Promoting a Healthy Housing Environment for the Older People Article

Promoting a Healthy Housing Environment for the Older People - Article Example The government visualizes the board as the main point in making decisions concerning the wellbeing and the local health (Cattan & Tilford, 2013, p. 3). Â  Houses developed for the older people in Devon are designed to enable older people to achieve greater independence and for their wellbeing. This is because they live freely in their own apartments that are designed and suitable for their conditions. There are a variety of housing options and care option. Some of the features that are prone in those houses are; the houses are accessible, this is because they designed for independent older people including those that are not physically fit. They are provided with Tele-care services where the majority of them run for twenty-four hours. These people do ensure and monitor their steps to ensure their wellbeing. There are provided with benefits such as communal services (Cattan & Tilford, 2013, p. 3). Â  The houses are usually cost friendly and so the majority of older people is able to afford them. There are also located in secure places. Their landlords, managing the house are professionals that are able to handle their business with the old people. The service providers, the tenant, the landlord and the Tele-care people are all human friendly, therefore, the environment is usually friendly for the elderly (Wahl, 2010, p. 13). Â  

Consent Essay Example | Topics and Well Written Essays - 2000 words - 1

Consent - Essay Example In this sense it is meant that the review should be constructed in such a manner that the researcher/researchers can either use it as a stand-alone published material that can assist other researchers with similar topical considerations in their own research or the researcher/researchers can use the review exclusively as an assisting part of their own research on the topic. This brings out the importance of literature review that is possibly more important a part of the initial research strategy than any other part of such initial research. Choice of Topic - The first consideration that a researcher or group of researchers must take into account is the choice of topic. For this, they may first scrutinize the subject they are studying and thresh out a topical part that has scope for research the researchers themselves feel they are competent enough to carry out. It is notable that this is the very initial step to beginning an earnest literature review (Reed, Undated). The choice of a suitable topic may constitute the following steps - Brainstorming - This is process with which the researcher scans the subject he/she is studying currently to pinpoint a topic within that subject he/she may want to research upon. Within this process the subject is broken down into its topical components and basic keywords are derived from these. For example, as in this case, where the subject is taken to be 'information within the healthcare system', some of the topical interests that may get unearthed are as follows: Acquisition of such information; Storage of such information; Retrieval of such information; Ethics regarding retrieval; etc. Next, books, journal articles and other resources that have been recommended for these topics within the main study subject are next scrutinized and studied to ensure which of these topics may be the most suitable for research. Some of the criteria with which this is judged may be as follows: if there is any scope for further research within any of these topics; and whether such research can be easily carried out with the resources and facilities that are presently available with the researchers. In this manner a topic is settled upon on which the researchers decide they want to do research. The topic has scope for further research and the research type is suitable for being conducted by the researchers, who consider themselves capable of it. The topic settled upon in this case is 'Ethical issues regarding retrieval of healthcare information available from storage'. Consultation with the Instructor/Professor - Another adjacent step to brainstorming to settle upon the research topic is consultation with the instructor ort professor in charge of the discipline the researchers are incumbent upon. Often, these able persons are capable of making valuable suggestions that allow the research to proceed smoothly and efficiently. Researching Background Information - After a topic suitable for conducting research upon has been selected, the next logical step is to find background information on it from resources like

Anthropology peoples sothesat asia Essay Example | Topics and Well Written Essays - 750 words

Anthropology peoples sothesat asia - Essay Example These two opposing clans are in constant conflict; but the advent of slavery will merge the steadfast differences of these two into one similarity: Borneans in Southeast Asia who are duty-bound under the guise of civilization and the economic vitality of slavery. In the end of the eighteenth century, western superpowers, specifically the Spanish, Dutch, and British in Borneo, commenced in subjugating Southeast Asia for the purpose of monopolizing sought after products and raw materials in the developing marketplace in the region and Europe. Borneo, endowed with the gift of a lucrative geographical position, intensified its trading relations with other Southeast Asian countries. This move improved the island’s economy and commercial power. The growth of trade perverted through notching a better market for slave traders and raiders owing to the demand for laborers to accumulate riches and resources. Asian slavery is different from the western slavery structure. Slavery in Asia w as of two types: the debt slaves and the bondage slaves. The former type were people who paid large debts through elicited free labor whereas the latter type were captives from raids and wars who were sold to slavery. In the seventeenth and the eighteenth centuries, the population of Southeast Asia remained at a very low level hence troubling the region with problems of labor shortage. This reality aggravated the lust for money of the raiders who traded slaves as laborers in a naturally bountiful but populace- deprived environment. Nevertheless, even before the Dutch colonized Borneo it had already been the fortress of slave trade in which slaves were awarded as tributes or trading commodities. Although, it is important to note that western concept of slavery in Asia which is characterized by slaves in chains and those who were frequently chastised and maltreated, is not coherent

