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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524141EnglishN-0001November30General SciencesFACTORS AFFECTING JOB SATISFACTION IN UNIVERSITY FACULTY MEMBERS
English1521Mohammad FarzanjouEnglishThis paper have been produced and developed to study the factors affecting job satisfaction of university faculty members. Various concepts and sometimes conflicting views have been formed and developed concerning the definition of “job satisfaction”. Some experts such as Herzberg believes that it has two dimensions: one group is the factors and conditions that their absence may lead to dissatisfaction; however, they do not cause a strong motivation if provided, but only prevents the occurrence of dissatisfaction, that is the health factors or the ones influencing in maintaining the status quo or survival factor. According to Herzberg, these factors include: staff attitudes and perceptions, methods of administration, organization policies, nature and extent of supervision, job security, working conditions, status, salary level, the establishment of bilateral relations, supervisors, homogeneous, subordinate staff?s personal life. The lack of these factors may lead staff to dissatisfaction to the extent that employees leave the organization and endanger its existence. Hence, Herzberg maintains that these factors are necessary to provide and maintain the organization?s health. The second factors are the ones affecting in creating motivation which are lead to individual?s motivation and satisfaction, but their absence only makes a poor satisfaction. Therefore, the absence of the second group factors is the same as not having attitude. According to Herzberg, the factors affecting in creating motivation are: business success, recognition and appreciation of the people and their work, job and career development, personal growth and the nature of one?s work are called motivational factors.
Englishsatisfaction, dissatisfaction, job satisfaction, needs, factors affecting job satisfactionINTRODUCTION
From the early 1920s, the discussion about job satisfaction has been paid attention to different management schools and as Locke (1972) predicted, at least 3,350 articles were published in this field up to 1978, and over hundreds of articles are annually published on job satisfaction now. Robbins defines job satisfaction as person?s general attitude towards his job. Shertzer maintains that job satisfaction is considered as having interest the tasks required to have a job, the conditions where a job is performed and the reward obtained for. Graham conducted a research concerning job satisfaction.
Needs Theory
One of the most important theories in the field of human motivation proposed by Abraham Maslow entitled “ hierarchy of needs” including: physiological needs, safety needs, need to love, esteem needs, self-actualization needs, the needs of research and freedom of speech and the need to acquire knowledge and understanding. Maslow maintained that once one level of needs is met they are no longer motivational and higher level of motivation are initiated for person?s motivation. In the end, Maslow did not maintain that his hierarchy of needs are comprehensive and total and the same goes for everyone and everywhere.
Health-Motivation Theory
One of the controversial theories in human nature was offered y “Frederick Herzberg” which is called Herzberg?s two-factor theory. Herzberg conducted a study of approximately 200 accountants and engineers employed in institutions located in the preview of Petersburg, Pennsylvania. He made use of “ expressing the critical events” to present a substantive theory, then the responses have been analyzed and the reasons of staff?s satisfaction and dissatisfaction are derived, and came to this conclusion that bad feelings often related to job environment and/or job satisfaction is concerned with job content and dissatisfaction is the one with job environment. Satisfying factors are called “stimulators” and dissatisfaction factors are “health factors”. Health factors or the ones maintain the status quo which their absence lead to dissatisfaction, but they are not lead to strong and powerful motivation if not existed provided and. Providing these factors only prevents dissatisfaction, but they are not lead to motivation if existed. Existing these factors maintain staff in the organization, otherwise they are affected to extreme satisfaction and may leave the organization. There are other factors affecting in creating motivation and are lead to individual?s satisfaction and motivation if existed, while they are lead to weak dissatisfaction if not existed. Therefore, their absence is lead to lack of motivation. Herzberg maintains that the factors affecting in creating motivation are business success, recognition and appreciation of people and their works, job development and advancement. He called these factors “motivational factors” . According to Herzberg, job satisfaction may be increased without job dissatisfaction decreased, and vice versa. Table (1) shows the theoretical relationship between motivational and health factors in job environment in Herzberg?s model.
Factors Affecting Job Satisfaction
1. Five dimensions of job are as follows: 1. Automatic or nature of work: the amount where by a job provides interesting tasks, opportunities to learn and the possibility to accept responsibilities for the person.
2. Promotion opportunities: opportunities and chances for advancement in the organizational hierarchy.
3. Payment: the rewards and wages someone receives and the amount considers the payment as just and equitable against other staff?s wages.
4. Monitoring and control: the ability to supervise in providing supportive behaviors and technical assistance.
5. Partners: the extent to which partners are technically efficient and are socially supportive to the person.
Concerning job satisfaction, Snider (1975) maintains that job satisfaction is one?s assessment of the current conditions and positions in the job and the results achieved from having the job. Poti and Khan maintain that one?s emotional aspects of working in an organization is called job satisfaction.
In 1960s, Mrs. Smith and colleagues mentioned the following factors affecting job satisfaction:
1. Wages (income)
2. The nature of job
3. Opportunities for job promotion
4. Monitoring and supervision
5. The peer group
Wage Sand Income
It has a determining role on staff?s job satisfaction. In a research conducted on more than 2000manager, Porter and Lowler found out that there is a relatively strong relationship between income and job satisfaction. Similar reports are offered by Smith and Kindal. They reckoned that there is a strong relationship between annual incomes of industrial workers in 21 factories and job satisfaction [2]. Automatic or nature of job: having published Herzberg?s research results in the book “Motivation at Work” in 1959, the role of automaticity in job satisfaction has been considered. Three aspects of nature of job affecting job satisfaction are as follows:
1. High control on methods of work performed and its rapid measurement.
2. Variation in scientific management school, high emphasis on specialization and division of responsibilities and jobs to enhance efficiency.
3. Using skills and abilities.
Instruments to Measure Job Satisfaction
People do not clearly express their views to what they reveal, but preserve their attitudes towards the policies, laws and other issues which are widely relevant to them. The attitudes associated with the work are no exception. Most of these people reveal their attitudes related to job towards their close friends and relatives, but do not make it clear to their own supervisors and chiefs. Thus, contrary to initial impressions, assessing job satisfaction is a difficult task. However, there are several ways to evaluate job satisfaction that one is briefly explicated in this section[3]. Job Description Index (JDI): one of the most accurate and common means of measuring job satisfaction is Job Description Index which has been developed by Smith at Cornell University (Nanchian, Tavakoli, Mousavi and Trameshlou). In this index, respondents respond to some descriptive and short sentences about each five aspects of job position. These include regarded nature of work, supervision, coworkers, salary and wage and job promotions.
Research on Job Satisfaction
The research conducted by Frederick Herzberg in 1966 on 200 accountants and engineers of industrial and commercial organizations in Petersburg City, in Philadelphia State of America may be considered as one of the first ones performed n this field. Having studied in this regard, he concluded that job satisfaction depends on five factors including job success, job identification, job attractiveness, job responsibilities and career advancement, and the lack of employee?s job satisfaction depends on some factors as below:
1. Regulations, policies and administrative regulations.
2. Proper monitoring and control.
3. Wage rights.
4. Private relations of corporate individuals
From the beginning of human relations movement, all comments were concerned with the relationship between performance and satisfaction. Content theories are absolutely assumed that satisfaction causes improving performance; and on the contrary, dissatisfaction causes the lack of attention to performance. However, Porter and Lowler maintain that motivation is not the same as satisfaction and performance. Satisfaction, motivation and performance are separate variables that are related to each other differently, and there is a complex relationship between motivation, satisfaction and performance. Porter and Lawler maintain that the rewards that are consecrated and how they received is determined by satisfaction and indicated that satisfaction is lead to performance and suggested that in practice, managers must step beyond what are traditionally thought and measure variables such as the values of possible rewards, understanding possible effort-reward and understanding the roles; these variables certainly helps management have a better understanding staff?s efforts and performance [4].
Woroum came to the following conclusions in his research:
There is a negative relationship between job satisfaction and retirement from work.
There is a negative relationship between job satisfaction and job absence.
There is a negative relationship between job satisfaction, the extent of injuries and accidents resulted from working.
There is a positive relationship between job satisfaction and performance.
Kimbel Wilez maintains, in his research lasted four years, that the following factors are the ones affecting job satisfaction among faculty members:
1. Assuredness and comfort in life
2. Fair and equitable treatment
3. Feeling love and attachment
4. Participating in determining working policy
5. Desirable working conditions
6. Support and assistance of managers towards personnel
7. Appreciation and acknowledgments to the services performed
8. Feeling of success and development
The above mentioned research hypotheses was confirmed by various correlation coefficients. Good satisfaction will increase the level of job satisfaction. Faculty member?s job dissatisfaction is more in men than women and there is an inverse relationship between job satisfaction and level of education. Having reviewed the scientific writings, Ratsoury concluded that job satisfaction among faculty members in bureaucratic universities is low and there is also a correlation between job satisfaction and motivation [5]. Hersy and Blanchard found out in their studies that there is a positive relationship between leadership styles (ordering, understanding, participative and delegating personality) and job satisfaction of faculty members. They found out that faculty members are less satisfied with wages and facilities and are more pleased with cooperation and participation in leadership.
Kenly and Lerin Stone (1993) reported that experienced faculty member?s participation in re-designing job cause their job satisfaction and this has less effect on new faculty members. But increasing salary is effective in increasing job satisfaction in both groups. Kahn also points out four independent factors in his investigations that indicate employee?s satisfaction:
Usefulness and satisfaction of duty.
Certain elements of the task that are mutually beneficial, such as
Being satisfied by job?s physical value, the present salaries and those of the future.
Job satisfaction, having interest and enthusiasm to job and the dignity that it provides for the owner.
Being satisfied by organization, working conditions and operation of device.
Being satisfied by professional?s competency in the role of supervisors and leaders[6]. Roll and Kani (1993), Barry Foldokrouk Vadani. etc. conducted some studies concerning job satisfaction and as others, they discussed the factors affecting the employee?s job satisfaction and considered it as an effective factor in organizational efficiency and effectiveness and implicitly indicated that regarding it is one of administrative needs. Maghaze found out in his study that faculty members are satisfied with their jobs but are not satisfied with the ones such as authorities? performance, how to administrate university, working conditions, inadequate facilities and amounts of salaries[7]. Esmailei (2008) came to this general conclusion, in a research conducted in Tehran, that there is a positive and significant relationship between faculty member?s job satisfaction and their academic performance. That is to say, the academic satisfaction is increased when job satisfaction is increased and their performance is reduced when their satisfaction is declined[8]. Moradi (2010) came to a conclusion, regarding job satisfaction that was almost different from the ones obtained in other studies. He concluded that men have more job satisfaction than women[9]. Kontenz maintains that job satisfaction is lead to organizational effectiveness and efficiency[10]. According to the aforementioned studies, it seems that factors as nature of work, colleagues, opportunities for promotion and advancement, salaries and facilities and management is effective on faculty member?s job satisfaction. Generally speaking, it can be said that people who are motivated to work are more required into high level needs of hierarchy of needs, such as the need to be respected, the need to be independent and the need for self-actualization. Regarding other?s needs is resulted to their satisfaction sand satisfaction is lead to consistency and commitment.
Suggestions
As the above-mentioned studies indicated, job satisfactions are among the factors that can be resulted into organizational effectiveness and efficiency and prevents from negative consequences of its absence. Therefore, the following cases are suggested to headmasters, assistants and administrators in universities and higher education:
1. Faculty members are often dissatisfied their jobs and currently do not involve in their working.
Being away from workplace help dissatisfied people avoid from unpleasant aspects of their workplace. On the contrary, satisfied faculty members are less inclined to making complain; they are more acquired with health and longevity and quickly learn the tasks associated with their job. Therefore, it is required the workplace to be pleasant and desirable for faculty members to be benefited from increasing satisfaction as well as other consequences.
2. Manager?s support and is an important factor in faculty member?s satisfaction which can be effective in developing a good understanding of organization?s environment. If faculty members are assured they are acquired with managers and co-worker?s support, they will continue their work with greater confidence and interest and will be more satisfied.
CONCLUSION
Being satisfied with university?s goals and performances can increase people?s motivation and morale in higher education, and on the contrary, if someone does not satisfied with one?s workplace, hell will not be satisfied and his working motivation will be reduced. An appropriate and desirable atmosphere is so effective in creating motivation and satisfaction in people. It would be diligent to create such an atmosphere. One of the important factors in creating motivation and an appropriate atmosphere is workplace satisfaction. If the workplace is inappropriate and people are not satisfied within, their motivation will be weakened. There are many factors involved in creating a suitable and satisfactory environment, including: Creating intimacy and cordiality among people, establishing security, regarding rest times and classroom, the time to handle personal affairs and being reasonable. Establishing facilities and healthy recreation: healthy and appropriate recreation must be arranged to create diversity and increasing work and life motivation with taking into account appropriate time and duration. Creating a sense of mutual respect among people, regarding respect and considerate attention. People are required to the opportunities for advancement and promotion in workplace, having rights and adequate facilities. Establishing these factors is highly significant in the profession of faculty and higher education members. Creating a rewarding and encouraging system which is appropriate with providing objectives and organization?s missions is tremendously significant in employee?s advancement and personal development as well as creating job satisfaction within.
ACKNOWLEDGMENTS
The author wish to tanks from Prof. M.H. Pardakhtchi and Prof. K. Fathi due to their efforts, also tanks to assistants for cooperation in editing this paper.
Englishhttp://ijcrr.com/abstract.php?article_id=1980http://ijcrr.com/article_html.php?did=19801. Muhammadzade, A, and Mehrojan, “Contingency Approach to Organizational Behavior”, Tehran, University of Alamme Tabatabaie Publications, 1996.
2. Rezaie, Doulatabadi, “Study of the relationship between administrative atmosphere and employee?s job satisfaction in Zoub Ahan Company”, Esfahan, 1994.
3. Abbaszadeh, M, “General Management Training”, Oroumie University Publications, 1995.
4. Boroumand, Z, “Organizational Behavior Management”, Tehran, Payame Nour Publications, 1995.
5. Hevi, V and Miskel, S “Practical Theory and Research in Administrative Management”, translated by M. Abbaszade, Oriumieh, Nozouli Publications, 1992.
6. Kourmen, A. K. “Industrial and Administrative Psychology”, translated by Hussein Shokr Shekan, Tehran, Roshd Publications, 1991.
7. Kamp, A “Applied Psychology”, translated by F. Maher, Tehran, Astan Qods Razavi Publications, 1991.
8. Houman, Heydarali, “Preparing and Measuring Job Satisfaction Assessment Scale”, 2002.
9. Robins, A, P, “Organizational Behavior Management”, translated by A, Parsaiian and M, Arabi, Tehran, Institute of Business Studies and Experiments, 1989.
