Sunday, January 26, 2020

Viable But Non Culturable Bacteria Biology Essay

Viable But Non Culturable Bacteria Biology Essay Viable but Non-Culturable state is a unique survival strategy of many bacteria in environment in response to adverse environmental conditions. VBNC bacteria cant be cultured on routine microbiological media but they remain viable and retain their virulent capacity too. VBNC bacteria can be resuscitated when provided with appropriate conditions. A good number of bacteria including many human pathogens have been reported to enter VBNC state. Though, there was disputes on the existence of VBNC in past, extensive molecular studies have resolved most of them and VBNC has been accepted as distinct survival state by all. VBNC bacteria are considered as threats to public health and food safety due to their non-detectability and virulence as food and water have been reported to be contaminated with pathogens at VBNC state though conventional methods declared them as safe and clean. A number of outbreaks have also been reported where VBNC bacteria has been implicated as causative agent. Furthe r molecular and combinatorial research in conjunction with predictive modeling are needed to elucidate the mechanisms and to identify the critical points to tackle the threat posed by VBNC bacteria with regard to public health and food safety. Key words: VBNC, Pathogen, public health, Food safety, Detection Introduction The cells that form colony in specific media are the culturable cells. Viable means metabolically or physiologically active. So the cells those are metabolically or physiologically active but cant be cultured on specific media are the viable but non-culturable cells (VBNC) (Bogosian Bourneuf, 2001). Most microorganisms growing in nature have yet to be cultured in the laboratory. In fact less than 1% of the microorganisms in natural water and soil samples are cultured in viable count procedures (Barcian Arana, 2009). In 1982, Prof. Rita Colwell and co-workers introduced the term Viable But Non-Culturable Bacterial Cells (VBNC) to distinguish particular cells that could not form colonies on solid media but obtained metabolic activity and the ability to elongate after the administration of nutrients (Xu et al., 1982). According to Oliver (1995), VBNC can be defined as a metabolically active bacterial cell that crossed a threshold in this way, for known or unknown reasons and become unable to multiply in or on a medium normally supporting its growth. Most of the bacteria that enter VBNC state are gram negative species belonging to the gamma subclass of the Proteobacteria branch, except for Rhizobium, Agrobacterium and Helicobacter-Campylobacter species (Oliver, 2000). History Debra Bashford and colleagues announced that they had recovered Vibrio cholerae from streams and drainage ditches, including sites with negligible chance of sewage contamination. Around the same lime, Rita Colwell was also finding Vibrio cholerae in Maryland. She and her coworkers showed that both this bacterium and E. coli, incubated in artificial sea water remained viable but lost the capacity to form colonies on culture media (Colwell Grimes, 2000). Soon Salmonella enteritidis, Shigella sonnie and Legionella pneumophila joined the list of organisms known to be capable of entering a state in which they failed to show up on nutrient agar yet took up substrates and signaled in other ways that they were certainly not dead. The use of laboratory media to recover and enumerate bacteria and lo link them with or absolve them from pathological and other activities became obsolete by the new discoveries and a term VBNC (viable but non-culturable) came (McDougald et al., 1998). VBNC Microorganisms that do not grow in culture methods, but which are still metabolically active and capable of causing infections in animals and plants are said to be in VBNC state. The conditions for these organisms to resume growth are not being met using the normal laboratory culture conditions (Yamamoto, 2000). Bacteria that have been semi-starved will immediately resume growth when provided with the appropriate nutrients and conditions. Viable but non-culturable cells will not resume growth even when nutrients are provided (NystrÃÆ'-m, 2001). VBNC cells exhibit active metabolism in the form of respiration or fermentation, incorporate radioactive substrates, and have active protein synthesis but cannot be cultured or grown on conventional laboratory media. They have been detected by observing discrepancies between plate count enumeration of bacterial population and direct staining and microscopic counts (Sachidanandham Gin, 2009). These cells may be of particular problems in the e nvironment if they are pathogens, for example, viable but non-culturable cells of Vibrio cholerae, Enteropathogenic E. coli, Legionella pneumophila and various other bacteria have been shown to regain culturability after they have entered the intestinal tracts of animals (Colwell et al., 1996). The VBNC state is defined as a state of dormancy triggered by environmental harsh conditions, such as nutrient starvation (Cook Bolster, 2007), temperature (Besnard et al., 2002), osmotic stress (Asakura et al., 2008), oxygen availability (Kana et al., 2008), several food preservatives (Quirà ³s et al., 2009), heavy metals (Ghezzi Steck, 1999), exposure to white light (Gourmelon et al., 1994) and decontaminating processes, as pasteurization of milk (Gunasekera et al., 2002) and chlorination of wastewater (Oliver, 2005). VBNC state is believed to be a unique survival strategy of bacteria in response to environmental stresses (Oliver, 2010). It is also considered as an important reservoir of many human pathogens in the environment (Lleo et al., 2007). VBNC state has been a matter of dispute for ling since its inception, due to the difficulty of differentiation of VBNC cells dormant cells through resuscitation phenotypic studies, recent molecular studies, data of which supported the existence of VBNC state, the dispute has mostly been put to rest (Barer and Harwood, 1999). VBNC Pathogens Following list includes but not limited to pathogenic bacteria that can enter VBNC state (Oliver, 2010)- Aeromonas hydrophila, Agrobacterium tumefaciens, Burkholderia cepacia, Campylobacter jejuni, Enterobacter aerogenes, Enterobacter cloacae, Enterococcus faecalis, Escherichia coli (including EHEC), Helicobacter pylori, Klebsiella pneumoniae, Legionella pneumophila, Listeria monocytogenes, Mycobacterium tuberculosis, Pseudomonas aeruginosa, Salmonella