Marketing Assignment Example | Topics and Well Written Essays - 6500 words

Marketing - Assignment Example ISO works process framework would be used to show how SIX Sigma might be laid out and what information will be needed to implement this tool. There are many tools that make up this collective and they need to be fully researched in the first instance for early stages and consultants/experts in the implementation of these tools are required to avoid pitfalls caused by redundant data. One problem that is required to be addressed with these tools is that of measuring potential work flows and bottle necks in the production process. "Six Sigma seeks to improve the quality of process outputs by identifying and removing the causes of defects (errors) and minimizing  variability  in manufacturing  and  business processes." (one six sigma web) The 'Qualicoat' status requires us to do much of the processes of Six Sigma would and we run a zero defects policy with any defects being reworked. New innovation such as Quality control measures before the Powder coating process will mean that many of the issues will be taken care of on the production line. Where Six Sigma and learn thinking tools will come into their own will be more visible in our data processing and customer liaising as well as in our order processing and sales activities. Sales personnel will be able to predict work flows with new centralised communications and above mentioned tools will assist in that role facilitating negotiations at first contact with customer queries. Six Sigma and Total Quality Management (TQM) are tools that are going to be used to push the changes forward and improve on employee participation towards a culture of due diligence. Various tools are now being introduced to engage the workforce in dialogue to identify problem areas, provide training and walk-through them in meeting new goals while actually improving their working environment. Brief look would be given to the questionnaires and language that are going to be used to encourage change. One problem is the resistance to c hange that has been experienced in the past because of people being stuck in the old ways of carrying out processes thus introduction of Six Sigma terminology will create confusion and animosity. These terminologies would only be used within management to avoid unnecessary chaos. It is to be assured that there is the balance of communication therefore new responsibilities are properly defined along with the changes in processes. In order to enhance data collection and processing new management positions would be introduced. There are several objectives of TQM that include process improvement, observing behaviour changes over a time period, defect prevention, developing operational definitions, priority of effort, developing improvement check forms and check lists, developing relationships of cause-effect, measuring capacity of the system, assisting teams in making better decisions and separating trivial from significant needs. TQM on the whole revolves around a few main things that include systems for facilitating improvement, techniques and tools for improvement, Senior Management and all employees’ commitment, team work, customers - Supplier relationships, effective vision, mission, strategy and goals, communication and trust. A systematic approach towards quality management involves components like modification of the processes and the inputs, examining of the processes performance, planning of the inputs

Organizational performance Essay Example for Free

Organizational performance Essay Todays society is diverse and rapidly changing and it is the organisations ability to adapt or be in fit with the external environment that will determine its overall performance. Ethical issues arise throughout all organisations daily activities but it is the continual debate about what is right or wrong that will shape ethical decision making now and for generations to come. This essay aims to investigate culture within the internal environment and how culture plays a role in an organisations ability to fit in with the environment. According to (Samson and Daft, 2003:80) the internal environment is composed of present employees, management and business culture. This essay will investigate how culture plays a role in the organisations overall success. In addressing the issue it will be shown how the external environment has changed in terms of how organisations are evaluated. How changing an organisations culture can help the organisation be in fit with the external environment and how the success of changing culture may depend on the level of employee moral development. It is argued that the organisations performance depends on a fit between the organisation and its external environment. Body Key point 1: How the external environment has changed in terms of how organisations are evaluated. The external environment has changed with respect to how organisations are evaluated today. According to ( Samson and Daft, 2003:79) the external environment is all elements existing outside the organisations limitations that have the possibility to affect the organisation. Companies around the world have started to realise that investors are not concerned exclusively with financial performance (Tschopp, 2003). The days of companies being  evaluated on their financial performance are gone and companies are now finding that they are being evaluated on a more overall perspective. Increasing the ethical obligations can help an organisation when adapting to the external environment. According to ( Samson and Daft, 2003:147) ethics is