Leptospira Cultures Maintenance

Leptospira Cultures Maintenance RESULTS The Leptospira serovars maintained in the Department of Veterinary Microbiology were used in the present study. For maintenance, EMJH medium (Difco) with albumin supplement was employed and subcultured at seven day intervals and incubation was carried out at 28-30Â °C. In addition, the stock cultures were maintained in semi-solid medium with subculturing at one month interval. IDENTIFICATION OF LEPTOSPIRES Under dark field microscope, the live leptospiral organisms were found tightly coiled and actively motile. The motility observed was of both spinning and bending. In highly concentrated cultures, the organisms formed entangled masses. No contaminants were observed in most of the time when streaked on blood agar plates for purity checking of the cultures. In case of contamination they were purified by filtration. RECOMBINANT PROTEIN PRODUCTION Preparation of template DNA from Leptospira The genomic DNA isolated from Leptospira interrogans serovar Australis had a DNA concentration ranging from 40- 60Â µg/ml. The purity of the extracted DNA was checked by measuring the ratio of absorbance (O.D of DNA preparation at 260 and 280 nm). The value of the ratio obtained was found to be in the range of 1.8 to 1.93, indicating that the preparations were almost free of proteins.(in mat and methods). Amplification of lipl21 gene and lipl32 genes The amplification of lipl21and lipl32 genes were carried out and observed amplicons of 507 bp and 767 bp, respectively. (Fig 1) Cloning of the lipl21 and lipl32 gene The colonies of E.coli Dh5∞ cells transformed with lipl21 and lipl32 genes was picked up separately and tested for the presence of the two genes. It was observed that lipl21 clones yielded an amplicon size of 507 bp and lipl32 with 767 bp. These confirmed clones were preserved for further studies (Fig 2) Induction of recombinant protein The above clones were subcultured in LB broth containing Ampicillin (100Â µg/ml) and expression was optimized with 2 mM IPTG for LipL21 and 1mM IPTG for LipL32. The induced recombinant cells were harvested after six hours and five hours for LipL21 and LipL32, respectively. Uninduced controls were set for each. The cells were then pelleted and lysed. The expression of recombinant 21 kDa (rLipL21) and 32 kDa outer membrane proteins (rLipL32) were confirmed in comparison with that of the uninduced cells where there was no significant protein expression (Fig 3) Purification of recombinant lipl21 and lipl32 proteins The rLipL21 and rLipL32 proteins were purified by Nickel chelating affinity chromatography without any contamination. The protein concentrations were estimated to be 0.69 mg/ml and 2.07mg/ml for rLipL21 and rLipL32, respectively. Immunoreactivity of the proteins The immunogenicity of both rLipL21 and rLipL32 proteins were tested using MAT positive canine sera and observed that both the proteins were reacting. Further the protein did not react when blotted with hyper immune serum raised against the different bacteria namely E.coli, Staphylococcus aureus and Pasteurella multocida. DIAGNOSIS Microscopic Agglutination Test A total of 124 canine serum samples from Leptospira suspected dogs were tested using MAT and among this 22 (17.74 per cent) were found to be positive for leptospirosis. Serum samples having a titre of 1:400 and above were considered as positive. The infecting serovars identified with canine leptospirosis are depicted in Table 3 Enzyme Linked Immunosorbent Assay Indirect ELISA was done in separate microtitre plates employing rLipL21 and rLipL32 as antigens and the results were compared with that of MAT. Checker board analysis Using checker board analysis the optimum concentration of antigen, antibody and conjugate were estimated. The optimum concentration of antigen was found to be 50 ng/well and 150 ng/well for rLipL21 and rLipL32, respectively. The rabbit anti-canine IgG HRP conjugate concentration estimated was 1:2500 and 1:2000 for rLipL21 and rLipL32, respectively. A 1:50 dilution of test serum was used as optimum working dilution in both ELISAs. Determination of cut off values In IgG ELISA, the mean OD and standard deviation for the negative sera samples (n=44) was 0.49 and 0.11 for rLipL21 and 0.59 and 0.09 for rLipL32, respectively. The cut off value obtained was 0.82 for rLipL21 and 0.86 for rLipL32. Test proper The results of