typhi, Salmonella typhimurium, Shigella dysenteriae, Shigella flexneri, Shigella sonnei, Streptococcus faecalis, Vibrio alginolyticus, Vibrio cholerae, Vibrio harveyi, Vibrio parahaemolyticus, Vibrio vulnificus (types 1 and 2) Characteristics of Bacteria in VBNC state 1. Maintain apparent cell integrity; 2. possession of some form of measurable cellular activity (Lai et al., 2009); 3. possess apparent capacity to regain culturability (Anuchin et al., 2009); 4. respond to external stimulus by specific gene expression (kell et al., 1998); 5. low metabolic activity (oliver, 2005); 6. Exhibit dwarfing (Costa et al., 1999); 7. reduced nutrient transport; 8. High ATP level and high membrane potential (Signoretto et al., 2000); 9. extensive modifications in cytoplasmic membrane fatty acid compositions (Day Oliver, 2004); 10. Changes in cell wall peptidoglycan such as increasing cross linking, increasing muropeptides bearing covalently bound lipoprotein shortening of average length of glycan strands (signoretto et al., 2002); 11. Higher autolytic capability than exponentially growing cells; 12. Plasmids are retained; 13. changed antibiotic sensitivity as metabolic activity is lower, most bacteria at VBNC state demonstrate high antibiotic resistance (Ol iver, 2010); 14. Changes in outer-membrane protein profile (Muela et al., 2008); 15. Continuous gene expression (Maalej et al., 2004) etc. Conditions stimulating VBNC state In the environment, bacterial cells can enter VBNC state may be due to- 1. Lack of nutrients; 2. Lack of temperatures; 3. High pressure; 4. Sharp changes in pH or salinity (Cunningham et al., 2009); 5.damage to or lack of an essential cellular component; 6. DNA damage; 7. activation of lysogenic phages or suicide genes such as sok/hak or autolysins (Aizenman et al., 1996); 8. Nutrient starvation; 9. incubation outside the normal temperature range of growth; 10. elevated or lower osmotic concentrations; 11. oxygen concentrations (Mascher et al., 2000); 12. food preservatives; 13. Heavy metals (Del Campo et al., 2009); 14. exposure to white light; 15. pasteurization of milk (Gunasekara et al., 2002); 16. chlorination of wastewater (Oliver, 2005) etc. Public health significance of VBNC Though virulence of bacteria in VBNC state is still not very clear, many believed that pathogens in VBNC state are unable to induce infection/disease but still retain their virulent properties has potential to cause disease infection following resuscitation and resume of active metabolic state, which occurs when they pass through host animal (Baffone et al., 2003). The VBNC state appears to be the common to many bacteria especially those which have aquatic habitats, and may represent a mechanism to survive adverse environmental factors as temperature, salinity etc. or have a means of inducing cross protection against other adverse factors (Du et al., 2007). Among these bacteria entering this state are many significant human pathogens and indicator bacteria of these pathogens; such cells may represent a public health hazard and may be a factor in human health and/or disease (Rivers Steck, 2001). Even today, it is still not possible to cultivate most bacterial species directly from the environmental samples or after exposure of previously culturable cells to environmental conditions unfavorable for growth and multiplication in vitro. The passage of VBNC through an appropriate animal host will induce return of culturability. Even these VBNC bacteria retain their pathogenicity and may trigger life in vivo and thus cause severe disease (Sardessai, 2005). Under normal condition it is not possible to culture or detect VBNC. Many diagnostic laboratory set up does not have sufficient molecular facilities to detect VBNC. In case of food and water quality control test, such VBNC may not be detected. Even some indicator of some pathogenic bacteria undergoes VBNC state and may remain undetected (Signoretto et al., 2004). Upon consuming such food or after drinking such water, one may be infected by those VBNC that can trigger life as well as pathogenicity (Adams et al., 2003). Thus, environmental and clinical samples no longer can be considered free from pathogens if culturing yields negative results. For the general public, the presence of VBNC in water and food may be related to low-grade infections or so called aseptic infection. For example, Vibrio cholerae O1 in the surface water remain as non-culturable state. These water sources are used for domestic purpose regularly and posed a risk of infection (Edwards, 2000). When conditions are not favorable for growth then it transforms to the non-culturable state in association with crustacean copepods. Persistence of Vibrio cholerae in water in the VBNC state is an important public health factor, since detection will not be successful if only conventional cultural methods are used (Barer et al., 1993). Similarly, Shigella can undergo VBNC state in water but become a threat when enter in human body. Thus it is important to recognize that non-culturable bacteria are capable of producing diseases. The first evidence of pathogenicity of non-culturable cells was the demonstration of fluid accumulation in rabbit ileal loop assay (RICA) by VBNC Vibrio cholerae O1, followed by human volunteer experiments (Amel et al., 2008). VBNC E. coli non-culturable cells were re-isolated after passage through rabbit ileal loops 4 days post inoculation and chick embryos died when injected with non-culturable cells of Legionella pneumophila, led to the conclusion that VBNC pathogens remain potentially pathogenic. So, VBNC has a huge significance in public health care (Cappelier et al., 2007). Reports indicate that many potentially harmful bacteria survive treatment and persist in processed food, pasteurized milk, potable water and in the environment (Colwell et al., 2000). Many evidences suggested that recurrent urinary tract infections in many individuals are caused by uropathogenic E. coli cells which remain in VBNC state (Anderson et al., 2004) thus resistant to antibiotic treatment cause reinfection when resuscitate back to active metabolic state (Steck, 2001; Mulvey et al., 2001). Studies also showed that uropathogenic E. coli retain enteropathogenicity at VBNC state through continued production of enterotoxin (Pommepuy et al., 1996). Nilsson et al. (2002) showed that VBNC Helicobacter pylori cells can express virulence factors such as cagA, vacA and vreA. All these above evidence proved that many deathly pathogenic strains not only enter but also