An aseptic technique

An aseptic technique Introduction Aseptic means to be free from microorganisms. Aseptic Technique is the procedure that is performed under sterile condition to prevent the growth of other microorganisms on the growth medium such as the Petri dishes containing the nutrient agar or the pure culture. If the growth medium or the pure culture is contaminated with microorganisms from the environment, it will results in confusion and inaccurate data. Hence, it is important to reduce the risks of these microorganisms to come in contact with the experimental materials. In addition, by maintaining a clean environment when transferring the culture of microorganism onto the nutrient agar is part of the aseptic technique. This is usually done by disinfecting the table before and after working with microorganism using alcohol. Flaming the experimental materials such as bacteriological loops, bottle or flask necks can help in sterilizing. It must be done for several seconds so as to raise the temperature to kill the contaminants; however, the bacteriological loop must be cooled for a while before it is used to pick up the microorganism as picking up microorganism with a hot tool will kill the cells. When removing caps from bottles, it is important to keep the cap in the hand as by putting them on the table, it will be contaminated. Flaming is also required before replacing the cap onto the bottle. It is important to handle open tubes at an angle so that airborne and other microorganisms will not fall into the tube and cause contamination. During streakin g, it is important to keep the lids of the Petri dish over it to prevent contamination. Lastly, try to avoid breathing, coughing, sneezing and talking while transferring the culture so as to reduce the risk on contaminating. Apart from this general aseptic technique, there also several other methods to ensure that destruction of living microorganisms in materials and apparatus. One of the methods is by using dry heat which is sterilising using naked flame or hot air. Sterilising materials or apparatus by a naked flame is usually heated to redness and allowed to cool. They are usually made of metal. Exposure to hot air helps to destroy microorganisms in glass and porcelain apparatus. The other method is sterilizing using moist heat which can be used in three different ways which are heating in water or steam at 100oC, heating in steam under pressure and discontinue heating at low temperature. The different ways are employed according to the different materials or apparatus used. The last method is by using chemical, it can be either in liquid or gaseous state. They are often used in the disposal of contaminated materials and apparatus after a laboratory session. Microorganisms worked with in a lab should not be released into the environment as these strains may contain genetic markers such as antibiotic resistance. Therefore, they must be discarded properly. In addition, aseptic technique is not only applied in laboratory, it is also applied in clinical and surgical setting. Aims There are two aims in this experiment. The first aim is to show that a large number of microorganisms exist on the surface of our hands. The second aim is carry out the aseptic technique properly by transferring pure culture and inoculating them onto an agar plate. Materials and Methods Bacteria on skin Please refer to the Laboratory Manual unless otherwise stated of changes made. Streak Plate Please refer to the Laboratory Manual unless otherwise stated of changes made. A disposable sterile bacteriological loop is use instead of the metal sterile bacteriological loop so no heating is required. Discussion Bacteria on skin The human skins surface do carry a large number of microorganism and that by washing hands, individual can reduce the number of microorganism noticeably. However, even after a hand wash, microorganisms are still present on the surface. Streak Plate By employing the streaking technique on an agar plate correctly, a single colony can be obtained. Furthermore, it can be used to separate colonies of mixed culture. Hence, this pure colony can be picked up and to be grown in large quantity. From the result above, it can be observed that single colonies of the S.aureus are found. Due to the colour and morphology, it can be noted that the S.aureus is of a pure culture. Conclusion Aseptic technique is a basic laboratory technique that must be employed especially during Microbiology laboratory session so as to prevent any contamination and affecting the accuracy of the result. Since microorganism can replicated rapidly, disposal of contaminants must be done properly so as to protect both the equipments and the health of individuals. B- Gram Staining Introduction Gram stain is also known as differential stain in which it will divide bacteria into two large groups, mainly Gram Positive and Gram Negative. This difference is due to the chemical and physical structure of the cell wall called peptidoglycan. During solvent treatment, if the peptidoglycan is able to retain the crystal violet