rLipL21 and rLipL32 based IgG ELISA are given in Table 4. Among 47 positive samples obtained by rLipL21 ELISA, only 20 found positive with MAT. In case of rLipL32 ELISA, 40 samples were recorded positive out of which 18 found positive with MAT. Comparison of MAT and rLipL21IgG ELISA The results of rLipL21 IgG ELISA were compared with that of MAT. Among 124 canine sera examined, 47 (37.90 per cent) showed OD more than the cut off value i.e. 0.82 and were considered positive for leptospirosis by rLipL21 IgG ELISA. The sensitivity, specificity and accuracy of rLipL21 IgG ELISA as relative to MAT was calculated to be 90.90 per cent, 73.52 per cent and 76.61 per cent, respectively (Table 5). On statistical analysis, it was found that there exists a significant difference between the two tests, ie, rLipL21 ELISA and MAT even at 1 per cent level of significance. Comparison of MAT and rLipL32 IgG ELISA The results of the IgG ELISA using rLipL32 as the antigen were compared with that of MAT. Among 124 canine sera examined, 40 (32.25 per cent) showed OD more than the cut off value i.e. 0.86 and were considered positive for leptospirosis by IgG ELISA. The sensitivity, specificity and accuracy of rLipL32 IgG ELISA as relative to MAT was calculated to be 81.81per cent, 78.43 per cent and 79.03 per cent, respectively (Table 6). Statistical analysis revealed that there exists a significant difference between the two tests, ie, rLipL32 ELISA and MAT at 1per cent level of significance. Comparison of rLipL21 and rLipL32 IgG ELISA On statistical analysis using Cochran’s Q test, at 1 per cent level of significance it was observed that there exists no significant difference between rLipL21 and rLipL32 ELISA, as the P value was found to be greater than 0.01. Table 3 Infecting serovars identified with MAT Table 4 Results of MAT, rLipL21 ELISA and rLipL32 ELISA Table 5 Comparison between rLipL21 ELISA and MAT Sensitivity = a/a+c = 90.90% Specificity = d/b+d = 73.52% Accuracy =a+d/a+b+c+d = 76.61%

Euthanasia of Pretamture Babies :: Free Euthanasia Essay

Amy gave birth to a baby boy after only 24 weeks gestation. The premature child weighed only 1 1/4 lbs and was only 13 inches. Statistically, the child has between a 5% and a 20% chance of survival and over a 50% chance of growing up with intellegence disabilities, cerebral palsy, blindness, and deafness. Should money be continually spent on medical treatment or should the parents and/ or doctor be given the rigt to take away the life support? What is euthanasia? Is it just a murder, or is it a helpful procedure to end people's suffering? It is the "act of helping a person end his or her life in a painless fashion to relieve unbearable suffering". It is the taking of a human life by another person to prevent further pain and misery. There are four types of euthanasia: Active, Passive, Voluntary and Involuntary. Active Euthanasia is when a second party, whether is be a doctor or someone else, discretely introduces a lethal amount of medication into the sick or dying person. Passive Euthanasia involves the removal of medical treatment from the suffering individual. Voluntary Euthanasia is when the patient asks his or her doctor to assist in the procedures to ensure the end of the patient's life. Involuntary Euthanasia is present in Amy's case, along with Passive Euthanasia. It is when the individual who is suffering is either in a coma, incompitent, or a premature child that cannot speak for itself, and instea d of the patient making the decision, the family and/ or doctor does. Prematurity is when a child is born within less than 36 weeks gestation. The child is usually born with low birth weight, some type of physical deformity and / or any kind of mental problems that can all lead to a future of being outcatsed. Around half the babies born at 24 weeks are at risk of growing up with severe disabilities. Minor disabilities dont usually occur until further in the child's life, approximately once the child begins attending school. Reading and learning problems may occur. Obviously, the earlier the child is born the less chance of survival and health there is. This is because the babies have not yet fully developed the lungs, causing sudden collapses with every breath the child takes. "Matters involving the most intimate and personal choices a person may take in a lifetime...are central to the liberty protected by the 14th amendment.