persist survive in VBNC state in environment most of them remain infectious as well. VBNC state of foodborne bacteria- a challenge in food safety Many evidences suggested presence of VBNC bacteria in food (Ordax et al., 2009). For example, in stored wine, acetic acid and lactic acid bacteria entered VBNC state as consequence of lack of oxygen and presence of sulphites, respectfully (Millet and Lonvaud-Funel, 2000). Food and its surrounding environment is a complex system, in which physic-chemical characterisitcs (pH, aw, chemical composition) and environmental factors (storage temperature and time, decontamination treatments, packaging under modified atmosphere) act simultaneously on contaminating bacteria (Sun et al., 2008). For example, it has been demonstrated that refrigerated pasteurized grapefruit juice induced VBNC state in E. coli O157:H7 and S. typhimurium within 24 hours of incubation (Nicolo et al., 2011). Again, Gunasekera et al. (2002) reported that in pasteurized milk which have undergone thermal treatment, contaminating bacteria such as E. coli and Pseudomonas putida enter into VBNC state but retained transcription and translation machineries. Several foodborne outbreaks has been reported in Japan, where pathogen such as Salmonella enterica subsp. enterica (Asakura et al., 2002) and E. coli O157 (Makino et al., 2000) in food in VBNC state were responsible for the outbreak. Therefore, the role of food and treatment for food preservation in induction of VBNC state has to be elucidated. Predicitve models offered by biomathematics and bioinformatics would be very helpful tools, in order to evaluate the possibility that, under certain conditions, pathogen bacteria contaminating a tipology of food may enter the VBNC state (Fakruddin et al., 2012). Methods of detection of VBNC bacteria 1. Bright Field Microscopy with Nalidixic acid For detection of Bright-field or light microscopic is usually used. Cell division inhibitor such as nalidixic acid (20-40 mg/L) is used to stop cell division. After such treatment the viable cells, which actively growing, will be appeared as lengthen and the non-viable/ metabolically inactive cell will remain as it is. The cells are then observed under microscope. Viable cells will be seen as elongated whereas VBNC/ dormant cells will be seen as oval and large. 2. Fluorescent Microscopy Various fluorescent staining procedures are used in combination with other procedure to determine VBNC organisms. Frequently used stains are Acridine orange, 4,6- Diamino-2-phenyl indole (DAPI), Fluorescein isothiocyanante (FITC), Indophenyl-nitrophenyl-phenyl tetrazolium chloride (INT), 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) etc (Villarino et al., 2000). Table: Fluorescent dyes used for detection of VBNC bacteria Dye Mechanism Reaction Acridine orange Acridine orange stain cells depending on the ratio of DNA to protein in the cells Actively reproducing cells appear green but slow-grower or non-reproducing cells at time of staining appear orange Di-amino-phenyl-indole (DAPI) Living cells look green under fluorescent microscope Indophenyl-nitrophenyl-phenyl tetrazolium chloride (INT) INT deposit red dye in cells that have active dehydrogenase and thus determine which of the observed cells are metabolically active INT react with dehydrogenase enzyme to produce formazone and red color, thus living cells appear red. Nalidixic acid (NA) Lengthen metabolically active cells, VBNC cells remain as it is Cells that are dividing appear to be longer in size than VBNC Fluorescein isothiocyanante (FITC) Enzyme activity in living cell FITC stain living cells violet or blue In recent years, a new differential staining assay, the BacLight ® Live/Dead assay, has been developed. The assay allows to simultaneously count total and viable (metabolically active) cells, by using two nucleic acid stains, that is green-fluorescent SYTO ® 9 stain and red-fluorescent propidium iodide stain. These stains differ in their ability to penetrate intact cell membranes. When used alone, SYTO ® 9 stain labels both live and dead bacteria. In contrast, propidium iodide penetrates only bacteria with damaged membranes, reducing SYTO ® 9 fluorescence when both dyes are present. Thus, live bacteria with intact membranes fluoresce green, while dead bacteria with damaged membranes fluoresce red (Rowan, 2011). 3. Gene probe / oligonucleotide probe / hybridization: Molecular analysis can also be used to study non-culturable microorganisms in nature. Oligonucleotide probes of l8-20 nucleotides are proving most useful because they hybridize rapidly to specific DNA sequences of target organisms. These gene probes can reveal closely related organisms or organisms with similar functional capabilities. Especially useful for the analyses of rRNA that demonstrate the presence of diverse microbial populations whose phylogenetic relationships can be ascertained by comparison with rRNA sequences from previously described microorganisms (Josephson et al., 1993). Blotting: There are different types of blotting such as colony blot, slot blot, dot blot and southern blot. The principle of blotting is the use of radio- or non-radioactive or fluorescence labeled probe (DNA/RNA/Antibody) to detect VBNC cells directly from the environmental samples. Fluorescent in situ Hybridization (FISH): In situ hybridization is an alternative format for hybridization probes in which fluorescence labeled DNA or RNA probes are hybridized with target nucleic acids in whole, permeabilized cells. The application of this method to the detection of single microbial cells by using rRNA-targeted probes in combination with epifluorescent microscopy has been developed. This is done through selective targeting of regions of rRNA, which consist of conserved and variable nucleotide regions. By choosing the appropriate rRNA probe sequence, FISH can be used to detect all bacterial cells (a universal probe) or a single population of cells (a strain specific probe) of VBNC. It has lower sensitivity and cannot distinguish live and dead cells. 