dye, the bacteria will be group as Gram Positive bacteria. However, if it is not able to retain the crystal violet, the bacteria will be group as Gram Negative bacteria and that it will be stained pink. Gram Positive bacteria has a thicker peptidoglycan (50-90% cell wall) as compared to the Gram Negative bacteria (10% cell wall). In addition, the Gram Negative bacteria has another layer which is make up of liposaccharides and proteins and is separated from the cell wall by the periplasm. In gram staining, there are four basic steps which include flooding the heat fixed smear with crystal violet stain, following by the addition of iodine solution to form complex, adding of alcohol for decolourisation and counterstaining with safranin. After flooding the peptidoglycan with crystal violet stain, the dye will enter the cells and all cells will turn purple. With the addition of iodine, a crystal violet-iodine complex will be form such that it will not be able to exit the cells easily. By decolourizing the cell with alcohol, the peptidoglycan of the Gram Negative bacteria will break down because the alcohol will dissolves the liposaccharides layer and hence, with the removal of the layer, the crystal violet-iodine complex will run off which will results in the loss of the crystal violet stain and the cells turn colourless. On the other hand, the alcohol will dehydrate the Gram Positive bacterias peptidoglycan, closing the pores as the peptidoglycan shrinks. As a result, the crystal violet-iodine complex will not be able to run off as the exits will be blocked and they remained stained. By counterstaining with safranin, the Gram Negative cell will turn pink and the Gram Positive cells will remain violet. With gram staining, one is able to differentiate if the culture is a pure or a mixed, the morphological details of the bacteria and the arrangement of the bacteria. Aims The aim is to prepare smears for staining, observe the morphological details of the bacteria and to be able to differentiate between Gram Positive and Gram Negative bacteria. Materials and Methods Preparation of Smears for staining Please refer to the Laboratory Manual unless otherwise stated of changes made. A disposable sterile bacteriological loop is use instead of the metal sterile bacteriological loop so no heating is required. Gram Staining Method Please refer to the Laboratory Manual unless otherwise stated of changes made. Discussion According to the result observed, Bacillus subtilis is rod shaped (bacillus). They are stained purple which suggests that they are Gram Positive bacteria. They are arranged in singles. Although, endospore cannot be observe in this experiment, they can also be found on Bacillus subtilis. The endospore enables the bacteria to tolerate harsh environmental condition such as high temperature. Bacillus subtilis can also be known as a single bacillus bacterium. Escherichia coli is stained pink and thereby, it is a Gram Negative bacterium. The cells are also rod shaped but they do not have any particular cell arrangement. They are found in singles, pairs and even clusters. Proteus vulgaris is also stained pink and hence, a Gram Negative bacterium. Its morphology rod shaped and is arranged in singles. They can also be known as a single bacillus bacterium. Staphylococcus aureus is a Gram Positive bacterium as it is stained dark purple after gram staining. It has a spherical shaped, otherwise known cocci and they are usually arranged in grape-like clusters. Therefore, they are known as a staphylococci bacterium. There were no differences in the shape and colour observed for each of the bacteria, hence, they can be known as a pure culture. Conclusion The Gram staining method is a useful tool used in most laboratories as it helps individual to visualise the bacteria accurately and effectively such as the shape, arrangement and even whether the culture is a pure or mixed. However, it should be noted that not all bacteria will give a gram reaction as some of them are gram variable, otherwise known as gram indeterminate. Therefore, they will give a mix of pink and purple cells after gram staining. For some of the Gram Positive bacteria, their peptidoglycan breaks easily during cell division, hence, after staining, they will give pink cells instead of purple. In addition, the duration of a culture can also affect the gram stain. C- Cell Counting Introduction Cells counting is the accurate and precise counting of cells. They are usually carried out manually or electronically. By counting cells manually, a counting chamber, otherwise known as the haemocytometer is used. The counting chamber is used to determine the number of cells per unit volume of a suspension. On the other hand, a coulter counter is used to count cells electronically. There are two approaches to count the number of cells, mainly total cell counts and the viable counts. Total cell counts are counted directly using the microscope and that both living and dead cells are counted. This is normally accompanied by the use of the counting chamber or coulter counter. Another approach is the