Competition in the Movie Rental Industry in 2008

Competition in the Movie Rental Industry in 2008: Netflix and Blockbuster Battle for Market Leadership Although the corporate strategies implemented by Netflix and Blockbuster have allowed them to become leaders of competitive advantage in the movie rental industry, they sometimes encounter strategic issues that slow down their product and services process. My research of Netflix and Blockbuster will enable me to present a SWOT analysis and recommendations for each company. Netflix, founded in 1997 by Reed Hastings, has achieved its goal of becoming the largest online movie rental service in the world. By the end of 2007, Netflix recorded revenues of $1. 2 billion. With a library of 100,000 movie titles and a subscriber base of over seven million, they had become the leaders of the movie rental industry (Gamble & Thompson, 2011). Netflix’s business model of internet subscription enabled them to compete in the movie industry. Consumers love going to the movies, but with increasing theatre prices found it too expensive to attend public viewings. Netflix provided an inexpensive way to view movies which could be done from the comforts of home. According to the text, (Essentials of Strategic Management, 2010), â€Å"Netflix’s success is due to its six-pronged strategy of providing comprehensive selection of DVDs, easy way to choose movies, fast delivery, no return due dates, and convenient drop in mail movie returns† (Gamble & Thompson, 2010). In an online survey by Nielsen Online, Netflix was rated number one for three years and for nine consecutive periods by Forsee/FGI Research (Netflix, 2009). Netflix Strategic Issues Blockbuster, Netflix’s fiercest competitor, experience many rental issues until 2007 when they regained market shares forcing Netflix to reduce subscription prices. Not only did Blockbuster gain presence, other competitors like Redbox also gained presence in the market due to new technologies (VOD & DVR) that are influencing the business environment. The business model used by Netflix caused a stir in the market industry. However, the damage control strategies of competitors and competitor recapture of market shares is threatening Netflix’s competitive advantage. Netflix has to change with the times. Competitive Forces Netflix and Blockbuster are affected by the five forces of competition which are potential new entrants, bargaining power of buyers, bargaining power of suppliers, threat of substitutes, and rivalry among existing competitors (Gamble & Thompson, 2010). The companies must understand how these forces work and affect their operation. Threats of New Entrants -In the home video/game industries, new entrants must own large amounts of movies/games for rental or sale to fulfill customers' demands. Meanwhile they have to build up various distribution channels for products to reach customers in a very quick way (Xie & Lin, 2008). Bargaining Power of Suppliers -The inputs of suppliers in the home video/game industries are very important and since there are only a few qualified suppliers in the industry, their bargaining power is high. Netflix acquires its movies from movie studios and distributors, buying DVDs on a fee-per-DVD basis, paying license content fees, and signing revenue sharing agreements. Blockbuster also has revenue sharing agreements with its suppliers. To some extent, these agreements reduce the bargaining power of suppliers. In terms of the computer system, Blockbuster is using Provia's Viaware warehouse management system (packaging, sorting, and distributing rental products) in its supply chain management to keep costs down (Xie & Lin, 2008). Threats of substitutes -Substitutes include movie theaters, satellite TV, and cable TV. Customers can go to movie theaters and enjoy the vivid atmosphere. Alternatively, they can order â€Å"pay-per-view† or subscribe â€Å"on-demand† from satellite TV and/or cable TV providers to watch movies at home. Users can watch anytime they want. Satellite and cable TV offer sufficient selections of new releases and are is easy to watch with just click on remote. Netflix and Blockbuster compete for customers by offering various kinds of movies and internet access (Xie & Lin, 2008). Rivalry among industry competitors – Netflix and Blockbuster are in a highly competitive industry. Competitors include merchant retailers, such as Wal-Mart, Best Buy, and Target; video and game store like Hollywood Video, Movie Gallery, and Game Stop; supermarkets, convenient stores such as Publix, and McDonald's. The significant rival is Hollywood Video which offers movie and game rentals (Xie & Lin, 2008). Bargaining power of buyers -Buyers are not