10. Kontenz, H, Oudanol, S and Wihrokh, H, “Principles of Management”, translated by M. Tousi and V. Alavi, Tehran, Administrative Management Training Center, 1996.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524141EnglishN-0001November30General SciencesIN VITRO EVALUATION OF AQUEOUS AND ETHANOLIC EXTRACTS OF VERNONIA COLORATA AS
AN ANTIBACTERIAL AGENT
English2228OseniEnglish Lateef AdebayoEnglish BerkohEnglish Eric AsamoahEnglish Mills-RobertsonEnglish Felix CharlesEnglishRecent reports have shown the increased emergence of bacteria resistance to many existing antimicrobial drugs. This has prompted the need to find alternative remedies, and plant products have proven to be vital in this search. Vernonia colorata has been reported to be active against syphilis, pneumonia, measles, dysentery and several skin infections in traditional medical practices. In the present study, aqueous and ethanolic extracts from the leaves of Vernonia colorata were evaluated in vitro for growth inhibitory activity on Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Staphylococcus aureus using Agar diffusion method. Phytochemical analysis revealed the presence of reducing sugars, saponins, polyphenols, tannins, phlobatannins, alkaloids, sterols and triterpenes in both extracts. These classes of phytochemicals have widely been reported for their antibacterial properties. Of the several bacteria tested, only S. aureus and P. aeruginosa showed significant susceptibility to both ethanolic and aqueous extracts with concentrations range between 2.00 to 5.00mg/ml. The aqueous extract also showed the highest activity on S. aureus at concentration of 5mg/ml. The Minimum Inhibitory Concentration of the aqueous extract ranged between 4.00 and 6.00mg/ml while that of the ethanolic extract ranged between 5.00 and 6.00mg/ml. Following the results from the current study, it can be concluded that V. colorata has significant antibacterial activity and will be very useful in the discovery of novel antibiotics against S. aureus and P. aeruginosa
EnglishVernonia colorata, Antibacterial activity, Minimum Inhibition Concentration, Phytochemical screening, Agar diffusion methodINTRODUCTION
Both ancient and modern men of all cultures have widely used medicinal plants for treating different ailments. Scientific discoveries have shown that plants produce a wide range of complex compounds (secondary metabolites) as part of their normal metabolic process. Several secondary metabolites have been reported to have significant therapeutic properties. Therefore, plants are model source of medicines as they contain many chemical agents with therapeutic properties. Despite increasing advancement in the field of medicine and molecular diagnosis, reports indicate that close to 80% of the world population still dependant on plant derived pharmaceuticals. There are also reports that suggest that nearly 28% of drugs available in the market are plant based products and its derivatives. (Newman et al., 2003). In recent times, scientists have extensively reported on bio-assays of several plants of nutritional and medicinal values. Furthermore,a large proportion of compounds used as lead molecules in drug discovery are plant based compounds. This suggests that plant based compounds play a vital role in diversity oriented synthesis of natural product-like pharmaceuticals. (Marcaurelle and Johannes, 2008). Bacterial infection treatment options include chemotherapy, radiation therapy and surgery. In chemotherapy, antibacterial drugs are usually employed in the treatment of various forms of bacterial infections. However, there are reports on increased emergence of multiresistant bacterial strains of clinically important pathogens. This development has fetched the interest of scientist to develop newer broad spectrum antimicrobial agents. Due to the high cost and the less availability of new generation antibiotics, it is imperative to look for the substances from alternative medicines with claimed antimicrobial activity. A significant number of medicinal plants with significant antimicrobial activity have been reported in different traditional literatures. Vernonia is a genus of about 1000 species of forbs and shrubs in the family Asteraceae. The uses of several species of Vernonia, including Vernonia amygdalina, Vernonia auriculifera, Vernonia colorata, Vernonia galamensis, and Vernonia hymenolepis in traditional health care have been reported. There are many convergence in the usage of Vernonia spp. in its traditional use throughout West and Central Africa and North America as anti-inflammatory, analgesic, antibacteria, anticancer, antidiabetic, antifungal, antimalaria and antioxidant.(Abosi and Raseroka, 2003; Phillipson et al., 1993). Previous studies on Vernonia spp have largely been confined to Vernonia amygdalina. This species has been reported to contain glycosides, tannins, steroidal saponins, sesquiterpenes lactones flavonoids and vitamin C. (Ifeoma and Chukwunonso., 2011). V. Amygdalina is also known for its activity against syphilis, malaria, measles, dysentery and yellow fever. (Oluwalana and Adekunle.,1998). V. colorata has also been reported to be used in traditional herbal medicine across many African countries for the treatment of bacteria, fungal, parasitic and inflammatory disorders. Despite the traditional uses of V. colorata in primary health care, reports on the phytochemical profile and bio-assay of V. colorata are limited. In this regard, we aim to explore scientifically, the antibacterial potential of V. colorata to substantiate the reported claims. The current research seeks to screen for phytochemicals in the leaves of Vernonia colorata and also evaluate its potential as an antibacterial agent by analysing its growth inhibitory activity on E. coli, K. Pneumoniae, P. aeruginosa and S. aureus.
MATERIALS AND METHODS
Materials
Plant material
Fresh leaves of vernonia colorata were collected from a piece of land at MampongAkuapim in the Eastern Region of Ghana. The leaves were later taken to the herbarium Department of the Centre for Scientific Research into Plant Medicine, MampongAkuapim, for botanical identification.
Reagents
Ethanol, Fehling?s solutions, Chloroform, Acetone, Sodium Picrate paper, Ferric chloride, Ammonia, Chloramphenicol test paper, Dimethyl sulphoxide (DMSO), Hydrochloric acid (HCl), and Sulphuric acid(H2SO4) were of analytical grade and from BDH, UK. Other reagents used were of analytical grades and water used was glass distilled.
Bacteria strains
Four different microbes of standard strains were purchased at the Komfo Anokye Teaching Hospital (KATH) Kumasi, with standard codes. The microbes were; E.coli (ATCC25922), K. Pneumoniae (ATCC 33495), P. aeruginosa (ATCC 27853) and S. aureus (ATCC 25923).
Methods
Extraction from plant material
Adequate quantity of the leaves were collected, sun dried and then milled. The fine powder was then divided into two portions. Preparation of ethanolic extract About 500g of the powder were soaked in 5L of 70% ethanol for 24 hours. The suspension was then filtered. The filtrate was concentrated using rotary evaporator and then freeze dried. Preparation of aqueous extract Another 500g sample of the powdered leaves was soaked in 5L of distilled water and then boiled for 15minutes after which the temperature was lowered to 60oC for another 15 minutes. This was then filtered and the filtrate evaporated using rotary evaporator at 60oC and then freeze dried.
Phytochemical analysis
About 5g of each freeze dried sample of crude extract was dissolved to 100ml and portions analyzed for phytochemical constituents using standard methods.(Stahl , 1969; Harborne, 1973)
Microbiology
Media
Muller Hilton Agar and Peptone Agar were used.
Sterilization
All other materials used in the microbiological work were sterilized before usage.
Preparation of Muller Hilton agar
About 15.2g of Muller Hilton agar was weighed and added to 400ml of distilled water, and then heated to dissolve. The media was sterilized at 121oC in an autoclave for 20 minutes and then cooled to about 60oC. About 20ml of the media was poured gently into each plate and left to cool and then stored in an oven at 370C for 16 hours.
Peptone water
Peptone agar was used as the broth for the culture of the microbes About 0.5g of peptone agar was weighed and added to 50ml of distilled water and then heated to dissolve. This was sterilised at 121°C for 20 minutes and cool to about 60°C. 5ml portions of the broth were pipetted into test tubes and then stored in an oven at 370C for 16 hours.
Antimicrobial susceptibility test
The antibacterial test was performed using the agar diffusion method of Collins et al. (1995). The test microorganisms were inoculated on nutrient agar plates and spread uniformly using a sterile glass spreader. Wells of 5 mm in diameter were made on the nutrient agar using a sterile cork borer. The cut agar disks were carefully removed by the use of forceps sterilized by flaming. Different concentrations of the freeze extracts were prepared by dissolving various masses in 20% dimethyl sulphoxide (DMSO). To each well was introduced different concentrations (1.0, 2.0, 3.0, 4.0, 5.0 mg/ml) of plant extracts. Control experiments were set up using Chloramphenicol and DMSO as positive and negative controls respectively.
The plates were allowed to stand for one hour at room temperature for diffusion of the substances to proceed before the growth of microorganisms commenced. The plates were made in triplicate and were incubated at 37°C for 24 h. Diameters of zones of inhibition in the triplicate plates were measured by calculating the difference between cork borer (5mm) and the diameters of inhibition (Hewett and Vincent, 1989; Singh et al., 2002; Adebayo and Adegoke; 2009). The zones of inhibition were then recorded.
Determination of minimum inhibitory concentration (MIC)
Various concentrations of both aqueous and ethanolic extracts ranging between 4.0 and 6.0 mg/ml were introduced into different test tubes; each tube was inoculated with an overnight culture of S. aureus, P. aeruginosa, E. coli and K. pneumonia diluted to give a final concentration of 106 cells per ml. The tubes were incubated at 37°C for 24 h. The least concentration of extract that did not permit any visible growth of the inoculated test organism in broth culture was regarded as the minimum inhibitory concentration (MIC) in each case (Collins et al., 1995).
RESULTS AND DISCUSSION
Phytochemistry of plant extracts
Preliminary phytochemical screening of crude aqueous and ethanolic extracts from leaves of V. colorata revealed the presence of triterpenes, sterols, alkaloids, reducing sugars, polyphenols, tannins, phlobatannins and saponins. Cynogenic glycosides, polyuronoids, anthracenocides and flavonoids were absent.
(Table1).
Antimicrobial activity
Both crude ethanolic and aqueous forms of the extracts of V. colorata exhibited varying degree of antimicrobial activities against the test organisms. (Table 2a,b) E. Coli and K. pneumoniae showed significant resistance against both aqueous and ethanolic extracts of V. colorata at concentrations below 5.00mg/ml. However, S. aureus and P. aerugenosa showed susceptibility to both extracts at concentrations between 4.00 and 5.00mg/ml. (Table 2a,b). It was observed that antibacterial effectiveness increased with increasing concentration of extracts. This supports previous work by Kurosaki and Nishi (1983) who reported that higher concentrations of antimicrobial substances showed appreciable growth inhibition to microorganisms. Although, phytochemical screening revealed similar results for both aqueous and ethanolic extracts, the compounds found in the classes of phytochemicals present in each extract may differ and this could account for their varying inhibitory activities. (Table 2a,b). Another possible reason for the varying degree of inhibition by the extracts could be the presence of other classes of phytochemicals that were not tested for in the ethanolic extract. Further to this, the boiling of leaves during the preparation of the aqueous extract may render some compounds inactive in the aqueous extract and this could also contribute to the observed variation in inhibitory activities. Although the positive control (Chloramphenicol) showed significant growth inhibitory activity on all the bacteria tested, the aqueous extract was found to be more effective on S. Aureus at concentration of 5mg/ml while the ethanolic extract was found to more effective on P. Pneumonia at the same concentration than the standard.(Table 2c). The MIC of the aqueous extract in this study against the test organisms ranged between 4.0 and 6.0 mg/ml while those of the ethanolic extract ranged between 5.0 and 6.0 mg/ml. (Table 3). Antimicrobial agents with low activity against an organism have a high MIC while a highly active antimicrobial agent gives a low MIC. The present result shows that the aqueous extract is slightly more effective than the ethanolic extract. Tannins, alkaloids saponins and phlobatannins have been reported for their antibacterial and antiviral activity (Enzo, 2007). Furthermore, alkaloids and saponins are classes of compounds that are known to be effective for the treatment of syphilis and other venereal diseases. (Sofowara, 1993). Steroids in modern clinical studies are known for their antiinflammatory and analgesic properties. (Pithayanukul et al., 2007). The antibacterial activities demonstrated by extracts from V. colorata may be attributed to the presence of these phytochemicals and this supports the use of the plant for the treatment of syphilis, pneumonia and skin infections as reported in traditional folk medicine.
CONCLUSION
Although, a large number of medicinal plants are constantly screened for their antimicrobial effects, many plant species with potent antimicrobial properties are yet to be discovered. The present study reveals the antibacterial potential of leaves of V. colorata. The antibacterial activities of the extracts against S. aureus and P. aeruginosa were comparable to those of the standard (Chloramphenicol). These results seem to justify the continued use of the plant in the treatment of microbial infections such as pneumonia, syphilis and skin diseases. In addition, the inhibition of growth of the test organisms (S. aureus and P. aeruginosa) that are known to display multidrug resistance to most antibiotics and nonantibiotic antimicrobial agents justify the continued use of these plants in folk and traditional medical practice.
Englishhttp://ijcrr.com/abstract.php?article_id=1981http://ijcrr.com/article_html.php?did=19811. Abosi, A.O. and Raseroka, B. H. (2003). In vivo anti-malarial activity of Vernonia amygdalina. Br. J. Biomedical Sci. 60: 89 -91.
2. Adegoke, Anthony A. and Adebayo-Tayo, Bukola C. (2009). African Journal of Biotechnology Vol. 8 (1), pp. 077-080
3. Collins GH, Lynes PM, Grange JM (1995). Microbiological Methods (7th edn) Butterwort – Heinemann Ltd, Britain pp. 175–190.
4. Enzo, A.P., (2007). Traditional plants and herbal remedies used in the treatment of diarrheal diseases. Mode of action, quality, efficacy and considerations. In: Ahmad I, Aqul F, Qwaiss M, Modern Phytomedicine Turning Medicinal Plants into Drugs. WILEY-VCH Verlag GMBH and Co. KGQA. Weinheim, pp: 248-260.
5. Harborne J.B. Phytochemical methods (1984). 2nd ed. Chapman and Hall, New York, 3:100-117.
6. Harborne J.B. Phytochemical methods (1984). 2nd ed. Chapman and Hall, New York, 1:4-7.
7. Harborne JB. Phytochemical Methods. Chapman and Hall, London (1973) 172- 278.
8. Hewitt W, Vincent S (1989). In:Theory and application of microbiological assay. Academic Press, San Diego, p. 39.
9. Ifeoma I. Ijeh and Chukwunonso E. C. C. Ejike (2011). Current perspectives on the medicinal potentials of Vernonia amygdalina Del. Journal of Medicinal Plants Research Vol. 5(7), pp. 1051-1061
10. Kurosoki F, Nishi A (1983). Isolation and antimicrobial activity of the phytoalexin– 6-methoxymellein from cultured carrot cells. Phytochemistry 22(3): 669-672.
11. Marcaurelle LA and Johannes CW (2008) Application of natural product-inspired diversity-oriented synthesis to drug discovery. Prog Drug Res. 66(187):89- 216.
12. Newman DJ, Cragg GM and Snader KM (2003) Natural products as sources of new drugs over the period 1981-2002. J Nat Prod 6:1022-37.
13. Oluwalana SA, Adekunle MF (1998). Forest plant roots in household nutrition and health care J. Trop. For. Resources 14(1): 120 - 136.
14. Phillipson JD, Wright CW, Kirby GC, Warhurst DC (1993).Phytochemistry of some plants used in traditional medicine for the treatment of protozoal diseases. Abstracts, Int Symposium of the Phytochem Soc of Europe; University of Lausanne, Switzerland, p.L3.
15. Pithayanukul P., Tubprasert J., Wuthiudomlert M (2007). I n vitro antimicrobial activity of Zingiber cassumunar (Plai) oil and a 5% Plai oil gel. Phytotherapy Research, 21:164-169.
16. Singh B, Sahu PM, Sharma MK (2002). Anti-inflammatory and Antimicrobial activities of triterpenoids from Strobilanthes callosus Nees. Phytomed. 9:355-359.
17. Stahl E. (1969) Thin Layer Chromatography. 2nd ed. MRF Ashworth Springer-Verlag, Heidelberg 421-470.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524141EnglishN-0001November30HealthcareA DIFFUSE CORONARY SPASM - A VARIANT OF A VARIANT?
English2933A.NoelEnglish B. AmirthaganeshEnglishA 55 year old mild hypertensive presented with typical angina pain associated with giddiness and profuse sweating. His initial Electrocardiogram showed „Junctional rhythm? with diffuse „T? wave changes in leads II, III, avF, V4 to V6. 2D Echocardiogram, on the same day, showed regional wall motion abnormality of Left Anterior Descending (LAD) and Left circumflex (LCX) /or Right Coronary Artery (RCA) territories with moderate Left Ventricular dysfunction. Subsequently, Coronary Angiogram done two weeks later showed normal epicardial coronaries.
EnglishCoronary Spasm; Prinzmetal; ECG abnormalities; Regional Wall Motion Abnormality (RWMA); Acute coronary syndromeINTRODUCTION
Prinzmetal variant angina has always been a medical curiosity and thought to be a rare entity, nonetheless increasing number of cases are being reported every year. It was first described as "A variant form of angina pectoris" in 1959 by the American cardiologist Dr. Malcolm Prinzmetal. [1] Prinzmetal angina most commonly affects a single site (usually the RCA) and presents as a focal spasm, which may vary for each attack (migratory spasm). [2,3] We report a 55 year old man who presented with multivessel involvement as evidenced by his Electrocardiogram and Echocardiogram findings.
Case Report
A 55 year old man was brought with history of typical anginal pain associated with giddiness and profuse sweating in the early morning hours lasting for half an hour. He was euglycemic and a non smoker diagnosed to have hypertension 6 months prior to the present hospitalization, which was well controlled on Enalapril Maleate (5mg). His lipid profile was normal and the family history was non-contributory. As he could reach the hospital only four hours later, his electrocardiogram that time showed „Junctional rhythm? at rate of 50/min with deep “T” inversion in leads II, III, avF, V4 to V6.
