text: logging in or signing up sterilisation and Disinfection By Prof. Satish Polshettiwar, MIP Pune satishmip Download Post to : URL : Related Presentations : Let's Connect Share Add to Flag Embed Email Add to Channel Copy embed code: Embed: Flash iPad Dynamic Copy Does not support media & animations Automatically changes to Flash or non-Flash embed WordPress Embed Customize Embed Use HTTPS URL: Copy Thumbnail: Copy The presentation is successfully added In Your Favorites. Views: 3509 Category: Education License: All Rights Reserved Like it (4) Dislike it (0) Added: March 18, 2011 This Presentation is Public Favorites: 1 Presentation Description No description available. Comments Posting comment... Premium member Presentation Transcript Slide 1: MAEER'S, MAHARASHTRA INSTITUTE OF PHARMACY, PUNE-38. CONTROL OF MICROBIAL GROWTH Prof. S. A. Polshettiwar M. Pharm, D.I.T., PhD Slide 2: Control of Microbial Growth: Introduction Early civilizations practiced salting, smoking, pickling, drying, and exposure of food and clothing to sunlight to control microbial growth. Use of spices in cooking was to mask taste of spoiled food. Some spices prevented spoilage. In mid 1800s Semmelweiss and Lister helped developed aseptic techniques to prevent contamination of surgical wounds. Before then: Nosocomial infections caused death in 10% of surgeries. Up to 25% mothers delivering in hospitals died due to infection Reasons for controlling microorganism : Reasons for controlling microorganism To prevent contamination in sterile product To prevent transmission of microorganisms responsible to cause disease. To prevent decompostion,spoilage of food and food product. To prevent contamination of unwanted microbes in pure culture To prevent contamination in aseptic area. Sterilization : Sterilization Sterilization (or sterilisation, see spelling differences) is a term referring to any process that eliminates (removes) or kills all forms of life, including transmissible agents (such as fungi, bacteria, viruses, spore forms, etc.) present on a surface, contained in a fluid, in medication, or in a compound such as biological culture media.Sterilization can be achieved by applying the proper combinations of heat, chemicals, irradiation, high pressure, and filtration. Slide 5: Control of Microbial Growth: D efinitions Sterilization: Killing or removing all forms of microbial life (including endospores) in a material or an object. Heating is the most commonly used method of sterilization. Commercial Sterilization: Heat treatment that kills endospores of Clostridium botulinum the causative agent of botulism, in canned food. Does not kill endospores of thermophiles, which are not pathogens and may grow at temperatures above 45oC. Slide 6: Control of Microbial Growth: D efinitions Disinfection: Reducing the number of pathogenic microorganisms to the point where they no longer cause diseases. Usually involves the removal of vegetative or non-endospore forming pathogens. May use physical or chemical methods. Disinfectant: Applied to inanimate objects. Antiseptic: Applied to living tissue (antisepsis). Degerming: Mechanical removal of most microbes in a limited area. Example: Alcohol swab on skin. Sanitization: Use of chemical agent on food-handling equipment to meet public health standards and minimize chances of disease transmission. E.g: Hot soap & water. Slide 7: Control of Microbial Growth: D efinitions Sepsis: Comes from Greek for decay or putrid. Indicates bacterial contamination. Asepsis: Absence of significant contamination. Aseptic techniques are used to prevent contamination of surgical instruments, medical personnel, and the patient during surgery. Aseptic techniques are also used to prevent bacterial contamination in food industry. Slide 8: Control of Microbial Growth: D efinitions Bacteriostatic Agent: An agent that inhibits the growth of bacteria, but does not necessarily kill them. Suffix stasis: To stop or steady. Germicide: An agent that kills certain microorganisms. Bactericide: An agent that kills bacteria. Most do not kill endospores. Viricide: An agent that inactivates viruses. Fungicide: An agent that kills fungi. Sporicide: An agent that kills bacterial endospores of fungal spores. Slide 9: Control of Microbial Growth: Rate of Microbial Death When bacterial populations are heated or treated antimicrobial chemicals, they usually die at a constant rate. Slide 10: Control of Microbial Growth: Rate of Microbial Death Several factors influence the effectiveness of antimicrobial treatment. 1. Number of Microbes: The more microbes present, the more time it takes to eliminate population. 2. Type of Microbes: Endospores are very difficult to destroy. Vegetative pathogens vary widely in susceptibility to different methods of microbial control. 3. Environmental influences: Presence of organic material (blood, feces, saliva) tends to inhibit antimicrobials, pH etc. 4. Time of Exposure: Chemical antimicrobials and radiation treatments are more effective at longer times. In heat treatments, longer exposure compensates for lower temperatures. Slide 11: Physical Methods of Microbial Control: Heat: Kills microorganisms by denaturing their enzymes and other proteins. Heat resistance varies widely among microbes. Thermal Death Point (TDP): Lowest temperature at which all of the microbes in a liquid suspension will be killed in ten minutes. Thermal Death Time (TDT): Minimal length of time in which all bacteria will be killed at a given temperature. Decimal Reduction Time (DRT): Time in minutes at which 90% of bacteria at a given temperature will be killed. Used in canning industry. Slide 12: Physical Methods of Microbial Control: Moist Heat: Kills microorganisms by coagulating their proteins. In general, moist heat is much more effective than dry heat. Boiling: Heat to 100oC or more at sea level. Kills vegetative forms of bacterial pathogens, almost all viruses, and fungi and their spores within 10 minutes or less. Endospores and some viruses are not destroyed this quickly. However brief boiling will kill most pathogens. Hepatitis virus: Can survive up to 30 minutes of boiling. Endospores: Can survive up to 20 hours or more of boiling. STERILIZATION METHODS : STERILIZATION METHODS Physical Methods: 1. Moist Heat Sterilization a. Autoclave b. Boiling at 100 deg. C c. Pasteurization d. heating with bactericide e. Tydallization 2. Dry heat sterilization a. Hot air oven b. Flaming c. Red hot technique d. Incineration 3. Filtration or Mechanical Method: Membrane filter 4. Chemical method Gaseous sterilization or by using disinfectant 5. Irradiation : UV., Gamma etc NOTE: End products must pass sterility tests. Autoclave: Closed Chamber with High Temperature and Pressure : Autoclave: Closed Chamber with High Temperature and Pressure Slide 16: Laboratory autoclave Industrial autoclave Autoclave: : Autoclave: An autoclave is a device to sterilize equipment and supplies by subjecting them to high pressure saturated steam at 121 °C or more, typically for 15-20 minutes depending on the size of the load and the contents. It was invented by Charles Chamberland in 1879 Mechanism: : Mechanism: Saturated steam condensate on the material or microbes and gives up heat and the material or microbes get heated to 121oC or more depending upon the pressure. While it condenses, it hydrate the microbes by proving latent heat from vapor thereby making them vulnerable to killing. Moreover, when the steam condensate it contract and vacuum is created. Fresh store of steam takes its place and pushes I under pressure, penetrating the load and killing the microorganism. Killing is done by : Coagulation of protein Denaturation of protien Sterilization cycle or steps: Loading and packing of the autoclave Raising temp and pressure Holding the load at specific temp and pressure Cooling and unloading Slide 19: Physical Methods of Microbial Control: Moist Heat (Continued): Reliable sterilization with moist heat requires temperatures above that of boiling water. Autoclave: Chamber which is filled with hot steam under pressure. Preferred method of sterilization, unless material is damaged by heat, moisture, or high pressure. Temperature of steam reaches 121oC at twice atmospheric pressure. Most effective when organisms contact steam directly or are contained in a small volume of liquid. All organisms and endospores are killed within 15 minutes. Require more time to reach center of solid or large volumes of liquid. STERILIZATION : STERILIZATION 10 lb Pressure (115.5 deg. C)...30 minutes 15 lb Pressure (121.5 deg. C)...20 minutes 20 lb Pressure (126.5 deg. C)... 15 minutes STERILIZATION : STERILIZATION Application: 1. Solutions sealed in containers ampuls, vials 2. Bulk Solutions 3. Glassware 4. Surgical Dressing 5. Instruments 6. Culture media STERILIZATION : STERILIZATION Advantages: Rapid, Inexpensive, Effective, Large volumes, Steam has Better penetrating power and applicable to wide variety of materials Shorter exposure of sterilization Disadvantages: 1. Cannot use for oily preparation (oil base ointment) 2. Cannot use for moisture sensitive preparations Slide 23: Physical Methods of Microbial Control: Moist Heat (Continued): Pasteurization: Developed by Louis Pasteur to prevent the spoilage of beverages. Used to reduce microbes responsible for spoilage of beer, milk, wine, juices, etc. Classic Method of Pasteurization: Milk was exposed to 65oC for 30 minutes. High Temperature Short Time Pasteurization (HTST): Used today. Milk is exposed to 72oC for 15 seconds. Ultra High Temperature Pasteurization (UHT): Milk is treated at 140oC for 3 seconds and then cooled very quickly in a vacuum chamber. Advantage: Milk can be stored at room temperature for several months. Slide 24: Physical Methods of Microbial Control: Dry Heat: Kills by oxidation effects. Direct Flaming: Used to sterilize inoculating loops and needles. Heat metal until it has a red glow. Incineration: Effective way to sterilize disposable items (paper cups, dressings) and biological waste. Hot Air Sterilization: Place objects in an oven. Require 2 hours at 170oC for sterilization. Dry heat is transfers heat less effectively to a cool body, than moist heat. Hot air Oven : Hot air Oven Hot air Oven : Hot air Oven It works on the principle of killing of microorganism by dry heat at 150 to 170oC for 1 hrs Killes microorgnaism by oxidation of protie. Slide 27: Physical Methods of Microbial Control: Filtration: Removal of microbes by passage of a liquid or gas through a screen like material with small pores. Used to sterilize heat sensitive materials like vaccines, enzymes, antibiotics, and some culture media. High Efficiency Particulate Air Filters (HEPA): Used in operating rooms and burn units to remove bacteria from air. Membrane Filters: Uniform pore size. Used in industry and research. Different sizes: 0.22 and 0.45um Pores: Used to filter most bacteria. Don't retain spirochetes, mycoplasmas and viruses. 0.01 um Pores: Retain all viruses and some large proteins. Slide 28: Nitrocellulose filter Slide 30: A complete filtering set up Slide 31: Physical Methods of Microbial Control: Low Temperature: Effect depends on microbe and treatment applied. Refrigeration: Temperatures from 0 to 7oC. Bacteriostatic effect. Reduces metabolic rate of most microbes so they cannot reproduce or produce toxins. Freezing: Temperatures below 0oC. Flash Freezing: Does not kill most microbes. Slow Freezing: More harmful because ice crystals disrupt cell structure. Over a third of vegetative bacteria may survive 1 year. Most parasites are killed by a few days of freezing. Slide 33: Physical Methods of Microbial Control: Dessication: In the absence of water, microbes cannot grow or reproduce, but some may remain viable for years. After water becomes available, they start growing again. Susceptibility to dessication varies widely: Neisseria gonnorrhea: Only survives about one hour. Mycobacterium tuberculosis: May survive several months. Viruses are fairly resistant to dessication. Clostridium spp. and Bacillus spp.: May survive decades. Slide 34: Physical Methods of Microbial Control: Osmotic Pressure: The use of high concentrations of salts and sugars in foods is used to increase the osmotic pressure and create a hypertonic environment. Plasmolysis: As water leaves the cell, plasma membrane shrinks away from cell wall. Cell may not die, but usually stops growing. Yeasts and molds: More resistant to high osmotic pressures. Staphylococci spp. that live on skin are fairly resistant to high osmotic pressure. Slide 35: Physical Methods of Microbial Control: Radiation: Three types of radiation kill microbes: 1. Ionizing Radiation: Gamma rays, X rays, electron beams, or higher energy rays. Have short wavelengths (less than 1 nanometer). Dislodge electrons from atoms and form ions. Cause mutations in DNA and produce peroxides. Used to sterilize pharmaceuticals and disposable medical supplies. Food industry is interested in using ionizing radiation. Disadvantages: Penetrates human tissues. May cause genetic mutations in humans. Forms of Radiation : Forms of Radiation Radiation : Radiation Non- ionizing radiation (UV light) Ultraviolet light Damages DNA Adjacent thymines (pyrimidine base) form bonds Forms thymine dimers Inhibits correct replication of DNA UV lamps Germicidal lamps Disadvantage Rays do not penetrate, microbes on surfaces Cannot penetrate paper Prolonged exposure Eyes damage, burns, and skin cancer Slide 38: Physical Methods of Microbial Control: Radiation: Three types of radiation kill microbes: 2. Ultraviolet light (Nonionizing Radiation): Wavelength is longer than 1 nanometer. Damages DNA by producing thymine dimers, which cause mutations. Used to disinfect operating rooms, nurseries, cafeterias. Disadvantages: Damages skin, eyes. Doesn't penetrate paper, glass, and cloth. Slide 39: Physical Methods of Microbial Control: Radiation: Three types of radiation kill microbes: 3. Microwave Radiation: Wavelength ranges from 1 millimeter to 1 meter. Heat is absorbed by water molecules. May kill vegetative cells in moist foods. Bacterial endospores, which do not contain water, are not damaged by microwave radiation. Solid foods are unevenly