viable counts which only count the living cells. The small volume of culture, otherwise known as the dilution of the culture is applied to the surface of an agar plate. After incubating, the colonies are counted, normally colonies between 30-300 are chosen to be used for the calculation of concentration of the given sample. The units given is colony forming units (CFU) per ml. The haemocytometer is a modified glass slides with two count chamber of known area. Each chamber grid is composed of nine squares which are known as subgrid, each square is 1mm2. Within each large square, there are further sub divisions that help in counting. When the coverslip is placed over the grooves of the slide, there will be a thickness of 0.1mm. Hence, the volume is 0.1mm3 or 1 x 10-4ml. Therefore, the cell concentration will be calculated as the number of cells multiply by 1 x 104ml and again, multiplying the dilution factor. Since cells are very small and they can be observed in a very high number, the suspensions should be diluted enough so that the cells are able to distribute uniformly in the counting chamber. Aims There are two aims in this experiment. Firstly, to be able to determine the cell count in different biological species and secondly, to be able to determine the viable count of a live bacteria, Staphylococcus aureus. Materials and Methods Cell Counting using Counting Chamber Please refer to the Laboratory Manual unless otherwise stated of changes made. Serial Dilution is carried out before the sample is loaded into the Neubauer Manual Counting Chamber. Normal saline (0.9% NaCl) is used to dilute the blood and broth medium is used to dilute the brewers yeast ( Saccharomyces cervisiae). Both blood and brewers yeast are dilute in the ratio of 1:10 and 1:100. The 1:10 dilution is prepared by diluting the 10Â µL of whole blood or yeast with 90Â µL normal saline or broth medium respectively. The 1:100 dilution is prepared by extracting 10Â µL from the respective sample from 1:10 and adding 90Â µL of normal saline and yeast into the respective sample. Cell Counting of Live Bacteria, S. aureus (after Serial Dilution) Please refer to the Laboratory Manual unless otherwise stated of changes made. Two changes were made in Step 1 and Step 12 respectively. Only three nutrient agar plates , 10-3, 10-4 and 10-5 were labelled and culture in these dilutions were spread on the respective agar plates. A new spreader and pipette tip were used everything a different dilution culture was spread on the agar plate. Count the number of colonies on the three different agar plates. Choose the agar plate with colonies between 30-300 to calculate the concentration of the original sample. Discussion Using the counter chamber, individual is able to give a quick assessment on the number of cells given that all the procedure on preparing and loading the sample onto it. One of it is that suspension/sample is not mixed before loading. This is due to the fact that cells tend to settle at the bottom of the tube and hence, while pipetting the sample out from the tube, individual do not have the actual or accurate number of cells. Therefore, to get a uniform suspension for a more accurate result, mixing the tube before pipetting is recommended. In addition, it can also help in reducing the clumping of cells. Furthermore, improper filling of chambers can lead to inaccurate volume of suspension in the chamber and leading to inaccurate cell concentration. Improper filling of chamber includes overfilling or under filling of sample. Moreover, there must be a consistency in counting cells which is in contact with the boundary lines (ie. the three lines just outside the grid) or when the cells are clump together. Individual will have to determine which cells to count and which not to count especially a cell which is situated on a border such as if a cell has half of its area outside the border, individual do not count those cells. The other method of cell counting is the viable count where a single cell will give rise to a colony which is visible to the naked eyes on the agar plate. Therefore, by counting the number of colonies on the agar plate, individual is able calculate the cell concentration. However, only plates which have 30 to 300 colonies are used to calculate the cell concentration. In the result for viable count of S.aureus, the plate chosen was 10-5 because there was 82 colonies in one quadrant which is equivalent to 328 (82 x 4) colonies on the agar plate. Although, the number of colonies (328) exceed the number of colonies of 300 that we were supposed to chose, this 10-5 dilution plate has the closest number to 300. However, we should dilute even further because a single colony can have clumps or chain of cells in it and hence, resulting in inaccurate number of colonies/cells in which the actual number of cells should actually be more than the calculated number of cells. The advantage of viable cell counting is that the organism counted will be a positive one (ie. S.aureus) instead of any other organism as if there is contaminant, the morphology or colour will be