Consequent 2D Echocardiogram showed regional wall motion abnormality involving the both Left coronary artery and Left Circumflex /or Right coronary artery domains with moderate LV dysfunction (Ejection Fraction ~ 38%). A provisional diagnosis of Acute Coronary syndrome was made and was managed conservatively with low molecular weight Heparin, Statins and Anti-platelet drugs before he was referred to our hospital. On arrival at our hospital, he was totally asymptomatic, his ECG was within normal limits (Figure: 2) and his ECHO showed normal LV systolic function with no RWMA. A coronary Angiogram was done which showed normal epicardial coronaries with left dominant system (Figures: 3 and 4). Provocative tests could not be done due to ethical and safety concerns. Intra Vascular Ultra sonogram (IVUS) could not be performed on him due to lack of technical expertise and economical consideration. He was managed with Diltiazem hydrochloride 90 mg (sustained release) along with Asprin 75 mg and Rosuvastatin 10 mg. His hospital course was uneventful and there were no further angina attacks till date (on 10 months follow up).
DISCUSSION
The typical presentations of Prinzmetal?s variant angina (PVA) are: Pain at rest, not related to any physical or emotional stress [4] and associated ST segment elevation. [3, 5] Some studies say that not all ECG changes are accompanied by symptoms, sometimes there are ECG changes even in the absence of symptoms. [6] Moreover the attacks tend to have a circadian rhythm usually between 12:00 am and 8:00 am [2] and occur in clusters. They generally are not associated with any classical risk factors (except for heavy smoking). [2] They are also found to be associated with other vasospastic disorders such as Migraine and Raynaud?s Phenomena. [2,3] In our study we found that the subject had involvement of both coronary artery territories. The symptoms were present during the early morning hours and were accompanied by ECG and ECHO changes. In some cases, PVA can take a benign course without any complications but in others it can present with dreaded complications such as syncope, AV block, asystole, ventricular tachy-arrhythmias [5] or MI. [2,3] Although numerous reports are available on variant angina, very little is known about spasm involving multiple vessels. Various theories have been proposed in the pathophysiology of Prinzmetal angina such as eNOS gene mutation,[7] increased Phospholipase C activity etc.[2] Basically it is an endothelial dysfunction causing increased vasomotor tone or vasospasm and repetitive vasospasm causes injury to the vasculature, which in turn leads to coronary stenosis.[2] There are few provocative tests [1] using Ergonovine and Acetylcholine that can induce coronary vasospasm. The test induced spasm can be relieved by intracoronary nitrates or calcium antagonists. This helps in diagnosing PVA. Apart from these, 24 hours ECG monitoring can show episodic ST segment elevation, any associated arrhythmias [5] or a silent MI. [6] Exercise stress testing [4] is not very contributory. Since Prinzmetal is not a "demand" induced symptom but rather a vasospastic abnormality, it can not be induced by exercise. [3] Diagnostic hallmark of this disease has always been Angiogram; which may even be normal during an asymptomatic period. Nitrates and Calcium channel blockers (CCBs) are the mainstay of treatment for Variant angina. Nitroglycerin (in any route) effectively treats an episode of angina within minutes and CCB can be used as a prophylaxis to prevent future attacks. [8, 9] Diltiazem in particular produces coronary dilation but is a less potent peripheral vasodilator. Accordingly our patient was also started on Diltiazem hydrochloride 90 mg (sustained release) along with Asprin 75 mg and Rosuvastatin 10 mg. And there were no further anginal episodes in him at ten months follow up confirming the efficacy of the drugs on multi vessel coronary spasm. [10]
CONCLUSION
A variant of Prinzmetal Variant Angina with features of multi-vessel involvement involving normal or near normal coronary arteries, although transient and seemingly benign, can be life threatening. A multivessel presentation should not deter a physician from diagnosing a variant Prinzmetal angina because the condition can be successfully treated and prevented with Calcium channel blockers.
ACKNOWLEDGEMENT Authors acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors are also grateful to authors/ editors/ publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed
Englishhttp://ijcrr.com/abstract.php?article_id=1982http://ijcrr.com/article_html.php?did=19821. Maile S, Moarof I, Michot M. Early morning recurrent chest pain. Cardiovascular Medicine 2011; 14(1): 24–26.
2. Braunwald E, ed. Heart Disease: A Textbook of Cardiovascular Medicine. 8th ed. Philadelphia, Pa: WB Saunders; 2008. P. 1337-40.
3. Keller KB, Lemberg L. Prinzmetal’s angina. Am J Crit Care 2004; 13: 350- 354.
4. Gubbay ER. Prinzmetal’s variant angina. Can Med Assoc J 1960; 83(4): 164–166.
5. Huckell VF, McLaughlin PR, Morch JE, Wigle ED, Adelman AG. Prinzmetal's angina with documented coronaryartery spasm: Treatment and follow-up. British Heart Journal 1981; 45: 649-55.
6. Guazzi M, Fiorentini C, Polese A, and Magrini F. Continuous electrocardiographic recording in Prinzmetal's variant angina pectoris. British Heart journal 1970; 32: 611- 616.
7. Valentin Fuster, Robert O?Rouke. Hurst’s the Heart. 12th ed. McGrawHill Professional; 2007. P. 1370-72.
8. Gomez FP, De Dious RM, Rey J and Aguado G. Prinzmetal's angina: reflex cardiovascular response during episode of pain. British Heart Journal 1979; 42: 81-87.
9. Madias JE. The syndrome of variant angina culminating in acute myocardial infarction. Circulation 1979; 59:297-306
10. Harper R, Peter T, Hunt D. Syncope in association with Prinzmetal variant angina. British Heart Journal 1975; 37: 771-774.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524141EnglishN-0001November30HealthcareANTINOCICEPTIVE EFFECTS OF DIAZOXIDE IN RODENT MODEL OF CHRONIC PAIN INDUCED BY
CHRONIC CONSTRICTION NERVE INJURY
English3443Deshmukh A. B.English Patel J. K.English Prajapati A. R.English Patel K. S.English Jadav R. J.EnglishAim: Previous studies show that neuropathic pain is refractory against conventional analgesics and thus novel medicaments are desired for the treatment. Activated K+ channels are associated with reducing inappropriate or excessive neuronal activity. The aim of this study was to investigate the possible analgesic effects of potassium channel opening on neuropathic pain. Therefore, the present study was designed to investigate whether potassium channel activator can generate qualitative analgesic effects on the acute pain induced by thermal and mechanical stimulation. Methods: The effect of diazoxide at the dose of 200 mg/kg on acute thermal and mechanical nociception were assessed by sensory testing like spontaneous pain, mechanical hyperalgesia, tactile as well as cold allodynia in chronic constriction injury (CCI) induced pain in rat. Results: After CCI surgery, the rats developed neuropathic pain syndrome. Behavioral studies demonstrated that rats with the CCI experienced spontaneous pain, dynamic allodynia and mechanical hyperalgesia which were significantly different from the sham group. Treatment with diazoxide decreased significantly the withdrawal durations in all sensory tests. Conclusion: The present study indicates that activity of K+ channel may contribute significantly to the development of central sensitization-mediated pain and suggests that K+
openers may be an important molecular target for the treatment of chronic pain of neuropathic origin.
EnglishNeuropathic pain, Diazoxide, Potassium channels, Chronic constriction injury, sensory testINTRODUCTION
Neuropathic pain is one of the most significant health problems in the world. Neuropathic (neurogenic) pain is defined by IASP as pain caused by a lesion or dysfunction of the nervous system (1). It is characterized by inappropriate spontaneous or excessive neuronal activity in response to physiological stimuli. Chronic pain is one of most common reasons for hospital visits so it should be considered to be a disease rather than just a symptom. Recent advances in molecular biology techniques and the subsequent discoveries of key molecules involved in pain production, have clearly contributed to better understanding acute pain (2-5), by which the molecular multiple mechanisms underlying chronic pain can be fully clarified. Proper diagnosis and early treatment are often found to be difficult in neuropathic pain, because it is quite different from other types of pain, such as nociceptive (or physiological) or inflammatory pain and it is irreversible, even when the underlying cause has been rectified (3). Also, the occurrence of neuropathic pain is commonly as a secondary symptom in diseases (e.g. diabetes, cancer, and herpes zoster infection) or as a side effect of chemotherapeutic treatments (4, 6-8). The management of this disorder is achieved by various classes of drugs that are capable of dampening neuronal excitability. Examples may be voltagegated sodium channel blockers (carbamazepine, phenytoin, lamotrigine and topiramate), voltage-operated calcium channel modulators (ethosuximide, gabapentin, levetiracetam) and modulators of inhibitory GABAergic neurotransmission (benzodiazepines, vigabatrin and tiagabine). Of these, the approved drugs for the treatment of neuropathic pain are gabapentin, and carbamazepine and lamotrigine has demonstrated efficacy for neuropathic pain in clinical trials (9). Various drugs with sodium channel blocking actions preferentially suppress thermal nociception which may be partly explained by the local anesthetic action of sodium channel blocking agents and differential sensitivities to local anesthetics of the fibers activated by thermal and mechanical nociception (10). However, a number of issues regarding this treatment, including the effective, meaningful drug dose range, the durability of pain-relief effects leading a poor treatment to patient with currently available drugs. So there is a need for new agents with their novel mechanism for the treatment of neuropathic pain. Potassium (K+ ) channel opening is one potential mechanism that has not yet been exploited for neuropathic pain. Activated K+ channels are associated with reducing inappropriate or excessive neuronal activity (11). Therefore, the present study was designed to investigate whether potassium channel activation generate qualitative analgesic effects on the acute pain induced by thermal and mechanical stimulation.
MATERIALS AND METHODS
Animals
Wistar-kyoto male rats of 8 weeks age with the body weight range from 250-300 gms were procured from Central Animal Facility, Nootan pharmacy college, Visnagar, India. They were maintained in essential condition of controlled temperature (Englishhttp://ijcrr.com/abstract.php?article_id=1983http://ijcrr.com/article_html.php?did=19831. Cruccu G, Anand P, Attal N, GarciaLarrea L, Haanp M, Jorum E et al. EFNS guidelines on neuropathic pain assessment. European Journal of Neurology 2004; 11 (3): 153–62
2. Woolf CJ, Ma Q. Nociceptors--noxious stimulus detectors. Neuron 2007; 55: 353-64.
3. Scholz J, Woolf CJ. Can we conquer pain? Nat Neurosci 2002; 5 Suppl: 1062-67.
4. Ueda H. Molecular mechanisms of neuropathic pain-phenotypic switch and initiation mechanisms. Pharmacol Ther 2006; 109: 57-77.
5. Hucho T, Levine JD. Signaling pathways in sensitization: toward a nociceptor cell biology. Neuron 2007; 55: 365-76.
6. Campbell JN, Meyer RA. Mechanisms of neuropathic pain. Neuron 2006; 52: 77-92.
7. Woolf CJ, Mannion RJ. Neuropathic pain: aetiology, symptoms, mechanisms, and management. Lancet 1999; 353: 1959-64.
8. Matsumoto M, Inoue M, Hald A, Xie W, Ueda H. Inhibition of paclitaxelinduced A-fiber hypersensitization by gabapentin. J Pharmacol Exp Ther 2006; 318: 735-40.
9. Eisenberg E, Lurie Y, Braker C, Daoud D, Ishay A. Lamotrigine reduces painful diabetic neuropathy: a randomized, controlled study. Neurology 2001; 57: 505-9
10. Sakaue A, Honda M, Tanabe M, Ono H. Antinociceptive Effects of Sodium Channel-Blocking Agents on Acute Pain in Mice. J Pharmacol Sci 2004; 95: 181–88.
11. Wickenden AD, Roeloffs R, McNaughton-Smith G, Rigdon GC. KCNQ potassium channels: drug targets for the treatment of epilepsy and pain. Expert Opin. Ther. Patents 2004; 14(4): 1-13.
12. Bennett GJ, Xie YK. A peripheral mononeuropathy in rat that produces disorders of pain sensation like those seen in man. Pain 1988; 33: 87- 107.
13. Choi Y, Yoon YW, Na HS, Kim SH, Chung JM. Behavioral signs of ongoing pain and cold allodynia in a rat model of neuropathic pain. Pain 1994; 59: 369–76.
14. Field M J, Scott M, Hughes J, Singh L. Gabapentin and pregabalin, but not morphine and amitriptyline, block both static and dynamic components of mechanical allodynia induced by streptozocin in the rat. Pain 1999; 80: 391–98.
15. Decosterd I, Woolf CJ. Spared nerve injury: an animal model of persistent peripheral neuropathic pain. Pain 2000; 87: 149–58.
16. Field MJ, Bramwell S, Hughes J and Singh L. Detection of static and dynamic components of mechanical allodynia in rat models of neuropathic pain: are they signalled by distinct primary sensory neurones? Pain 1999; 83: 303–11.
17. Hunt SP, Mantyh PW. The molecular dynamics of pain control. Nat. Rev. Neurosci. 2001; 2(2): 83-91.
18. Kawano T, Zoga V, Mccallum JB, Wu HE, Gemes G, Liang MY et al. ATPsensitive potassium currents in rat primary afferent neurons: biophysical, pharmacological properties, and alterations by painful nerve injury. Neuroscience 2009; 162(2): 431-43.
19. Sachs D, Cunha FQ, Ferreira SH. Peripheral analgesic blockade of hypernociception: Activation of arginine/NO/cGMP/protein kinase G/ATP-sensitive K+ channel pathway. Proc Natl Acad Sci USA 2004; 101: 3680–85.
20. Sawynok J, Esser MJ, Reid AR. Peripheral antinociceptive actions of desipramine and fluoxetine in an inflammatory and neuropathic pain test in the rat. Pain 1999; 82: 149–58.
21. Tiraboschi E, Giambelli R, D'Urso G, Galietta A, Barbon A, de Bartolomeis A. Antidepressants activate CaMKII in neuron cell body by Thr286 phosphorylation. Neuroreport 2004; 15: 2393–96.
22. Attal N, Jazat F, Kayser V, Guilbaud G. Further evidence for „pain-related? behaviours in a model of unilateral peripheral mononeuropathy, Pain 1990; 41: 235–51.
23. Ossipov MH, Lai J, Malan Jr TP, Porreca F. Spinal and supraspinal mechanisms of neuropathic pain. Ann NY Acad Sci 2000; 909: 12–24.
24. Yeomans DC, Pirec V, Proudfit HK. Nociceptive responses to high and low rates of noxious cutaneous heating are mediated by different nociceptors in the rat: behavioral evidence. Pain 1996; 68: 133–40.
25. Ossipov MH, Bian D, Malan Jr TP, Lai J, Porreca F. Lack of involvement of capsaicin sensitive primary afferents in nerve-ligation injury induced tactile allodynia in rats. Pain 1999; 79: 127–33.
26. Willis WD, Al-Chaer ED, Quast MJ, Westlund KN. A visceral pain pathway in the dorsal column of the spinal cord. Proc Natl Acad Sci USA
Fig 1: Comparison of withdrawal duration (sec) in sham group and CCI group at the basal, day 1, 3, 5, and 7 days post surgery by sensory tests like (a) Spontaneous pain, (b) Dynamic allodynia, (c) Cold allodynia and (d) Mechanical hyperalgesia. Each data point represents the mean ± S.E.M. The significance of differences between the sham group and CCI group values was determined by unpaired t-test. *PRadiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524141EnglishN-0001November30HealthcareISOLATION OF SALMONELLA TYPHI FROM DIFFERENT WATER SOURCES AND IDENTIFICATION
OF DRUG RESISTANT GENES CAT P AND TEM.
English4450Sita Lakshmi TEnglish Geetha R.VEnglish Devi.GEnglish Anitha RoyEnglishThe aim of the present study was to isolate Salmonella typhi from various water sources and identification of drug resistant genes cat P and tem using Multiplex PCR technique. Enteric fever is prevalent world over and continues to be a major public health problem in developing countries. Infection with Salmonella typhi, the causative organism of this disease, requires effective antimicrobial chemotherapy in order to reduce mortality. Antibiotic-resistant strains of Salmonella are now encountered frequently and the rates of multidrug-resistance have increased considerably in recent years. In the present study, 30 samples were collected from various water sources, of which Salmonella typhi was isolated from 12 samples. All the isolates were subjected to antimicrobial susceptibility testing by standard disc diffusion method against 10 antibiotics. Isolates resistant to chloramphenicol and ampicillin were selected and the drug
resistant genes were identified as cat P and tem respectively by Multiplex PCR technique. There was a very good correlation between the genotypic analysis by PCR and the phenotype determined by standard methods of susceptibility testing.