in concentrated groups and do not buy in large amounts. However, within the entertainment industry, customers have a lot of alternatives and have no switching cost. However the introduction of DVDs, influenced customers to purchase DVDs since the cost is almost the same cost of rentals. This makes buyer power moderate (Xie & Lin, 2008). The five forces of competition of the movie rental industry presents little force against a competitor’s market position based on buyer power, supplier power, and new entrant threats. However, threat of substitutes and rivalry among competitors can affect the amount of profits a company will gain and retain. Netflix SWOT Analysis The presence of Netflix and Blockbuster in the movie rental industry has assisted me in developing this analysis of each corporation’s strength, weaknesses, opportunities, and threats as followed: Netflix’s strengths are: Good reputation, unlimited movie rental subscription, no due dates or late fees, easy website access, and large movie selection. Netflix’s weaknesses are: Limited to internet rental access, and long waiting period for new releases. Netflix’s opportunities are: To provide Video on Demand media access, music and movie download, and acquire partnership with gaming industry. Netflix’s threats are: Low DVD rentals, government regulation of rental distribution, high level of competition, and new movie media technology. Blockbuster SWOT Analysis Blockbuster’s strengths are: national and international recognition provides movie and game rentals, various vices for rental access (i. e. stores, kiosk, and internet), offers snacks and beverages for purchase in stores, and accommodates all customer types. Blockbuster’s weaknesses are: Poor distribution process, increased operating expense, decreased reputation, limited new released video games, unable to compete with game distributors (i. e. GameStop), and constant rental policy changes. Blockbuster opportunities are: Unlimited access to downloadable media libraries, increased production of brand movie rentals, and partnership with gaming system industry. Blockbuster’s threats are: New video formats, decreased DVD movie rentals, increased competition, and government regulation of movie rentals. The SWOT analysis shows that Netflix and Blockbuster have strong strengths and opportunities, however in order to Netflix to maintain its competitive advantage there needs to be an upgrade in innovation and technology that will strengthen their reputation and reliability. Blockbuster’s loss in reputation will continually plague their ability to remain competitive and maintain the customer base that they have. Blockbuster will continually face challenges until they upgrade their innovation and technology to level where they will enter into recovery mode. As I studied the financial information presented in the text, it appears that Netflix has been able to offer customers the benefit of low rental fees because they have been able to maintain a low level of expenses. The benefits offered to customers have increased the amounts of net income and movie rentals. Netflix prides itself on promoting customer satisfaction and meeting demands of its internet base. According to the numbers presented by Internet World Stats, the percentage of internet penetration by the North American population was 74. 2 percent in 2009, which was a 134 percent increase from 2000. At this rate, Netflix could secure an internet rental customer base of 74. 2 percent which would strengthen its success (Miniwatts Marketing Group, 2009) Blockbuster’s biggest challenge is development of a viable strategic approach to price setting. When Netflix entered the game, Blockbuster began to lose profits. In an attempt to recoup some of its losses, Blockbuster presented campaign where the rental policy constantly changed leaving customers confused and unsure of the rental process. Although Blockbuster has a large selection of movie titles and global presence, its reputation of dissatisfied customers and inefficient distribution has caused its overall cost to rise significantly. Blockbuster’s competitive advantage is that it offers various channels by which customers can access entertainment media. According to Blockbuster, â€Å"The same customer can choose different ways to access media entertainment on different nights† (Blockbuster, 2010). Recommendations In order for Netflix to maintain its competitive advantage there must be an adjustment of their strategy. Customer satisfaction should be a key factor of its strategy. I recommend that they base their marketing promotions on increasing customer awareness of the variety of accessible content offered. They should also