the code of honourable principles and standards that governs the behaviours of an individual or group with respect to what is correct or incorrect. If ethics is incorporated as part of the organisation this can lead to improvements in the workplace and towards society. The triple bottom line approach has been introduced as a way of achieving overall success both internally and externally throughout the organisation. Triple bottom line entails reporting on economic, social, and environmental issues. Corporate success should be considered not just by the traditional financial bottom line, but also by its social/ethical and environmental performance ( Samson and Daft, 2003). Triple bottom line has not only put the emphasis on managers to not only make a profit but to also consider the surrounding external environment that they could be affecting. Businesses now report annually on social and environmental performance as well as their financial performance because they know it provides a more complete measure of long-term value creation and strategic opportunity (Tuchman. J, 2004). Key Point 2: Organisations need to change to fit. They can do this by changing an organisations culture to fit in with the external environment. The external environment has altered and its the organisations ability to change to keep in touch with the external environment that determines its performance. One way an organisation can change to keep in contact with the environment is by altering its culture. According to (Samson and Daft. 2003: 94) culture is the knowledge, beliefs, values, behaviours and ways of viewing shared among members of a society. Organisational culture has been defined, in very simple but intuitive words, as the way we do things around here (Domenec, 2003). The notion that we can make others do what we want them to do by persuading them to want to do  it is one that has a long pedigree. This notion became formalised as an integrative view of organisation culture and became more ingrained after the publishing of the book In Search of Excellence in 1982 (Thomas J. Peters and Robert H. Waterman, Jr., 1982) Moving towards greater corporate responsibility may require culture change or at least cultural re-enforcement. The fundamental values that exemplify cultures at these and other organisations can be understood through the noticeable manifestations of symbols, stories, heroes, slogans and ceremonies. Any organisations culture can be interpreted by observing these factors (Samson and Daft, 2003:95). By incorporating symbols, stories, heroes, slogans and ceremonies into an organisations culture they will be able to adapt to the changing external environment. According to (Samson and Daft, 2003:97) slogan is a phrase or sentence that succinctly expresses a key corporate value. Hungry Jacks for example has the slogan The burgers are better at Hungry Jacks. They have incorporated this slogan as part of their culture to try and separate them from the other competitors. By incorporating the different types of culture into and organisation, may change the way managers and employees think to incorporate social and natural environmental responsibilities into the workforce. Key Point 3: The success of changing culture may depend on the level of employee ethical development. The success of changing culture can depend on numerous factors but the level of employee ethical development plays a role. The three levels of personal moral development could pose a problem for employee ethical development. The theory developed by Kohlberg goes through the different stages of employee development from pre-conventional to post conventional. Starting at the pre-conventional level which focuses on right vs. wrong and the behaviour is on ones self to the conventional level which focuses on the  group rather then ones self. Then lastly Kohlbergs post-conventional level of individual development which focuses on abstract and self-chosen principles (Arnold and Lampe, 1999). Kohlbergs theory brings attention to the fact that if employees are going in different directions it can hinder the success of an organisation. If there are employees who are focusing on what is right vs. wrong and other employees who are following self chosen principles even though they know people hold different views. In this sense, the greatest danger to modern organisations is the betrayal of ambitious, selfish, untrustworthy people who care more for their own progression than the mission of the organisation (Domenec, 2003). Since each person is unique, each one can focus on personal accomplishment in very different ways (Domenec, 2003). Conclusion This essay investigated culture within the internal environment and how culture plays a role in an organisations ability to fit in with the environment. In support of this argument there has been evidence supported to show how organisations incorporate culture to be in fit with the changes of external environment. Organisations are finding that they are being evaluated not only on their financial status but also their social and environmental performance. This has meant that organisations have had to change their culture to compensate in the change in evaluation. An organisation culture can be observed through such factors as slogans and symbols which now have to coincide with the external environment. The success of the changing culture can depend on the level of employee ethical development and at what stage everyone is at. If an organisation is united and following the same path or views they may find greater overall success. Views that link an organisations culture with its performance seek to shape managers and employees understandings in a common and coherent direction (Kolter and Keskett, 1992). Bibliography: 1. Samson, D., Daft, R.L. (2003) Management: Pacific rim edition. Victoria: Thomson. 2. Thomas J. Peters and Robert H. Waterman, Jr., In Search of Excellence: Lessons from Americas Best-Run Companies (New York: Warner Books, 1982), 3. Tuchman, J. 2004, Big Owners Balance Triple Bottom Line [online], Available from URL: http://www.enr.com/news/bizlabor/archives/040809-1.asp 4.