EnglishMultiple drug resistance, Salmonella typhi, Disc diffusion technique, DNA extraction, Agarose gel electrophoresis, Multiplex PCRINTRODUCTION
Water meant for human consumption should be free from pollution and should be safe and acceptable. Indeed the microbial quality of potable water should not exceed limits specified in the water quality guidelines (APHA – 1998).1 The presence of bacteria and pathogenic (diseasecausing) organisms is a concern when considering the safety of drinking water.2 Human and animal wastes are the primary source of bacteria in water. These sources of bacterial contamination include runoff from feedlots, pastures, dog runs, and other land areas where animal wastes are deposited.3,4 Additional sources include seepage or discharge from septic tanks, sewage treatment facilities and natural soil or plant bacteria. Bacteria from these sources can enter wells that are either open at the land surface, or do not have watertight casings or caps. Pathogenic organisms can cause intestinal infections, dysentery, hepatitis, typhoid fever, cholera, and other illnesses.5
Salmonella typhi is the primary aetiological agent of typhoid fever which is responsible for significant morbidity and mortality, particularly in the developing countries. S. typhi is an obligate human pathogen. It causes infection by the faeco-oral route. Typhoid fever is typically acquired by ingesting food or water that has been contaminated by faeces of typhoid-infected individuals. Many outbreaks, caused either by consumption of contaminated water or food have been report.6 Effective antimicrobial therapy is required for the treatment of typhoid fever to reduce morbidity and mortality. Historically, the drugs of choice were chloramphenicol, ampicillin, and co-trimoxazole. Antibioticresistant strains of Salmonella are now encountered frequently and the rates of multidrug-resistance have increased considerably in recent years. Chloramphenicol resistance is known in Salmonella Typhi since 1972, when plasmids of incompatibility group Inc H, coding for chloramphenicol resistance were found in S. typhi. Multi drug resistance, (defined as resistance to all the first line antibiotics used to treat typhoid fever, i.e chloramphenicol, ampicillin, cotrimoxazole and tetracycline) has been endemic in India since 1984. MDR S. typhi have emerged as the newer challenges to treatment of typhoid fever.7-10 So in this study ,we have made an attempt to isolate Salmonella typhi from the water samples collected from in and around Chennai and to identify the resistant gene ,responsible for resistance to the conventional drug therapy of typhoid,which is a life threatening endemic infection.
MATERIALS AND METHODS
A total of 30 water samples were collected from various water bodies like sewage, stagnant, marine, well and ponds from different areas in and around Chennai. Six different samples were collected from above mentioned water sources and it was brought to the laboratory for further processing.
ISOLATION OF SALMONELLA TYPHI
For the isolation of Salmonella typhi, 1ml water samples were inoculated in 9ml selenite-F-broth and incubated for 18hrs at 37oC for enrichment. The enriched samples were plated onto Salmonella-Shigella Agar (Oxoid) and incubated for 24hrs at 37oC. Small colorless colonies suggestive of S.typhi were subjected to preliminary tests like Gram staining and motility and biochemical reactions like Indole, Methyl red, Voges prausker, Citrate utilization, Urease test, Triple sugar iron test and further confirmed by slide agglutination test with specific antisera.10,11 The confirmed colonies of Salmonella typhi were stored at 40C in nutrient agar slant.
ANTIBIOTIC SUSCEPTIBILITY TESTING
The antibiotic susceptibility pattern of the isolated stains were found by disc diffusion technique [Kirby Bauer method]12, against the following antibiotics like Ampicillin(A30mcg/disc), Chloramphenicol(C30 mcg), Tetracycline(TE 30 mcg), Co-trimoxazole( CO 23.75 mcg), Ciproflaxacin(CIP 5mcg), Gentamycin(G 10 mcg), Ceftriazone (Ci 30 mcg) ,Nalidixic acid (Na 30 mcg), Nitrofurantoin (NIT 300 mcg), Amoxycillin (AMC 30mcg). The antibiotic discs were purchased from Hi- media laboratories Pvt limited, Mumbai. Broth culture of the bacterial strains compared to Mac Farland?s standard 11,13 0.5 were prepared. Lawn culture of the test organisms were made on the Muller Hinton agar [MHA-Hi media M1084] plates using sterile cotton swab and the plates were dried for 15 minutes. Filter paper discs impregnated with different antibiotics were placed on the plates. The plates were incubated at 37°C overnight and the zone of inhibition of growth was measured in millimeter diameter.
EXTRACTION OF DNA
Overnight broth cultures of Salmonella typhi was taken for DNA extraction following manufacturer?s instruction using LTRC kit MSS 12. The broth culture was centrifuged at 10,000 rpm for 10 minutes. The supernatant was discarded and the pellet obtained after centrifugation was used for DNA isolation. The pellet was suspended in 300µl of solution A ( 200 µl 1 x TE buffer and of SDS) at room temperature and vortexed completely. It was then incubated at 60° C for 20 minutes and then solution B (300 µl of Phenol: Chloroform: Isoamyl alcohol) was added and completely vortexed. It was centrifuged at 10,000 rpm for 10 minutes. About 500µl of the aqueous supernatant solution was taken in a fresh vial and an equal volume of isopropanol was added and mixed throughly by inverting the vials. It was again kept for centrifugation at 10,000 rpm for 10 minutes. About 200µl of ethyl alcohol was added to the vial and mixed by inverting the tube till the white strands of DNA precipitation were seen. It was then centrifuged at 10000 rpm for 10 minutes and the supernatant was discarded. Then the alcohol was decanted without dislodging the pellet. It was completely air dried to remove ethyl alcohol smell from the vial. To the final pellet, about 20µl of TE buffer was added and mixed by tapping the tube till the solution settle at the bottom. The isolated DNA was separated and visualized with the help of agarose gel electrophoreses and viewed in the UV transilluminator. 15,16
AGAROSE GEL ELECTROPHORESIS
Agarose gel electrophoresis is the easiest and commonest way of separating and analyzing DNA. The DNA is visualized in the gel by addition of ethidium bromide. This binds strongly to DNA by intercalating between the bases and is fluorescent.17 1% agarose gel, was prepared by weighing and mixing1g agarose in 100ml of 1 x TBE buffer in a conical flask. The solution was heated until the agarose was completely dissolved. The gel casting tray was prepared by sealing ends of gel chamber with tape and appropriate number of combs were placed in gel tray. 5 ul of ethidium bromide was added to cooled gel and poured into gel tray and allowed to cool for 15-30 min at room temperature. The combs were removed and the gel is placed in electrophoresis chamber and covered with TAE buffer. 20 µl of digested DNA was loaded along with 3 µl of gel loading buffer onto gel and electrophoresed at 100V for 1 h. The DNA bands were visualized using UV transilluminator.18,19
MULTIPLEX PCR
The multiplex PCR assay offers a rapid, simple and accurate identification of antibiotic resistance profiles and could be used in clinical diagnosis as well as for the surveillance of the spread of antibiotic resistance determinants in epidemiological studies. 20,21 . The protocol developed by Asma Haque et al., 2005 was followed for the amplification of DNA. 22 The procedure was carried with the following PCR temperature cycling parameters: Initial denaturation at 95°C for 45 sec followed by 35 cycles of denaturation at 95°C for 45 s; Primer annealing at 51°C for 45 s, primer extension at 72°C for 1 min and 30 s and the final extension at 72°C for 10 min. After the reaction, 5 µl of loading dye was added to amplify PCR products and mixed well. Then, 20 µl of total sample was loaded on 1% Agarose Gel Electrophoresis.
RESULTS AND DISCUSSION
A total of 30 samples were collected from various water bodies. Out of this, Salmonella typhi was isolated from 12 samples. The isolated organisms were confirmed by subjecting to preliminary and various biochemical tests, the results of which are given in table I. Confirmed organisms were subjected to antibiotic susceptibility testing using disc diffusion technique and the zone of inhibition was measured in mm diameter. The results are interpreted as sensitive(S) and resistant(R) and given in table II. Out of 12 isolates, six were resistant to chlorampenicaol. Among this, six strains resistant to chloramphenicol, five were resistant to ampicillin also. The DNA from the samples was extracted and bands, were observed by performing Agarose gel electrophoresis. The isolated DNA was used as template in the PCR study. The protocol developed by Asma Haque et al., 2005 was followed for the amplification of DNA. Multiplex PCR was employed to generate genomic amplification products of Salmonella species. The results showed that there was a very good correlation between the genotypic analysis by PCR and the phenotype determined by standard methods of susceptibility testing and identification of salmonella species:
Table II : Results of Antibiotic susceptibility testing of Salmonella typhi A 30- Ampicillin, C 30- Chloramphenicol, TE 30- Tetracycline, CO 25- Co-trimoxazole, CIP 5- Ciproflaxacin, , Na 30- Nalidixic acid, G 10- Gentamycin, NF300- Nitrofurantoin , Ci30- Ceftriazone , AMC 30- Amoxycillin.
Salmonellae are primarily intestinal parasites of humans and many other animals. They are found frequently in sewage, river and other waters, and soil. The presence of Salmonella in other habitats like water, food, natural environment is due to faecal contamination. Under suitable environmental conditions, they may survive for weeks in waters and for years in soils. The prevalence of salmonellosis depends on the water supply, waste disposal, food production and preparation practices, and climate.
Treatment with an appropriate antibiotic is essential for Salmonellosis. Multi drug resistant S. typhi are now endemic in many developing countries and are responsible for significant morbidity and mortality. As a result of the proliferation of such strains, the use of chloramphenicol has been compromised, and that of ampicillin and trimethoprim similarly impaired. Our present study was to isolate Salmonella typhi from various water sources. Isolates resistant to chloramphenicol and ampicillin were selected and the drug resistant genes were identified as cat P and tem respectively by Multiplex PCR technique.
CONCLUSION
Though public health, sanitation and vaccines do have a role to play in control of typhoid fever, it is the antimicrobial therapy which plays a key role in management of typhoid fever. It is likely that S. typhi will continue to acquire genes responsible for antibiotic resistance. Restrictions on the irrational use of antibiotics, and public awareness activities should be undertaken to alert the public to the risks of the unnecessary use of antibiotics .The timely detection of these genes and prevention of their spread is the need of the hour to control antibiotic resistance among S. typhi. More research could also be directed towards factors responsible for their spread and means to prevent dissemination of such plasmids so as to limit drug resistance.
ACKNOWLEDGEMENT
The authors are grateful to Dr.I Seetha Lakshmi,Director of Life Tech Research Centre, Chennai-26 and the Principal ,Valliammal Ammal College for Women,Chennai-102 for their kind support to carry out this project work.
Englishhttp://ijcrr.com/abstract.php?article_id=1984http://ijcrr.com/article_html.php?did=19841. APHA (1998). Standard Methods for the Examination of Water and Wastewater, 20th edition. American Public Health Association, Washington.
2. Dufour, A. and Ballentine, R. (1986). Ambient Water Quality Criteria for Bacteria 1986. USEPA, Washington DC
3. Jim Wright, Stephen Gundry and Ronan Conroy, Household drinking water in developing countries: a systematic review of microbiological contamination between source and point-of-use. Tropical Medicine and International Health volume 9 no 1 pp 106–117 January 2004
4. Health Risks From Microbial Growth and Biofilms in Drinking Water Distribution Systems U.S. Environmental Protection Agency Office of Ground Water and Drinking Water Standards and Risk Management Division 1200 Pennsylvania Ave., NW Washington DC 20004 [2002]
5. J.A. Cotruvo, A. Dufour, G. Rees, J. Bartram, R. Carr, D.O. Cliver, G.F. Craun, R. Fayer, and V.P.J. Gannon. World Health Organization (WHO). Waterborne Zoonoses: Identification, Causes and Control.Published by IWA Publishing, London, UK.
6. S Kumar, K Balakrishna, G.P. Singh and H.V. Batra ,Rapid detection of Salmonella typhi in foods by combination of immunomagnetic separation and polymerase chain reaction World Journal of Microbiology and Biotechnology Volume 21, Number 5, 625-628.
7. Threfall EJ, Antimicrobial drug resistance in Salmonella: problems and perspectives in food- and water-borne infections. FEMS Microbiol Rev. 2002 Jun;26(2):141-8
8. Shrikala Baliga. Drug Resistance in Salmonella Typhi: Tip of the Iceberg Online J Health Allied Scs.2004;4:1
9. Cooke FJ, Wain J. The Emergence of Antibiotic Resistance in Typhoid Fever. Travel Medicine and Infectious Disease 2004; 2:67-74
10. Collins,CH and Lyne, P.M 1976.Microbiological methods, London, Butterworths and co.288p
11. J.G.Colle., A.G.Faster., B.P.Marmion., A.Simmons. Practical Microbiology [Mackie and Mc Cartney] 14th edition page 851 – 85
12. Betty A.Forbes., Daniel F.Sahm., Alice S.Weissfeld. Bailey and Scott?s Diagnostic Microbiology 11th edition Mosby page 229 – 257
13. Connie R.Mahon., George Manuselis., Saunder?s Diagnostic Microbiology 2nd Edition
14. L.P. Samaranayake.,Brain M.Jones., Essential Microbiology for Dentistry. Second edition page 217 – 223.
15. Rui Huang, Shuyan Wu, Xueguang Zhang and Yanyun Zhang.,Molecular Analysis and Identification of Virulence Gene on pRST98 from Multi-Drug Resistant Salmonella typhi Cellular and Molecular Immunology, Volume 2 Number 2 April 2005
16. Alessandra Carattoli, PlasmidMediated Antimicrobial Resistance in Salmonella enterica. Curr. Issues Mol. Biol. (2003) 5: 113-122.
17. Sambrook J, Russel DW (2001). Molecular Cloning: A Laboratory Manual 3rd Ed. Cold Spring Harbor Laboratory Press. Cold Spring Harbor, NY. 18. Brody, J.R., Kern, S.E. (2004): History and principles of conductive media for standard DNA electrophoresis. Anal Biochem. 333(1):1-13.
19. David Freifelder, Molecular Biology 2 nd edition 2006:pg 72-76
20. Ali Karami, Zeynab Ahmadi, Zahra Safiri, Fateme Pourali, Detection of Salmonella strain by rapid-cycle multiplex PCR , Junidishapur journal of microbiology (2011); 4(2): 91-98.
21. Cheng-Hsun Chiu And Jonathan T. Rapid Identification Of Salmonella Serovars In Feces By Specific detection Of Virulence Genes, Inva And Spvc, By An Enrichment broth Culture-Multiplex PCR Combination Assay, Journal Of Clinical Microbiology, Oct. 1996, P. 2619– 2622 Vol. 34, No. 10
22. Asma Haque, Abdul Haque, Yasra Sarwar, Aamir Ali, Saira Bashir, Mushkoor Moshin.2010. Identification of drug resistance genes in cinical isolates of Salmonella typhi for development of diagnostic Multiplex PCR. Medical Sciences,vol.21,N(4),P.402-407.
23. Deepa Anbazhagan etal., Multiplex polymerase chain reaction (PCR) assays for the detection of Enterobacteriaceae in clinical samples, African Journal of Microbiology Research Vol. 4(11), pp. 1186-1191, 4 June, 2010
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524141EnglishN-0001November30General SciencesPOLYANILINE COATED EXPANDED GRAPHITE ACTS AS A NEW ELECTRODE MATERIAL FOR ETHANOL
FUEL CELL
English5155Abhik ChatterjeeEnglish I BasumallickEnglishAn expanded graphite electrode coated with a thin layer of polyaniline and loaded with platinum has been used to study electro-oxidation of ethanol. The graphite plate electrode was expanded by doping with potassium (K)-vapour using vapour incorporation technique developed at our laboratory. The expanded graphite was platinised by electro-deposition
technique and its electrochemical behaviour was examined using a laboratory model fuel cell. The open-circuit potential (OCP) and shortcircuit current (SCC) values of laboratory model fuel cell in 1M H2SO4 comprising of normal and expanded graphite electrodes having same catalytic loading as working electrode were observed. The expanded graphite electrode was also coated with a thin film of polyaniline and then loaded with Pt catalyst as before. Cyclic voltammogram of polyaniline coated electrodes in H2SO4 solution was investigated. Electrocatalytic activity of the modified electrode is better than Pt deposited electrode due to the higher reaction area of Pt particles.