implement the availability of music content and digital media to their library. The development of an award program for customers with lengthy memberships would influence word of mouth recommendations. Most of all, Netflix must continue to offer their customers the benefit of more titles at modest cost subscription memberships. In order for Blockbuster to regain its competitive advantage and reputation there must be the development of a strategy that will increase their profit. They have to become more innovation in internet and in-store rental process. There should be monitoring devices placed on scanner registers and mailed movie rentals to better track returns in order to keep their cost at a minimum. Staff should be trained and monitored on a constant basis to ensure that customer satisfaction is the key priority. Blockbuster must implement policies that will be enforced and understood by customers. They should also team up with gaming system producers to ensure that they will be supplied with current games and possibly offer gaming system rentals. Most of all they should find their own niche market which will provide them with a specialized competitive advantage. The information researched on Netflix and Blockbuster has enabled me to conclude that competition in the movie rental industry is highly competitive. There are new companies entering the market that have a competitive advantage whether it consist of internet rental, vending machine rental access, or in-store rental access. Netflix and Blockbuster have taken the lead in providing thousands of movie title to be viewed in the comfort of our own homes without the expense of movie theatres. Although both companies have a competitive advantage, they each have a following of dedicated customers who will stay with them because of familiarity. Through my research I have learned that consumers are the controllers in the movie rental industry. If consumers dislike the way they are serviced or the cost of movie rentals they will most certainly move on to a company which will cater to their movie wants and needs. However, the movie rental industry must stay up on new innovations and technologies in order to remain competitive, profitable, and maintain modest levels of overall costs.

Should Marijuana Be Legalized - 1617 Words

Aashish Pandey Dr. William Carney English Comp. 1113 2 December 2014 Legalization of marijuana According to Wikipedia, marijuana is a name for cannabis plant and a drug preparation made from it. Commonly it is named as cannabis, bud or weed, pot or ganja. I have found marijuana to be one of the most controversial things in United States. Not necessarily is marijuana controversial, the views related with the legalization of this thing are truly dubious. It is true that different persons have different views to see every object. So the view of peoples towards the legalization of marijuana varies with the person. In the United States, I have found that some of the people think that marijuana should be legalized because it is not much harmful as people’s belief and also there are lots of therapeutic values. Others believe that it is a drug and gateway to other drugs so it should not be legalized. And some believe that as it has both good and bad effects it should be legalized with certain condition. In this essay I am going to talk about the views of different people on the legalization of marijuana. â€Å"You don’t need to be cool to smoke weed, but if you smoke weed you are really cool†. I found this statement in one of the page in Facebook. †(I love smoking weed)†. This statement surely encourages the teenagers to be attracted towards marijuana. Each and every thing related with marijuana increases the curiosity ofShow MoreRelatedShould Marijuana Be Legalized?849 Words   |  4 Pageswhether marijuana should be legalized. Around 23 states have legalized marijuana for medical and recreational use. In the state of Illinois, medicinal use of marijuana has been passed on April 17, 2013. Since January 2014, patients are able to obtain marijuana with a doctor s recommendation. The new debate is whether marijuana should be legalized for the general public as a recreational drug. Although some believe that marijuana is harmless, and that it has beneficial medicinal uses, marijuana shouldR ead MoreShould Marijuana Be Legalized?1715 Words   |  7 PagesMarijuana in Society Cannabis, formally known as marijuana is a drug obtained from the tops, stems and leaves of the hemp plant cannabis. The drug is one of the most commonly used drugs in the world. Only substances like caffeine, nicotine and alcohol are used more (â€Å"Marijuana† 1). In the U. S. where some use it to feel â€Å"high† or