penetrated by microwaves. Trichinosis outbreaks have been associated with pork cooked in microwaves Comparison of Methods : Comparison of Methods Slide 41: Chemical Methods of Microbial Control Types of Disinfectants 1. Phenols and Phenolics: Phenol (carbolic acid) was first used by Lister as a disinfectant. Rarely used today because it is a skin irritant and has strong odor. Used in some throat sprays and lozenges. Acts as local anesthetic. Phenolics are chemical derivatives of phenol Cresols: Derived from coal tar (Lysol). Biphenols (pHisoHex): Effective against gram-positive staphylococci and streptococci. Used in nurseries. Excessive use in infants may cause neurological damage. Destroy plasma membranes and denature proteins. Advantages: Stable, persist for long times after applied, and remain active in the presence of organic compounds. Slide 42: Chemical Methods of Microbial Control Types of Disinfectants 2. Halogens: Effective alone or in compounds. A. Iodine: Tincture of iodine (alcohol solution) was one of first antiseptics used. Combines with amino acid tyrosine in proteins and denatures proteins. Stains skin and clothes, somewhat irritating. Iodophors: Compounds with iodine that are slow releasing, take several minutes to act. Used as skin antiseptic in surgery. Not effective against bacterial endospores. Betadine Isodine Slide 43: Chemical Methods of Microbial Control Types of Disinfectants 2. Halogens: Effective alone or in compounds. B. Chlorine: When mixed in water forms hypochlorous acid: Cl2 + H2O ------> H+ + Cl- + HOCl Hypochlorous acid Used to disinfect drinking water, pools, and sewage. Chlorine is easily inactivated by organic materials. Sodium hypochlorite (NaOCl): Is active ingredient of bleach. Chloramines: Consist of chlorine and ammonia. Less effective as germicides. Slide 44: Chemical Methods of Control Types of Disinfectants 3. Alcohols: Kill bacteria, fungi, but not endospores or naked viruses. Act by denaturing proteins and disrupting cell membranes. Evaporate, leaving no residue. Used to mechanically wipe microbes off skin before injections or blood drawing. Not good for open wounds, because cause proteins to coagulate. Ethanol: Drinking alcohol. Optimum concentration is 70%. Isopropanol: Rubbing alcohol. Better disinfectant than ethanol. Also cheaper and less volatile. Slide 45: Chemical Methods of Control Types of Disinfectants 4. Heavy Metals: Include copper, selenium, mercury, silver, and zinc. Oligodynamic action: Very tiny amounts are effective. A. Silver: 1% silver nitrate used to protect infants against gonorrheal eye infections until recently. B. Mercury Organic mercury compounds like merthiolate and mercurochrome are used to disinfect skin wounds. C. Copper Copper sulfate is used to kill algae in pools and fish tanks. Slide 46: Chemical Methods of Control Types of Disinfectants 4. Heavy Metals: D. Selenium Kills fungi and their spores. Used for fungal infections. Also used in dandruff shampoos. E. Zinc Zinc chloride is used in mouthwashes. Zinc oxide is used as antifungal agent in paints. Slide 47: Chemical Methods of Control Types of Disinfectants 5. Quaternary Ammonium Compounds (Quats): Widely used surface active agents. Cationic (positively charge) detergents. Effective against gram positive bacteria, less effective against gram-negative bacteria. Also destroy fungi, amoebas, and enveloped viruses. Zephiran, Cepacol, also found in our lab spray bottles. Pseudomonas strains that are resistant and can grow in presence of Quats are a big concern in hospitals. Advantages: Strong antimicrobial action, colorless, odorless, tasteless, stable, and nontoxic. Diasadvantages: Form foam. Organic matter interferes with effectiveness. Neutralized by soaps and anionic detergents. Slide 48: Chemical Methods of Control Types of Disinfectants 6. Aldehydes: Include some of the most effective antimicrobials. Inactivate proteins by forming covalent crosslinks with several functional groups. A. Formaldehyde gas: Excellent disinfectant. Commonly used as formalin, a 37% aqueous solution. Formalin was used extensively to preserve biological specimens and inactivate viruses and bacteria in vaccines. Irritates mucous membranes, strong odor. Also used in mortuaries for embalming. Slide 49: Chemical Methods of Control Types of Disinfectants 6. Aldehydes: B. Glutaraldehyde: Less irritating and more effective than formaldehyde. One of the few chemical disinfectants that is a sterilizing agent. A 2% solution of glutaraldehyde (Cidex) is: Bactericidal, tuberculocidal, and viricidal in 10 minutes. Sporicidal in 3 to 10 hours. Commonly used to disinfect hospital instruments. Also used in mortuaries for embalming. Slide 50: Chemical Methods of Control Types of Disinfectants 7. Gaseous Sterilizers: Chemicals that sterilize in a chamber similar to an autoclave. Denature proteins, by replacing functional groups with alkyl groups. A. Ethylene Oxide: Kills all microbes and endospores, but requires exposure of 4 to 18 hours. Toxic and explosive in pure form. Highly penetrating. Most hospitals have ethylene oxide chambers to sterilize mattresses and large equipment. Slide 51: Chemical Methods of Control Types of Disinfectants 8. Peroxygens (Oxidizing Agents): Oxidize cellular components of treated microbes. Disrupt membranes and proteins. A. Ozone: Used along with chlorine to disinfect water. Helps neutralize unpleasant tastes and odors. More effective killing agent than chlorine, but less stable and more expensive. Highly reactive form of oxygen. Made by exposing oxygen to electricity or UV light. Slide 52: Chemical Methods of Control Types of Disinfectants 8. Peroxygens (Oxidizing Agents): B. Hydrogen Peroxide: Used as an antiseptic. Not good for open wounds because quickly broken down by catalase present in human cells. Effective in disinfection of inanimate objects. Sporicidal at higher temperatures. Used by food industry and to disinfect contact lenses. C. Benzoyl Peroxide: Used in acne medications. Slide 53: Chemical Methods of Control Types of Disinfectants 8. Peroxygens (Oxidizing Agents): D. Peracetic Acid: One of the most effective liquid sporicides available. Sterilant : Kills bacteria and fungi in less than 5 minutes. Kills endospores and viruses within 30 minutes. Used widely in disinfection of food and medical instruments because it does not leave toxic residues. Efficiency of Different Chemical Antimicrobial Agents : Efficiency of Different Chemical Antimicrobial Agents The Bioburden : The Bioburden Initial number of microorganisms present in a given product. Sensitivity of Microorganism : Sensitivity of Microorganism Ideally , the sterilizing treatment to be used should be decided for each specific load with prior knowledge of the type and number of contaminating M.O. present. Sensitivity of Microorganism : Sensitivity of Microorganism Microorganism show their varying degrees of resistance to heat, radiation and chemicals. The vegetative forms of bacteria and fungi are most sensitive. The thermophilic bacteria, smaller viruses and mold spores are killed at temperature between 70 to 90 deg., while bacterial spores may be destroyed at 90 t o 120 deg. Temperature. Slide 58: SITES OF ANTIMICROBIAL ATTACK DNA replication Cytoplasm Death curves or Survivor curve : Death curves or Survivor curve Death in a microbial population is determined by assessing the reduction in the number of viable microorganism resulting from contact with a given destructive force. This can be represented graphically with a "survivor curve" drawn from the exposure time or dose. The death of a population of cells exposed to heat, radiation or toxic chemicals is often found to follow first order kinetics. Slide 60: Heating time or radiation dose Survivor Curve Fraction of survivors 10-1 10-2 10-3 10-4 10-8 A C B linear Slide 61: dN/dt = -kN where: N = number of viable organisms at time t k = thermal inactivation constant, or specific death rate Slide 62: Nt / No = e-kt Where: No = the number of original organisms present Nt = number of organisms remaining after time t Advantages : Advantages 1. The determination of death rates provides the facility to compare the resistance of the same microorganisms at different temperatures or to compare the resistance of different microorganism to the same lethal agent. E.g. temperature, radiation, chemical etc. 2. It may also be used to give quantitative measure of the effect of environmental factors such as pH, osmolarity and presence of various chemicals on the sterilization process. Physical Methods: Moist Heat : Physical Methods: Moist Heat Calculations using D and z values Given: For Clostridium botulinum spores suspended in phosphate buffer, D121 = 0.204 min How long would it take to reduce a population of C. botulinum spores in phosphate buffer from 1012 spores to 100 spores (1 spore) at 121°C?Answer: Since 1012 to 100 is 12 decimal reductions, then the time required is 12 x 0.204 min = 2.45 min Physical Methods: Moist Heat : Physical Methods: Moist Heat Calculations using D and z values (cont.) Given the D value at one temperature and the z value, we can derive an equation to predict the D value at a different temperature: Physical Methods: Moist Heat : Physical Methods: Moist Heat Calculations using D and z values (cont.) First, write an equation for this line. Since the y axis is on a log scale, then y = log (D). The slope of the line is -1/z; we'll let the y intercept be equal to c. Therefore: At a given temperature Ta, D = Da, so we can eliminate the "c" term Physical Methods: Moist Heat : Physical Methods: Moist Heat Calculations using D and z values (cont.) We can explicitly refer to the second temperature and D value as Tb and Db, so: Physical Methods: Moist Heat : Physical Methods: Moist Heat Calculations using D and z values (cont.) Given: For Clostridium botulinum spores suspended in phosphate buffer, D121 = 0.204 min and z = 10°C How long would it take to reduce a population of C. botulinum spores in phosphate buffer from 1012 spores to 100 spores (1 spore) at 111°C?Answer: To answer the question we need to know D111, which we can calculate from the formula:log(D111/0.204) = (121-111) /10D111 = 0.204(10) = 2.04 min12D111= 24.5 min Physical Methods: Moist Heat : Physical Methods: Moist Heat Calculations using D and z values (cont.) Given: For Staph. aureus in turkey stuffing, D60 = 15.4 min and z = 6.8°C How long would it take to reduce a population of Staph. aureus in turkey stuffing from 105 cells to 100 cells at 55°C, 60°C, and 65°C?Answers: Work it out for yourself. Here are the answers.At 55°C: 419 minAt 60°C: 77 minAt 65°C: 14.2 min (((! : (((! The time taken under defined conditions to reduce a population by 90% or --------------- for the survivor curve to traverse 1 log cycle Written as: Dtemp: x min D121 : 2 min Slide 71: Determination of D value (thermal death rate) DECIMAL REDUCTION TIME (D value) : DECIMAL REDUCTION TIME (D value) log10No Time (mins) 1log cycle D value (mins) Thermal Death Time D value as a function of temperature for Clostridium sporogenes PA 3679 : D value as a function of temperature for Clostridium sporogenes PA 3679 Temp (oC) D (min) 110 19.65 113 10.46 115 4.92 118 2.56 121 1.23 Values for k and D for various bacterial spores at 121oC : Values for k and D for various bacterial spores at 121oC B.subtilis 2.6 0.9 B.stearothermophilus 0.77 3 Clostridium sporogenes 1.8 1.8 K min-1 D min Z value : Z value The temperature change that will bring about a 10-fold change in D value written as: z = yoC Bacterial spores have a Z-value in the range 10 to 15 deg. while most Non-sporing organism have Z-values of 4 to 6 deg. Z value : Z value log10D Temp (oC) 1 log cycle Z value (oC) Inactivation Factor (IF) : Inactivation Factor (IF) IF = 10t/D F - value : F - value F = D121(log a - log b) a = initial population b = final population Factors affecting heat resistance of microbial population : Factors affecting heat resistance of microbial population Type & Species Nature of cell Age Number Physico-chemical environment Heat : Heat DRY MOISTPRESSURE BATCH CONTINUOUS Heat resistance of a variety of microorganisms : Heat resistance of a variety of microorganisms D value z value B.stearothermophilus (115) 22.6 7.1 B.megaterium (100) 1 8.8 Salmonella typhimurium 0.5 4.2 (55.5) Yeast spores (55) 0.9 Adenovirus (55) 17 2.5 Batch Sterilizations curve : Batch Sterilizations curve time temp Heating Holding Cooling STERILIZATION : STERILIZATION Different organisms for different methods of sterilization. The organisms that are resistant to a particular sterilization method should be chosen as the marker organism STERILIZATION MONITORS : STERILIZATION MONITORS STERILIZATION MONITORS : STERILIZATION MONITORS It is essential that strict controls are carried out on products to be labelled "STERILE". Such control must then ensure , the absence of viable microorganism from these product. There are basically two types of control: Controls on the process of sterilization i.e. sterilization indicators. Sterility testing of the products. Slide 87: Monitoring of the sterilization process can be achieved by the use of physical, chemical or biological indicators of sterilizer performance. PHYSICAL INDICATORS : PHYSICAL INDICATORS MOIST HEAT: A Master process record (MPR) is prepared as part of the validation procedure for a particular autoclave and for each specified product and load. Thermocouple is placed in each load. 2. DRY HEAT: Temperature record chart is made of each sterilization cycle and is compared against a master temperature record. Slide 89: 3. FILTRATION: Bubble point pressure test is a technique employed for determining the pore size of filters device immediately after use. 4. RADIATION: A Plastic dosimeter gives an accurate measure of the radiation dose absorbed and is considered to be the best technique currently available for the radiation sterilization. CHEMICAL INDICATORS: : CHEMICAL INDICATORS: Chemical monitoring of a sterilization process is based on the ability of heat, steam, sterilant gases and ionizing radiation to alter the chemical or physical characteristics of a variety of chemical substances. i) Browne's tubes: containing reaction mixture