different. Another disadvantage of viable cell counting, other than cells that clump together or have chains which will form a single colony, is that organism will only grow in condition which is suitable for their growth on the agar plate. Cell counting usually is accompanied by serial dilution as it is impossible to count the number of cells if the concentration is too high as it will lead to a very high number of cells. Conclusion There are several other methods, other than using counting chamber and viable cell count, to count cells in a suspension. However, they are the least expensive and is able to give accurate result in a very short period of time. References Abcam, 2009. Cell counts using a haemocytometer. Abcam plc. Source: http://www.abcam.com/index.html?pageconfig=resourcerid=11454 Accessed: 1 October 2009 George Xu, 2007. History of the Gram Stain and How it Works. University of Pennsylvania. Source: http://www.uphs.upenn.edu/bugdrug/antibiotic_manual/Gram1.htm Accessed: 1 October 2009 H. Kayser, A. Bienz, Johannes Eckert. M. Zinkernagel. 2005. Medical Microbiology. Thieme Stuttgart., New York. p. 264-270. Kenneth Todar, 2008. The Growth of Bacterial Population. Todars Online Textbook of Bacteriology. Source: http://textbookofbacteriology.net/growth_2.html Accessed: 6 October 2009 Linda B, Mary R, 2007. Aseptic Transfer. Austin Community College. Source: http://www.austincc.edu/microbugz/aseptic_technique.php Accessed: 3 October 2009 Steve Hogg, 2008. The Gram Stain. Newcastle University. Source: http://www.ncl.ac.uk/dental/oralbiol/oralenv/tutorials/gramstain.htm Accessed: 6 October 2009

All Quiet on the Western Front by Erich Remarque

All Quiet on the Western Front In the book â€Å"All Quiet on the Western Front† by Erich Remarque, the author uses nature, and comradeship, to describe what the characters are going through. Erich uses nature in several ways, such as describing how the soldiers are facing terrible hardships, also it reflects on their sadness, and provides a contrast to the unnatural world of war. The author also uses the theme Comradeship through all the horrifying pictures of death and inhumanity, he talks about when Paul and his friends pick on Himmelstoss and beat him.We think it’s funny because Himmelstoss deserves it for being rude to them, and Paul and his friends are just giving him what he deserves. As we start going farther into the book, we start to realize that beating on someone isn’t funny anymore. We read the how the soldiers feel after assaulting and killing other people, it gives us a disturbing thought about war. Erich shows the theme Nature in many parts of the book. In chapter 2, when Kemmerich dies Paul takes his identification tags and walks outside.He then says â€Å"I breathe as deep as I can, and feel the breeze in my face, warm and soft as never before. † (Remarque 33) This is one of many times, when nature has helped the men go through bad experiences, and help them move on. Nature also reflects the terrible sadness of the lost generation. In Chapter 4, Paul's company sustains heavy losses and a recruit is wounded so badly Paul and Kat consider killing him to end his suffering. The Lorries and medics arrive too quickly, and they are forced to rethink their decision.Paul watches the rain fall and says: â€Å"It falls on our heads and on the heads of the dead, up in the line, on the body of the little recruit with the wound that is so much too big for his hip; it falls on Kemmerich grave; it falls in our hearts. † (Remarque The cleansing rain falls upon the hopelessness of Paul's life and the lives of those around him. Throughout Remarque's book, we also see a strong affinity between nature and lost dreams and memories. When Paul is on sentry duty in Chapter 6, he remembers his childhood and thinks about the poplar avenue where such a long time ago they sat beneath the trees and put their feet in the stream.Back then the water was fragrant, the wind melodious; these memories of nature cause a powerful calmness and awaken a remembrance of what was but sadly, will never be again. Finally, butterflies play gracefully and settle on the teeth of a skull; birds fly through the air in a carefree pattern. This is nature in the midst of death and destruction. While men kill each other and wonder why, the butterflies, birds, and breeze flutter though the killing fields and carry on as if mankind were quite insignificant.Even at the end when Paul knows there is so little time until the armistice, he reflects on the beauty of life and hopes that he can stay alive until the laws of nature once again prevail a nd the actions of men bring peace. He describes the red poppies, meadows, beetles, grass, trees at twilight, and the stars. How can such beauty go on in the midst of such heartache? Remarque says that this novel â€Å"will try simply to tell of a generation of men who, even though they may have escaped shells, were destroyed by the war. If words can touch what men hold to be dear in their hearts and so cause them to change the world, this book with its words of a lost generation, lost