EnglishPolyaniline, ethanol, electrocatalyst, expanded graphiteINTRODUCTION
Metal microparticles dispersed in polymer modified electrodes have been widely used as electro-catalyst. There are various polymeric films, such as polypyrrole (PPY), polyaniline (PANI), poly(3- methylethiophene) (PMT), poly(3,4- ethylenedioxythiophene) (PEDOT), and so forth, have been investigated as conducting catalyst supports 1,2 . These polymers are usually used as matrix to incorporate noble metal catalysts in the application for electro-oxidation of small molecules such as hydrogen, methanol and formic acid, etc 3,4,5 . Among the various conducting polymers, polyaniline (PANI) is one of them most interesting material because of its moderately high conductivity, well behave electrochemistry, easy preparation and possible applications as electro-catalyst towards various electro-oxidation reactions. A thin film of a conducting polymer (CP) improves the interfacial properties between the electrode and the electrolyte. Conducting polymer can allow a facile flow of electronic charge during the electrochemical oxidation of alcohol on Pt. Generally, an electrochemically deposited conducting polymer develops three dimensionally on a substrate 6 . Therefore, it introduces a high porosity and roughness; as a result it generates a large surface area for electrochemical reactions. The major problems of fuel cells are poisoning of the electro-catalyst even with Pt as the catalyst and its crossover from anode to cathode compartment. Besides elelectro-catalyst, the carbon material used as catalytic support also plays an important role in dictating fuel cell efficiency. In the present research an attempt has been made to improve catalytic activity of ethanol (EtOH) fuel cell using polyaniline coated expanded graphite as catalytic support material.
MATERIAL AND METHODS
H2SO4 (Merck), H2PtCl6,6H2O (Arora Matthey Limited) were used as supplied. Aniline (Merck) and EtOH (Bengal chemicals) were distilled before use. The distillate was collected rejecting head and tail fractions. The anode was a 1cm2 graphite plate of thickness 3.8 mm obtained from the R&D of BHEL, India. Graphite plates were expanded using a simple technique developed at our laboratory. The experimental arrangement for expansion is shown in figure 1 7 . The graphite plate was expanded to an extent of 20% by potassium vapour. Pt particles were deposited on the working electrode (2 cm2 ) under galvanostatic condition using +10 mA.cm-2 current for 30 minutes from chloroplatinic acid solution (0.01M) in 0.5M H2SO4. The air cathode was fabricated by loading a 1cm2 graphite plate with Pt. For the sake of comparison a Pt loaded anode without expansion was used. All experiments were carried out at 30oC. Polyaniline was prepared galvanostatically. Electrolyte solution was 0.5 M aniline in 1M H2SO4 solution. The solution was taken in a one compartment cell in which supporting material was used as working electrode and counter and reference electrode merged to Pt wire. -1 mA.cm-2 current under galvanostatic condition was passed for 100seconds to deposit a film of PANI. After polymerization the polymer coated electrodes were washed repeatedly with distilled water and used for electrochemical studies. Cyclic voltammogram of PANI coated electrodes in H2SO4 solution was investigated using a potentiostat-galvanostat (PAR Versastat TM II). Laboratory model fuel cell Laboratory model fuel cells were housed in two glass rectangular chambers (50ml capacity) connected by an inverted Ushaped bridge. Cathode and anode, as fabricated above, were inserted into the respective chambers through the openings of the lid. H2SO4(1M) were poured into cathode and anode chambers, respectively. Alcohol was added to the anode chamber to obtain the desired concentration. The U tube was plugged with foam, which had been soaked in the acid to avoid crossover of alcohol. Air was bubbled slowly through cathode chamber using an air pump.
RESULTS
Figure 2 shows the galvanostatic polymerization curve of aniline on support material surface from a deposition bath of 0.5M aniline in 1M H2SO4. From the figure, it is clear that the polymerization starts at about potential of 0.95 volts and continues in the potential range of 0.9 to 1.0 V for a period of 100 seconds. Figure 3 shows the cyclic voltammogram of expanded graphite-PANI electrode in H2SO4 solution (blank) at a scan rate of 50 mV.s-1 . It is found that within the potential limits – 0.2 to 1.3 V vs. SCE, the response changed on cycling 8 . After about the five cycles, the PANI film shows a steady response and then it was used as an anode material for deposition of Pt. The open-circuit potential (OCP) and shortcircuit current (SCC) values of laboratory model fuel cell in 1M H2SO4 comprising of normal and expanded graphite electrodes having same catalytic loading as working electrode are shown in table1. Steady open-circuit potentials (OCP) were reached within 30 minutes. The potential remained undisturbed for an appreciable duration which indicates that these electrodes are stable in acidic medium. The shortcircuit current (SCC) values were measured after attainment of a steady OCP value. These data demonstrates the role of support material on the activity of catalyst. To compare the performances of laboratory model fuel cells with different anodes, cells were discharged under 0.20 mA current for 180 minutes and profiles are presented in figure 4. The observed discharge profiles indicate that the cell with expanded graphite electrode exhibits better performances.
DISCUSSION
The CV of PANI coated electrode shows two peaks (figure 3). In the anodic scan, the peaks at about 0.57 V is due to SO4 2- anion up taking. In the reverse scan, the peaks at about 0.23 V is due to SO4 2- anion expulsion 8 . Generally, graphite electrodes are utilised for providing support only, no electrocatalytic activity has been observed for alcohol fuel cell. Pt deposited on support material is known as an efficient electrocatalyst for alcohol oxidation. But the catalytic activity of the electrode can be enhanced if the support material is modified. It is seen from the table that OCP value is higher for Pt loaded expanded graphite material electrode. This indicates better catalytic effect of expanded graphite.
Electro-oxidation of EtOH takes place via an initial absorption onto the anode surface followed by deprotonation. For expanded graphite the nanographite channels act as better absorption sites 7 and the electrocatalytic activity of Pt within these channels is also enhanced as is reflectd in their SCC values (table1). Basically, potassium metal as dopant changes the interlayer distance between graphite layers. Table1 also indicates the relative improvement of catalytic effect of normal and expanded graphite when loaded with Polyaniline and Pt. It is very interesting that when Pt particles were deposited onto PANI coated electrode, catalytic effect is further improved. This is due to high dispersion of Pt particles. As a result the specific reaction area of these electrodes is increased and thus improves catalytic efficiency. The observed discharge profiles (figure 4) indicate that the cell with expanded graphite electrode exhibits better performance. The reason is the higher tolerance of the platinum particles to poisoning effect, in comparison with the serious problem of poisoning effect on bulk platinum electrodes. PANI may adsorb some of the poisonous intermediates and thus the adsorption prevents the dispersed Pt particles from becoming deactivated 9 . Thus this research introduces the use of polyaniline modified expanded graphite as an efficient electrode material in fuel cell.
ACKNOWLEDGEMENTAuthors acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors are also grateful to authors / editors /publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed.
Englishhttp://ijcrr.com/abstract.php?article_id=1985http://ijcrr.com/article_html.php?did=19851. Patra S, Munichandraiah N. Electrooxidation of methanol on Ptmodified conductive polymer PEDOT. Langmuir 2009; 25 (3): 1732–1738.
2. Swathirajan S, Mikhali YM. Methanol oxidation on Platinum-Tin catalysts dispersed on Poly(3-methyl) thiophene conducting polymer. J Electrochem Soc 1992; 139( 8) :2105-2110.
3. Kitani A, Akashi T, Sugimoto K, Ito S. Electrocatalytic oxidation of methanol on platinum modified polyaniline electrodes. Synth Met 2001; 121(1- 3):1301-1302.
4. Liu F, Huang L, Wen T, Gopalan A. Large-area network of polyaniline nanowires supported platinum nanocatalysts for methanol oxidation. Synth Met 2007; 157(16-17):651-658.
5. Kost KM, Bartak DE, Kazee B, Kuwana T. Electrodeposition of platinum microparticles into polyaniline films with electrocatalytic applications. Anal Chem 1988; 60(21): 2379-2384.
6. Patra S, Barai K, Munichandraiah N. Scanning electron microscopy studies of PEDOT prepared by various electrochemical routes. Synth Met 2008; 158(10): 430–435.
7. Bhattacharya A, Hazra A, Chatterjee S, Sen P, Laha S, Basumallick I. Expanded graphite as an electrode material for an alcohol fuel cell. J Power sources 2004; 136: 208-210.
8. Hatchett DW, Josowicz M , Janata J. Acid Doping of Polyaniline: Spectroscopic and Electrochemical Studies. J Phys Chem B 1999; 103(50): 10992-10998.
9. Yano J, Shiraga T, Kitani A. Dispersed platinum and tin polyaniline film electrodes for the anodes of the direct methanol fuel cell. J Solid State Electrochem 2008; 12:1179-1182.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524141EnglishN-0001November30General SciencesANTIMICROBIAL ACTIVITY OF NIGELLA SATIVA, ACORUS CALAMUS, MYRISTICA FRAGRANS AND
HEMIDESMUS INDICUS AND ITS SYNERGISTIC EFFECT WITH ANTIBIOTICS
English5664A.VidhyaEnglish V.GopikrishnanEnglish M.RadhakrishnanEnglish R. BalagurunathanEnglishMethanol and aqueous extracts of four plant species namely Nigella sativa, Acorus calamus, Myristica fragrans and Hemidesmus indicus traditionally used in Indian folklore medicine for the treatment of various bacterial and fungal infections were investigated for antimicrobial activity against pathogens viz Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus sp., Escherichia coli and Candida albicans by well diffusion and minimum inhibitory concentration
(MIC) methods. Comparative study of the antimicrobial activity of the plant extracts and its synergistic effect with antibiotics was carriedout. Maximum zone of inhibition was observed against S. aureus, E.coli, Bacillus sp and P. aeruginosa by methanol extracts of Nigella sativa and Hemidesmus indicus. C albicans showed susceptibility to methanol extracts of Acorus calamus and Hemidesmus indicus. Phytochemical analysis of methanol and aqueous extracts of Nigella sativa and Hemidesmus indicus revealed the presence of alkaloids, flavonoids, triterpenes, polyphenols and sterols. Both the active extracts were separated by adopting analytical thin layer chromatography. Totally two spots were observed for Hemidesmus indicus and Nigella sativa showed only one spot, when chloroform: methanol (70:30) used as a solvent system. The active fraction was determined by bioautography by using S. aureus as test organism. Only Nigella sativa extract alone showed synergistic effect with antibiotics and the positive results were showed by S. aureus and E. coli. The results provide justification for the use of the plants in folk medicine to treat various infectious diseases.
EnglishNigella sativa, Hemidesmus indicus, Antimicrobial Activity, Synergistic effect, BioautographyINTRODUCTION
India is endowed with a rich wealth of medicinal plants which have been a valuable source of natural products for maintaining human health. A large number of these medicinal plants are used in several formulations for the treatment of various diseases caused by microbes. According to World Health Organization, medicinal plants would be the source of obtaining a variety of drugs. Various societies across the world have shown great interest in curing diseases using plants/ plant based drugs. Microbes are closely associated with the health and welfare of human beings. Some are beneficial and some are detrimental. As preventive and curative measures, plants and their products are used in the treatment of infections for many centuries ago. WHO estimated that 80% of the people worldwide rely on plant based medicines for their primary healthcare1,2 and India happens to be the largest user of traditional medical cure, using 7000 plant species. The increasing failure of chemotherapies and antibiotic resistance exhibited by pathogenic microbial infections agents have led to the screening of several medicinal plants for their potential antimicrobial activity 1,3, 4 . Antibacterial properties of various plants parts, such as leaves, seeds and fruits have been well documented for some of the medicinal plants for the past two decades1,5 . Antibiotic principles are distributed widely among angiospermic plants. A variety of compounds are accumulated in plant parts accounting for their constitutive antimicrobial activities 1,6 . Within the recent years, infections have increased to a great extent and antibiotic resistance effects become an ever-increasing therapeutic problem. Natural products of higher plants may possess a new source of antimicrobial agents with possibly novel mechanisms of action. They are effective in the treatment of infectious diseases while simultaneously mitigating many of the side effects that are often associated with synthetic antimicrobials. Therefore, it is of great interest to carry out a screening of some plant materials in order to validate their use in folk medicine and to reveal the active principles by isolation and characterization of their constituents7 . The present study aimed to screening and evaluating the antimicrobial assay of Nigella sativa, Acorus calamus, Myristica fragrans and Hemidesmus indicus against five pathogenic microorganisms.
MATERIALS AND METHODS
Collection of plant Material
Nigella sativa seeds, Rhizome of Acorus calamus, Root of Hemidesmus indicus and Myristica fragrans seeds were collected from nearby areas in Vellore (Latitude „12° 55' N? North: Longitude 79° 11' E? East) Tamilnadu.
Preparation of plant extracts
Plant samples were air dried at room temperature and dried samples were coarsely powdered with pestle and mortar. Each 20 gms of powdered material were completely soaked in 100ml of sterile water and 70% methanol and then covered with aluminum foil. The Extraction was allowed to proceed for 48h. The extract was decanted and the solvents were removed by evaporation in vacuom by rotary evaporator. The air dried extracts were used for the antimicrobial and phytochemical analysis8 .
Microorganisms used
A total of 5 clinical isolates of bacteria and a fungi belonging to 5 genera comprising of S. aureus, P. aeruginosa, Bacillus sp., E .coli and C. albicans isolated from various clinical samples were obtained from Government hospital, Vellore, Tamilnadu. The clinical isolates were identified by biochemical methods as recommended by Bergey?s Manual of Systemic Bacteriology9 .
SCREENING FOR ANTIMICROBIAL ACTIVITY
(i)Well diffusion method
The 18 hrs bacterial and fungal suspensions were prepared with help of McFarland standard. The aliquot was spread evenly on Muller Hinton agar and Sabouraud?s Dextrose agar. On each plate, equidistant wells were made with a 6 mm diameter with help of sterilized cork borer. Then fifty micro liter of each plant extract containing 0.25mg was aseptically introduced into a respective agar well. Ciprofloxacin (5 g/ml) and amoxicillin (25 g /ml) were used as positive controls and nystatin (5 g/ml) and amphotericin B (25 g /ml) were used for fungal isolates. The methanol and distilled water were used as negative controls and incubated at 37 oC for 24-48 hrs for bacteria and fungus. The formations of clear inhibition zone of ≥12 mm diameters around the wells were regarded as significant susceptibility of the organisms to the extract. The experiment was performed with duplicate10 .
(ii) Phytochemical Analysis
The solvent extracts which showed maximum antibacterial activity was subjected to qualitative test for the identification of various plant constituents such as carbohydrates, proteins, phytosterols and flavonoids11
(iii) Separation of active compound by TLC
The methanol extract of both plants which showed promising activity against the test pathogens were subjected to separation by thin layer chromatography (TLC) using silica gel coated TLC plates 12. About 3µl of active crude extract was spotted at the bottom of the sheet using capillary tube. To find out the better solvent system for separation, the sheet was placed in a beaker containing the different solvent systems, such as methanol, chloroform, acetic acid, n-butanol, n-hexane and water with following proportions: n-butanol: acetic acid: water (60:30:10), (70:20:10), (75:20:5) chloroform: methanol (70:30), (60:40), (40:60), (50:50), and nhexane : chloroform (60:40). After running the chromatogram the sheet was kept in a developing chamber containing potassium iodide crystals. The compound present in the crude extracts were appeared as brown spots 13.
(iv) Detection of active compound by bioautography:
The bioautography method used for the detection of active compound separated in TLC14. Chromatogram developed as described above was placed in a sterile bioassay petridish and overlaid with 10ml of molten nutrient agar seeded with 0.2ml of S aureus as test organism and incubated overnight at 370C for 24 hours.
(v) Minimum inhibitory concentration
The minimum inhibitory concentration (MIC) of the extracts was estimated for each of the test organisms in triplicates. 0.5ml of varying concentrations of the extracts (50-800µg/ml) are added with 2ml of nutrientbroth and Sabouraud?s Dextrose broth?s and then a loopful of the test organism previously diluted to 0.5 McFarland turbidity standard for bacterial isolates and 106 cfu/ml for fungal isolates were introduced to the tubes. The procedure was repeated on the test organisms using the standard antibiotics (ciprofloxacin and cotrimoxazole for bacteria and nystatin and ampothericin B for fungal isolates). A tube containing nutrient broth and Sabouraud?s Dextrose broth only were seeded with the test organisms as described above to serve as control. Tubes containing bacterial cultures were then incubated at 37oC for 24 h while tubes containing fungal spore cultures were incubated for 48 h at room temperature (30 – 32oC). After incubation the tubes were then examined for microbial growth by observing for turbidity15 .