and an indicator. Types of Browne's tubes : Types of Browne's tubes BIOLOGICAL INDICATORS: : BIOLOGICAL INDICATORS: BIs Are the Most accepted means of monitoring the sterization process because they directely determine the most resistant microorganisms. STERILIZATION : STERILIZATION Sterilization Method Marker organisms Steam sterilization Bacillus stearothermophyilus Dry-heat sterilization Bacillus subtilis Ethylene oxide Bacillus subtilis sterilization Ionizing radiation Bacillus pumilus sterilization Sterility Testing as per IP : Sterility Testing as per IP The test for sterility are intended for detecting the presence of viable forms of micro-organisms in or on pharmacopoeial preparations. The test must be carried out under conditions designed to avoid accidental contamination of the product during the test. Precautions taken for this purpose should not adversely affect any micro-organisms which should be revealed in the test. The tests are based upon the principle that if micro-organisms are placed in a medium which provides nutritive material and water, and kept at a favourable temperature, the organisms will grow and their presence can be indicated by a turbidity in the originally clear medium. Slide 97: A laminar flow biological safety cabinet ☺ Method of Sterilization for the accessories to beused : ☺ Method of Sterilization for the accessories to beused ☺ Steam sterilization ☺ (121°C, 15 lbs Pressure for 15 min) ☺ Dry Heat sterilization ☺ (170°C/1 Hr or 140°C /3 hrs or Suitable temp. & times) ☺ Gas sterilization ☺ (Ethylene Oxide, Chlorine Dioxide) ☺ Sterilization by Radiation ☺ (Gamma sterilization, UV sterilization) ☺ By filtration ☺ (Membrane filtration & Cartridge filtration) Slide 99: A sterility test may be defined as — 'a test that critically assesses whether a sterilized pharmaceutical product is free from contaminating microorganisms'. Selection of Test Method : Selection of Test Method ☺ Direct inoculation ☺ Membrane filtration method Open funnel method (Manifold method) Closed method Culture Media : Culture Media 1.Fluid thioglycollate media - For use with clear fluid products. Used for detection of aerobic bacteria and fungi 2. Alternative thioglycollate medium – For use with turbid and viscid products and for devices having tubes with small lumina Used for detection of anaerobic bacteria and fungi 3. Soyabean- casein digest medium : universal media The media used should comply with the following tests carried out before or in parallel with the test on the preparation being examined : The media used should comply with the following tests carried out before or in parallel with the test on the preparation being examined Sterility : Incubate portions of the (a) fluid thioglycollate medium / alternate Thioglycolate medium at 30º to 35º and (b) soyabean casein digest medium at 20º to 25º for not less than 7 days; no growth of micro-organisms occurs. Growth promotion test : Test each autoclaved load of each lot of the medium for its growth-promoting qualities by separately inoculating duplicate test containers of each medium with about 100 viable micro-organisms or each of the strains listed in Table-2 and incubating according to the conditions specified. The test media are satisfactory if clear evidence of growth appears in all inoculated media containers within 7 days. The tests may be conducted simultaneously with the use of the test media for sterility test purposes. The sterility test is considered invalid if the test medium shows inadequate growth response. If freshly prepared media are not used within 2 days, store them in dark, preferably at 2º to 25º Finished media, if stored in unsealed containers, may be used for not more than one month provided that they are tested within one week of the time use. Method A : Membrane Filtration : Method A : Membrane Filtration The method needs exceptional skill and special knowledge; it also calls for the routine use of positive and negative controls. A suitable positive control is the occasional use of a known contaminated solution containing a few micro-organisms of different types (approximately 10 microbial cells in the total volumes employed). (a) Apparatus A suitable unit consists of a closed reservoir and a receptacle between which a properly supported membrane of appropriate porosity is placed. A membrane generally suitable for sterility testing has a nominal pore size not greater than 0.45 µm and diameter of approximately 47 mm, the effectiveness of which in retaining micro-organisms has been established. Preferably assemble and sterilise the entire unit with the membrane in place prior to use. Where the sample to be tested is an oil, sterilise the membrane separately and, after thorough drying, assemble the unit, using aseptic precautions. Diluting fluids Fluid A : Dissolve 1 g of peptic digest of animal tissue (such as bacteriological peptone) or its equivalent in water to make 1 litre, filter or centrifuge to clarify, adjust to pH 7.1 ± 0.2, dispense into flasks in 100 ml quantities and sterilise at 121° for 20 minutes. Method B : Direct Inoculation : Method B : Direct Inoculation Quantities of sample to be used The quantity of the substance or preparation being examined which is to be used for inoculation in the culture media varies according to the quantity in each container and is given in Table 3 along with the volume of medium to be used (b) Method of test : (b) Method of test For aqueous solutions and suspensions : The tests for microbial contamination are carried out on the same sample of the preparations being examined using the above stated media. When the quantity in a single container is insufficient to carry out the tests, the combined contents of the two or more containers are used to inoculate the above stated media. Remove the liquid from the test containers with a sterile pipette or with a sterile syringe or a needle. Aseptically transfer the specified volume of the material from each container to a vessel of the culture medium. Mix the liquid with the medium but not aerate excessively. Incubate the inoculated media for not less than 14 days, unless otherwise specified in the monograph, at 30° to 35° in the case of fluid thioglycollate medium and at 20° to 25° in the case of soyabean-casein digest medium. Observation and Interpretation of Results : Observation and Interpretation of Results Slide 109: At intervals during the incubation period, and at its conclusion, examine the media for macroscopic evidence of microbial growth. If no evidence of growth is found, the preparation being examined passes the test for sterility. If evidence of microbial growth is found, reserve the containers showing this and, unless it is demonstrated by any other means that their presence is due to causes unrelated to the preparation being examined and hence that the tests for sterility are invalid and may therefore be recommenced, perform a retest using the same number of samples, volumes to be tested and the media as in the original test. If no evidence of microbial growth is then found, the preparation being examined passes the tests for sterility. If evidence of microbial growth is found, isolate and identify the organisms. If they are not readily distinguishable from those growing in the containers reserved in the first test, the preperation being examined fails the tests for sterility. If they are readily distinguishable from those growing in the containers reserved in the first test, perform a second retest using twice the number of samples. If no evidence of microbial growth is found in the second retest, the preparation being examined passes the tests for sterility. If evidence of growth of any micro-organisms is found in the second retest, the preparation being examined fails the tests for sterility. Objectives, by the end of these lectures you should be able to: : Objectives, by the end of these lectures you should be able to: Define Sterilisation and Sterility State the reasons why the above is necessary Give examples of materials and objects requiring sterilisation Describe the methods used to achieve sterility and the principles upon which they are based State the factors that have to be considered when setting up a sterilisation protocol Expected questions???? : Expected questions???? 1. Define the following term i. D-value(2007,2009) ii. Z-value (2007,2009,2010) iii. Antiseptic iv. Sterilization v. Preservatives 2. Differentiate the following : i. Sterilization and Disinfection (2006) ii. Dry heat sterilization and steam sterilization. 3. Define sterilization. List the different methods used for sterilization with suitable examples. (2004,2007,2008) 4. Explain the different sterilization monitors or indicators. (2003,2007) 5. Whether sterility testing for product containing antimicrobial agent is necessary. 6. Name the biological indicator used for monitoring Moist heat sterilization process. 7. How is milk sterilized? 8. Write note on Pasturization. (2003) 9. Explain in details sterility test as per I.P. (2002,2004,2007,2009) 10. Write a Classification of sterilization method. What do you mean by Sterilization cycle? How will you monitor Sterilization process? (2006,2010) 11. Enlist the advantage and disadvantages of radiation sterilization.(2010) 12. Suggest suitable methods of sterilization for the following(2007) i. Culture media ii. Powders iii. Oils iv. Official aq. Injection. 13. What do you mean by survivor curve? (2004) 14. Name the biological indicator used for monitoring Moist heat sterilization process.(2006) Definitions : Definitions Page 184 Control of Microbial Growth: Chemical Antimicrobials• Previously described physical factors (e.g., temperature) affecting microbialgrowth. Here focus on chemical antimicrobials affecting microbial growth. : Control of Microbial Growth: Chemical Antimicrobials• Previously described physical factors (e.g., temperature) affecting microbialgrowth. Here focus on chemical antimicrobials affecting microbial growth. Two main categories of antimicrobial agents: • non-chemotherapeutic agents: (e.g., antiseptics, disinfectants); used to inhibit or kill microbes on living sufaces (topically) or inanimate objects, but are too toxic for internal use inside the human body. • chemotherapeutic agents: (growth factor analogs, antibiotics) can be used internally to control microbes that cause infectious diseases. • Based on degree of selective toxicity to the human or domestic animal host. • Some Important (suffix) Terminology: • "-static": antimicrobials that inhibit microbial growth without killing them, e.g., bacteriostatic, fungistatic. • "-cidal": antimicrobials that kill microbes, e.g., bacteriocidal, fungicidal. • "-lytic": antimicrobials that kill microbes by lysing them, e.g., bacteriolytic. Distinguish these 3 different categories by examining the effect of theantimicrobial agent on a growing bacterial culture: : Distinguish these 3 different categories by examining the effect of theantimicrobial agent on a growing bacterial culture: 3 types of action of antimicrobial agents. At the time indicated by the arrow, a growth-inhibitory concentration of the agent was added to the exponentially growing culture. Note that the bacteriostatic agent inhibits growth without killing cells, bacteriocidal and bacteriolytic agents kill. Cell # decline when lysed. Rate of Microbial Death : Rate of Microbial Death Bacterial death occurs at a constant rate I.e. 90% killed every minute of contact Rate of Microbial Death : Rate of Microbial Death Plotting microbial death Death curve Logarithmically Linear Arithmetically Exponential curve Factors Affecting Antimicrobials : Factors Affecting Antimicrobials 1- Number of microbes present More microbes = longer time to kill Also called load Factors Affecting Antimicrobials : Factors Affecting Antimicrobials 2 – Environmental factors Organic matter Often inhibits antimicrobials Feces Vomit Blood Temperature Temperature dependent reactions Warm temperatures are preferred Factors Affecting Antimicrobials : Factors Affecting Antimicrobials 3-Time of exposure "contact time" Extended times for endospores Longer times can offset lower temperatures Milk pasteurization 4-Microbial characteristics Virus vs gram+ vs gram - Actions of Microbial Agents : Actions of Microbial Agents Actions of microbial agents Alter membrane permeability Damage phospholipids or proteins in plasma membrane Cellular contents leak out Interferes with growth Actions of Microbial Agents : Actions of Microbial Agents Damage to proteins and nucleic acids Denatures proteins Enzyme Proteins necessary for bacteria metabolism Shape necessary for function Hydrogen bonds broken shape changes Covalent bonds are broken Sulfhydryl bonds – SH All can be broken Nucleic Acids DNA and RNA Can no longer replicate or synthesize proteins DISINFECTION : DISINFECTION It is the process of destruction or removal of microorganisms and reducing them to a level not harmful to health. Generally kill the sensitive vegetative cells but not the heat resistance endospores. Applied on inanimate (nonliving) object such as working areas,floor,dishes,benchs etc. Main difference with sterilization = the lack of sporocidal activity • Categorized into 3 levels: – High, – Intermediate – Low: Difference between Sterilization and Disinfection : Difference between Sterilization and Disinfection Killing or removing all forms of microbial life (including endospores) in a material or an object. Bacterial spore are destroyed Sterilization is done by any of the Physical, Chemical and Mechanical method. Done for animate and inanimate object Eg. Autoclave, Hot air oven etc.. It is the process of destruction or removal of microorganisms and reducing them to a level not harmful to health. bacterial spore are not destroyed Disinfection is done by using any of the chemical disinfectant such as phenol, formaldehyde Applied on inanimate object Phenol, bezalkonium chloride etc. Slide 126: Resistance of different pathogens to chemical and/or physical methods Ideal Properties required : Ideal Properties required Broad spectrum Non toxic Fast acting Odorless Surface compatibility Economical Easy to use Solubility and miscibility Not affected by