values, and lost humanity is surely one that should be required reading for all generations. . When Paul and his friends waylay Himmelstoss and beat on him, we laugh because he deserves it and they are only giving him his due. As time goes by, however, the pictures of camaraderie relieve the terrible descriptions of front line assaults and death, and they provide a bright light in a place of such terrible darkness.A young recruit becomes gun-shy in his first battle when a rocket fires and explosions b egin. He creeps over to Paul and buries his head in Paul's chest and arms, and Paul kindly, gently, tells him that he will get used to it (Chapter 4). Perhaps the two most amazing scenes of humanity and caring can be found in the story of the goose roasting and the battle where his comrades' voices cause Paul to regain his nerve. In Chapter 5, Paul and Kat have captured a goose and are roasting it late at night.Paul says, â€Å"We don't talk much, but I believe we have a more complete communion with one another than even lovers have. We are two men, two minute sparks of life; outside is the night and the circle of death. † As he watches Kat roasting the goose and hears his voice, it brings Paul peace and reassurance. Over and over again, in scenes of battle and scenes of rest, we see the comradeship of this tiny group of men. Even though Paul counts their losses at various points, he always considers their close relationship and attempts to keep them together to help each oth er.In Chapter 9, when Paul is alone in the trench, he loses his nerve and his direction and is afraid he will die. Instead, he hears the voices of his friends: â€Å"I belong to them and they to me; we all share the same fear and the same life; we are nearer than lovers, in a simpler, a harder way; I could bury my face in them in these voices, these words that have saved me and will stand by me. † There is a grace here, in the face of all sorrow and hopelessness, a grace that occurs when men realize their humanity and their reliance on others.Through thick and thin, battle and rest, horror and hopelessness, these men hold each other up. Finally, Paul has only Kat and he loses even this friend and father-figure in Chapter 11. Kat's death is so overwhelming and so final that we do not hear Paul's reaction; we only see him break down in the face of it. There is such final irony in the medic's question about whether they are related. This man, this hero, this father, this life â €” has been closer to Paul than his own blood relatives and yet Paul must say, â€Å"No, we are not related. † It is the final stunning blow before Paul must go on alone.

The Advantages of Ethical Retail

Retailing The advantages of ethical retail are as follows: Higher revenues. Improved Brand, Business Awareness and Recognition. Better employee Motivation and Recruitment. There are several ethical issues to be considered in retailing. To be a throughly ethical-concerned company, we are going to take some actions to guarantee everyone from the owners to employees in our company know and understand what is ethic and make sure they act ethically during the daily selling practice. 1. Mutual integrity A mutual integrity environment is expected to establish in our company.Employees are required to be honest when dealing with the business and with their co-workers. While the business,its owners and management should be honest in dealing with them. And our company, including owners and employee are required to be honest and ethical when dealing with customers. 2. Development of ethical standards Retail salespeople need guidelines on ethical issues. An ethical standards will be developed and posted by the ethical department. Something simple which commits the business and its employees to certain behavior ought to be clearly and complete to provide a guide to day to day decisions.Because it is found that some salespeople are not aware what is ethical issues, what is the the right thing and what is not. Besides some common sense about ethical issues, there are some misconducts will be listed that they may be not realized. Charge full price for a sale item without the customers’ knowledge. Don’t tell the complete truth to a customer about the characteristics of a product. Sell more expensive product when a less expensive product would be better for the customer. Don’t offer information to the customer about an upcoming sale.Make excuses to customers about unavailable merchandise when merchandise is not in stock or is sold out. Take return from customers when you believe the item should not be accepted. Give preferential treatment to certain customers . Give your employee discount to your friends Sell merchandise that is not of good quality. Don’t Use the Customer Information privately or in other business activities. Employees are required to remember it and act it accordingly. 3. Fair workload employers in our company also need to provide a good working situation for the employees.No excessive pressure and workload of the job are allowed to put on salespeople, which place them in uncomfortable situation. 