Topshop vs Urban Outfitters Essay Example for Free

Topshop vs Urban Outfitters Essay Topshop was established in 1964 within a Sheffield department store called Peter Robinson in U. K. In 1974, Topshop was taken out of Peter Robinson and set up as a standalone retailer. It is a chain retail clothing store that operates in more than 20 countries. It is part of Arcadia Group, which was also owns a number of well-know outlets including Burton, Dorothy Perkins, Miss Selfridge and Wallis. It is mainly known for womens clothing and its primary sales come from clothing and fashion accessories. Their womenswear carry dresses, tops, bottoms, bags, shoes, accessories, cosmetics and lingeries. Their mission is to bring in high street fashion at affordable price, their clothing style is very edgy, chic and classy. Their target customers are students to upper-middle class, demographic from 18 to 30s. Urban Outfitters is an apparel company that was incorporated in 1976. The company was originated as â€Å"Free Peoples Store† in Philadelphia in 1970, with concentration of â€Å"funky† household items and clothing. The founder were two Philadelphians, Scott Hayne and Scott Belair. The companys womenswear carry dresses, swimwear, tops, bottoms, bags, outerwear, accessories and intimates. Their products are urban hipster style, vintage feel with a modern twist, something that you can wear everyday. They target at young adult market, age from teens to 20s. Both Urban Outfitters and Topshop are specialty retail store. Topshop s visual merchandising at the front door is quite eye-catching. I think the clothes on the mannequins give a clear picture of what their product style is and what they are selling –cutting edge style. Their recent exterior is displaying mannequins which are facing inside the store rather than outside, this stimulates ones curiosity to enter the store and find out how the front view of the mannequins look like. The merchandise that are featured in the window are positioned on the ground floor, mostly in the center of the store where the themed merchandised items are located. Urban Outfitterss store window decor is not as appealing compare with Topshop. However, they are able to give out a message of their style—earthy, hip and unconventional style. Most of their merchandise that are featured in the window are located on the ground floor but they are not easy to be found as products are placed in different section. Merchandise of Topshop is organized by category, theme and clearance with different size range while Urban Outfitters is organized by category, designers and clearance with different size range. Sale merchandise in Topshop is positioned on the top floor of the store. They are placed according to category and located in a different area. Clothing from 10% to 30% off and shoes from 30% to 60% off. Sale merchandise of Urban outfitters is also positioned on the top floor of the store. However, they are grouped in a sale zone. Clothing and shoes from 30% to 40% off. In Topshop, all clothing are hanging on the rack, part of the accessories are placed on the table but mostly are hanging on the rack. Shoes are displayed on the wall mounted shoe rack while a few of them like boots are placed on the table within the same category section. On the other hand, Urban outfitters have their clothing both hung on the rack and folded on the table. Shoes are displayed on the wall mounted shoe rack and some are placed on the table. Accessories are displayed both on the rack and table. Books and toys are placed on either the table and the shell. Topshop is a UK national brand, their vendor assortment include collaboration with other designers. Recent collaboration is the JW Andersons collection which was launched last friday September 14, 2012. JW Anderson is a london fashion week star designer. The Topshop store in Soho has a special designed partition located by the entrance door to showcase the whole JW Andersons collection. Topshop also collaborated with starts like British supermodel Kate Moss and British artist Stella Vine in 2007. Urban Outfitters carry private label brands, all label products are mix together and placed by category. However, expensive products are organized by label brands in a particular section. In Topshop, there is a big sign of directory on the mirror wall with backlight letters next to the escalator which is hard to miss. In addition, clear signage underneath the escalator shows you the directory of the next floor while you are taking the escalator up. There are posters on the wall shows message like items on sales and student discount. In contrast, Urban Outfitters has no signage for direction used inside the store, but there are signage above the racks that indicates a particular priced group of items. Topshop is not publicly traded while Urban Outfitters is. Urban Outfitters is one of the retail brands under Urban Outfitters Inc. It is a publicly traded American company and is owned and chaired by Robert Hayne. The company is also held by several large investors, such as Fidelity Management Research Company, Maverick Capital Ltd, Vanguard Group, and Capital Guardian Trust Company. According to NASDAQ website posted on September 21, 2012. The total net sales during the second quarter of Urban Outfitters Inc. has increased increase 11% to $676. 3 million, the net sales by brands grew 14. 1% to $310. 7 million at Urban Outfitters. Topshop prices vary from line to line to accommodate all aspect of spectrum. One of their popular line â€Å"Mainline† offering classic style is marked at a reasonable pricepoint. On the other hand, â€Å"Premium† line is focus on shoppers of quality products who are looking for detailed, superior material apparel is on a higher pricepoint. Urban Outfitters prices does not vary much from label to label, products are marked at the same price range at a reasonable pricepoint. There are a few high-end label brands on a much higher pricepoint than the regular brands. In conclusion, both stores offers similar merchandise with the same merchandising philosophy. However, Topshop covers a wider range of target customers than Urban Outfitters. However, the only draw back within Topshop is its failure to provide prices that accommodate their large student market. Both of their style is very different and does not portray the same merchandising message. Topshop is more internationally recognized due to its market diversity and intense marketing strategies. I think Urban Outfitters should broaden their global threshold, for instance, expanding their market to Asia.