physical factor Mechanism of killing : : Mechanism of killing : DENATURATION 1. STRONG MINERAL ACIDS AND ALKALIES 2. ORGANIC ACIDS (lactic acid, citric acid, propionic acid) INHIBITION OF ENZYME ACTIVITY OXIDAZING AGENTS (-SH group oxidized to S-S /disulfid/) 1. HALOGENS: chlorine, hypochlorites (free chlorine), iodine 2. HYDROGENE PEROXIDE 3. DYES: brilliant green, crystal violet (triphenylmethanes - dermatology), acridine dyes (bactericidal) ALKYLATING AGENTS: formaldehyde, glutaraldehyde (spores are sensitive!) SOLUBLE SALTS OF HEAVY METALS (mercury, silver, and others) Slide 131: DAMAGE OF THE CELL MEMBRANE 1. ORGANIC SOLVENTS: disorganization of the cell membrane 2. ALKOHOLS: disorganization of the lipid structures (50-70% ethanol as skin disinfectant, isopropyl alcohol: higher toxic effect) 3. SURFACE ACTIVE AGENTS (detergents):reduction of the interface tension (wetting agents, emulsifiers (cationic compounds/quaternary ammonium compounds: benzalkonium chloride for skin, but spores and viruses are not sensitive) 4. PHENOLIC COMPOUNDS: lysis (bactericidal), irreversible inactivation of oxidases and dehydrogenases, alkyl derivates: cresol (tricresol) chloro derivates: hexachlorophene, chlorhexidine NOT EFFECTIVE ON SPORES AND VIRUSES! Chemical Agents : Chemical Agents Phenolics Alcohols Halogens Heavy metals Quaternary Ammonium Compounds Aldehydes Sterilizing Gases Evaluating Effectiveness of Chemical Agents Chemical Agents: Phenolics : Chemical Agents: Phenolics Aromatic organic compounds with attached -OH Denature protein & disrupt membranes Phenol, orthocresol, orthophenylphenol, hexachlorophene Commonly used as disinfectants (e.g. "Lysol"); are tuberculocidal, effective in presence of organic matter, remain on surfaces long after application Disagreeable odor & skin irritation; hexachlorophene once used as an antiseptic but its use is limited as it causes brain damage Phenols : Phenols Phenols First used by Lister Rarely used now Irritates skin Throat sprays and lozenges 1 % solution Antibacterial Phenolics : Phenolics Phenolics Derivatives of phenol Increased antibacterial activity Decrease irritation to tissue Often with soap or detergent Injure plasma membrane Active in presence of organic material Phenolics : Phenolics Phenolics Good for disinfecting pus, saliva and feces Effective against Mycobacterium Cell wall high lipid content Very effective Cresols O – phenylphenol Lysol Bisphenols : Bisphenols Phenol derivatives Hexachlorophene pHisoHex Prescription antibacterial lotion Gram + in newborns Staph Strep Bisphenol : Bisphenol Triclosan Anti-bacterial soaps Kitchen cutting boards Some cases of resistance Inhibits synthesis of fatty acids Effective against g+ and g- Pseudomonas aeruginosa Biguanides : Biguanides Chlorhexidine Broad spectrum Used on skin and mucus membranes Scrubs Washes Low toxicity Damaging to eyes Damages plasma membrane Mycobacteria, endospores and protozoa are resistant Effective on some viruses Lipohilic viruses Chemical Agents: Alcohols : Chemical Agents: Alcohols Ethanol; isopropanol; used at concentrations between 70 – 95% Denature proteins; disrupt membranes Kills vegetative cells of bacteria & fungi but not spores Used in disinfecting surfaces; thermometers; "ethanol-flaming" technique used to sterilize glass plate spreaders or dissecting instruments at the lab bench Chemical Agents: Halogens : Chemical Agents: Halogens Act as oxidizing agents; oxidize proteins & other cellular components Chlorine compounds Used in disinfecting municiple water supplies (as sodium hypochlorite, calcium hypochlorite, or chlorine gas) Sodium Hypochlorite (Chlorine Bleach) used at 10 - 20% dilution as benchtop disinfectant Halazone tablets (parasulfone dichloroamidobenzoic acid) used by campers to disinfect water for drinking Chemical Agents: Halogens : Chemical Agents: Halogens Iodine Compounds Tincture of iodine (iodine solution in alcohol) Potassium iodide in aqueous solution Iodophors: Iodine complexed to an organic carrier; e.g. Wescodyne, Betadyne Used as antiseptics for cleansing skin surfaces and wounds Halogens : Halogens Iodine (I2) Oldest Very effective Bacteria, endospores, various fungi, some viruses Exact mode is unknown Possibly combines with amino acids Tincture Iodine in an aqueous alcohol solution Iodophor Iodine attached to an organic molecule, slowly releases Iodine Do not stain like tinctures Chlorine : Chlorine Gas (Cl2) or in combination Hypochlorous acid (HOCl) forms in water Unknown mechanism of action NaOCl – sodium hypochlorite Clorox = bleach Used to disinfect drinking water, swimming pools, sewage Chlorine : Chlorine Chlorine dioxide (ClO2) Can kill endospores Anthrax Chloramines Chlorine and ammonia Release chlorine over long periods Slow acting Toxic to fish Chemical Agents: Heavy Metals : Chemical Agents: Heavy Metals Mercury, silver, zinc, arsenic, copper ions Form precipitates with cell proteins At one time were frequently used medically as antiseptics but much of their use has been replaced by less toxic alternatives Examples: 1% silver nitrate was used as opthalmic drops in newborn infants to prevent gonorrhea; has been replaced by erythromycin or other antibiotics; copper sulfate used as algicide in swimming pools Heavy Metals : Heavy Metals Inorganic mercury Mercuric chloride Mercurochrome Control mildew in paint Bacteriostatic Toxicity Heavy metals : Heavy metals Copper Copper sulfate Destroy algae Control mildew in paint Zinc Used to galvanize nails Zinc chloride Mouthwashes Zinc oxide Antifungal in paints, and adds pigment Chemical Agents: QuaternaryAmmonium Compounds : Chemical Agents: QuaternaryAmmonium Compounds Quaternary ammonium compounds are cationic detergents Amphipathic molecules that act as emulsifying agents Denature proteins and disrupt membranes Used as disinfectants and skin antiseptics Examples: cetylpyridinium chloride, benzalkonium chloride Chemical Agents: Aldehydes : Chemical Agents: Aldehydes Formaldehyde and gluteraldehyde React chemically with nucleic acid and protein, inactivating them Aqueous solutions can be used as disinfectants Quaternary Ammonium Compounds (Quats) : Quaternary Ammonium Compounds (Quats) Surfactants Surface-active agents Cationic detergents Strong bactericidal (alter plasma membrane) Gram + Gram – (less effective) Fungicidal Amoebicidal Virucidal (enveloped) Do not kill Endospores mycobacteria Quaternary Ammonium Compounds (Quats) : Quaternary Ammonium Compounds (Quats) Zephiran Benzalkonium chloride Cepacol Cetylphyridinium chloride Organic material interferes Rapidly broke down by soaps Pseudomonas can actually grow in quats Surfactants : Surfactants Decrease surface tension Soaps and detergents Soap breaks up oil film into tiny droplets Emulsification Acid anionic surfactants Used on dairy equipment EVALUATION OF ANTIMICROBIAL AGENT EFFECTIVENESS : EVALUATION OF ANTIMICROBIAL AGENT EFFECTIVENESS The best known disinfectant screening test is PHENOL COEFFICIENT TEST in which the potency of the disinfectant is compared with that of phenol. The higher the phenol coefficient value the more effective the disinfectant under these test conditions. A value greater than 1 indicate that the disinfectant is more effective than phenol. Table. Phenol coefficient (PC) of some disinfectants : Table. Phenol coefficient (PC) of some disinfectants Chemical Agents:Evaluating the Effectiveness : Chemical Agents:Evaluating the Effectiveness Phenol Coefficient Test A series of dilutions of phenol and the experimental disinfectant are inoculated with Salmonella typhi and Staphylococcus aureus and incubated at either 20°C or 37°C Samples are removed at 5 min intervals and inoculated into fresh broth The cultures are incubated at 37°C for 2 days The highest dilution that kills the bacteria after a 10 min exposure, but not after 5 min, is used to calculate the phenol coefficient Evaluation of Disinfectant : Evaluation of Disinfectant 1. Agar well diffusion method 2. Tube Dilution Methods 3. Phenol Coefficient test 4. Kelsey sykes method Slide 162: Tube dilution Method: . A series of increasing concentrations of the antimicrobial agent are prepared in the culture broth medium. Each tube is equally inoculated and incubated to allow microbial growth to proceed. Growth (turbidity) occurs in those tubes containing the antimicrobial at concentrations below the MIC. Tube cultures are non-turbid (clear, no growth) at MIC and higher concentrations of the antimicrobial agent. Disc agar diffusion method: : Disc agar diffusion method: The test organism is spread on the culture medium in an agar plate and then sterile antibiotic discs are applied. After incubation, the organism produces a confluent "lawn" of growth except in zones of inhibition around discs containing antibiotics to which the organism is susceptible. When a filter paper disc impregnated with a chemical is placed on agar the chemical will diffuse from the disc into the agar. This diffusion will place the chemical in the agar only around the disc. The solubility of the chemical and its molecular size will determine the size of the area of chemical infiltration around the disc. If an organism is placed on the agar it will not grow in the area around the disc if it is susceptible to the chemical. This area of no growth around the disc is known as a "zone of inhibition". Chemical Agents:Evaluating the Effectiveness : Chemical Agents:Evaluating the Effectiveness Phenol Coefficient Test (cont.) The reciprocal of the maximum effective dilution for the test disinfectant is divided by the reciprocal of the maximum effective dilution for phenol to get the phenol coefficient For example:Suppose that, on the test with Salmonella typhiThe maximum effective dilution for phenol is 1/90The maximum effective dilution for "Disinfectant X" is 1/450The phenol coefficient for "Disinfectant X" with S. typhi = 450/90 = 5 Slide 168: Phenol coefficient test: standardizes the potency of bacteriocidal agents in comparison with phenol using specific test microorganisms. 1. A series of dilutions of phenol and the experimental antimicrobial agent are prepared in sterile tubes. 2. Viable test bacteria (Staphylococcus aureus and Salmonella typhi) are inoculated into each tube and incubated at 37 ーC. 3. After 10 minutes, samples of inoculated tubes are transferred into fresh culture medium without disinfectant and incubated for several more days. 4. The broth culture tubes are evaluated for growth by turbidity. 5. The highest dilutions of the test antimicrobial agent and of phenol that effectively kill the test organisms within 10 minute exposure are used to compute the phenol coefficient as follows: 1 / [Effective dilution of test antimicrobial agent] 1/ [Effective dilution of phenol] Example: If the highest effective dilution of the test antimicrobial = 1/500 and that of phenol = 1/100, the phenol coefficient of the test antimicrobial = 500/100 = 5. Agents with a phenol coefficient > 1 are more effective than phenol. The higher the phenol coefficient value, the more effective the test antimicrobial is in killing bacteria under the test conditions. Note that phenol coefficient tests are only valid for bacteriocidal (not bacteriostatic) antimicrobials, since cells inhibited (but not killed) by the latter would resume growth when transferred to fresh medium. Investigation of disinfectant activity : Investigation of disinfectant activity Conditions Influencing Antimicrobial Activity : Conditions Influencing Antimicrobial Activity Several critical factors play key roles in determining the effectiveness of an antimicrobial agent, including: Population size Types of organisms Concentration of the antimicrobial agent Duration of exposure Temperature pH Organic matter Biofilm formation Kelsey-Sykes test : Kelsey-Sykes test Kelsey-Sykes test is a triple challenge test, designed to determine concentrations of disinfectant that will be effective in clean and dirty conditions. The disinfectant is challenged by three successive additions of a bacterial suspension during the course of the test. The duration of test takes over 30 minutes to perform. The concentration of the disinfectant is reduced by half by the addition of organic matter (autoclaved yeast cells), which builds up to a final concentration of 0.5%. Depending on the type of disinfectant, a single test organism is selected from S. aureus, P. aeruginosa, P. vulgaris and E. coli. The method can be carried out under 'clean' or 'dirty' conditions. The dilutions of the disinfectant are made in hard water for clean conditions and in yeast suspension for dirty conditions. Test organism alone or with yeast is added at 0, 10 and 20 minutes interval. The contact time of disinfectant and test organism is 8 min. The three sets of five replicate cultures corresponding to each challenge are incubated at 32oC for 48 hours and growth is assessed by turbidity. The disinfectant is evaluated on its ability to kill microorganisms or lack of it and the result is reported as a pass or a fail and not as a coefficient. Sets that contain two or more negative cultures are recorded as a negative result. The disinfectant passes at the dilution tested if negative results are obtained after the first and second challenges. The third challenge is not included in the pass/fail criterion but positive cultures serve as inbuilt controls. If there are no positive cultures after the third challenge, a lower concentration of the disinfectant may be tested. Slide 173: Interpretation of result: No growth occurs in 2 or more of the 5 tubes of the 18 min samples Factors Affecting Effectiveness ofDisinfection : Factors Affecting Effectiveness ofDisinfection • Cleaning – Residual particles harbor & shelter from disinfectant – Organic load restrict disinfectants effectiveness of alcohol, phenols, chlorine & iodines • Nature of object: crevices, hinges, lumens more difficult to disinfect. • Concentration of disinfectant: – Diluted during application – Lose potency with time • Time of contact • Physical and chemical environment: temperature, water hardness, pH. Formulation You do not have the permission to view this presentation. In order to view it, please contact the author of the presentation.