get an escape from reality. The drug is referred to in many ways; weed, grass, pot, and or reefer are some common names used to describe the drug (â€Å"Marijuana† 1). Like mostRead MoreShould Marijuana Be Legalized?1489 Words   |  6 Pagescannabis plant or marijuana is intended for use of a psychoactive drug or medicine. It is used for recreational or medical uses. In some religions, marijuana is predominantly used for spiritual purposes. Cannabis is indigenous to central and south Asia. Cannabis has been scientifically proven that you can not die from smoking marijuana. Marijuana should be legalized to help people with medical benefits, econo mic benefits, and criminal benefits. In eight states, marijuana was legalized for recreationalRead MoreShould Marijuana Be Legalized?1245 Words   |  5 PagesMarijuana is a highly debatable topic that is rapidly gaining attention in society today.   Legalizing marijuana can benefit the economy of this nation through the creation of jobs, increased tax revenue, and a decrease in taxpayer money spent on law enforcement.   Ã‚  Many people would outlaw alcohol, cigarettes, fast food, gambling, and tanning beds because of the harmful effects they have on members of a society, but this is the United States of America; the land of the free and we should give peopleRead MoreShould Marijuana Be Legalized?1010 Words   |  5 PagesThe legalization of marijuana became a heated political subject in the last few years. Twenty-one states in America have legalized medical marijuana. Colorado and Washington are the only states where marijuana can be purchased recreationally. Marijuana is the high THC level part of the cannabis plant, which gives users the â€Å"high† feeling. There is ample evidence that supports the argument that marijuana is beneficial. The government should legalize marijuana recreationally for three main reasonsRead MoreShould Marijuana Be Legalized?1350 Words   |  6 Pagespolitics in the past decade would have to be the legalization of marijuana. The sale and production of marijuana have been legalized for medicinal uses in over twenty states and has been legalized for recreational uses in seven states. Despite the ongoing support for marijuana, it has yet to be fully legalized in the federal level due to cultural bias against â€Å"pot† smoking and the focus over its negative effects. However, legalizing marijuana has been proven to decrease the rate of incrimination in AmericaRead MoreShould Marijuana Be Legalized?1231 Words   |  5 Pagesshows the positive benefits of marijuana, it remains illegal under federal law. In recent years, numerous states have defied federal law and legalized marijuana for both recreational and medicinal use. Arizona has legalized marijuana for medical use, but it still remains illegal to use recreationally. This is absurd, as the evidence gathered over the last few decades strongly supports the notion that it is safer than alcohol, a widely available substance. Marijuana being listed as a Schedule I drugRead MoreShould Marijuana Be Legalized? Essay1457 Words   |  6 PagesSHOULD MARIJUANA BE LEGALIZED? Marijuana is a drug that has sparked much controversy over the past decade as to whether or not it should be legalized. People once thought of marijuana as a bad, mind-altering drug which changes a person’s personality which can lead to crime and violence through selling and buying it. In the past, the majority of citizens believed that marijuana is a harmful drug that should be kept off the market and out of the hands of the public. However, a recent study conductedRead MoreShould Marijuana Be Legalized?1596 Words   |  7 Pages But what needs to be known before a user can safely and completely make the decision if trying Marijuana is a good idea? Many do not want the drug to be legalized because they claim that Cannabis is a â€Å"gateway drug†, meaning it will cause people to try harder drugs once their body builds up a resistance to Marijuana, because a stronger drug will be needed to reach a high state. This argument is often falsely related to the medical si de of the debate over legalization. It is claimed that this wouldRead MoreShould Marijuana Be Legalized?985 Words   |  4 PagesLegalize Marijuana Despite what people believe about marijuana, it hasn’t once proved to be the cause of any real issue. It makes you wonder what the reason as to why there is a war on drugs. Why is marijuana the main concern? Since the time that alcohol and tobacco became legal, people wonder why marijuana isn’t legal yet. The fact that marijuana is illegal is mainly caused by the amount of money, jobs, and pride invested in the drug war. Once the government starts anything, they stick to it. At