4. Molecular techniques: Hybridization probes and DNA/RNA amplification: Hybridization probes are nucleic acids (DNA/RNA) which have been (a) chemically or radioactively labeled and are used to detect complementary target DNA/RNA. Hybridization assay DNA/ RNA probes form a stable double stranded structure with target nucleic acid via H-bonding between complementary bases. Amplification of targets: DNA Based methods: Specific amplification of DNA targets in bulk DNA extracts from environmental and clinical samples permits detection of specific organisms or groups of related organisms without the need to cultivate them. DNA recovery procedures do not discriminate between culturable and non-culturable forms of the target organisms- all cells with intact amplification targets will be detected. Confocal laser microscopy in combination with fluorescence-based hybridization assays, also provide a more sensitive method for detecting and identifying VBNC organisms. RNA based methods: Due to the failure of distinguishing between dead or live cells by DNA-based methods, the mRNA level may be a valuable estimate of gene expression and/or cell viability under different conditions (Lleo et al., 2000). RT-PCR: RT-PCR (Reverse Transcriptase PCR) can distinguish between Live and dead cell. This is possible because this is an mRNA based method and mRNA is short lived (half-life less than 1 minute), mRNA is only present in metabolically active cells, not found in nature after the cell death. By this method we can study community relationship and can also detect non-culturable but active or live cells. DNase enzyme is used during the isolation of RNA from environmental samples. Reverse transcriptase and random primers are added to the reaction mixture and the RNA in the sample (both RNA and rRNA) is transcribed into DNA. PCR is then use to amplify the specific sequence of interest (Pai et al., 2000). Is the concept of VBNC is a misnomer? By extending the concept of bacterial self-suicide scientists tried to explain what happens when cells are exposed to chemical and physical injury (Forsman et al., 2000). Thus VBNC organisms came alongside with those, which do not grow in ordinary media but which do grow when offered selective or enrichment media. They said, Such cells are not un-culturable they wrote We are simply failing to provide appropriate conditions to support culture (Sinton, 2006). The reasons, which made the term, VBNC a misnomer are as follow: 1. Bacteria that have been semi-starved will immediately resume growth when provided with the appropriate nutrients and conditions. But viable but non-culturable bacteria do not resume growth when nutrients are provided. Evidence suggests that these cells have become too starved to be able to grow on rich medium initially; this phenomenon was observed over three decades ago and was known then as substrate accelerated death (Heim et al., 2002). It was found that substrate that normally supported growth of certain Gram-negative bacteria hastened their death when introduced into starved populations of these bacteria. These substrates were considered lethal substrates. These starved cells require an adjustment period to allow phenotypic adaptation back to the normal growth state (Epstein, 2009). A sudden shift to nutrient rich agar creates a metabolic imbalance that results in the formation of super-oxide and free radicals, causing DNA damage that can result in cell death (Barer Harwood , 1997). 2. There is yet no complete and perfect media to isolate arid culture all the organisms from environment. 3. Cells are usually-injured or stressed or starved condition in natural environment. So complete system has been devised to enrich or resuscitated the VBNC cells. Culture condition that can be applied in laboratory is not sufficient to recover all microorganisms i.e. yet it is not possible to provide or stimulate exact environmental conditions in the laboratory. Conclusion Regardless of the role that the VBNC state plays, it is clear that a large number of non-spore-forming bacteria, most notably a large number of human pathogens, are capable of entering this state, maintaining cellular structure and biology and continuing significant gene expression while otherwise non-culturable by standard laboratory methods. That they can exit from this state, and become culturable again, is also undeniable. Finally, it can no longer be questioned that the VBNC state plays a critical role in the survival of important human (and other) pathogens, and possibly in their ability to produce disease.

Saturday, January 18, 2020

Vacation Destination Preferencees

CHAPTER 1: The Problem and its Background Introduction Vacation destinations are places where someone can go or take a visit to free from work and away from home. These places may be beaches, landmarks, forests, and others in type. Visiting these places requires no age but still, it is better that the place is suitable or appropriate to anyone who visit it. Summer is approaching. Many families or people will surely find a place for their vacation, a place where they can spend their times together. Somewhere which is stress-free and will give them the peaceful feeling they want.Wherever place it is, they’ll definitely need some money to enjoy and in able to reach their desired destination. The Philippines contains many islands that will definitely give the qualities people want from vacation places, especially the amenities that can be found on it. The purpose of this research is to identify the preferred vacation destinations of the constituents and to know their plans on visi ting the places. Moreover, the researcher is interested in taking a more serious look into the most preferable vacation destinations for the residents or constituents of a particular barangay.Statement of the Problem This study was undertaken to find out the most preferable vacation destinations of the constituents of Barangay 502 Sampaloc, Manila and their plans regarding to their visit on the place. Specifically, this study sought answers to the following research questions: 1. What type of place do the constituents prefer for a vacation? 2. Whom do the constituents like to go with for a vacation? 3. How much money do the constituents willing to spend for a vacation? 4. What particular vacation destination/s here in the Philippines do the constituents prefer for a vacation? . 1 In Luzon? 5. 2 In Visayas? 5. 