Pharmaceutical Microbiology Notes PDF

Page 1 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 1 of 22 search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise ...

Page 1 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 1 of 22 search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 2 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 2 of 22 O search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 3 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 3 of 22 uysay 1 H search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 4 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 4 of 22 6 6 search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 5 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 5 of 22 tiit B search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 6 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 6 of 22 ii search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 7 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 7 of 22 V search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 8 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 8 of 22 E search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 9 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 9 of 22 search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 10 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE 5 search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 11 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE * search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 12 of 29 Carewell Pharma SUBSCRIBE us,on YOUTUBE A 3 search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 13 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE F V IE search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 14 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE + L E i search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 15 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE # search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 16 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Muh (Advauragu) ve effcehve quiceer n heading up t expossd aahids(moleuial). T povid greater lehal achon o MoisF he Duuts (Disadvantogs) NoF Suable o anydrous Maleiod Such powdtrs, oils, fal el NoF Sutabl for har Jaleilu substac, which Can mo u1h stand hlatnq a aud above fAppliationg usud usd -fo seilizaion bacteioloqia udia, haa stobla Liqud, saliut solution, hea Hcisant equipmaLils aud instrunuii glasnares, illersubbn poducs eh search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 17 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 10 of 22 search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 18 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 11 of 22 search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 19 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 12 of 22 ,i 3 search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 20 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 13 of 22 F search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 21 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 14 of 22 RovG DIeGRAM search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 22 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 15 of 22 (Asbestos filter (Seitz filter): They are disposable, single-use discs made up of asbestos (magnesium trisilicate). It is supported on a perforated metal disc within a metal funnel (Fig. 74). It is then fitted on to a sterile flask through a silicone rubber bung. The fluid to be sterilized is put into the funnel and flask is connected to the exhaust pump. After completion of fitration, the filter is discarded and the entire unit sterilized. The pore size of iters range from 0.01 to 5 microns. Sintefed glass filters (fritted glass filter/morton filters): Borosilicate glass is finely Powdered in a ball mill and packed into disc moulds and heated until suitable adhesion Takesplace between the granules. The sintered discs are finally fused into funnels of a Sutable size (Fig. and shape 7.5). Sintered glass filters are available in several different Orosties but for filtration sterilization a number or grade 5 or 5 on 3 must be used. They a low adsorptive property and can be cleaned easily. They are brittle and expensive and have a small area of filtration. search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 23 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 16 of 22 Sintered dlsc Funnel Asbestos pad (Flterlng medla) Wing nut Perforated plate Fig. 7.5: Sintered glass filter Fig.7.4: Seitz filter These are manufactured in different (ii) Filter candles (ceramic/Berkefield filter): purification of water for industrial and grades of porosity and have been used widely for drinking purposes. They are made of either porous porcelain or kieselguhr. They are usual walls (Fig. 7.6). These are depth encountered as cylindrical candles with comparatively thick filters with cellular walls and are available in various sizes. Fig.7.6: Filter candles The filter is fixed to the filter assembly and placed in a mantle. The liquid to be filtered is poured into the mantle where vacuum forces it through the filter. After filteration, filter candle is removed from the assembly and filterate istransferred toa sterile container. These filters are inexpensive and available in different sizes. They are easily clogged and blocked and require high pressure for filtration. search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 24 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 17 of 22 av) Membrane filter (millipore/ultra filter): These are made up of various types of lklose and cellulose esters. They are 150 um thick and contain millions of microscopic cellu fro pores ranging from 0.01 to 10 um in diameter. The pore sizes most often used for rilization are 045 um t 0.02 pm (Millipore grade, HA) or 0.22 steri m t 0.02 um (Millipore de, GS), particularly for very small bacterial contaminants. They are sterilized by utoclaving, in the holder or packed between thick flter pads to prevent curling. They are 5o available at ready sterilised form (by ethylene oxide or ionizing radiation), Membrane Fiters are supported on a rigid base of perforated metal, plastic or coarse sintered glass Eia. 7.7). The HA grade filiters are approximately 65 ml/min./sq.cm. (GS 22 ml/min/sq.cm) with a differential pressure of 70 cm mercury across the membrane. -Funnel Filter Filter holder -Cotton To suction pump Flask -Fltrate (Sterile) (a) Components of filter (b) Membrane filtration assembly Fig. 7.7: Membrane filter Advantages of membrane filters are as follows 1 microorganisms are separated by process of sieving. All 2 Membranes have a high and uniform porosity permitting a rapid rate of filtration. 3. Membranes are disposable. Hence, there is no cross contamination between filtered products. 