(vi) Evaluation of synergistic effect of antibiotics and plant extracts on resistant bacterial samples
Aliquots of 100 μl of resistant bacterial cultures (0.5 MacFarland Standard) grown in 10 ml of nutrient broth for 6 h were inoculated in nutrient broth supplemented with the respective antibiotics (50μg/ml) and 106 cells/ml fungal cultures grown in Sabouraud?s Dextrose broth supplemented with 100 μg /ml coltrimazole with different concentrations of plant extracts. The concentration for plant extracts ranged from 10 to 500 μg/ml, based on MIC values that had previously been evaluated. Only streptomycin or gentamicin was used as the sub-inhibitory concentration (50 g / ml) and incubated at 37oC for 48 hours. After 48 h, the optical density of each sample was recorded and compared to those of MIC to verify the synergistic effect of the tested compounds16 .
RESULTS
(i) Well diffusion method In the present study among the 4 plants screened, the methanolic extracts of N.sativa and H.indicus showed promising activity and found to be more effective than aqueous extract except for Nigella sativa. Maximum zone of inhibition were observed against S.aureus, P.aeruginosa and E.coli by aqueous and methanol extracts of N.sativa and H.indicus. Bacillus sp. Showed susceptibility to N. sativa, M. fragrans and H.indicus extract. C.albicans inhibited by methanol extracts of A.calamus and H.indicus. The results of the antimicrobial activity of plant extracts tested against microorganisms by well diffusion method are shown in Table -1 & 2,2A.
(ii) Separation of active compound by TLC
Methanol extract of H.indicus has showed two clear spots, when chloroform: methanol (70:30) used as solvent system. Their Rf values are calculated as 0.56 and 0.88. Aqueous extract of N. sativa showed only one clear spot, when chloroform: methanol (70:30) used as a solvent system and its Rf value was calculated as 0.75.
(iii) Detection of active compounds by bioautography:
In bioautography, two spots of H.indicus and one spot of N sativa, totally three different spots separated in TLC were taken. In this the first spots of H.indicus and N sativa showed good activity against S. aureus with 15-26 mm of inhibition zone.
(iv) Phytochemical Analysis
Methanolic extract of H indicus showed presence of flavonoids, triterpenes, polyphenols and sterols. Aqueous and methanolic extracts of N.sativa only comprised of alkaloids (Table 3). The result will help in identification of various compounds through gas chromatography in the further studies.
(v) Minimum inhibitory concentration method
Aqueous extract of N. sativa and methanol extract of H. indicus was taken for MIC assay. N.sativa extract showed minimum inhibitory concentration to S aureus, E coli, Bacillus sp. and P. aeruginosa at the concentrations of 20µg/ml, 80µg/ml, 20µg/ml and 40µg/ml respectively (Table 4). A. calamus extract showed minimum inhibitory concentration to C. albicans and Bacillus sp. at concentrations of 40 µg/ml and 80 µg/ml respectively (Table 5). The extract of M fragrans exhibited MIC to Bacillus sp alone at concentration of 20mg/ml (Table 6). H. indicus extract showed minimum inhibitory concentration to S aureus, Bacillus sp, E coli, P. aeruginosa and C. albicans at concentrations of 40µg/ml, 80µg/ml, 20µg/ml 160µg/ml and 80µg/ml respectively (Table 7).
(vi) Synergistic effects of antibiotics with plant extract
The comparative studies of the synergistic effect of antibiotics with plant extracts alone were studied. The effect of association of N.sativa extract with streptomycin and gentamicin on S aureus and E coli were shown in figure 1. S aureus showed MIC at concentration of 8µg/ml, when N sativa extract combined with streptomycin and Gentamicin, whereas N.sativa alone showed MIC only at 20µg/ml. E coli showed MIC at 40µg/ml concentration, When N sativa extract combined with streptomycin and gentamicin, whereas N.sativa extract alone showed MIC only at 80µg/ml. No other plant extracts showed synergistic effects with antibiotics to any other test organisms.
DISCUSSION
In the last few decades, there has been particular interest in the use of abundant naturally occurring antimicrobials (herbs, spices and plants) 17 . N.sativa, A.calamus, M. fragrans and H. indicus are traditionally used in Indian folklore medicine. So far only a limited data is available regarding its efficacy against pathogenic microorganisms. Hence the present study was therefore designed to evaluate it?s antimicrobial activity. Methanolic extracts, hot and cold water extracts of N. sativa showed excellent antibacterial activity against pathogenic microbes18. Chloroform extract of H.indicus showed promising activity against the clinical isolates of Helicobacter pylori19. In the present study the methanolic extracts of N.sativa and H.indicus also showed promising activity than the standard antibiotics and methanol extracts found to be more effective than aqueous extract except for N.sativa. In TLC Petroleum-ether extract of N.sativa using benzene: ethyl acetate (6:1), showed five spots. In the chloroform extract, using benzene: ethyl acetate (4:1), five spots and in ethanol extract, using chloroform: methanol (93:7), six spots were observed13. Methanol extract of H.indicus has showed two clear spots and N. sativa showed only one clear spot, when chloroform: methanol (70:30) used as a solvent system. The assay for bioautography demonstrated that the strong inhibition zones of H. indicus and N.sativa against the growth of S aureus. The clear zones were located in separate places on the TLC plate, suggesting the potent antimicrobial effect. In phytochemical analysis, H indicus showed presence of flavonoids, triterpenes, polyphenols and sterols. Aqueous and methanolic extracts of N. sativa only comprised of alkaloids. the antimicrobial activity of black pepper is due to the presence of essential oil (3%), whose aroma is dominated by monoterpenes hydrocarbons: sabinene, β-pinene and limonene. The mechanism of action of terpene is not fully understood but is speculated to involve membrane disruption by the lipophilic compounds20 . In MIC assay, N.sativa extract showed minimum inhibitory concentration against S aureus and Bacillus sp. at the concentrations of 20 µg/ml respectively. H.indicus extract showed minimum inhibitory concentration against S aureus and E.coli at the concentrations of 40µg/ml and 20 µg/ml respectively. This antimicrobial activity may be due to the presence of alkaloids in plant extracts. Synergistic effect was found by S aureus and E coli in the combination of N sativa with streptomycin. and gentamicin. No other plant extracts showed synergistic effects with antibiotics to any other test organisms. In earlier studies, N. sativa seed extract has antimicrobial activity against MRSA21 and it was found to be active against ESBL producers22 . H.indicus showed good activity against Propionibacterium acnes 23. The antimicrobial activity and phytochemical analysis also carried out for N. sativa and H.indicus extracts .But the present study focused on purification, bioautography and comparative study of N.sativa and H. indicus synergestic effects. The results of present investigation clearly indicate that the antibacterial and antifungal activity vary with the species of the plants and plant material used. Thus, the study ascertains the value of plants used in folk medicine, which could be of considerable interest to the development of new drugs.
CONCLUSION
Based on the results obtained in this study, it may be concluded that out of 4 plants screened, plant extracts of N. sativa and H. indicus have a stronger and broader spectrum of antimicrobial activity against pathogenic microorganisms and the extracts may be used to discover bioactive natural products that may serve as basic source for the development of new antimicrobial compounds to overcome the problem of increasing resistance to known traditional antibiotics. The further purification through HPLC and spectral analysis will be worthy study to identify the nature of compound present.
ACKNOWLEDGEMENT
The authors are thankful to Management of DKM College for Women, Vellore, Tamilnadu, India for providing all the facilities and also thankful to Vice Chancellor and Registrar , Periyar University, Salem, Tamilnadu, India . Authors acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript.
Englishhttp://ijcrr.com/abstract.php?article_id=1986http://ijcrr.com/article_html.php?did=19861. Alagesaboopathi C, Antimicrobial screening of selected medicinal plants inTamilnadu, India, African Journal of Microbiology Research 2011:5(6); 617-621.
2. Farnsworth NR. Ethnopharmacology and drug development, In Prance GT (Ed.). Ethnobotany and the Search for New Drugs. Wiley, Chichester (Ciba Foundation Symposium 185). 1994. 42–59.
3. Ritch-Krc EM, Turner NJ and Towers GH. Carrier herbal medicine an evaluation of the antimicrobial and anticancer activity in some frequently used remedies. J. Ethnopharmacol. 1996; 52:152-156.
4. Martins AP, Salgueiro L, Goncalves MJ, Proenca cunha V, Vila R, Canigueral S and Mazzoni V. Essential oil composition and antimicrobial activity of three Zingiberaceae from S. Tomee principe. J. Planta Med. 2001; 67: 580-584
5. Leven M, Vannen Berghe DA and Mertens F. Medicinal Plants and its importance in antimicrobial activity. J. Planta Med. 1979;36:311-321
6. Callow JA. Biochemical plant phology. A Wiley. Interscience Pub. 1983
7. Mohamed S. Hansi H and Kavitha D. Antimicrobial activity and phytochemical analysis of selected Indian folk medicinal plants. International Journal of Pharma Sciences and Research, 2010;1(10):430-434.
8. Okogun JI. Methods of Medicinal Plant Extract Preparation. National Institute for Pharmaceutical Research and Development (NIPRD) . Idu – Abuja, Nigeria. 2000.
9. Brenner DJ, Krieg NR, Garrity GM and James T. Staley Bergey?s Manual of Systemic Bacteriology. Springer. 2005
10. Biruhalem T, Mirutse G, Abebe A and Jemal S., Antibacterial activities of selected medicinal plants in traditional treatment of human wounds in Ethiopia. Asian Pacific Journal of Tropical Biomedicine. (2011),370-375
11. Ramalan A, Harrax, FM and Ezmougy SA. Fitoterapia .1998; 65(5): 418-422.
12. Balagurunathan R and Subramanian A. Isolation and purification of γ- lactone antibiotic from Streptomyces grisobrunneus. International symposium on bioproducts processing Technologies for the Tropic, University of Malaya, Malaysia, 1994, 315-316.
13. Neeraj K, Dheeraj A, Sandeep G and Deenanath J., Pharmacognostic standardization, physico and phytochemical evaluation of Nigella sativa linn. Seed. ijpsr. 2011; 2(3):713-718.
14. Selvameenal L, Radhakrishnan M and Balagurunathan R. Antibiotic pigment from desert soil actinomycetes; biological activity, purification and chemical screening. Indian Journal of Pharmaceutical Science. 2009; 71(5): 499–504.
15. Doughari JH. Antimicrobial Activity of Tamarindus indica Linn. Tropical Journal of Pharmaceutical Research.2006; 5(2):597-603.
16. Doughari, JH, Mahmood El and Tyoyina I. Antimicrobial activity of leaf extracts of Senna obtusifolia (L). African Journal of Pharmacy and Pharmacology. 2008 ;2(1):007-013
17. Burt, S. Essential oils: their antibacterial properties and potential applications in foods-a review Int. J. Food Microbiol. 2004, 94: 223– 253.
18. Anwar K, Uzair R, Ayesha S and Sonia K. Antimicrobial activity analysis of extracts of Acacia modesta, Artimisia absinthium, Nigella sativa and Saussurea lappa against Gram positive and Gram negative microorganisms, African Journal of Biotechnology. 2011 ;10(22): 4574-4580,
19. Anoop A, Jegadeesan M and Gowrishanker R. Antimicrobial activity of Hemidesmus indicus var R.Br. Human isolates of Helicobacter pylori, Natural product science. 2003;9(1): 1-3
20. Ali MA, Alam NM , Yeasmin MS , Khan AM and Sayeed A. Antimicrobial screening of different extracts of Piper longum Linn. Res. J. Agri. Biol. Sci. 2007; 3: 852-857.
21. Abdul H, Sidrah S, Saadia C, Muhammad B and Muhammad A. Anti bacterial activity of nigella sativa against clinical Isolates of methicillin resistant staphylococcus aureus. J Ayub Med Coll Abbottabad . 2008;20(3): 72-74.
22. Zuridah H, fairuz ARM, zakri AHZ and Rahim MNA. Invitro antibacterial activity of Nigella sativa against S.aureus, P.aeruginosa, K.pneumoniae, E.coli and B.cereus. Asian journal of plant science. 2008,1-4.
23. Kumar GS , Jayaveera KN , Ashok Kumar CK , Umachigi PS, Vrushabendra Swamy BM and Kishore Kumar DV. Antimicrobial effects of Indian medicinal plants against acne-inducing bacteria. Tropical Journal of Pharmaceutical Research, 2007; 6 (2): 717-723.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524141EnglishN-0001November30HealthcarePHOTODYNAMIC THERAPY - A NOVEL APPROACH IN POCKET STERILIZATION
English6573Jannani MuthuEnglish Jaideep MahendraEnglishAnti microbial photodynamic therapy is one of the most upcoming treatment approaches in field on dentistry. Applications of PDT in dentistry are growing rapidly: the treatment of oral cancer, bacterial and fungal infection therapies, and the photodynamic diagnosis (PDD) of the malignant transformation of oral lesions. Periodontitis as known is polymicrobial disease and this novel therapeutic approach sounds promising in eliminating the microorganisms when usedin adjunct to conventional debridement methods. This technique involves application of a photosensitizer and activating it with a light source of specific wavelength. This system in presence of oxygen creates reactive oxygen species that exerts the classic photodynamic reaction. The advantage of this new approach includes rapid bacterial elimination, minimal chance of resistance development and safety of adjacent host tissue and normal microflora. This review discusses about the general principles, mechanism of photodynamic therapy with additional highlights on its application in periodontal diseases.
EnglishAntimicrobial photodynamic therapy, periodontitis, photosensitizers, lasers.INTRODUCTION
There have been many changes and developments in dentistry over the past decade than in the previous hundred years combined, and the pace is accelerating! Oral cavity is an abode of variety of microorganisms and periodontitis is an infectious disease caused by number of these organisms. Current treatment techniques involve either periodic mechanical disruption of oral microbial biofilms or maintaining therapeutic concentrations of antimicrobials in the oral cavity, both of which are fraught with limitations. The development of alternative antibacterial therapeutic strategies therefore becomes important in the evolution of methods to control microbial growth in the oral cavity. Numerous adjunct conventional antimicrobial therapies have been attempted. One disadvantage of these therapies is development of resistant strains. So alternate antimicrobial treatment modalities have been researched and one such promising alternative is antimicrobial photodynamic therapy1 . The history dates back over 3000 years when Indians used psoralens in the treatment of vitiligo and the Egyptians employed it in the treatment of leucoderma.
Later It was rediscovered by Western civilization at the beginning of the twentieth century. In 1834, Kalbrunner isolated the chemical bergapten from bergamot oil but did not use it in any therapeutic application2 . In 1900 Prime, a French neurologist, used eosin orally in the treatment of epilepsy. He discovered that this induced dermatitis in sun-exposed areas of skin. This discovery then led to the first medical application of an interaction between a fluorescent compound and light3 . It was further developed by the Danish physician, Niels Finsen, who at the turn of the last century described the successful treatment of smallpox using red light. He then went on to use ultraviolet light to treat cutaneous tuberculosis and developed the use of carbon arc phototherapy in the treatment of this condition for which he was awarded a Nobel Prize in 19033 . History of photodynamic therapy began when Von Tappeiner,who along with dermatologist Jesionek, used a combination of topical eosin and white light to treat skin tumors. They demonstrated the requirement of oxygen in photosensitization reactions and in 1907 they introduced the term "photodynamic action" to describe this phenomenon. The German physician Friedrich Meyer–Betz performed the first study with what was first called photoradiation therapy (PRT) with porphyrins in humans in 1913. Meyer. In 1989, First Paper on photodynamic therapy was presented at the Congress on photodynamic therapy of Tumours4 . This review elucidates the evolution and the current position of photodynamic therapy, its applications in dentistry, especially in periodontal treatments and its likely impact in future.
COMPONENTS OF PHOTODYNAMIC THERAPY
Photodynamic reaction is a physicochemical reaction that basically involves 2 components: Photo sensitizer and the activating light source.
Photosensitizers
These are natural or synthetic photoactive compounds that have photosensitizing potential. They function by trapping the light in the form of photons and transferring the energy to other molecules resulting in the liberation of short-lived energetic species that interact with biological systems and produce tissue damage32 . Ideal properties of photosensitizers – this includes photo-physical, chemical, and biological characteristics5 .
> An ideal photosensitizer must be biologically stable.
> Photochemically efficient.
> Selectively retained in the target tissue.
> Low toxicity and fast elimination from the skin and epithelium.
> Absorption peaks in the low-loss transmission window of biological tissues.
> High quantum yield of singlet oxygen production in vivo.
> Cost-effectiveness and commercial availability
> High solubility in water, injection solutions, and blood substitutes. > Storage and application light stability.