Growth Characteristics of Bacteriophage

Growth Characteristics of Bacteriophage CHAPTER 4 GROWTH CHARACTERISTICS OF BACTERIOPHAGE INFECTING AQUACULTURE BACTERIAL PATHOGENS 4.1  Introduction Bacteriophage are naturally occurring viruses that predated on bacteria (Clokie et al., 2011). They self-replicate exponentially and leave the commensal flora unaffected makes them useful for industrial application (Tsonos et al., 2014). However, the high number of bacteriophage in environment (Clokie et al., 2011) provide the challenges to the discovery of the most effective phage in treating bacterial pathogens (Lindberg et al., 2014). Even there were many extensive reports on bacteriophages, the clinical outcome of therapy trials are variable (Tsonos et al., 2014). This indicates that there are still many parameters which are unclear that may contributed to efficacy of the phage therapy. Previously, the most common practice to evaluate the therapeutic efficacy of phages was from in vivo studies. However, Lindberg et al. (2014) provide the alternative to evaluate the efficacy the phage treatment. The information could be assessed from the important phage traits such as adsorption, lysis time and burst size (Ackermann et al., 2004). Besides that, there are various physical and chemical factors like temperature, pH and salinity which could determine the occurrence and stability of bacteriophage (JoÅ„czyk et al., 2011). These factors reported to cause the inactivation of phage through damage of the phage structure (head, tail or envelope) or DNA structural changes (Ackermann et al., 2004). Therefore, the next section in this study is aimed to characterize the bacteriophage isolates (VALLPKK3, VHLPKM4 and VPLPKK5) based on their adsorption profile, one step growth profile and stability to various range of temperature, pH and bile salt concentration. 4.2  Materials and Methods 4.2.1  Bacteriophage Isolates The bacteriophage isolates that were used in the third chapter were further characterized in this chapter. The bacteriophage isolates were designated as VALLPKK3, VHLPKM4 and VPLPKK5. 4.2.2  Bacteriophage Adsorption Assay The bacteriophage adsorption assay was carried out following the method described by Hsieh et al. (2011) with few modification. In adsorption test, the host bacteria was first grown to OD600 1.0 or equivalent to ~108 cfu/ml and diluted to ~105 cfu/ml with TSB media. About nine ml of the host bacteria was mixed with one ml of phage lysate (~103 pfu/ml) to MOI of 0.001. Then, 100  µl of the bacteria-phage mixture was taken to determine the initial phage titer. The mixture was then incubated at 28 °C with no agitation. After 10 min, one ml of the samples was collected and centrifuged at 16,000 xg for 2 min to precipitate the absorbed phages. The same was repeated every 10 min for a period 50 min. The count of unabsorbed free phages in the supernatant was determined. Then, the free phage particles over the initial phage particles was calculated and expressed in percentage. The accuracy of the free phage count was improved by triplicate separate experiments. 4.2.3  Bacteriophage One Step Growth The one step growth assay was determined following method described Hsieh et al. (2011) with slight modification. First, host bacteria (OD600 1.0) was diluted to ~106 cfu/ml. Then, 100  µl of bacterial suspension was mixed with 100  µl of phage lysate (~103 pfu/ml) to a 1 ml of final volume with sterile TSB media (MOI 0.001). Second, the phage was allowed to adsorb into bacterial cells for 30 min at 28 °C. Then the bacterial cells were precipitated by centrifugation at 16,000 xg for 2 min. Third, the bacterial-phage pellet was suspended in 50 ml sterile TSB. Subsequently, 1 ml of the bacterial-phage suspension was precipitated by centrifugation and plated to determine the initial phage count. Then, two sets of bacterial-phage suspension concurrently collected every 12 min for a period of 84 min (for VALLPKK3 and VPLPKK5) and 132 minutes (VHLPKM4) for the determination of latent period, eclipsed period and burst size. The first set was subjected to above treatment to determine t he latent period and burst size while the second set was added with 40  µl chloroform, mixed and incubated at 28 °C for 5 min before centrifugation to determine the eclipse period. The free phage count in the supernatant was determined in triplicate. The latent period and burst size was determined according to Middleboe et al. (2010). The eclipse period was determine according to Sillankorva et al. (2008). The accuracy of the free phage count was improved by triplicate separate experiments. 4.2.4  Bacteriophage Tolerance Test The stability of the bacteriophage isolates was test in different range of temperature, pH and bile salt concentrations. The temperature test was conducted for one hour, while, the pH and bile salt concentration tests were conducted for 24 hours. a.  