3 In Mindanao? Scope and Limitations This study looked into the preferable vacation destinations here in the Philippines, specifically in Luzon, Visayas, and Mindanao and their plans regarding to their visit on the place. Only 50 constituents of Barangay 502 Sampaloc, Manila were involved. No attempt was made to compare and correlate these vacation destinations with any other places in any other countries. Definition of Terms Preference the choice of one thing or person over another (Webster’s Student Dictionary)Vacationa time set aside from work (Webster’s Student Dictionary) Amenityagreeableness; pleasantness (Webster’s Student Dictionary) Constituentserving to form or compose as a necessary part (Webster’s Student Dictionary) Relaxto rest (Webster’s Student Dictionary) Research Methodology Research Design The Descriptive method was used to achieve the objectives of this study. This method seeks to describe a present existing condition, which in this study were the preferable vacation destinations of the constituents of a particular barangay and their plans regarding on the visit to the place.Respondents The target pop ulation consisted of 50 constituents of Barangay 502 Sampaloc, Manila, 32 males and 18 females to be specific. Research Instrument The researcher prepared a checklist which was used for the survey. Data-Gathering Procedure The data were drawn from the 50 constituents of Barangay 502 Sampaloc, Manila who were given a checklist-type of questionnaires for the survey. The researcher gave the checklist individually and it lasted for 3 days. Statistical Treatment Individual responses were tallied before they were put in tables ready for the statistical treatment.Percentage was utilized to quantify the data gathered for subproblems 1, 2, 3, 4. 1, 4. 2 and 4. 3. The formula for percentage is: P= f/N x 100 Where: P= percentage f= frequency N= total number of respondents CHAPTER 2: Presentation, Analysis, and Interpretation of Data Gathered This chapter presents the data gathered regarding the most preferred vacation destinations of the constituents of a particular barangay and their plans re garding on visit to the place, and proceeds to the analysis and interpretation of said data. Subproblem 1What type of place do the constituents prefer for a vacation?Table 1 Type of Place of the Constituents for a Vacation Type of Place| Male| %| Female| %| Total| %| Rank| a. Beach| 24| 75%| 13| 73%| 37| 74%| 1| b. Landmark| 2| 6%| 2| 12%| 4| 8%| 3| c. Amusement Park| 2| 6%| 3| 17%| 5| 10%| 2| d. Mountain| 2| 6%| 2| 12%| 4| 8%| 3| e. Forest| 2| 6%| 3| 17%| 5| 10%| 2| Table 1 presents the preferred type of place of the constituents for a vacation. Among the male constituents, 24 or 75% preferred to go to Beach for a vacation, while the Landmark, Amusement Park, Mountain, and Forest each got a 2 or 6% from the male constituents.Among the female constituents, 13 or 73% preferred to go to beach for a vacation, Amusement Park and Forest each got 3 or 17%, and both Landmark and Mountain got 2 or 12% from the female constituents. As a whole, 74% of the constituents preferred the Beach as t heir type of place for vacation, 10% for Amusement Park and Forest, and 8% for Landmark and Mountain. Subproblem 2With whom do the constituents like to go for a vacation? Table 2 Companion of the Constituents for a Vacation Companion| Male| %| Female| %| Total| %| Rank| a. Family| 18| 56%| 10| 56%| 28| 56%| 1| . Friends| 6| 19%| 3| 17%| 9| 18%| 3| c. Special Friend| 5| 16%| 5| 28%| 10| 20%| 2| d. Alone| 3| 9%| 2| 12%| 5| 10%| 4| Table 2 presents the preferred companion of the constituents for a vacation. Among the male constituents, 18 or 56% like to go with their families for a vacation, 6 or 19% with their friends, 5 or 16% with a special friend and 3 or 9% like to go by themselves. Among the female constituents, 10 or 56% want to go with their families for a vacation, 5 or 28% with a special friend, 3 or 17% with their friends and 2 or 12% like to go alone.As a whole, 56% of the constituents like to go with their families for a vacation, 20% with their special friends and 10% wan t to go by themselves. Subproblem 3How much money do the constituents willing to spend for a vacation? Table 3 Amount Money of the Constituents to Spend for a Vacation Amount| Male| %| Female| %| Total| %| Rank| a. P1,000-P3,500| 6| 19%| 4| 23%| 10| 20%| 2| b. P3,500-P7,000| 2| 6%| 4| 23%| 6| 12%| 3| c. P7,000-P12,000| 8| 25%| 2| 12%| 10| 20%| 2| d. P12,000-P20,000| 15| 47%| 7| 39%| 22| 44%| 1| Table 3 presents the amount that the constituents willing to spend for a vacation.Among the male constituents, 15 or 47% were willing to spend P12,000-P20,000 for a vacation, 8 or 25% for an amount of P7,000-P12,000, 6 or 19% for P1,000-P3,500, and 2 or 6% were willing to spend P3,500-P7,000. Among the female constituents, 7 or 39% were willing to spend P12,000-P20,000, while an amount of P1,000-P3,500 and P3,500-P7,000 each got 4 or 23% from the female constituents and 2 or 12% for P7,000-P12,000. As a whole, 44% were willing to spend P12,000-P20,000 for a vacation, 20% for an amount of P1,0 00-P3,500 and P7,000-P12,000, and 12% for P3,500-P7,000.Subproblem 4What vacation destination/s here in the Philippines do the constituents prefer for a vacation? Subproblem 4. 1 in Luzon? Table 4. 1 Vacation Destinations of the Constituents in Luzon Luzon| Male| %| Female| %| Total| %| Rank| Baguio City| 18| 56%| 8| 45%| 26| 52%| 1| Puerto Galera| 10| 31%| 2| 12%| 12| 24%| 4| Puerto Princesa| 12| 38%| 4| 23%| 16| 32%| 2| Tagaytay| 7| 22%| 5| 28%| 12| 24%| 4| Makati City| 2| 6%| 0| 0%| 2| 4%| 8| Caramoan Peninsula| 7| 22%| 1| 6%| 8| 16%| 5| Camarines Sur| 11| 34%| 4| 23%| 15| 30%| 3| Subic| 11| 34%| 5| 28%| 16| 32%| 2| La Union| 2| 6%| 3| 17%| 5| 10%| 6|Banaue| 1| 3%| 3| 17%| 4| 8%| 7| Table 4. 1 presents the preferred vacation destinations in Luzon of the constituents for a vacation. Among the male constituents, 18 or 56% want to go to Baguio City, 12 or 38% to Camarines Sur and Subic, 10 or 31% to Puerto Galera, 7 or 22% to Tagaytay and Caramoan Peninsula, 2 or 6% to Makati City a nd La Union, and only 1 or 3% want to go to Banaue. Among the female constituents, 8 or 45% want to go to Baguio City, 5 or 28% to Tagaytay and Subic, 4 or 23% to Puerto Princesa and Camarines Sur, 3 or 17% to La Union and Banaue, 2 or 12% to Puerto Galera and 1 or 6% to Caramoan Peninsula.As a whole, 52% of the constituents want to go to Baguio City for a vacation, 32% to Puerto Princesa and Subic, 30% to Camarines Sur, 24% to Puerto Galera and Tagaytay, 16% to Caramoan Peninsula, 10% to La Union, 8% to Banaue and only 4% want to go to Makati City for vacation. Subproblem 4. 2in Visayas? Table 4. 2 Vacation Destinations of the Constituents in Visayas Visayas| Male| %| Female| %| Total| %| Rank | Aklan| 3| 9%| 2| 12%| 5| 10%| 4| Boracay| 25| 78%| 8| 45%| 33| 66%| 1| Bohol| 5| 16%| 7| 39%| 12| 24%| 2| Guimaras Island| 4| 13%| 1| 6%| 5| 10%| 4|Cebu City| 7| 22%| 5| 28%| 12| 24%| 2| Antique| 3| 9%| 1| 6%| 4| 8%| 5| Bacolod City| 5| 16%| 1| 6%| 6| 12%| 3| Valencia, Negros Oriental| 0| 1 0%| 0| 0%| 0| 0%| 8| Tacloban City| 1| 3%| 0| 0%| 1| 2%| 7| Iloilo City| 2| 6%| 0| 0%| 2| 4%| 6| Table 4. 