Pharmaceutical Microbiology Chapter 1

History of Microbiology

• John Tyndall showed that open broth tubes were free of bacteria when air was free of dust.
• Louis Pasteur is known as the Father of Microbiology.
• Lord Lister demonstrated the antiseptic method.
• Edward Jenner discovered the small pox vaccine.
• Antonie van Leeuwenhoek is known as the Father of Medical Microbiology.

Microscope

• In a fluorescent microscope, the objective lens is made of polythene.
• The resolution power of the compound microscope is 0.2 micron.

Prokaryotic Organism

• Prokaryotic organisms are characterized by the absence of nuclear envelope.

Antiseptic Method

• Lord Lister introduced the antiseptic method.

Microbiology

• Plasmids are responsible for motility.
• Endotoxin is present in the lipopolysaccharide of gram-negative bacteria.
• Gram-negative bacteria have a thin cell wall.
• Fimbriae are used for cell attachment.
• Catalase production is negative in Streptococcus.

Gram Staining

• Crystal violet is the primary stain of Gram's method.
• Gram's decolouriser is alcohol.
• Gram-positive bacteria stain purple, while gram-negative bacteria stain pink.
• The order of reagents used in the Gram stain is crystal violet, iodine, decolorizer, and safranin.

Sterilization and Disinfection

• Autoclave is used to sterilize heat-stable materials.
• Membrane filtration is used to sterilize heat-labile solutions.
• The D value is the time required to kill 90% of the microorganisms in a sample at a specific temperature.
• Pasteurization is used to preserve food, and it involves heating at 63°C for 30 minutes.
• Dry heat sterilization is used to sterilize glass pipettes.
• Ionizing radiation is used to sterilize and disinfect.
• Sterilization is the process of making an object free from living organisms, including bacteria, fungal spores, and viruses.

Disinfectants

• Cationic detergents act by disrupting microbial membranes.
• Silver nitrate and mercuric chloride are disinfectants containing heavy metals.
• Ethylene oxide is a sterilant that can be used to sterilize.
• Bactericidal concentration of phenol is 1%.
• Iodophores are a mixture of iodine and surface active agents.
• Halogens are commonly used as general antiseptics to treat cuts and scratches.

Virology

• For a virus to replicate, the genome must be released in the cytoplasm.### Viruses
• Viruses have been successfully grown in pure cultures in test tubes.
• All viruses are obligatory intracellular parasites.
• All viruses have either DNA or RNA as their genetic material.
• Viruses probably arose from small fragments of cellular chromosomes.
• The size of viruses is usually measured in nanometers.

Viral Capsid

• The function of a viral capsid is to provide protection against the viral genome from physical and enzymatic destruction.
• It also provides binding sites that enable the virus to attach to specific receptor sites on the host cell.

Fungi

• Fungi that lack partitions (septa) are called coenocytic.
• Dimorphic fungi exhibit yeast-like growth at human body temperatures and mold-like growth at room temperature.
• Slime molds produce motile sexual and asexual spores.
• Fungi secrete extracellular enzymes to break down nutrients.
• Fungi in the division Deuteromycota are characterized by the fact that a method of sexual reproduction has not been identified.
• Black bread molds produce zygospores.

Laboratory Safety

• The desire to maintain a safe laboratory environment begins with prevention.
• When a chemical splashes in the eye, rinse for 15 minutes.
• Personal Protective Equipment (PPE) such as safety glasses, lab coats, and gloves are frequently used.
• Chemical, reagents, or broth cultures should not be pipetted by mouth.
• Good work practices include confining long hair and loose clothing, and not using damaged equipment and glassware.

Microbiological Assay

• The name of the procedure performed under sterile conditions to eliminate contamination in hopes to obtain a pure culture of one type of microorganism is aseptic technique.
• After a biohazard spill, the area must sit for 30 minutes before cleaning.
• Media is needed as a source of nutrient for the growth and reproduction of microbes.
• To prevent the contamination of microscopes and surrounding areas, disinfect/clean used slides with 70% ethanol and lens paper.

Antibiotics

• Bacteriostatic antibiotics stop bacteria from growing without killing them.
• One level assay of antibiotics is used to determine concentration.
• Two level assay of antibiotics is used to determine potency.

Clean Room Technology

• Cleanrooms maintain particulate-free air through the use of HEPA filters.
• Cleanrooms are classified by how clean the air is.
• A cleanroom is a controlled environment that has a low level of airborne microbes.
• Following is the lowest level of cleanroom standards: ISO 9.

Microbiological Assay (continued)

• Zone of inhibition measured in microbiological assay of antibiotics.
• Zone of growth measured in microbiological assay of vitamins.
• Median concentration in microbial assay is 1 µg/ml.
• Disadvantage of Agar diffusion method except dilution.

Food Processing Technology

• Most spoilage bacteria grow at neutral pH.### Food Processing Technology
• Acetic acid, Tartaric acid, Citric acid, and Maleic acid are types of acids used in food processing.
• Salmonella, E. coli, Staphylococcus, and Cyano bacteria are types of bacteria that can be present in raw or undercooked meat, eggs, seafood, and unpasteurized milk.
• Milk and curry left over can spoil at room temperature, high temperature, very low temperature, or constant temperature.

Microbiology

• Spoilage in food can be prevented or delayed by hindering the activity of micro-organisms, physical removal of dust, or cleaning the environment.
• Aerobic food spoilage and pathogenic microorganisms can be suppressed by removing nutrients from food or adding antibodies.
• In spore-forming bacteria, maximum resistance occurs at pH 4, 5, 6, or 7.
• Any change that renders food unfit for human consumption is called deterioration or spoilage.

Food Intoxication

• Food intoxication occurs through the ingestion of toxins produced by microorganisms, toxin-producing additives, chemical interaction, or toxins produced by the body.
• Clostridium Botulinum is a type of bacteria that can cause food intoxication.

Microorganisms

• Yeast is used for alcoholic fermentation, and Saccharomyces Cerevisiae is a type of yeast used for this process.
• CO2 concentration required for culturing animal cells is between 2-5% or 1-10%.
• The human fibroblast is a classical example of stable primary cell lines.
• pH of culture medium is initially controlled by the presence of CO2, bicarbonate buffer, or addition of bases.

Cell Culture

• In animal cell culture, transformation means uptake of new genetic material, phenotypic modifications of cells in culture, increased division of the cell, or release of genetic information.
• Cells that have undergone transformation frequently become anchorage independent or anchorage dependent.
• Lactic acid, Creatine phosphate, ATP, and Glucose are sources of energy in active muscle cells.

Microbiology History

• The history of microbiology involves the study of bacteria, microscopy, and staining techniques.
• Sterilization and disinfectant methods are used to control microorganisms.
• Study of viruses, fungi, and bacteria involves understanding their growth curves, staining techniques, and sterilization methods.

Aseptic Area Design

• Designing an aseptic area involves understanding general aspects of environmental cleanliness.
• Assessment of a new antibiotic involves understanding its effects on microorganisms.

Spoilage

• Types of spoilage include physical, chemical, and microbiological spoilage.

Animal Cell Culture

• Growth of animal cells in culture involves understanding the constituents of culture medium, pH control, and sources of energy for active muscle cells.