4. Good ethics demonstration Employers are needed to be ethical-concerned firstly. Employees may follow the behaviors of employers. Poor ethics demonstrated by ourselves or senior management can educate others that the business is prepared to cut corners or deal in areas of grey between what is right and what is wrong. This leads to employees themselves following this behavior and acting against the business but doing no worse than they have seen a more senior person do.When comes to social responsibility and environment concerned,we are going take some actions in our daily retailing practice. 1. Better inventory management Inventory management is the process of efficiently overseeing the constant flow of units into and out of an existing inventory. It helps in controlling the costs associated with the inventory. Since our warehouse where our inventories placed are not located near our retailing store, it needs vehicles to transfer the products when they needed.Through a better inventory management, we can lessen the times of transportation, so as to reduce the pollution of the environment and cost at the same time. 2. Shorten opening time reasonable As it known to all,electricity and water are used during opening time of retailing store. However, electricity are wasted during low consumer flow when only few people in the store. An investigation is conducted to show the consumer flow statistics and we are going to rearrange our opening time accordingly to reduce the electricity and water consumptio n.Environmental friendly packaging Over-packaging or packaging with no-environment friendly material are also needed to be eliminated concerning of environment. Therefor, switching over to environment friendly packaging materials is our first step. Recycle Bags are going to sell in low price in our retailing store instead of plastic bags. Consumers who bring the bag back can have 10% discount at all items. It is a good way to avoid plastic pollution, in the other hand, it is also a good promotion way. 4. Undertaking activities that are beneficial to the societyTo build good ethical-concerned image of our company, we plan to undertake activities that are beneficial to the society. For example, this season, our theme is about pets. We consider to join a series of activities held by PETA, People for the Ethical Treatment of Animals, it is the largest animal rights organization in the world, with more than 3 million members and supporters. To show our company is animal-friendly, we are going to join â€Å"Shopping Guide to Compassionate Clothing: Vegan Companies† of PETA, which defines as company sells only animal-friendly, cruelty-free products. Promotion 1.Public-interested ad Public-interested advertisements about every season’s ethical emphasis are going to made. This season,considering our brand is sportswear and the ethical emphasis is animal rights, we are going to place our advertisements at the paths and space specially for pets and their owners, for instance Wan Dog Park, Pet World, Peel Rise, Discovery Bay, Sai Kung, Clearwater Bay and so on. Considering our target customers who are animal-lovers , they play with their pets in casual wear. They may be attracted by our public-interested ad: a man in our clothing playing with dogs harmoniously.

The Persuasive Power of Television in the 1960’s Essay

For Americans, the 1960’s were a time of both unnerving turmoil and exciting change. Following on the heels of the 1950’s themes of tradition and conformity, the contrasting events and attitudes in the sixties constituted a perfect storm leading to a reconstruction of American social, cultural, and political ideals. Although each decade has experienced identifying features, events occurring during the sixties provided for a definitive coming of age era for the United States. While much of this revolution can be attributed to the events themselves, the medium used for disseminating these ideas bears some of the responsibility. Throughout the decade television replaced radio and newspaper as the primary source of news and entertainment.†¦show more content†¦The popularity of television developed in the 1950’s, but the power of television as a medium secured its place in the 1960’s. This is demonstrated statistically in a Roper research poll aimed to investigate American television habits and related attitudes (Small 12). Adult participants were asked how they received most of their news about world current events and were then given options including television, newspapers, magazines or radio. In 1959, 57% identified newspapers over television; by 1969, television had taken the lead as the preferred source as stated by 64% of individuals (13). Participants were also asked which source they would be more inclined to believe if they experienced conflicting reports of the same news story. In the 1959 poll, the results showed television slightly behind newspapers, but the 1969 responses showed an increase of 44% choosing television as a more trusted source over 21% identifying newspapers. 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