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A study on the occurrence of pharmaceuticals and personal care products (PPCPs), drug resistant bacteria and their removal efficacy of PPCPs in the wastewater treatment plants
Course work- 4: Basic microbial techniques for resistance detection
Microorganisms require nutrients to grow which is supplied either by solid or liquid media. Nutrient agar is a solid media, gelatinous substance derived from seaweed. The
basic liquid medium is nutrient broth, typically a mix of water, meat extract peptone, and sodium chloride. Some of the microorganisms require enriched media with growth promoting ingredients such as animal blood, glucose or egg. MacConkey agar is a differential media contains chemicals designed to distinguish microorganisms by the appearance of their colonies.
Based on the composition the media has been classified: Defined media classified as selective medium or differential media. Complex media which contains extracts of beef (peptone), milk (tryptone), soyabean meal (soytone) or yeast. Complex media and define media can also be called as selective media. For example, MacConkey agar is selective for gram negative bacteria.
There are different types of media used for culturing and isolating bacteria. Some of them are
Blood agar, MacConkey agar, Nutrient agar and Chromocult coliform agar.
Culturing of microbes is a method for multiplying microorganisms by allowing them to reproduce in the determined culture media under lab conditions. These cultures are used to determine the type of organism. It is one of the important diagnostic method of microbiology and also used as a tool to identify the infectious disease.
Staining is an auxiliary technique used in microscopic techniques to enhance the clarity of the microscopic image. Stains and dyes are widely used to explore the structure of the biological specimens, cells, tissues etc.
Simple staining is a technique that microorganisms gives the color characteristic of the staining solution which reveals size and shape. The basic stains that are used are methylene blue, basic fuschin, malachite green etc. The bacteria present in the smear takes stain and appears in the color of stain. It is basically used to identify morphology and arrangement of bacteria.
Gram staining is a technique that differentiates bacteria into two groups: gram positives and gram negatives. It is based on the ability of microorganisms to retain color the stains used during the reaction. Gram negative bacteria can be easily decolorized by the alcohol, losing the color of the primary stain, it will appear in purple color. Gram positive bacteria are not decolorized by the alcohol and will remains purple, after decolorization step, a counterstain is used to impart a pink
Chemicals may be liquids, gases, solids or powders have the tendency to harm health when it used or handled improperly, those can be corrosive, toxic agents, reactive or flammables can generate fumes, vapors, dust, and mists.
The usage, handling, storage and the disposal of chemicals have been increased. For estimating the benefits and dangers, one should consider the whole lifecycle of the chemical products. The accidents which have happened through the chemicals have a limited effect. These effects may not be the workers itself but also in homes by accidents or by the consumer products through foods. (1)
Radiation exposure more than permissible limits can result in serious biological damage which can be transfer to future generations. This results for the proper care and adoption of safe work practices while working with radioactive isotopes. The use of radioactive sources has become very wide quick after the discovery of radioactivity. It expanded from academic laboratories where physical characteristics of radiation as well as effects of radiation on humans, animals, plants or materials were studied to the broad use in industry, medicine, and research. Sources used today span from nuclear fuel in NPPs to smoke detectors used in homes. (2)
The hazardous chemicals have to be stored according to their conditions and properties. Chemicals have the tendency to react together to become unstable or toxic. Liquids which are flammable should be stored separately to avoid fire.
Waste chemicals should be handled properly to minimize the personal hazards. Disposal of chemicals should be in a specifically labeled containers. The non-returnable cylinders may contain toxic and the reactive gas which should be disposed of properly. Chemicals which contain toxic agents should be decontaminated before the disposal.
Chemical splash goggles are to be used while handling the chemicals. Non-flammable, non-porous aprons are the least expensive and protective instead of lab jacket.
There is a direct relationship between the dose of exposure and the time duration of the exposure.
To reduce the exposure maintaining the maximum distance helps to reduce the number of photons per unit time to an acceptable level.
A barrier or shield before handling the radioactive isotope is used to protect against exposures. Shielding can be either container with heavy walls, concrete walls or cladding on radioactive material. (7)
The area should be posted with a caution sign where the radioactive elements are used. The levels of radiation intensity should be labelled on the doors of the rooms. The indicator tape should be marked to the containers of the materials containing radioactive
materials. The beakers, tubes and forceps should be marked with the caution tape. Lab coat or apron, eye protection and gloves should wear compulsorily before unsealing and while handling the radioactive materials. Pipetting the solutions with mouth is extremely dangerous and shouldn’t be done at any circumstances. Applying cosmetics, storing food or smoking near working area is dangerous.
Radioactive waste should be disposed only according to the safety rules. No waste should be thrown to the ordinary trash. No liquid waste may be disposed of into the drains or any sewer openings. For the mixed waste one should request radioactive safety for the proper disposal. (8)
Every laboratory should be equipped with the first aid kit and the instructor should teach the usage of the kit. The lab should include fire extinguishers, wash fountains, eye wash and first aid supplies. Reporting must be done compulsorily for all the minor and major accidents.
Follow the necessary rules before operating the high voltage equipment and maintain the proper wiring and electrical connections. Extension cords should be electrically grounded with fuse. Never modify the high voltage equipment and make sure all the capacitors are discharged. (12)
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