Temperature Tolerance Test The stability of bacteriophage in different temperature was done following method described by Phumkhachorn and Rattanachaikunsopon (2010) with slight modification. The bacteriophage solution was set to approximately 105 pfu ml-1 in sterile phage buffer. About 900  µl of sterile phage buffer was distributed into sterile empty 1.5 ml microfuge tube. The tube was incubated in the dry bath at desirable temperature (40, 50, 60, 70, 80, 90 and 100 °C) at least for 30 minutes. After 30 minutes, about 100  µl of bacteriophage solution (~104 pfu) was added into the preheated tube and mixed immediately. The tube was incubated again at desirable temperature for an hour. After incubation, the tube was placed in ice-warm bath to cool the bacteriophage solution. The titer of the survival phage was calculated by double layer method. The percentage of surviving phage was calculated by dividing the number of survival phage over initial phage count. b.  pH Tolerance Test The stability of bacteriophage in different pH was done following method described by Hsieh et al. (2011) with slight modification. The pH of phage buffer was adjusted into desirable pH (2, 3, 4, 5, 6, 7, 8 and 9) using pH meter (brand). The phage buffer was sterilized using autoclave machine at 121 °C for 15 minutes. The bacteriophage was set to approximately 107 pfu ml-1 in sterile phage buffer. The bacteriophage suspension was diluted to 105 pfu ml-1 (1/100) in phage buffer with different pH. The initial phage count was calculated and the bacteriophage solution was incubated at room temperature for 24 hours. After incubation, the bacteriophage solution was diluted using normal phage buffer and plating to calculate the survival phage by double layer method. The percentage of surviving phage was calculated by dividing the number of survival phage over initial phage count. c.  Bile Salt Tolerance Test The stability of bacteriophage in different bile salt concentration was done following method described by previous. The stock of bile salt (Brand) in phage buffer was prepared by filter sterilized to final concentration of 5 %. Then, the phage buffer was adjusted into desirable bile concentration (5000 ppm, 6000 ppm, 7000 ppm, 8000 ppm and 9000 pm). The phage buffer which used for the dilution of bile concentration was presterilized using autoclave machine at 121 °C for 15 minutes. The bacteriophage was set to approximately 107 pfu ml-1 in sterile phage buffer. The bacteriophage suspension was diluted to 105 pfu ml-1 (1/100) in phage buffer with different bile concentration. The initial phage count was calculated by serial dilution in normal phage buffer. The treated bacteriophage solution was incubated at room temperature for 24 hours. After incubation, the bacteriophage solution was diluted again using normal phage buffer and plated to calculate the survival phage by double laye r method. The percentage of surviving phage was calculated by dividing the number of survival phage over initial phage count. 4.3  Result 4.3.1  Bacteriophage Adsorption Assay In the adsorption analysis, all isolates have two adsorption phases, rapid and slow adsorptions. The rapid adsorption of VALLPKK3 was occurred within 10 minutes where almost 80% of the phage adsorb to the host (Figure 4.1). This result was similar to the VHLPKM4 (Figure 4.2). Meanwhile, the rapid adsorption of VPLPKK5 showed that around 60 % of the phage adsorbed to the host (Figure 4.3). After 10 minutes, the slow rate was occurred to all isolates. The number of unadsorbed phages was approximately below 20% within 40 minutes in all phages. The increase of phage count in VPLPKK5 was occurred after 40 minutes. The increase in free phages after 50 minutes indicates that the newly formed phages are being release from the infected cells (Figure 4.3). Figure 4.1: Adsorption of VALLPKK3 to V. alginolyticus ATCC ® 17749TM Figure 4.2: Adsorption of VHLPKM4 to V. harveyi VHJR7 Figure 4.3: Adsorption of VPLPKK5 to V. parahaemolyticus VPHG1 4.3.2Bacteriophage One Step Growth The one step growth was performed to identify different phases of the phage infection process. During the initial stage, the phage-bacteria cell was separated from the free phage during the adsorption since the adsorption result showed the availability of free phage after 30 minutes of incubation. After the infection, the phage growth parameters (latent period, eclipse period and burst size) were determine from the average of three independent curves. The analysis showed that the latency and eclipse periods of VALLPKK3 (Figure 4.4), VHLPKM4 (Figure 4.5) and VPLPKK5 (Figure 4.6) were 48 and 36 minutes, 60 and 36 minutes and, 36 and 24 minutes, respectively. The latent period of VHLPKM4 was longer compared to VALLPKK3 and VPLPKK5. Meanwhile, the eclipse period of VALLPKK3 and VHLPKM4 was similar, while, the eclipse period of VPLPKK5 were shorter than those two isolates. The