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BOOK REVIEW: WE SHOULD ALL BE FEMINISTS by CHIMAMANDA NGOZI ADICHIE
BOOK: We Should All Be feminists
AUTHOR: Chimamanda Ngozi Adichie
GENRE: Non Fiction/Essay
PUBLISHER: Vintage Books
LANGUAGE: English
Feminism. This is a subject that is often misunderstood. Vocal feminists are sometimes mocked and teased, and called all manner of names by folks who don't get the concept behind feminism. Feminists advocate for women empowerment. They are pro-equality. Feminism has helped a lot of girls and women rise world over. It has helped the girl child to beat stereotypes, and made her achieve what society might have initially thought impossible.
This book, "We Should All Be Feminists" by the accomplished Nigerian writer, Chimamanda Ngozi Adichie, is a modified version of a talk she gave in December 2012 at the TEDxEuston .
Chimamanda is a story teller. A very gifted story teller. She is the type of author who makes one addicted to their works. You read one of her books and are forced to look for any other material published under her name. She is that good. She starts the book sharing how she first came across the word "Feminist". It was Okoloma, a childhood friend of hers who had called her that. Fourteen year old Chimamanda didn't know who or what a feminist was then. she later looked up the word feminist in a dictionary when she went home.
Chimamanda uses examples in her life to expound why we need feminism. She tells of a story when she was in primary school. Her teacher had given a test and promised that whoever scored highest in the test, would be made class monitor. Chimamanda scored the highest in the test, but to her surprise, the teacher said that the class monitor had to be a boy. Chimamanda wanted to be the class monitor, but the teacher didn't allow that. Why? despite scoring the highest, she was a female, and so could not be made a monitor. The boy was made the class monitor. That was unfair, but it was normal. "If we do something over and over again, it becomes normal… If we keep seeing only men as heads of corporations, it starts to seem 'natural' that only men should be heads of corporations." She says.
Misconceptions Of Feminism
In her book/speech, Chimamanda mentioned some misconceptions about feminism. They are not true. They are often peddled by people who don't understand what feminism is about.
- That feminists are unhappy because they can't find husbands
- feminism is un-African. Feminism in Africa has been influenced by western books.
- Feminists hate men.
- Feminists hate bras (ridiculous thinking).
- Feminists hate African culture.
- Feminists think women should always be in charge.
- Feminists don't wear make up, don't shave and are always angry (Haha.. What?).
- Feminists don't use deodorant.
"A thousand years ago. Because human beings lived then in a world in which physical strength was the most important attribute for survival; the physically stronger person was more likely to lead. And men in general are physically stronger. (There are of course many exceptions.) Today, we live in a vastly different world. The person more qualified to lead is not the physically stronger person. It is the more intelligent, the more knowledgeable, the more creative, more innovative. And there are no hormones for those attributes. A man is as likely as a woman to be intelligent, innovative, creative. We have evolved. But our ideas of gender have not evolved very much." Reads a paragraph in the book.
This book is such a short read. It's that which can be completed in hours, and re-read over and over because it helps those who had no idea of what feminism is, open their eyes. Chimamanda breaks down everything that even the youngest of minds can understand. The author helps one picture the whole ideology of feminism. She helps us understand why preaching about and practicing equality should be a norm. Girls shouldn't be fighting for their rights, society should recognize that the girl child has the same capabilities as the boy. Women should embrace their femininity, and not live in fear like slaves.
Feminism plays a significant role in today's world. It helps women be aware that yes, I am a woman, but that does not make me weak or lesser than men. I know my rights and freedoms, I know I can achieve this and no one should stop me from aiming higher. That's what feminism teaches us. Feminism is for equal opportunities for both genders, not putting the both boy child down while only concentrating with the girl. I've heard of this crazy fallacy that feminists are rebels. I ask, how? That feminists are bitter and always plan for the downfall of men. Really? These are lies. Real feminists want to see a world where both men and women work to better the living of everyone. Some people confuse Feminism with misandry. Maybe out of ignorance, or misinformation. Misandry is very different from feminism. And no, it's not just women who preach and practice feminism, men too can be feminists. We have men who are pro equality, and they support feminism, and all its ideas. Like the great Chimamanda says; we should all be feminists :-).

