tetanus.

Shibasaburo Kitasato and Sakahiro Hata o o

Important contribution in serum therapy Discovery of salvasaran for the treatment of syphilis

Patrick Manson o

Confired the presence of microscopic parasite in mosquito vector of filiarisis.

Ronald Ross o

Prove the parasitic nature of malaria and transmission by mosquitoes (1896)

Oswald Avaery, Maclyn McCarty and Colin MacLeod o

identified the “holy grain” of genetics (1944)

Organization, classifying, and naming of living things Formal system originated by Carl von Linné Concerned with: • Classification – orderly arrangement of organisms into groups • Nomenclature – assigning names • Identification – determining and recording traits of organisms for placement into taxonomic schemes Levels of Classifications: • Domain – Archaea, Bacteria, and Eukarya • Kingdom • Phylum or Division • Class • Order • Family • Genus Assigning Specific Names • Binomial (scientific) nomenclature • Gives each microbe 2 names: o Genus – capitalized. o Species – lowercase • Both italicized / underlined o Staphylococcus aureus (S. aureus) • Inspiration for names is extremely varied and often imaginative

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Three Domains of Life • Bacteria – true bacteria • Archaea – odd bacteria that live in extreme environments, high salt, heat, etc. • Eukarya – have a nucleus and organelles

Major References: S. Opal (n.d) A Brief History of Microbiology and Immunology. Retrieved March 18,2021 from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7176178 / Madigan, Mantiko, Dunlap and Clark Brock Biology of Microorganisms 12 th Edition

Review Key terms Aseptic technique

Basic dyes

Differential stains Enrichment culture Technique

Koch’s postulate Nutrient agar

Pure culture

Septic Sterile Sterilization

Using procedures contamination.

to

avoid

Ex. Kapag mag subculture ng bacteria, gagamit ka ng Laminar flow hood para maprotect ang mga samples for airborne contaminations, also gagamit ng alcohol lamp/Bunsen burner para mapatay ang other microbes habang nagiinoculate. (sa lab malalaman nyo ang procedures). Positively charged dyes, kaya nilang magbind strongly sa negatively charged cellular constituents tulad ng nucleic acid and acidic polyssacharides. Ex. Methylene blue, crystal violet and safranin. Usually ginagamit to in gram staining method to check if ang organism is gram (+) or gram (-). Stains that render different kinds of cells different color. A method of isolating specific microorganisms from nature using specific culture media and incubation conditions. According to E. Salud when we say ECT bibigay mo lang yung optimum nutrients na need ng microbes mo, while sa subculture is ililipat mo sila sa another petri plates for viability because may different phases ang microbial growth. A set of criteria for proving that a given microorganism causes a given disease. Is a polysaccharide derived from red algae. It was used widely in 19 th century as a gelling agent. A culture containing a single kind of microorganism. Kapag ibat ibang microbes na ang nasa plate, mixed culture na ang tawag doon. Putrefying; infected by microbes Free of all living organisms and viruses A process in killing all the bacteria or other microorganisms in or on objects.

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PLM-BS. BIOLOGY BATCH 2023 | MICROBIOLOGY 22 1

Characterization of Microorganisms Modul 1 Lecture 2 b Prof. Emmanuel Divinagracia Legend Book

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