VALLPKK3, VHLPKM4 and VPLPKK5 showed a burst size of ~174, ~52 and ~180 phage per infected cell, respectively, at the 28 °C. Figure 4.4: One step growth curve of VALLPKK3 infected with Vibrio alginolyticus ATCC ® 17749TM at MOI of 0.001. The number of PFU per infected cell in untreated culture () and chloroform-treated culture () are also shown. The burst size, latent period and eclipse are indicated as B, L and E, respectively. Figure 4.5: One step growth curve of VALLPKK3 infected with Vibrio harveyi VHJR7 at MOI of 0.001. The number of PFU per infected cell in untreated culture () and chloroform-treated culture () are also shown. The burst size, latent period and eclipse are indicated as B, L and E, respectively. Figure 4.6: One step growth curve of VPLPKK5 infected with V. parahaemolyticus VPHG1 at MOI of 0.001. The number of PFU per infected cell in untreated culture () and chloroform-treated culture () are also shown. The burst size, latent period and eclipse are indicated as B, L and E, respectively. 4.3.3  Bacteriophage Tolerance Test The activity of all phage isolates was stable at 40 °C and declined at 50 °C following heating for 60 minutes. The activity was disappeared entirely when heated at more than 60 °C for 1 hour (Figure 4.7). When compared among the isolates, the activity of VHLPKM4 were decline dramatically to less than 20 % when incubated at 50 °C. The activity of VALLPKK3 and VPLPKK5 were dropped to 80% and 40%, respectively. The activity of bacteriophages VALLPKK3, VHLPKM4 and VPLPKK5 can be measured after incubation at pH 4 to pH 9, but disappear completely at pH 2 and pH 3 (Figure 4.8). When compared among isolates, the VALLPKK3 was sensitive to wide range of pH. Almost all of the VALLPKK3 activity was drop to 20 to 40 % after 24 hours incubation. Meanwhile, the activity of VHLPKM4 was decline to 60 % at pH 4 and 5, relatively stable at pH 6 to pH 8 and decline again to less than 60 % at pH 9. However, the activity of VPLPKK5 relatively stable at wide range of pH (pH 4 to pH 9). Meanwhile, the activity of VALLPKK3, VHLPKM4 and VPLPKK5 can be detected after incubated at bile salt concentration from 5000 ppm to 9000 ppm (Figure 4.9). Among the isolates, VALLPKK3 was more sensitive to the bile compared to VHLPKM4 and VPLPKK5. Figure 4.7:The temperature stability of VALLPKK3, VHLPKM4 and VPLPKK5. All isolates were incubated at various range of temperature (40 °C, 50 °C, 60 °C, 70 °C, 80 °C, 90 °C and 100 °C) for 1 hour. Data are the means from three independent experiments + SD. Figure 4.8:The temperature stability of VALLPKK3, VHLPKM4 and VPLPKK5. All isolates were incubated at various range of pH (2, 3, 4, 5, 6, 7, 8 and 9) for 24 hours. Data are the means from three independent experiments + SD. Figure 4.9:The bile salt stability of VALLPKK3, VHLPKM4 and VPLPKK5. All isolates were incubated at various range of bile salt concentration (5000, 6000, 7000, 8000 and 9000 ppm) for 24 hours. Data are the means from three independent experiments + SD. 4.4  Discussion The phage adsorption of VALLPKK3 and VHLPKM4 was fast (more than 80% after 10 minutes) compared to Vibrio phage PW2 (60% after 10 minutes) (Phumkhachorn and Rattanachaikunsopon, 2010). Meanwhile, the adsorption of VPLPKK5 was comparable to PW2. This might due to both phages were belonged to same family (Sipboviridae). However, the phage adsorption was reported dependent on various condition. According to Binetti et al. (2002), the phage adsorption was shown to be affected by the presence of ion calcium, physiological state of the cell, pH and temperature. The one step growth is a method to assess the life cycle of the phage (Middleboe et al., (2010). The latent period was the time from adsorption to the release of new progeny from host cell, and the burst size was the number of new virus particles liberated from a single bacterial cell (Bao et al., 2011). When compared to other vibriophage infecting same host species, there were difference in term of the burst size of the phages Ñ„As51 and Ñ„A318 (Liu et al., 2014). The V. alginolyticus phage VALLPKK3 showed high burst size compared to those two (72 and 10 PFU/infected cell). Similar finding with VPLPKK5. This V. parahaemolyticus phage was different to other V. parahaemolyticus phage VP-2 (15 PFU per infected cell) (Silva et al., 2014) where it showed bigger burst size (180 PFU per infected cell). Meanwhile, the VHLPKM4 showed different finding. This study showed smaller burst size and longer latent period compared to previous report on V. harveyi phages цžH17-7b and Ñ„H17-8b (Okano et al., 2007). They reported that where the latent period and burst size of Ñ„H17-7b and Ñ„H17-8b were 35 minutes and 100 particles, and 40 minutes and 170 particles, respectively. Thus, the findings showed that the life cycle of each phage isolates was different from each other. However, the significant of the differences was unclear since