subset: commoncrawl


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Vampire, and by extension the World of Darkness, has had a massive effect on my life as well as how I developed my nickname. Given the power of nicknames it is no surprise that Vampire is more than a hobby to me, it lead to work and could even be argued to have become part of my identity; although crucially I do not think I am a literal Vampire.
A quick clarification: I love all sorts of Role-Playing Games (RPGs), whatever the system or genre, no matter the RPG Type. Whilst I think systems matter, I think players & implementation matter more.
I was sixteen when I first played Vampire the Masquerade; it had only been out for a few months. I was fortunate enough to have been invited in to a group of much older role players, so I was able to get a lot of experience from more worldly people. This proved helpful when I later started running Vampire for my long running group. At the time there were only a few supplements for the game, and there were none of the other games set in the World of Darkness had been released. Given the limited information and the newness of the game, it all felt very refreshing. This links back to the point about age, since our Storyteller Dale was in his 30s, he brought in all sorts of different cultures, not just USA or European ideas. Over many sessions our characters met all sorts of unusual antagonists; a deranged yogi was my favourite. All in all quite different from my experiences playing games like D&D, Cyberpunk or Warhammer. I quickly knew Vampire was going to be something I was going to play a lot, plus another thing I wanted to collect.
I was fortunate in that I had been brought up loving all sorts of music. I had become obsessed with metal and particularly thrash in 1989, but by 1991 I was broadening back out again. Vampire played a big part in influencing what music to investigate; I am sure it did for many gamers.
In the summer of 1992 I was working at a computer shop, and the boss insisted that everyone had to have a nickname. One of the young staff members couldn't think of anything, so since he was slightly annoying the boss nicknamed him Snot! The computer shop was next door to the local games shop, and this led to the boss asking me about my visits to that shop during my breaks, and my role-playing obsession, and a brief explanation about Vampire the Masquerade. Since I did a lot of long-distance running, generally late at night, I was very pale skinned and surprisingly strong for my slender frame, which led to the boss deciding that I should be called something vampire related, and this led to me being given the nickname of Bat. When I got to college the new large social group I spent most of my time with had other people called Richard, so my nickname was used instead.
Unfortunately this Bat nickname has resulted in numerous conversations with new people about where the nickname comes from, and they always guess it's to do with Batman. I've even had to deal with some odd enquiries like whether I can do the Batdance(Batusi), or my opinion on Batman versus Superman, shish. My family have purchased me several Batman t-shirts and jumpers to help rub this in. Eventually I embraced (yep, pun intended) this Batman tie in, after all given the cultural significance and number of Batman stories it is an iconic character. So far I have avoided any Batman cosplay as Vampire-Batman, but I have done an undead-Batman.
The Batjutsu nickname came from wanting a unique handle for using online, as well as email address. Given that my obsession with martial arts matched that of role-playing, I decided on the jutsu addition. This was also a way of differentiating from Batman, although Batman is a black belt in jujitsu, of course.
All in all, I am quite thankfully that I was given the nickname Bat, especially considering what my old boss could have come up with!


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It's been a long time since we've upholstered a bench seat in fabric (velour style). It's more common to get orders for vinyl or leather or combo vinyl/tweed. I'm so glad we got this chance to make a new bench seat for our Canadian customer Ryan and his 53 Ford F-150. It turned out so great that we'll be making another one to show at the LA Roadster Show next month at the Pomona Fairgrounds. We're really looking forward to seeing this baby installed.
Update from customer:
"I got to take the first look at my seats today. Let me say wow I'm impressed!! They look great! Such beautiful work! I can't wait to install them in my truck. Your assistance and customer service was phenomenal. I would recommend you guys to anyone looking for great looking seats with awesome customer service. "
"Thanks so much!"


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Although I have not lived in Washington for more than thirty years it has remained home to my heart, and always draws me back… sometimes with a deep longing that is almost hard to describe. It is the place of … Continue reading
Tag Archives: Seattle Washington
Seattle and the Olympic Peninsula
After an amazing day in Mount Rainier National Park I headed north toward Seattle. Three of my brothers and several of my friends live in the vicinity and I hoped to spend time with as many of them as possible. … Continue reading


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Just 29 percent of the 1,506 adults surveyed by the Pew Research Center for the People and the Press between July 22-26 said news organizations generally get the facts straight.
Sixty-three percent said news stories are often inaccurate, up from 34 percent in a 1985 study, Pew said.
Sixty percent of those polled said the press is biased, up from 45 percent in 1985. Just 26 percent in the latest survey said that news organizations are careful their reporting is not politically biased.
This in light of recent arguments by outlets, such as the San Francisco Chronicle, which assert that our republic depends on the press. That argument is based on two invalid premises, the first being that what mainstream outlets are doing is objective journalism; and the second which assumes that what MSM is doing benefits the republic. Neither of these are valid, which is why the Pew study confirms what we've all known for quite some time: media, the watchdog of the people, does not exist. Traditional media does more in a fascistic sense for the government than it does for the people. Need more proof? When NBC does an expose on Van Jones, ACORN, SEIU, Organizing for America, or any other questionable surrogates of this White House, we can redefine the role of the MSM.


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As our program expands, we are seeking exceptional teachers who share our philosophy and commitment to progressive education, developmentally appropriate practice, and community-based learning. Our teachers provide instruction at the Workshop (104 1st Place, 2nd Floor).
Competitive hourly rates.
Specific areas of expertise:
Elementary, Middle, & High School Public school curricula (Common Core State Standards)
- math, ELA (TC Reading & Writing Project), science
French Dual Language Curricula
- sequential bi-literacy
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Test Prep
- Common Core State Tests
- Private School Admissions: ISEE (all levels)
- High School Admissions: SHSAT, TACHS, HPST
French language instructors
Please send your resume to [email protected] to apply.


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(UPDATE: Due to illness, the October 30th meeting had to be canceled; our new presentation night date is January 8th, 2012. Thanks for your patience!)
Many people here at home have been waiting to hear a live report from my last trip to Mozambique, and I can now say that we've finally got a date!
On October 30th, I'll be making a very short presentation in both morning services at Summerland Baptist Church, but the main event will be that evening at 7pm, 2nd floor of the church in the Fireside Room. So mark it down on your calender!
I'll be showing photos and video from the trip, giving a recap of what we accomplished during our time there in August, and sharing some thoughts about what's in the works for the future.
Best of all, it's Dessert and Coffee Night! So don't miss it; in fact, invite some friends. Let them see what God is doing when we share our skills with people in need.
PS If anyone would like to have me come and present the work of Mercy Tech Mission to their group or organization, then just send me an e-mail at email@example.com .


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Looking for your next event venue?
Do you need a venue that is different than all the others? Are you planning a fundraiser or personal party? Or are you simply looking for a space for people to gather, eat, and relax after an occasion welcoming out-of-town guests?
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Akrona is the perfect location for your next event! The large addition to the back of the building is a great main space for your affair. All whilst the areas to the front of the gallery are the perfect size for guests to gather as well as talk both amongst themselves and our many artworks and sculptures.
The combination of our beautiful artworks lining the walls and your wonderful guests makes for a perfect atmosphere for your event, no matter the reason!
Contact us for information on pricing and date availability.


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urbaMonde: Second North American Hub of the Co-Habitat Network
urbaMonde: Second North American Hub of the Co-Habitat Network
This October 1-4, over 500 affordable housing professionals and activists gathered in Pittsburgh, Pennsylvania for Grounded Solutions Network's 2018 Intersections conference. The conference included the largest gathering of CLTs worldwide, and for the second time it also served as the North American Hub of the Co-Habitat Network, which is a global network of community-led housing organizations and allies that are working together to implement housing rights through collective, non-speculative, and people-led solutions.
In connection with the Co-Habitat Network, several internationally focused events were added to the schedule in order to enhance understanding of the global affordable housing crisis as well as find joint solutions or ways to act in solidarity through peer exchange and partnership. International events included:
-Two lunch tables with the following topics:
· International Innovations with the CLT model, Co-Chaired by Theresa Williamson, Executive Director, CatComm (Brazil) and Marie-Sophie Banville, Development Coordinator, Vivacite (Canada)
· The Canadian National Housing Policy, Chaired by Thom Armstrong, Executive Director, CHF BC
-A Global Spotlight session. Panelists included:
· Robert Burns, Board Chair, Grounded Solutions Network (moderator)
· Theresa Williamson, Executive Director, CatComm
· Tiffany Duzita, Director, Community Land Trust Foundation of BC
· Maria E. Hernandez, Board President, Fideicomiso de la Tierra del Cano Martin Pena
· Bea Varnai, Program Manager, urbaMonde
-A gathering of the nascent National Canadian CLT Network. Approximately 20 Canadian CLT staff and support actors attended, with representation from CLTs in each province.
-A gathering of the newly established Center for Community Land Trust Innovation, which has a mission to help the CLT model grow globally- and particularly across the Global South, as well as apply the model to non-housing sectors including urban agriculture.
-An opportunity for winners of Grounded Solution's IGNITE! Community Pitch Fest and other interested organizations to learn about the World Habitat Awards and how to apply.
Support for these international activities came from Grounded Solutions Network, World Habitat, urbaMonde, Cooperative Housing International, Vancouver City Savings Credit Union, CHF BC, and Confédération québécoise des coopératives d'habitation.