2 presents the preferred vacation destinations of the constituents in Visayas. Among the male constituents, 25 or 78% want to go to Boracay, 7 or 22% to Cebu City, 5 or 16% to Bohol and Bacolod City, 4 or 13% to Guimaras Island, 3 or 9% to Aklan and Antique, 2 or 6% to Iloilo City, and 1 or 3% want to go to Tacloban City.Among the female constituents, 8 or 45% want to go to Boracay, 7 or 39% to Bohol, 5 or 28% to Cebu City, 2 or 12% to Aklan, and 1 or 6% want to go to Guimaras Island, Antique, and Bacolod City. As a whole, 66% of the constituents want to go to Boracay for a vacation, 24% to Bohol and Cebu City, 12% to Bacolod City, 10% to Aklan and Guimaras Island, 8% to Antique, 4% to Iloilo City, and 2% to Tacloban City. Subproblem 4. 3in Mindanao? Table 4. 3 Vacation Destinations of the Constituents in MindanaoMindanao| Male| %| Female| %| Total| %| Rank| Camiguin Island| 7| 22%| 4| 23%| 11| 22%| 1| Compostella Valley| 1| 3%| 2| 12%| 3| 6%| 7| Davao| 6| 19%| 5| 28%| 11| 22%| 1| Dinagat Island| 4| 13%| 1| 6%| 5| 10%| 5| Zamboanga| 6| 19%| 1| 6%| 7| 14%| 3| Butuan City| 1| 3%| 1| 6%| 2| 4%| 8| Dipolog City| 4| 13%| 2| 12%| 6| 12%| 4| Koronadal City| 2| 6%| 2| 12%| 4| 8%| 6| Bucas Grande Island| 5| 16%| 2| 12%| 7| 14%| 3| Tawi-tawi| 6| 19%| 4| 23%| 10| 20%| 2| Table 4. 3 presents the preferred vacation destinations of the constituents in Mindanao.Among the male constituents, 7 or 22% want to go to Camiguin Island for a vacation, 6 or 19% to Davao, Tawi-tawi, and Zamboanga, 5 or 16% to Bucas Grande Islands, 4 or 13% to Dinagat Island and Dipolog City, 2 or 6% to Koronadal City and 1 or 3% to Compostella Valley and Butuan City. Among the female constituents, 5 or 28% want to go to Davao, 4 or 23% to Camiguin Island and Tawi-tawi, 2 or 12% to Compostella Valley, Dipolog City, Koronadal City, and Bucas Grande Islands, and 1 or 6% want to go to Dinagat Island, Zamboanga, and Butuan City.As a whole, 22% of the constituents want to go to Camiguin Island and Davao for a vacation, 20% to Tawi-tawi, 14% to Zamboanga and Bucas Grande Islands, 12% to Dipolog City, 10% to Dinagat Island, 8% to Koronadal City, 6% to Compostella Valley and 4% want to go to Butuan City. CHAPTER 3: Summary of Findings, Conclusions, and Recommendations The purpose of this study was to know the most preferable vacation destinations here in the Philippines mainly, in Luzon, Visayas, and Mindanao for the constituents of a particular barangay and their plans regarding on visit to the place.Using the Descriptive method, particularly the use of a checklist for the survey, the researcher gathered data from the 50 constituents of Barangay 502 Sampaloc, Manila. The data were quantified using percentage. Summary of Findings Based on the analysis and interpretation of data gathered, the findings are: Subproblem 1Type of Place * 74% wanted Beach as a type of vacation place * 10% preferred to go to Amusement Parks and Forests * 8% chose Landmarks and Mountains as vacation place Subproblem 2Companion * 56% preferred to go with family * 20% liked to go with special friend 18% wanted to go with friends * 10% just wanted to go alone Subproblem 3Amount to Spend * 44% were willing to spend P12,000-P20,000 * 20% were willing to spend an amount of P1,000-P3500 and P7,000-P12,000 * 12% were willing to spend P3,500-P7,000 Subproblem 4. 1Places in Luzon * 52% desired to go to Baguio City * 32% desired Puerto Princesa and Subic as a vacation place * 30% desired Camarines Sur as a vacation place * 24% desired Puerto Galera and Tagaytay as a vacation place * 16% desired Caramoan Peninsula as a vacation place 10% desired La Union as a vacation place * 8% desired Banaue as a vacation place * 4% desired Makati City as a vacation place Subproblem 4. 2Places in Visayas * 66% wanted to go to Boracay * 24% wanted Bohol and Cebu City as a vacation place * 12% wanted Bacolod City as a vacation place * 10% wanted Aklan and Guimaras Island as a vacation place * 8% wanted Antique as a vacation place * 4% wanted Iloilo City as a vacation place * 2% wanted Tacloban City as a vacation place Subproblem 4. 3Places in Mindanao * 22% liked to go to Camiguin Island and Davao 20% liked Tawi-tawi as a vacation place * 14% liked Zamboanga and Bucas Grande Islands as a vacation place * 12% liked Dipolog City as a vacation place * 10% liked Dinagat Island as a vacation place * 8% liked Koronadal City as a vacation place * 6% liked Compostella Valley as a vacation place * 4% liked Butuan City as a vacation place Conclusions Based on the findings, the researcher arrived at these conclusions: 1. Type of place of the constituents for a vacation Majority of the constituents prefer the Beach for the type of vacation place. 2.Companion of the constituents for a vacation Majority of the constituents are going to a vacation with their families. 3. Amount of money the constituents wi lling to spend for a vacation Majority of the constituents are willing to spend an amount of P12,000-P20,000 for a vacation. 4. Vacation destinations here in the Philippines that the constituents want to visit. 5. 1 Luzon Majority of the constituents prefer to visit Baguio City for a vacation in Luzon. 5. 2 Visayas Majority of the constituents prefer to visit Boracay for a vacation in Visayas. 5. 3 MindanaoMajority of the constituents prefer to visit Camiguin Island and Davao for a vacation in Mindanao. Recommendations Based on the conclusions, the researcher endorses some ways to make the constituents enjoy and make their vacation as a memorable and an unforgettable experience of their lives. 1. In able to enjoy the vacation of the constituents, they must set aside first their problems and make the best things they could do to the place where they are taking vacation. This will surely make everyone on the vacation stress-free and free themselves from the things bothering them. 2. C herish every moment of the vacation.These moments will surely make the constituents smile every time they remember the happenings and experiences they had on that vacation. 3. Enjoy the place. The view, the people, and the amenities that can be seen on the place. These things will definitely fulfill the enjoyment and make the vacation one of a kind. 4. Know the culture, the dialect, and the people of the place. These will give an extra-ordinary experience and additional knowledge for the constituents from the place of vacation. 5. Make the vacation as the time of loved ones. Vacation is the best time to have a quality time with the loved ones.