Generations – Three generations of photosensitizers have evolved over period of time. I generation includes Hematoporphyrins and pthalocyanines, II generation photosensitizers are mesotetra(hydroxyphenyl)porphyrins marketed as foscan, 5-aminolevulinic acid , tin ethyl etiopurpurin, benzoporphyrin derivative monoacid ring (verteporfin, visudyne) and phenothiazine dyes5,6,7
Phenothiazine dyes includes toluidine blue and methylene blue. These are the recent major photosensitizers used in dental field. Both have similar chemical and physiochemical characteristics. (I).Toluidine blue – It?s a blue-violet solution. It stains granules within mast cells, proteoglycans and glycosaminoglycans. Used to detect mucosal tumors or atypical epithelia.
Englishhttp://ijcrr.com/abstract.php?article_id=1987http://ijcrr.com/article_html.php?did=19871. Raghavendra M, Bhola S, Koregol A. Photodynamic therapy: A target therapy in periodontics. Australian dental journal, 2009; 54(1); 102 – 109.
2. Epstein, J. M. (1990) Phototherapy and photochemotherapy, 1990; N. Engl. J. Med. 32, 1149-1151.
3. Rajvir M and Anish; PDT – A strategic review. Indian journal of dental research, 2010; 21 (2); 285 – 291.
4. Ackroyd R, Kelty C, Brown N. The History of Photodetection and Photodynamic Therapy, Journal of Photochemistry and Photobiology 2006; 74 (5); 656–669.
5. Bown, SG. Tralau, Smith C , Akdemir D, Wieman TJ. Photodynamic therapy with porphyrin and phthalocyanine sensitisation: quantitative studies in normal rat liver. Br. J. Cancer 1986; 54, 43-52.
6. Zheng Huang, M.D., Ph.D. A Review of Progress in Clinical Photodynamic Therapy. Technol Cancer Res Treat. 2005 June; 4(3): 283–293.
7. Solban N, Rizvi I, Hasan T. Targeted photodynamic therapy. Lasers Surg Med 2006; 38:522-531.
8. Komerik, Goslinski T, in vivo killing of p.gingivalis by toluidine blue mediated photosensitization, Antimicrobes agents chemother 2003 : 47:932-940.
9. Whitehurst C, Byrne B, Moore JV. Development of an alternative light source to lasers for photodynamic therapy - Comparative in vitro dose response characteristics. Lasers Med. Sci. 1993; 8, 259-267.
10. Takasaki AA, Aoki A, Mizutani K, Schwarz F, Sculean A. Application of antimicrobial photodynamic therapy in periodontal and peri-implant diseases. Perio 2000, vol-51, 109-140.
11. Sharman WM, Allen CM, Van Lier JE. Role of activated oxygen species in photodynamic therapy. Methods in Enzymology 2000; 319; 376-400.
12. Nikolaos s. Soukos j and Max goodson. photodynamic therapy in the control of oral biofilms. Periodontology 2000, vol. 55, 2011, 143–166
13. Busch TM, Local physiological changes during photodynamic therapy. Lasers Surg Med,2006, 38:494-399.
14. Hamblin MR and Hasan T. Photodynamic therapy: a new antimicrobial approach to infectious disease. Photochem . photobiol . Sci, 2004; 3, 436 – 450.
15. Wainwright M.J. Antimicrob. Chemother.1998; 42; 13-28.
16. Konopa K and Golinsky T; Photodynamic therapy in dentistry. J Dent Res, 2007; 86 (8); 694 – 704.
17. Maisch T, Bosl C, Szeimies RM. Photodynamic effects of novel XF porphyrin derivatives on prokaryotic and eukaryotic cells. Antimicrob Agents Chemother 2005;49(4):1542– 52.
18. Herbert L. Vascular Targeted Photodynamic Therapy for Localized Prostate Cancer, Rev Urol. 2008 Fall; 10(4): 254–261.
19. Haas R, Dörtbudak O, Mensdorff NP, Mailath G. Elimination of bacteria on different implant surfaces through photosensitization and soft laser - An in vitro study. Clinical Oral Implants Research 1997; 8 (4); 249 – 254.
20. Jamil AS, Martins MC, Nociti FH jr, Garcia VG. Treatment of ligatureinduced peri-implantitis by lethal photosensitization and guided bone regeneration: a preliminary histologic study in dogs. J Periodontol 2003; 74(3); 338 – 345
21. Hayek RR, Araújo NS, Gioso MA, Ferreira J, Baptista-Sobrinho CA, Yamada AM, Ribeiro MS. Comparative study between the effects of photodynamic therapy and conventional therapy on microbial reduction in ligature-induced periimplantitis in dogs. J Periodontol 2005; 76(5), 1275 – 1281.
22. Dobson J and Wilson M. Sensitization of oral bacteria in biofilms to killing by light from a low-power laser. Arch oral biol; 1992; 32; 883-887.
23. Wilson M, Burns T, Pratten J, Pearson GJ. Bacteria in supragingival plaque samples can be killed by low-power laser light in the presence of a photosensitizer. J Applied Bacteriol. 1995; 17(5); 569 – 574
24. Packer S, Batthi M, Burns T, Wilson M. Inactivation of Proteolytic Enzymes from Porphyromonas gingivalis Using Light-activated Agents. Lasers Med Sci 1999; 15(1);
25. Dörtbudak O, Haas R, Bernhart T, Matejka M. Photodynamic Therapy for Bacterial Reduction of Periodontal Microorganisms 2001; 1(2); 115 – 118.
26. Kömerik N, Nakanishi H, MacRobert AJ, B. Henderson. In Vivo Killing of Porphyromonas gingivalis by Toluidine Blue-Mediated Photosensitization in an Animal Model. Antimicrob Agents Chemother. 2003; 47(3): 932–940
27. Kömerik N, Nakanishi H, MacRobert AJ, B. Henderson. In Vivo Killing of Porphyromonas gingivalis by Toluidine Blue-Mediated Photosensitization in an Animal Model. Antimicrob Agents Chemother. 2003; 47(3): 932–940
28. Oliveira RR, Schwartz HO, Novaes BA Jr, and Taba M Jr. Antimicrobial Photodynamic Therapy in the NonSurgical Treatment of Aggressive Periodontitis: A Preliminary Randomized Controlled Clinical Study. J Periodontol 2007; 78(6), 965 – 973.
29. Qin YL, Luan XL, Sheng YO. Comparison of toluidine blue-mediated photodynamic therapy and conventional scaling treatment for periodontitis in rats. J periodontal research 2008; 43(2); 162 – 167.
29. Roger Andersen, MD, MPH Nicolas Loebel, PhD David Hammond, BS. Treatment of Periodontal Disease by Photodisinfection Compared to Scaling and Root Planing. J Clin Dent 2007; 18(2); 1 – 5.
30. Braun A, Dehn C, Krause F, Jepsen S. Short-term clinical effects of adjunctive antimicrobial photodynamic therapy in periodontal treatment: a randomized clinical trial. J Clin Periodontol 2008; 35(10); 877 – 874.
31.Christodoulides N, Nikolidakis D, Chondros P, Becker J, Schwarz F, Rössler R, Sculean A. Photodynamic therapy as an adjunct to non-surgical periodontal treatment: a randomized, controlled clinical trial. J Periodontol 2008; 79(9); 1638 – 1644.
32. Oliveira RR, Oswaldo HS, Arthur BN Jr, Garlet GP. Antimicrobial Photodynamic Therapy in the NonSurgical Treatment of Aggressive Periodontitis: Cytokine Profile in Gingival Crevicular Fluid, Preliminary Results. Journal of Periodontology 2009, 80(1); 98-105.
33. Sigusch BW, Engelbrecht M, Völpel A, Holletschke A, Pfister W, Schütze J. Full-mouth antimicrobial photodynamic therapy in Fusobacterium nucleatum-infected periodontitis patients. J Periodontol. 2010;81(7):975-81.
34. Romanos GE, Brink B. Photodynamic therapy in periodontal therapy: microbiological observations from a private practice. Gen Dent. 2010 ;58(2):68-73.
35. Lui J, Corbet EF, Jin L. Combined photodynamic and low-level laser therapies as an adjunct to nonsurgical treatment of chronic periodontitis. J Periodont Res 2011; 46: 89–96.
36. Allison NY. Photosensitizers in clinical PDT. Photodiagn Photodyn Ther 2004; 7; 27-42.
37. Augthun M, Tinschert J, Huber A. In vitro studies on the effect of cleaning methods on different implant surfaces. J Periodontol 1998: 69: 857–864.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524141EnglishN-0001November30General SciencesAPPLICATION OF OPERATIONS RESEARCH WITH SPECIAL REFERENCE TO TRANSPORTATION
PROBLEM
English7485V. P. DeshmukhEnglish S. P. ShindeEnglishDecision making in today?s social and business environment has become a complex task. High costs of technology, material, labor, competitive pressures, different economic, social as well as political factors and view points, greatly increase the difficulty of managerial decision making. For well handling of arising problem the decision maker must examine a problem from both qualitative as well as quantitative approach. In today?s global era Operations Research tools can help the decision maker to solve the arising problem. Operations Research represents the study of optimal resource allocation. In order to make the effective and efficient decisions, managers must have fundamental understanding of the decision science tools utilized in developing set of recommendations to choose from. The Operations research is usually the mathematical treatment, analysis of a process, problem, or Operations to determine its purpose and effectiveness and to gain maximum efficiency. Quantitative methods which comprises of Simulation, Linear and nonlinear programming, Queuing Theory and Stochastic Modeling are well-accepted techniques by both research and practice communities. Project scheduling techniques: PERT and CPM are efficient tools for scheduling and monitoring lengthy, complex and expensive projects of that time. This research Paper focuses mainly on the process of
Operations research and explains some of the applications of Operations Research, elaborates some of the applications and benefits that may be gained by incorporating Operations Research into the actual business framework. The researcher has a wide interest in the foresaid topic andtheir further research area is the same.
English1. INTRODUCTION
In the global age decision making in business environment has become a complex task. High costs of technology, materials, labor, competitive pressures and so many different economic, social as well as potential factors and view points, greatly increase the difficulty of managerial decision making [27]. Whenever some national crises emerges due to the emerges due to the impact of political , economic or cultural factors the talents from all walks of life join together to overcome the situation and solve the problems. These combined efforts always result in new discoveries and techniques. O.R is also outcome of such situations [28]. The term Operations Research describes the discipline that is focused on the application of information technology for informed decisionmaking. Operations Research represents the study of optimal resource allocation. In Operations Research much of the actual work is conducted by using analytical and numerical techniques to develop and manipulate mathematical models of organizational systems that are composed of people, machines, and procedures Some of the problems in the area of hospital management, energy conservation, environmental pollution, etc. have been solved by Operations Research specialists and this is an indication that Operations Research can also contribute towards the improvement in the social life and areas of global need.
1.1 History of Operations Research
Operations Research as a new field started in the late 1930's and has grown and expanded tremendously in the last 30 years. The British army conducted exercises on the radar system for detecting the aircrafts. In July 1938, the Superintendent of Bawdsey Research Station, announced that although the exercise had demonstrated the technical feasibility of the radar system for detecting aircraft, its Operational achievements were not up to what was required. He therefore proposed that a crash program of research into the Operational - as opposed to the technical - aspects of the system should begin. The term "Operational Research" was coined as a suitable description of this new branch of applied science. On 15th May 1940, with German forces advancing rapidly in France, Stanmore Research Section was asked to analyze a French request for ten additional fighter squadrons. They prepared graphs for Winston Churchill (British Prime Minister), based upon a study of current daily losses and replacement rates, indicating how rapidly such a move would deplete fighter strength. No aircrafts were sent and most of those currently in France were recalled. This is held by some to be the most strategic contribution to the course of the war made by Operations Research as the aircraft and pilots saved were consequently available for the successful air defense of Britain, the Battle of Britain. In 1941 Operational Research Section (ORS) was established in Coastal Command which was to carry out some of the most well-known Operations Research work in World War II. Thus Operations Research was born as a separate field of specialization. In order to make the effective and efficient decisions, managers must have fundamental understanding of the decision science tools utilized in developing set of recommendations to choose from. The Operations Research is usually the mathematical treatment, analysis of a process, problem, or Operations to determine its purpose and effectiveness and to gain maximum efficiency. The Operations technique is utilized by functional groups such as Industrial Engineering in effort to support Operations Managers to make economically feasible decisions on a range of systematic challenges. The main responsibilities of Operations management are to manage and operate as efficiently and effectively as possible with the given resources.
Quantitative methods which comprises of Simulation, Linear and nonlinear programming, Queuing Theory and Stochastic Modeling, are well-accepted techniques by both research and practice communities. Functional entities such as Industrial or Systems Engineering use methodologies to provide feasible alternatives for Operations s mangers to decide on. An important component of decision-making process is verifying and validating alternatives, which typically involve decision makers, engineers or analysts. Growth of Operations Research is to a large extent, the result of the widespread availability of computers. Most Operations Research involves carrying out a large number of numeric calculations and without computers this would simply not be possible. In India, Operations Research came into existence in 1949 when an Operations Research unit was established at Regional Research Laboratory, Hyderabad. Also Prof. R.S.Verma set up an Operations Research team at Defense Science Laboratory to solve problems of store, purchase and planning. During the 1950?s there was substantial progress in the application of Operations Research techniques for civilian activities along with a great interest in the professional development and education in OR. Many colleges and universities introduced Operations Research in their curricula. They were generally schools of engineering, public administration, business management, applied mathematics, economics, computer science etc. In 1953, Prof. P.C. Mahalanobis [8] established an Operations Research team in the Indian Statistical Institute, Calcutta to solve problems related to national planning and survey. In 1958, project scheduling techniques: PERT (Program Evaluation and Review Technique) and CPM (Critical Path Method) were developed as efficient tools for scheduling and monitoring lengthy, complex and expensive projects of that time. The real development of Operations Research in the national field was carried out by Prof. Mahalanobis in India when he used it in national planning. It is also being used in Railway, waiting or queuing problems of passengers for tickets at booking windows or trains queuing up in marshalling yard, waiting to be sorted out are tackled by various Operations Research technique.
2.1 Operations Research Process
The actual Operations Research process can be described in the following steps [15]:
Identification of problem
Data collection and mathematical model construction
Solving the mathematical model Identification of optimum solution
Validity of solution
Solution Implementation
Model modification
Establishing controls over the solution
Specific Application Area: Operations Research in Manufacturing The term Operations in Operations Research may suggests that the manufacturing application category represents the original home of Operations Research. That is not quite accurate, as the name originated from military Operations, not business Operations. Nevertheless, it is a true statement that Operations Research?s successes in contemporary business pervade manufacturing and service Operations, logistics, distribution, transportation, and telecommunication. The myriad applications include scheduling, routing, workflow improvements, elimination of bottlenecks, inventory control, business process reengineering, site selection, or facility and general Operational planning. Revenue and supply chain management reflect two growing applications that are distinguished by their use of several Operations Research methods to cover several functions. Revenue management entails first to accurately forecasting the demand, and secondly to adjust the price structure over time to more profitably allocate fixed capacity. Supply chain decisions describe the who, what, when, and where abstractions from purchasing and transporting raw materials and parts, through manufacturing actual products and goods, and finally distributing and delivering the items to the customers. The prime management goal here may be to reduce overall cost while processing customer orders more efficiently than before. The power of utilizing Operations Research methods allows examining this rather complex and convoluted chain in a comprehensive manner, and to search among a vast number of combinations for the resource. Optimization and allocation strategy are most effective and hence beneficial to the Operations.
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22. Dominique A. Heger, Fortuitous Technology, (dom@fortuitous.com), Austin, TX, 2006, An Introduction to Operations s Research – Benefits, Methods and Application
23. Hillier F., and Lieberman G. Introduction to Operations s Research. Holden-Day, 2001
24. Banks J., Carson II J., and Nelson B. Discrete-Event System Simulation. Prentice-Hall, 1995
25. Rardin, R.L., Optimization in Operations s Research. Englewood Cliffs: Prentice Hall, 1998
26. Taha, H.A., Operations s Research: An Introduction. Englewood Cliffs: Prentice Hall, 2002
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524141EnglishN-0001November30General SciencesCHENNAI'S REGIONAL TEMPERATURE VARIATION AND ITS CORRELATION WITH SUNSPOT NUMBERS
AND CO2 EMISSION
English8694R. Samuel SelvarajEnglish P.R. Uma raniEnglish N. MahalakshmiEnglish S.P. Vimal PriyaEnglishEarth's climate is determined by complex interactions between the Sun, oceans, atmosphere, land, and living things. The composition of the atmosphere is particularly important because certain gases (including water vapour, carbon dioxide, methane, ozone, and nitrous oxide) absorb heat radiated from the Earth's surface. As the atmosphere warms, it in turn radiates heat back to the surface, to create what is commonly called the "greenhouse effect." Changes in the composition of the atmosphere alter the intensity of the greenhouse effect.Variations of Chennai?s average mean temperature and diurnal temperature range for a period of ten years are investigated and is compared with past hundred years data. There is an observed variability of average mean temperature and diurnal temperature range. The variation in the temperature can be influenced by various climate forcing. In this paper we have correlated the sunspot numbers and Carbon dioxide emission with DTR and have estimated which influences the DTR more. Global warming over land can be characterized by faster warming at nights. This observed recent trend should lead to considerable decrease in the diurnal temperature range (DTR). The decrease of the diurnal temperature range is approximately equal to the increase of mean temperature1.