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Back to Stovesonline news and blog main screen
Rising fuel costs continue to be a problem in the UK: it has been estimated that fuel prices could go up by as much as 40% in the Winter of 2008. This could hit many households very hard and it is unlikely to get any better. Rising wholesale gas prices have been blamed and these are closely linked to the price of oil. The price of oil has also been rising steeply.
Reduce your heating costs: there has never been a better time to reduce your heating costs by switching to wood heating
Read the full bbc news story about rising fuel prices


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. . . sometimes go awry.
Of course, we are remodeling, so that is to be expected. The biggest hiccup we've run into to date is this: our (supposedly) custom front door we ordered more than 2 months ago arrived—not custom at all. The guys showed up to install it they didn't bring the windows we ordered with them, so they wasted their entire day. The door was okay, but the sidelights attached were not full glass as I ordered (they had fiberglass frames so the actual glass was only about 6 inches wide). On top of that, the whole unit was 2 inches too small (because they didn't custom build it like they said they would!). Big fail on their part. Now we are reevaluating and starting over—trying to decide if we will give this shop a chance to fix their mistake or start over with someone new.
We have been ordering a lot of things online for the kitchen. We have a big delivery from Overstock.com sitting our entry.
The big box in the back is our Cavaliere range hood. It was a great price (maybe too great—you know what they say, "you get what you pay for"—at least it looks nice).
The other two boxes are our new Ticor sinks. Great zero radius sinks at a great price also. I did some research before settling on these and felt like so many others you pay mostly for a name. They seem to be good quality in person. But . . .
Look a little closer:
The corner is bent. Luckily this is an under mount sink and the bent part will be under the counter top. We are going to try to bend it back ourselves, hopefully without damaging the visible part of the sink. It seems easier than dealing with the trouble of returning it.
We are still plugging away in our makeshift kitchen slash laundry room.
Not perfect, but so much better than washing dishes in the bathtub.
It does get a bit interesting when we have to do laundry in our "kitchen".
But at least we can.
Our living room is feeling a little cramped also. The fridge is off to the right here, the dining table and buffet are taking up more than their share of space, the piano is permanently draped with a sheet to protect it from construction dust, and the 2 main entrances are sheeted with plastic. Oh yeah, and the dog has claimed her residence in the corner also. We'll be quite glad to air out this room again when all is finished!
Plugging along and going with the flow . . . it wouldn't be remodeling without a few snafus! Thanks for following our adventure.


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Moorways Run Pass
Thu 16 May 2019
Enjoy running, but want somewhere safe to complete your distance?
Why not join us at Moorways Stadium on the floodlit running track?
Unlimited use for only £30 for 3 months
Monday 09:30 – 19:00
Tuesday 09:30 – 17:30 & 20:00 - 21:00
Wednesday 09:30 – 18:00
Thursday 09:30 17:30 & 20:00 - 21:00
Friday 09:30- 20:00
Saturday 08:30 – 15:00
Sunday 08:30-10:00 & 12:00-15:00
How to join
Step 1 - Join online and pay for your run pass.
You can also join at the Sales and Information Centre, Riverside Chambers, Market Place, Derby, DE1 3AH
Step 2 - Take your receipt with you to your first run at the Stadium and the Stadium staff will issue you a run pass
Step 3 - Enjoy your run!
Do you want more information?
Complete our online enquiry form below.
Terms and conditions
- This run pass is only valid for the person who purchased it.
- Following purchase you will need to collect your run card ID from Moorways Stadium or the Sales and Information Centre, Market Place, Derby. Please bring your receipt as proof of purchase, this can be a printed or shown on a mobile device.
- Track usage is subject to availability due to the site hosting events, please see inderby.org.uk/moorwaystadium website for event schedule.
- All runners must ring the site to check in with staff before using the track call 07812 300 709.
- There is no time limit for usage but must coincide with the site closing time.
- The management team may close the track for scheduled maintenance, we will give as must notice as possible displaying signage at the Stadium and on the website noticeboard inderby.org.uk
- This pass will give access for track usage only, any additional athletics equipment must be booked.
- The run pass includes parking at Moorways and changing facilities (subject to availability).
- This pass will include floodlighting for the winter months.
- Run passes are not transferable.
- Any questions regarding your membership email: email@example.com.


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After two long years of (Im)patiently waiting and saving for a new vehicle, my husband and I were FINALLY able to replace my tiny little, 10 year old car with 205,000 miles on it with an SUV! To say I'm excited is a huge understatement.
My previous mode of transportation went to my little sister, who was equally excited for her own wheels. It was a win-win situation 🙂 However, I left my previous "Emergency Kit" in the car for her to use, meaning I was without. I took this opportunity to really put some thought and research on the All-Knowing-Internet into building a truly useful Vehicle Emergency Kit for our family.
Misty Marsh writes at one of my favorite prepping sites, Your Own Home Store, and wrote a great post about her own Emergency Car Kit, so I kind of combined her ideas with things from my husband's Get Home Bag, and personalized it for Little Man.
When we put together my husband's Get Home Bag, we prepared it only for him. 99% of the time, he is the only one in that vehicle, and we took that into consideration. When designing my own car kit, we needed to take into account that if all three of us go somewhere, 99% of the time, we'll take my vehicle. If we have guests, we're taking mine. If it's just Little Man and I we are definitely taking my vehicle. Because of this, we have supplies for AT LEAST the three of us, but some supplies, like water, we have extra for guests as well. This is a fairly extensive kit, and it takes up some room, but in the time I've had it (2 week or so…) I've used at least one thing from each component. To me, it has already proved it's worth.
Like Misty, I separated my kit into three categories:
- First, I wanted to prepare for minor, everyday "emergencies." For example: Little Man screaming his head off for no apparent reason, me subsequently having a headache, being thirsty, leaving the diaper bag at home (which happens more often than I'd care to admit,) a boo boo at the park, etc.
- Second, I wanted to prepare for Emergencies involving the vehicle: The car not starting, having a flat tire, getting stuck in a ditch for whatever reason, overheating…things of that nature.
- Last, I wanted preparations for major emergencies: Natural disasters like a tornado or snowstorm, EMP (heaven forbid), etc. Mostly things that would require us to leave the car and seek shelter elsewhere. I realize that in MOST instances, our best option will be to stay in the car, and in MOST cases, that will be my first instinct, but just in case we have to leave the car, we will be prepared to do so.
So… What's in My Kit?
For Minor, everyday "Emergencies"
- Mini First Aid Kit plus extra meds- Just some bandaids of various sizes, alcohol wipes, and a pair of gloves along with the medications (Sunscreen, Ibuprofen, Benadryl, Asprin, Children's Motrin, Children's Benadryl, and Sting Relief gel)- I don't necessarily LOVE using conventional medicines, and I generally carry a few of my favorite essential oils in my purse; however, Benadryl and Asprin can save lives (think: allergic reactions and heart attacks), and Motrin/Ibuprofen/Advil (all the same product) is a miracle worker in my opinion haha so I certainly like to have all of those on hand. Even if I don't need them, someone else might. Quick medical tip: Benadryl in liquid form works faster and is easier to swallow if your throat is closing up because of an allergic reaction… So even if you don't have kids, its a good idea to have in an emergency kit.
- Feminine Products- Besides the obvious, "sanitary napkins" and tampons can come in handy for covering cuts and wounds since they are clean and "sanitary."
- Car Phone Charger
- Sippy Cup
- Paci- We're trying to get rid of this thing, but for now I keep one in the car
- Snack Cup with snacks for Little Man- Helps minimize crumbs 🙂 (…I said minimize, not eliminate haha)
- Diapers
- Baby Wipes- These have MILLIONS of uses besides their actual intended use
- Small toys or entertainment for Little Man- We rotate these often so he doesn't get bored with them
- Hand Sanitizer- Of all the things I keep in my car… this might be the most used and most important
- Water Bottle- I keep a filtered Camelback in my cup holder. It gets used frequently. I just have to remember to wash it out and fill it back up with fresh water
- Lotion- Lifted from a hotel 🙂
- Chapstick
- Hair Ties
- Hair Clip
- Rubber Bands
- Paper with Pen & Sharpie
- Flashlight– I love these Mini Cree lights! They're small and lightweight, but they're strong and bright. And only $4!
- Lighter
- Grocery sacks- For dirty clothes or shoes, vomit (heaven forbid), trash….you name it
- Tissues
- Cash and a roll of quarters– I try to keep about $50 in small bills (mostly ones) plus quarters for vending machines, pay phones (if you can find a working one), and the car wash… wait…that's not an emergency? 🙂
- Sunglasses for each family member
- Sun & Bug Spray– Because even in an emergency, the sun and mosquitos will still be out!
- Granola Bars- You never know when you'll need a little snack 🙂
- Napkins- I collect free ones whenever we order/eat out and stash them in the car
- Umbrella- Stays in the driver side door
Most of this fits into a little box inside the center console. I just covered a box in black duct tape so it wouldn't look as ghetto 🙂
For Car Emergencies:
I like to start with a Pre-Made Kit, and add things to it. Although you'll be hard pressed to find a kit that has EVERYTHING you need in it, It will have several useful things and will likely be cheaper than buying them separately. This kit made by AAA is the one I started with.
Along with the pre-made kit, I have also added:
- Tow Rope
- Road Atlas of the United States– We don't travel outside Oklahoma or Texas often, but we did take a trip over Christmas so I bought this just in case, and it stays in the kit
- Shop Towels- Paper Towels on steroids 🙂
- Fix A Flat
- Hand Crank Weather Radio– One with a two-way function would be a better option, but this works for now!
- Flashlight and extra batteries- I love these Dorcy lights when I need something a little bigger than the Mini Cree. We keep these in the car (obviously), on our nightstands, and with our severe weather emergency supplies.
- Lighter
- Fire Extinguisher– Be sure NOT to leave this loose in your car! Mine came with a bracket so it can be secured in the trunk or under a seat. I keep ours inside the tub so I didn't secure the bracket, but you definitely DON'T want one of these flying around in your car if you have an accident so just make sure it's not loose.
- Extra Snacks- As I mentioned above about medications, I don't necessarily LOVE using packaged snacks, but homemade ones don't hold up to the heat of the car so well, and these are only to be used in emergencies. Make sure you pick things your kids will actually eat. (AKA not so much the 3000 calorie emergency ration bars)
- Extra Water- I have a few of these Water Pouches that are lined with aluminum (think: BPA free and therefore NOT leaching cancer-causing chemicals into your water), but I would like to get several more of them. They're a tad on the expensive side, but they stand up to heat better than plastic bottles, and they have a 5 year shelf life vs. 1 year for your typical water bottle. Until I'm able to get some more pouches (hopefully within the next couple of weeks), I keep an entire case of plastic water bottles in the trunk. Not ideal, but they'll do in an emergency.
- Winter Items- Like my Husband's Get Home Bag, we rotate supplies at least every 6 months to accommodate for season changes and expired food/meds; however, there are a few things that will stay in the car year-round despite the weather.
- Heat Cells (2)
- Fleece Stadium Blanket
- Hand Warmers
- Knit hats and gloves (One set for each family member)
All of this stays inside an "under the bed" storage tub in the trunk.
In the Glovebox:
- Insurance Card
- Car Manual
- Family Emergency Information Sheet <– Free download!
- Tire Gauge
- Seatbelt Cutting/Window Breaking Tool
- Ice Scraper
Some of the most important things to have in your vehicle include a jack along with a spare tire (that is aired up and ready to go…Yes, I'm talking to you… You who has the nail in his "spare" from two years ago… Go get that replaced!)
I also believe that it is very important to have gas in your car at all times. I'm preaching to myself here, too, but please try to keep your gas tank AT LEAST half full. If you need to get out of town fast, you don't want to be waiting in line behind 15 other people waiting to get gas. And if you're worried about people stealing your gas, they make a nifty Locking Gas Cap to help prevent that. We have one on my husband's truck, and we'll likely put one on my new car as well.
For Major Emergencies:
This part of the kit is very similar to my husband's Get Home Bag. I keep it in a backpack so we can quickly leave if we need to. I have little piece of paper that lists emergency contact numbers on one side, and has a list of other things I need to grab before leaving the car on the other. This paper goes inside a plastic ID holder on the outside of the backpack.
- Hat for each family member- Protection from the elements and to hide mine and Little Man's white-blonde hair 🙂
- Pocket Knife
- Flashlight with extra battery
- Lighter
- Mini Multi-tool– Would like to upgrade to a full-size one soon
- Bandanas– Can serve as a mask in case of debris
- Rain Poncho
- Whistle
- Notepad/Pen/Sharpie
- Map/Compass
- First Aid Kit with extra meds (see Minor emergencies First Aid Kit above)
- Cash- Another $50 in small bills
- Paracord
- Glow Sticks– Some green ones for light and some red for flares
- Toilet Paper- the center stuffed with cotton balls rubbed in vaseline to act as a fire starter
- Work Gloves
- Dust Mask for myself and my husband- backup to the bandanas
- Change of clothes for each family member
- More diapers and a small package of wipes
- Sleeping Bag (SOL Emergency Bivvy)– We just have a single size one, but it's plenty big enough for all three of us to squeeze in if we needed to. We may upgrade to a double size in the future.
- Duct Tape
- Trash bags- Can double as extra rain ponchos or be used as a temporary shelter, another layer of protection under the Bivvy, etc.
- Emergency Mylar Blankets
- Water
- Snacks- I'm a big fan of granola bars if you can't tell. I also have a few cracker type things for Little Man to munch on, plus some Fruit and Veggie Pouches. While these tend to have more fruit than veggies, he LOVES them and they're a great way to get him to eat semi-nutritious food on the go, plus they're shelf-stable. Just check expiration dates every so often!
We also keep a pair of tennis shoes for each family member, and our ERGO Baby Carrier in the car. Although the backpack holds quite a bit of stuff, it only weighs about 10 pounds. In a worst-case-scenario emergency situation, I would be able to carry the backpack and all 22 pounds of Little Man to a safer location.
Things I would like to add:
- SteriPEN and/or Filter Water Bottle(s) for the Backpack
- Multi-tool (Full-size)
- Shovel to keep in the car
- Small Tool Kit
Things that are NOT in the kit:
You'll notice I haven't listed any hygiene items other than hand sanitizer, toilet paper, and baby wipes. This is because the purpose of the kit is to help us get somewhere safer, with more supplies and resources as quickly as possible, preferably home. I won't be stopping to shower or do my makeup, and in such a situation, and I don't think we'll be too worried that we stink a little bit….but that is a call you'll need to make on your own: keep a light pack or include deodorant, soap, toothbrush/toothpaste, etc for convenience. Update: I have included a contact case and small bottle of contact solution for my husband so he doesn't have to wear dry contacts for too long. Next time he gets a new pair of glasses we'll leave his old pair in the backpack.
I also have not included any type of shelter other than the SOL Bivvy because in MOST cases the car will act as our shelter, and otherwise we will be moving as quickly as possible to get to a permanent shelter. Absolute worst-case-scenario will be having to spend the night out in the elements, and the chances of that are slim to none.
What About You?
Do you have an emergency kit in your vehicle? What does it include? Do you have any suggestions of things to add/remove from mine?
Please see my Disclaimer about affiliate links.