Thursday, January 9, 2020

Gossip, Lies and Essay Topics Romeo and Juliet

Gossip, Lies and Essay Topics Romeo and Juliet Young men and women love each regardless of the feud between their families. She's found in the early hours, declared dead, and put in the vault. Students lead busy lives and frequently forget about a coming deadline. If you're grateful for the fantastic relationships with your family and friends, tell them. The Battle Over Essay Topics Romeo and Juliet and How to Win It There's among the most renowned play, and juliet. It was an incredible theatre event that someone might never experience again! Ambiguity appears fitting as what is seen among the best romances in literary history also ranks among the fantastic tales of tragedy. Talk about the expression star crossed lovers as it is related to the play. Some people think that the play is a romance. Specifically, the play delivers an assortment of versions of masculinity. It conveys the idea that people are powerless to change their fate. What You Should Do to Find Out About Essay Topics Romeo and Juliet Before You're Left Behind Great for students who must write Romeo and Juliet essays. From the very start, we're completely aware that the story of Romeo and Juliet is going to have a tragic end. If you're looking for help, here are a few argumentative essay topics on Romeo and Juliet. Think about the amount of the association between Romeo and Juliet. Romeo also demonstrates he believes in the ability of dreams that could also foretell the future. Romeo arrives, opening Juliet's coffin to take a look at his love one final time. As a consequence, Romeo is depressed. Still, Romeo and his buddies earn their way to the feast. Non-fiction prose does prose the point to an extent which you do not have to make facts so as to compose a meaningful story. Writing a comparative essay is a chance to pick sides and analyze either side of a problem or a character. You have to be at first really clear about what you would like to write, than you need to craft a topic in line with the topic of your essay on Shakespeare. The adventures of Huckleberry is a famed book that's frequently the subject for essay assignments for the students studying literature, thus, it ought to be written in the very best way possible to make sure that you get good grades in your essay exams, this is the reason why we gave you a sample outline for Huckleberry Finn essay outline so you can secure a glimpse of the idea on how best to create huck essay outline. Consequently, comparison and discussion about both of these interpretations can be readily achieved. There are various types of sonnets. Several themes present themselves in various parts of the story that cause you to wonder what the most important argument is. Why Almost Everything You've Learned About Essay Topics Romeo and Juliet Is Wrong The morning after, he's made to leave when Juliet's mother arrives. This distinction isn't just due to how the Lion King is meant for children. Lady Capulet tells her husband that it's late, but he says that he'll stay and be sure that everything is perfectly prepared for tomorrow. Romeo wonders how they are going to get in the feast without being recognized, regardless of the simple fact they are totally wearing masks. Wonderful rating and very good reviews should tell you everything that you need to understand about this excellent writing service. It's possible to write about whatever you want, but do make sure it's an excellent artic le. You have to use three quotes to back up your answer. It's important to get the suitable quantity of info in your article. The Chronicles of Essay Topics Romeo and Juliet Writing an essay on Romeo and Juliet is a great chance to check your critiquing abilities. To Romeo there is not any other like Juliet and Shakespeare indicates that by employing celestial imagery. Romeo and Juliet is among the most well-known works made by Shakespeare. The balcony scene in Romeo and Juliet is among the most well-known scenes in all Shakespeare's plays. Successful people search for the very best in others and recognize their specific talents and strengths. Students within this category are eager to receive their hands dirty and are quick to aid others. Too a lot of people simply don't have the ideal attitude. When you're viewed as having a fantastic attitude, many more people will naturally gravitate toward you and often will be prepared to assist you somehow. Essay Topics Romeo an d Juliet: the Ultimate Convenience! No one needs to be discouraged from taking action, though there are consequences. The option of treatment is dependent on the age and general state of the individual, the size of thyroid gland, and patient preference. Examples may be that you're grateful for not having a specific life threatening disease, or that you're not homeless. Many factors can influence an individual's susceptibility to Graves' disease. Megara is a rather intriguing heroine to me, particularly for a Disney character. You may always watch them twinkle however, you cannot shine and twinkle like them. It is thought that these characters were famous for at least 400 decades. The characters are also appealing since they are simple to relate to. The 5-Minute Rule for Essay Topics Romeo and Juliet Assuming that it's clear from the context of your paper which play you're quoting from, you don't need to recognize the job by its title. After 400 decades, today's avid rea ders can clearly recognize lots of the words and expressions commonly utilised in the current speech. This list, naturally, is extremely limited, only detailing a small percent of the abundance of topics possible. The two make plans to find each other again the following day.