the dissimilarity was influenced by the host, medium, temperature and its own growth rate (Carey-Smith et al., 2006). In this study, the bacteriophages VALLPKK3 and VPLPKK5 showed a short period of latent period and large burst size. The shorter latent period and large burst size showed that the bacteriophages replicated more quickly and the new virus particle release more efficiently (Bao et al., 2011). This characteristic showed good candidacy of phage therapy (Silva et al., 2014). Finally, both adsorption and one step growth of phage are important to determine the phage fitness (Wang, 2006) since the phage fitness would determine the efficacy of the phage therapy (Lindberg et al., 2014). The stability in various stress condition were useful for the application of bacteriophage to inhibit the target bacteria (Lee et al., 2014; Krasowska et al., in press). In this study, the resistance to heat, pH and bile was investigated to determine the efficacy of those phages for biocontrol of V. alginolyticus, V. harveyi and V. parahaemolyticus infections. Phage which can withstand various environmental stress may be useful for the application in aquaculture (Phumkhachorn and Rattanachaikunsopon, 2010). The temperature is a important factor that affects bacteriophage survivability (Olson et al., 2004). It plays important roles in the bacteriophage attachment, penetration and multiplication (JoÅ„czyk et al., 2011). In this study, the result showed that all phages were stable at 40 °C. However, the viability was reduced after one hour incubation at 50 °C. All phage were completely inactivated in temperature over 60 °C. The phage in this study showed that they are sensitive to high temperature. This findings was different to the findings by Phumkhachorn and Rattanachaikunsopon (2010) where the phage can withstand high temperature. However, in the natural environment, the temperature usually fluctuated at the range of 28 to 32 °C (Albert and Ransangan, 2013). Since the isolates in this study were stable at the temperature up to 40 °C, the isolates would survive when release to natural environment. Nevertheless, the period of viability of these isolates after release to natural environment was unknown. In the natural environment, the phage was also facing the other stress factor such as pH. According to Krasowska et al. (in press), the acidity and alkalinity of environment are other important factors influencing phage stability. It was also reported that low pH influences phage aggregation and reduce their adsorption on bacteria cell (Langlet et al., 2007). Therefore, it was important to access the stability of the current phage isolates in different pH. The VALLPKK3 and VHLPKM4 showed resistance to acid (pH 4) and alkaline (pH 9). This showed that the member of Myoviridae family stable at acid and alkaline condition (Krasowska et al., in press). Similar to the other isolates, VPLPKK5 was also showed resistance to acid and alkaline condition. This is similar to the finding by Lasobras et al. (60) where the member of family Siphoviridae were most resistant to adverse conditions. However, this finding was different to phage ARà ¯Ã¢â€š ¬Ã‚ , a member of Siphoviridae, which is only a ctive in a narrow pH range (Krasowska et al., in press). The result of the phage tolerance to pH indicated that they were tolerant to wide range of pH. In aquaculture, oral administration was the most practical delivery method for immunization (Yasumoto et al., 2006) due to low cost and less stress to fish (Pal et al., 2009). However, the viability of orally administered phage might be rapidly reduced the presence other digestive compounds such as bile (Joerger et al., 2003). In this study, the phage isolates were exposed to various concentration of bile concentrations and result showed that the phage were still survived after incubation. However, there were reduction on the viability of the phage isolates which might showed the adverse effect of bile. With the addition of pH and other enzymes, the phage might not persist for long time in gut environment (Ma et al., 2008). 4.5  Conclusion In summary, the VALLPKK3, VHLPKM4 and VPLPKK5 were characterized by the growth and tolerance. The life cycle of the current isolates might be different when conducted different time and with different media. Therefore, the optimization was required for optimum phage multiplication which generally required for large scale production. This optimization was also contributed to the development of phage therapy. All phages are inactivated at high temperature but showed stability at temperature 40 °C. They are also stable at wide range of pH but not low pH. But, they could tolerate normal fish bile content. However, the study need to be conducted to collect the information of the period of phage survival in fish body. This information would be beneficial for the phage administration of disease treatment.