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Sunday, February 13, 2011
Daniel is growing so big, as well as Michael!
Daniel goes in for his 4 month check up next weekend - time feels like it is flying by which I want to bottle every moment up and keep it forever.
Nothing really exciting has been going on here, except working, and keeping the kids fed. I DO wish that there were more hours in a day, because by the time I get home from work and getting the kids dinner I have no time for anything else. I feel my house is a wreck and I am never going to be able to catch up to the clean it needs to be.
All is good though :)
I just wanted to check in with my blogger nation and let yall know we are well :) we love you very much!
Sunday, January 9, 2011
New Addition!
I have not been on and blogged in a minute, Sorry about that...I have sorta had my hands full...we have a new family member to annouce, Daniel Jacob! He was born October 4th. He was 2 months premature, with low white blood cell counts, he is fine now and growing fast!
I will need to update my blog name from Casey & Michael in Texas to Casey, Michael & Daniel in Texas...Or I was thinking 2 Virginians and a Texan :) hahaha Since Daniel was born in Texas and Michael and I are born in VA...I am still brain storming ideas on the blog name change.
I am back to work, and whew what an adventure, I am up 3 times at night so when I have to get up and get going I am draggin!
I have to get me up, Daniel is already up at this point begging for formula and then I have to fuss with Michael to get him out of bed and ready for school. So our mornings are very eventful as you can imagine.
But I wouldnt trade it for anything in the world. Daniel will be over this hungry all the time hump and maybe momma can get some sleep at night.
I love my boys!
Monday, November 22, 2010
Mama's Holiday Wish List Meme
Mama's Holiday Wish List Meme
TodaysMama (link to: http://bit.ly/tmwishlist) and GameStop (link to: http://bit.ly/gamestop10) are giving away a sleighful of gifts this holiday season and to enter I'm sharing this meme with you.
1. What is your holiday wish for your family? My wish for my family is happiness and love, we are missing a lot of loved ones this year and we need to remember the good times and be thankful for the people we were blessed to have known and the ones we know.
2. What is your Christmas morning tradition? Usually wake up and wait til the kids wake up and once we hear the gasp and then the little patters of feet running to my room to yell that Santa has come it is blissful! We just relax and do church.
3. If you could ask Santa for one, completely decadent wish for yourself, what would it be?I would wish for my family to be complete and happy. (can Santa do that? I think more God who can then Santa)
4. How do you make the holidays special without spending any money? We decorate (with things we already have), go to church, help the homeless and needy.
5. What games did you play with your family growing up? We played all sorts of games - not partically board games, but i do remember playing games on the Nintendo! Old school style!
6. What holiday tradition have you carried on from your own childhood? Putting the christmas tree up near Thanksgiving and not taking it down til after New Years.
7. Where would you go for a Christmas-away-from-home trip? I would go to a tropical island some where - with palm trees
8. Check out GameStop (link to: http://bit.ly/gamestop10) and tell us, what are the three top items on your GameStop Wish List this year? As my 8 year old tells me everyone is asking for Kinect this season!
Also a DSi (although he already has a DS lite - but hey an upgrade is an upgrade right) and a Red wii!
Also he has been asking for Michael Jackson Experience which comes out November 23rd!
Monday, February 22, 2010
Home Make Over - Casey Edition!
Michael's room is done and I am so pleased with it! It is nice if I do say so myself :) And he loves it so that is a bonus! He has actually been sleeping in HIS bed and hasn't wandered into my room some time in the middle of the night, which is sorta exciting considering he KICKS me when he sleeps with me and let me tell you IT HURTS!
I have not taken pictures of EVERYTHING yet but stay tuned they will come :)
Michael's new bed :) Thanks Ikea! Oh and I put together all by myself!!! And it is still holding strong!
Michael's new bed sheets :) We have another set same but minus the skulls.
Michael's name on the wall, which was really easy to do! A trip to Michael's, some wooden letters, things to nail into the back so they could hang, and black spray paint!
Michael's book shelf! I love this thing! I love the baskets, I got them for a DEAL at Michael's (LOVE THAT PLACE!) and the newest is the shadow box with his football jersey in it!
Mommy also got a new bed too which I love!!!
The room is nearly done I need to take pictures of it - but here is my bed :)
This is my new bed sheets - love them! Love Laura Ashley :)
I added a new frame behind the bed and I put 3 of the pictures from Chera in them! It looks SO good!
Saturday, February 20, 2010
Michael's Snow Pictures 2-13-10
Hope you enjoy them as much as I do!
Sunday, February 14, 2010
Revamp of Michael's Room - 'Big Boy Room'
Michael picked out a gray, white and black camo sheet set with skulls, with a black and gray comforter!
Added his name on his wall...took wooden letters, spray painted black and hung on his wall.
He LOVES his room!


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24 hours a day. 365 days a year.
Pumps and Motors retain an extensive knowledge of the building services industry and have established themselves in recent years as the industry-leading company in the sector. We take great pride in delivering service excellence and have gained worked in partnership with a number of major clients over the 35 years we have been in business. As a service provider we always strive to offer business efficiency and value for money.
If you'd like to know more, feel free to get in touch firstname.lastname@example.com
Find out more about us, our 30+ years experience and how we have established a reputation as the foremost service provider, always striving to offer your business efficiency and value for money.
Take Action
Pumps and Motors have a dedicated contracts management team, enabling us to deliver immediate, informed and focused response to site requirements.


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[League City High School Class of 1947]
Description: Photograph of a group of students from the League City High School class of 1947; however, this photo was taken when the students were juniors. They are arranged in two rows on the steps just outside an unidentified building.
Date: 1946
Item Type: Refine your search to only Photograph
Partner: League City Helen Hall Library


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- Home
- About Us
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For the second year in a row Integrated Standard Solutions (ISS) signed the HP supplies agreement with the Arab bank. During the agreement period ISS is committed to deliver the toner cartridges to the Arab Bank upon request. Needless to say that Arab Bank is one of the largest HP Imaging and Printing and supplies customers in the region. Earlier this year ISS signed with the Arab Islamic bank, not to mention the supply of Housing bank, Orange, and UN institutes. ISS is also considered one of the strongest suppliers of HP Imaging and Printing in Jordan and its customers include Financial, government, educational, telecommunication and SMB sectors.