Wednesday, January 1, 2020

Communication Process - Free Essay Example

Sample details Pages: 5 Words: 1410 Downloads: 8 Date added: 2017/09/12 Category Advertising Essay Did you like this example? Communication can best be summarized as the transmission of a message from a sender to a receiver in an understandable manner. The importance of effective communication is immeasurable in the world of business and in personal life. From a business perspective, effective communication is an absolute must, because it commonly accounts for the difference between success and failure or profit and loss. It has become clear that effective business communication is critical to the successful operation of modern enterprise. Every business person needs to understand the fundamentals of effective communication. Currently, companies in the United States and abroad are working toward the realization of total quality management. Effective communication is the most critical component of total quality management. The manner in which individuals perceive and talk to each other at work about different issues is a major determinant of the business success. It has proven been proven that poor communication reduces quality, weakens productivity, and eventually leads to anger and a lack of trust among individuals within the organization. The communication process is the guide toward realizing effective communication. Don’t waste time! Our writers will create an original "Communication Process" essay for you Create order It is through the communication process that the sharing of a common meaning between the sender and the receiver takes place. Individuals that follow the communication process will have the opportunity to become more productive in every aspect of their profession. Effective communication leads to understanding. The communication process is made up of four key components. Those components include encoding, medium of transmission, decoding, and feedback. There are also two other factors in the process, and those two factors are present in the form of the sender and the receiver. The communication process begins with the sender and ends with the receiver. The sender is an individual, group, or organization who initiates the communication. This source is initially responsible for the success of the message. The senders experiences, attitudes, knowledge, skill, perceptions, and culture influence the message. The written words, spoken words, and nonverbal language selected are paramount in ensuring the receiver interprets the message as intended by the sender (Burnett Dollar, 1989). All communication begins with the sender. The first step the sender is faced with involves the encoding process. In order to convey meaning, the sender must begin encoding, which means translating information into a message in the form of symbols that represent ideas or concepts. This process translates the ideas or concepts into the coded message that will be communicated. The symbols can take on numerous forms such as, languages, words, or gestures. These symbols are used to encode ideas into messages that others can understand. When encoding a message, the sender has to begin by deciding what he/she wants to transmit. This decision by the sender is based on what he/she believes about the receivers knowledge and assumptions, along with what additional information he/she wants the receiver to have. It is important for the sender to use symbols that are familiar to the intended receiver. A good way for the sender to improve encoding their message, is to mentally visualize the communication from the receivers point of view. To begin transmitting the message, the sender uses some kind of channel (also called a medium). The channel is the means used to convey the message. Most channels are either oral or written, but currently visual channels are becoming more common as technology expands. Common channels include the telephone and a variety of written forms such as memos, letters, and reports. The effectiveness of the various channels fluctuates depending on the characteristics of the communication. For example, when immediate feedback is necessary, oral communication channels are more effective because any uncertainties can be cleared up on the spot. In a situation where the message must be delivered to more than a small group of people, written channels are often more effective. Although in many cases, both oral and written channels should be used because one supplements the other. If a sender relays a message through an inappropriate channel, its message may not reach the right receivers. That is why senders need to keep in mind that selecting the appropriate channel will greatly assist in the effectiveness of the receivers understanding. The senders decision to utilize either an oral or a written channel for communicating a message is influenced by several factors. The sender should ask him or herself different questions, so that they can select the appropriate channel. Is the message urgent? Is immediate feedback needed? Is documentation or a permanent record required? Is the content complicated, controversial, or private? Is the message going to someone inside or outside the organization? What oral and written communication skills does the receiver possess? Once the sender has answered all of these questions, they will be able to choose an effective channel. After the appropriate channel or channels are selected, the message enters the decoding stage of the communication process. Decoding is conducted by the receiver. Once the message is received and examined, the stimulus is sent to the brain for interpreting, in order to assign some type of meaning to it. It is this processing stage that constitutes decoding. The receiver begins to interpret the symbols sent by the sender, translating the message to their own set of experiences in order to make the symbols meaningful. Successful communication takes place when the receiver correctly interprets the senders message. The receiver is the individual or individuals to whom the message is directed. The extent to which this person comprehends the message will depend on a number of factors, which include the following: how much the individual or individuals know about the topic, their receptivity to the message, and the relationship and trust that exists between sender and receiver. All interpretations by the receiver are influenced by their experiences, attitudes, knowledge, skills, perceptions, and culture. It is similar to the senders relationship with encoding. Feedback is the final link in the chain of the communication process. After receiving a message, the receiver responds in some way and signals that response to the sender. The signal may take the form of a spoken comment, a long sigh, a written message, a smile, or some other action. Even a lack of response, is in a sense, a form of response (Bovee Thill, 1992). Without feedback, the sender cannot confirm that the receiver has interpreted the message correctly. Feedback is a key component in the communication process because it allows the sender to evaluate the effectiveness of the message. Feedback ultimately provides an opportunity for the sender to take corrective action to clarify a misunderstood message. Feedback plays an important role by indicating significant communication barriers: differences in background, different interpretations of words, and differing emotional reactions (Bovee Thill, 1992). The communication process is the perfect guide toward achieving effective communication. When followed properly, the process can usually assure that the senders message will be understood by the receiver. Although the communication process seems si mple, it in essence is not. Certain barriers present themselves throughout the process. Those barriers are factors that have a negative impact on the communication process. Some common barriers include the use of an inappropriate medium (channel), incorrect grammar, inflammatory words, words that conflict with body language, and technical jargon. Noise is also another common barrier. Noise can occur during any stage of the process. Noise essentially is anything that distorts a message by interfering with the communication process. Noise can take many forms, including a radio playing in the background, another person trying to enter your conversation, and any other distractions that prevent the receiver from paying attention. Successful and effective communication within an organization stems from the implementation of the communication process. All members within an organization will improve their communication skills if they follow the communication process, and stay away from the different barriers. It has been proven that individuals that understand the communication process will blossom into more effective communicators, and effective communicators have a greater opportunity for becoming a success. BIBLIOGRAPHY Burnett, M. J. , Dollar, A. (1989). Business Communication: Strategies for Success. Houston, Texas: Dane. Ivancevich, J. M. , Lorenzi, P. , Skinner, S. J. , Crosby, P. B. (1994). Management: Quality and Competitiveness. Burr Ridge, IL: Irwin. Gibson, J. W. , Hodgetts, R. M. (1990). Business Communication: Skills and Strategies. NY, NY: Harper Row. Bovee, C. L. , Thill, J. V. (1992). Business Communication Today. NY, NY: McGraw-Hill. Berko, R. M. , Wolvin, A. D. , Curtis, R. (1986). This Business of Communicating. Dubuque, IO: WCB. Wright, P. M. , Noe, R. A. , (1995). Management of Organizations. Chicago, IL: Irwin.