4.Adsorption is very less. Disadvantages of membrane filters are as follows: Prefilter is used before the membrane filter to avoid clogging and breaking. They have less chemical resistance to certain organic solvents such as chloroform, ketones and esters. sterilization, mbrane filters are routinely used in water purification and analysis, sterility They are also been and for the preparation of solutions for parenteral use. d for the identification and enumeration of microorganisms from water samples ana other materials. search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 25 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 18 of 22 E Sunuivina pach'm (No. of backua) N 3 search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 26 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 19 of 22 Temperatua (°c) 2 i N : search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 27 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 20 of 22 STERLT INDIGATORS that strict controls are carried out on products to be labelled 'sterile. Suc It is essential controls must then ensure, the absence of viable microorganisms from these product There are basically two types of controls: L Controls on the process of sterilization ie. sterilization monitors or sterilization indicators. 2. Sterility testing of the products. Monitoring of the sterilization process can be achieved by the use of physical, chemic or biological indicators of the sterilization performance. 1. 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 configuration. This may then be used as a reference for the process record btained from a single thermocouple placed in a strategic part of each load (batd process record, BPR). The MPR should be checked at annual intervals and whenever significant changes occur in the BPR when compared with the MPR Microprocessor controlled sterilization cycles are now a part of modem autoclaves. Pressure is measured by pressure gauges or through pressu transducers Dry heat: In dry heat sterilization processes, a temperature each sterilization cycle and is record chart is madeo compared against a master GD Radio sterilization: A temperature record. plastic dosimeter gives dose absorbed and is considered an accurate measure of the radiatio" to be tl.2 best technique the radio sterilisation process. currently available 0) Gaseous methods: For gaseous are monitored for each sterilizationsterilization procedures, elevated temperatu cycle by temperature probes and routine e search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 28 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 21 of 22 tests are performed to ensure gas-tight seals. Gas concentration is measurea independently of pressure rise, often by reference to the weight of gas used. Pressure and humidity measurements are recorded. Filtration: Bubble point pressure test is a technique employed for determining the () pore size of filters and may also be used to check the integrity of certain types of filter devices immediately after use. The principle of the test is that the filter is soaked in an appropriate fluid and pressure is applied to the filter. The pressure difference when the first bubble of air breaks away from the filter is equivalent to the maximum pore size. When the air pressure is further increased slowly, there is general eruption of bubbles over the entire surface. The pressure difference is equivalent to the mean pore size. 2 Chemical indicators: Chemical monitoring of a sterlization 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. Browne's tubes: The most commonly used chemical indicators for heat processes are Browne's tubes. These are small sealed tubes containing a reaction mixture and a an indicator. Exposure to high temperature completes the reaction producing change in the colour of the indicator (Table 7.5). All four types change from red through yellow brown to green, the latter colour only being achieved after a specified time at the given temperature. Table 7.5: Types of Browne's tubes Browne's Method of sterilization Temperature Colour of indicator tube (C) ype 126 Black spot Moist heat ype High vacuum moist heat 130 or more Yellow spot ype II Dry heat 160 Green spot ype IV conveyor oven 180 Blue spot |Dry heat infra-red CGi) Witness tubes: Witness tubes consist of single cnystalline substances of known melting point contained in glass tubes e.g. sulphur (115 C), succinic anhydride (120C), benzoic acid (121") etc. A dye may be included to show more clearly that the crystals have melted. Such a device only indicates that a certain temperature has been reached. Exposure time can be calculated by putting the crystals in one end of an 'hour-glass' tube, the volume of the crystals and the diameter of the constriction of the tube being adjusted so that the time for transfer of the melt is the same as that required for the sterilisation at the required temperature. search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise  Page 29 of 29 Carewell Pharma SUBSCRIBE us on YOUTUBE Page 22 of 22 (i) Heat-sensitive tape: Heat-sensitive tape is used quantitatively in the Bowie-Di test. This is a test to determine that all air has been removed from dressings and that subsequent steam penetration has been even and rapid. The tape is placed suitably wrapped at the centre of a test pack. All the bars on the tape shoul change colour to demonstrate full penetration of the steam. (iv) Royce sachet: The Royce sachet is a chemical indicator for ethylene oxide sterlization. This consists of a polythene sachet containing magnesium chloride, HCI and a bromophenol blue indicator. A given concentration-time exposure to ethylene oxide results in the formation of ethylene chlorohydrin and a colour change from yellow to purple. (v) Chemical dosimeters: Chemical dosimeters give an accurate measure of the radiation dose absorbed and are considered to be the best technique currently available for controlling radiation sterilization. Qualitative indicatorsS made of radiosensitive chemicals impregnated in plastic are also available. The indicator changes from yellow to red during irradiation. 3. Biological indicators: are Biological indicators consist of a suitable organism deposited on a carrier and process, the units distributed throughout the sterilizer load. At the end of the sterilization survivors. The are recovered and cultured to determine the presence or absence of is able to integrate all biological indicator measures sterilization processes directly and reproducible sterilization parameters. The selected organism should possess high and resistance to the sterilizing agent, should be genetically stable, readily characterizable and the non-pathogenic. The viability of the organisms, the storage conditions before use and The incubation and culture conditions after sterilization must be standardized for the results. organisms used as biological indicators are usually resistant bacterial spores (Table 7.6). Table 7.6: Biological indicators formonitoringsterilization processes Sterilization process Species D-value Autoclave at 121°C |Bacillusstearothermophilus 1.5 min Clostridium sporogenes 0.8 min 160°C Bacillus subtilis var. niger Dry heat at 5-10 min Ethylene oxide at 600 mg/lit. Bacillus subtilis var. niger 2.5 min Temperature 54 C 60% - relative humidity) Tonizing radiation Bacillus pumilus 3 kGy (0.3 M rad) Membrane filter (0.45 um pore size) Serratia marcescens Membrane filter(0.22 um poresize) Pseudomonas diminuta search CAREWELL PHARMA on YouTube Watch Videos complete Syllabus wise