EnglishINTRODUCTION
The work of the farmers, power company engineers, weather analysts and many others would be great deal more difficult without accurate information of careful recording of temperature data. The difference between the daily maximum and minimum temperature is called the diurnal temperature or range of temperature. The greatest variation in daily temperature occurs at the earth?s surface and becomes progressively smaller as we move away from the surface. This daily variation in temperature is larger on clear days than on cloudy days. The diurnal temperature range can be analyzed in four different categories like the deserts, plateau regions, cities and humid regions. The largest diurnal range of temperature occurs on high desserts, where the air is often cloud free, and there is less carbon dioxide and water vapour above to radiate much infrared radiation back to the space. An elevated plateau region like Reno which is located at 1350m above sea level will have 35degree Celsius in the day and will cool down to 15 degrees at night time.
Thus, showing a temperature range of 25 degrees. The city region can be split into two categories coastal and inland. Coastal cities usually have a smaller diurnal temperature range than the inlands as the water vapour will heat and cool more slowly than land. The city inlands which also have a decreased DTR are caused due to urban heat islands. The urban heat islands are due to industrial and urban development. In rural areas a large part of the incoming solar energy is used to evaporate water from vegetation and soil. In the cities where less vegetation and exposed soil exits, the majority of the solar energy is absorbed by urban structures and asphalt. Hence during warm daylight hours, less evaporative cooling in cities allows the surface temperature to rise higher than in rural areas. And during night time the temperature does not reduce much due to radiation emitted from the urban structures. Chennai is located on the thermal equator on the southeast coast (Coromandel Coast) of India at an average altitude of 6 metres from the sea level. The latitude and longitude of this city is 13°04 N 80°17 E respectively. It covers a total area of 174 sq km that is spread irregularly in the northeast corner of Tamil Nadu. Its proximity to the Bay of Bengal ensures a hot & humid climate for most of the year. The highest temperature is attained in late May and early June which is usually about 38 °C (100.4 °F) it exceeds 40 °C (104° °F) for a few days. Average daily temperature in Chennai during January is around 24 °C (75.2 °F), though the temperature rarely falls below 18 °C (64.4 °F). The lowest temperature recorded is 15.8 °C (60.44 °F) and highest 44.1 °C (111.38 °F) 2 . Thus Chennai falls under the fourth category of the diurnal temperature range In this paper we study the relationship between the solar activity, carbon dioxide emission and their influence on the daily temperature which may prove to be the evidence for the climate change over Chennai.
DATA AND METHODOLOGY
The maximum and minimum temperature data of Chennai for the month of March, April, May and June are taken in two parts. First, for a period of hundred years ( 1901- 2000). Next, for a period of ten years (2001-2010). The data for maximum and minimum temperature are obtained from the Indian Meteorological department. The sunspot number data for the corresponding months are obtained from National Geophysical Data Centre, Boulder, Colorado, USA3 . Similarly the carbon dioxide data are obtained for a period of 1980 to 2009. From the NOAA web site4 . Solar activity, green house gases and climate change are much of much interest these days. The statistical analysis of the examined data series involves a non parametric rank correlation technique (Spearman Rank Correlation)5 . The statistical reliability of the obtained correlations is evaluated by a significance test. The Diurnal Temperature Range(DTR) between the hundred years data and ten years temperature maximum and minimum data are determined. Followed by this a statistical correlation is worked out between diurnal temperature range and sun spot numbers. And the same is carried out for the carbon dioxide emission data. The anomaly in diurnal temperature is shown in Table 1. And the correlation values are shown in Table 3. A graph is plotted between the temperature range and sun spot numbers and between the temperature range and carbon dioxide for the months of March, April, May and June. It is shown in Figure 1 and Figure 2 respectively.
RESULTS
Comparing the value of maximum and minimum temperatures for March, April, May and June in column two and three of Table 1: we observe a slight rise in the maximum and minimum temperature individually. The diurnal temperature range shows a slight anomaly. The anomaly was expected to be more prominent as per the concept of global warming. As the global warming increases the night time temperature increases leading to decrease in the diurnal temperature range. Column 2 of Table 1 shows a decreased diurnal temperature range. But column 3 shows slightly higher values of diurnal temperature range. This result is contrary to several observed trends in the other regions of the world. A city is characterized by a decrease in the DTR. Thus the anomaly is investigated further. For this we have considered the DTR in six other coastal cities of Tamil Nadu and compared it with the DTR of six Inland cities in the state. The results are tabulated in Table 2. From the data it is inferred that all the coastal cities exhibit an increased DTR. The Inland cities exhibit a decreased DTR effect. Thus the anomaly in Chennai City can be attributed to the effect of its coast line. The change in DTR can be forced by natural and anthropogenic factors. Table 3 shows the correlation between the diurnal temperature range, sunspot numbers and the carbon dioxide emission. We can infer from table 3 the sunspot numbers shows a good correlation with the DTR than the carbon dioxide emission.The Figure 1 shows a prominent relevance between the sunspot numbers and the DTR than the carbon dioxide emission as in Figure 2.
DISCUSSION
The coastal parts of India often come under the warm and humid zones. The city of Chennai falls under this zone. The contradictory result obtained from the Table 1 can be due the increased aerosol concentration in the atmosphere in the city during the last ten years6 , apart for this the ocean currents, direction of prevailing winds, the El Nino effect, water vapour concentration all these factors affect the DTR in the Chennai region and thus we observe an increase in DTR. The city inland are affected by the Urban Heat Island Effect (UHIE)7 which increase the night time temperature thus increasing the minimum Temperature at a faster rate than the increase in the maximum temperature. This leads to a decreased DTR in these regions. In Table 3 the significance of correlation depends on the sample size. For large sample as in our case even the small relations between variables will be significant. Here we have r = 0.2 at a significant level pEnglishhttp://ijcrr.com/abstract.php?article_id=1989http://ijcrr.com/article_html.php?did=19891 Karl Braganza, David J. Karoly, J. M. Arblaster, Diurnal temperature range as an index of global climate change during the twentieth century, Geophysical Research Letters, Vol. 31, L13217, doi:10.1029/2004GL019998, 2004
2 Bansal N.K. and Minke G., Climatic zones and rural housing in India, Kernforschungsanlage, Juelich, Germany, 1988.
3 NOAA/NESDIS/NGDC/STP, Boulder-Sunspot Number Data from NGDC http://www.ngdc.noaa.gov/stp/solar/ssndata.html
4 Trends in CO2 - NOAA Earth System Research Laboratory ftp://ftp.cmdl.noaa.gov/ccg/co2/trends/co2_mm_gl.txt
5 Press, W. H., Teukolsky, Vetterling, W. T., Flannery, B. P. 1992, Numerical Recipes in C (2nd edn.), Cambridge: Cambridge University Press, p. 640.
6. S. Sarkar, R. Chokngamwong, G. Cervone, R.P. Singh *, M. Kafatos Variability of aerosol optical depth and aerosol forcing over India, Advances in space research, In press 2005
7 Monsingh D. Devadas, Lilly Rose A, Urban Factors and The Intensity of Heat Island In The City of Chennai, proceedings of The seventh International Conference on Urban Climate,29 June - 3 July 2009, Yokohama, Japan
8 Variability of climate change in india, s.k. dash and j.c.r. hunt, current science , vol.93,no.6,2007,pg782-788
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524141EnglishN-0001November30General SciencesImpact of Distillery Spentwash Irrigation on Sprouting and Growth of Himalayan Balsam (Balsaminaceae) and Crossandra (Acanthaceae) Flowering plants
English95103S. ChandrajuEnglish C. ThejovathiEnglish C. S. Chidan KumarEnglishGermination of Himalayan Balsam (Balsaminaceae) and Crossandra (Acanthaceae) seeds was made by irrigated with distillery spentwash of different concentrations. The spentwash i.e., primary treated spentwash (PTSW), 1:1, 1:2, and 1:3 spent wash were analyzed for their plant nutrients such as nitrogen, phosphorous, potassium and other physical and chemical characteristics. Experimental soil was tested for its chemical and physical parameters. Himalayan Balsam and crossandra seeds were sowed in different pots and irrigated with raw water (RW), 1:1, 1:2 and 1:3 spentwash. The nature of germination of seeds was studied. It was found that, the germination was very good (100%) in 1:3 SW irrigation, while very poor (25%) in 1:1 SW, moderate (80%) in 1:2 SW and 95% in RW irrigations.
EnglishDistillery spentwash, Himalayan Balsam , Crossandra , Germination, Growth, Irrigation, Soil.INTRODUCTION
Himalayan Balsam belongs to the family Balsaminaceae. It typically grows to 1 to 2 m high, with a soft green or red-tinged stem, and lanceolate leaves 5 to 23 cm long. The crushed foliage has a strong musty smell. The flowers are pink, with a hooded shape, 3 to 4 cm broad the flower shape has been compared to a policeman?s helmet, giving rise to the alternative common name Policeman?s Helmet. Although it does not range all over India and is by no means the only Impatiens native to that country, it is also known as Indian Balsam in countries where it is introduced. In India it is commonly called as Karnakundala.
Himalayan Balsam, Impatiens glandulifera, is a large annual plant, native to the Himalayans (resulting in its colloquial name of Kiss-me-on-themountain1 in the UK. After flowering between June and October, the plant forms seed pods 2 to 3 cm long and 8mm broad. Which explode when disturbed, scattering the seeds up to 7 meters. The green seed pods and seeds can be eaten, and also the young leaves and shoots. Which is a method of controlling the plant?s spread2- 4 . However, a recent study it may cause more harm than good. Destroying riparian stands of Himalayan Balsam can open up the habitat for more aggressive invasive plants. The Bionic Control of Invasive Weeds in Wiesbaden, Germany is trying to establish a self sufficient project to conserve their local biodiversity by developing several food products made from the Impatiens flowers. Eventually, if all goes well, this project will have the Himalayan Balsam financing its own eradication.
Crossandra is belongs to the family Acanthaceae, comprising 52 species that occur in Africa, Madagascar, Arabia and the Indian subcontinent. Some species, especially Crossandra infundibuliformis5,6, are cultivated for their brightly colored flowers. It is known as Kanakambara in Karnataka, southern states in India. Crossandra from the Greek, meaning fringed anthers. The male portion of the flower, the anthers, is distinctly fringed in this genus of plants. The firecracker flower, while relatively unknown to the general public as a houseplant, is just about the most prolifilic indoor flowering plant. A well tended specimen will bloom continuously for years. It is growing from four sided stalked spikes; the asymmetrical petals arise as a slender tube and then split in to their ends. Plant breeders, especially in Europe, have been hybridizing Crossandras. Cultivars with yellow and even red flowers are available. Crossandra is a sturdy, productive ornamental that should be more popular with indoor gardens.
Molasses (one of the important byproducts of sugar industry) is the chief source for the production of ethanol in distilleries by fermentation method. About 08 (eight) liters of wastewater is generated for every liter of ethanol production in distilleries, known as raw spentwash (RSW), which is known for high biological oxygen demand (BOD: 5000- 8000mg/L) and chemical oxygen demand (COD: 25000-30000mg/L), undesirable color and foul odor7 . Discharge of RSW into open field or nearby water bodies results in environmental, water and soil pollution including threat to plant and animal lives. The RSW is highly acidic and contains easily oxidisable organic matter with very high BOD and COD 8 . Also, spentwash contains high organic nitrogen and nutrients 9 . By installing biomethenation plant in distilleries, reduces the oxygen demand of RSW, the resulting spentwash is called primary treated spentwash (PTSW) and primary treatment to RSW increases the nitrogen (N), potassium (K), and phosphorous (P) contents and decreases calcium (Ca), magnesium (Mg), sodium (Na), chloride (Cl- ), and sulphate (SO4 2- ) 10. PTSW is rich in potassium (K), sulphur (S), nitrogen (N), phosphorous (P) as well as easily biodegradable organic matter and its application to soil has been reported to increase yield of sugar cane, wheat and rice 11, Quality of groundnut 12 and physiological response of soybean13. Diluted spentwash could be used for irrigation purpose without adversely affecting soil fertility14 , seed germination and crop productivity 15. The diluted spent wash irrigation improved the physical and chemical properties of the soil and further increased soil micro flora16, 17, 18 . Twelve pre-sowing irrigations with the diluted spent wash had no adverse effect on the germination of maize but improved the growth19. Diluted spentwash increases the growth of shoot length, leaf number per plant, leaf area and chlorophyll content of peas20 . Increased concentration of spent wash causes decreased seed germination, seedling growth and chlorophyll content in Sunflowers (Helianthus annuus) and the spentwash could safely used for irrigation purpose at lower concentration21. The spentwash contained an excess of various forms of cations and anions, which are injurious to plant growth and these constituents should be reduced to beneficial level by diluting spentwash, which can be used as a substitute for chemical fertilizer22 . The spent wash could be used as a complement to mineral fertilizer to sugarcane23. The spentwash contained N, P, K, Ca, Mg and S and thus valued as a fertilizer when applied to soil through irrigation with water24. The application of diluted spentwash increased the uptake of Zinc (Zn), Copper (Cu), Iron (Fe) and Manganese (Mn) in maize and wheat as compared to control and the highest total uptake of these were found at lower dilution levels than at higher dilution levels. Mineralization of organic material as well as nutrients present in the spentwash was responsible for increased availability of plant nutrients. Diluted spentwash increase the uptake of nutrients, height, growth and yield of leaves vegetables25 , nutrients of cabbage and mint leaf26, nutrients of top vegetable27 , pulses, condiments, root vegetables, of some root vegetables in untreated and spentwash treated soil, yields of top vegetables (creepers). However, no information is available on sprouting and growth of Himalayan Balsam and Crossandra flowering plant irrigated by distillery spentwash. Therefore, the present investigation was carried out to study the influence of different proportions of spent wash on the sprouting and growth35, 36, 37, 38 of Himalayan Balsam and Crossandra.
MATERIALS AND METHODS
Physico-chemical parameters and amount of nitrogen (N), potassium (K), phosphorous (P) and sulphur (S) present in the primary treated diluted spentwash (1:1, 1:2 and 1:3 SW) were analyzed by standard methods28 . The PTSW was used for irrigation with a dilution of 1:1, 1:2 and1:3. A composite soil sample collected prior to spentwash irrigation was air-dried, powdered and analyzed for physico-chemical properties29, 30, 31, 32, 33, 34 . Flowering plants selected for the present investigation were Himalayan Balsam and Crossandra. The sets were planted in different pots 30(h), 25(dia)] and irrigated (by applying 5-10mm/cm2 depends upon the climatic condition) with raw water (RW), 1:1 SW, 1:2 SW and 1:3 SW at the dosage of twice a week and rest of the period with raw water as required. Cultivation was conducted in triplicate, in each case sprouting, growth were recorded.
RESULTS AND DISCUSSION
Chemical composition of PTSW, 1:1, 1:2, and 1:3 SW such as pH, electrical conductivity, total solids (TS), total dissolved solids (TDS), total suspended solids (TSS), settelable solids (SS), chemical oxygen demand (COD), biological oxygen demand (BOD), carbonates, bicarbonates, total phosphorous (P), total potassium (K), ammonical nitrogen (N), calcium (Ca), magnesium (Mg), sulphur (S), sodium (Na), chlorides (Cl), iron (Fe), manganese (Mn), zinc (Zn), copper (Cu), cadmium (Cd), lead (Pb), chromium (Cr) and nickel (Ni) were analyzed and tabulated (Table-1). Amount of N, P, K and S contents are presented (Table- 2). Characteristics of experimental soils such as pH, electrical conductivity, the amount of organic carbon, available nitrogen (N), phosphorous (P), potassium (K), sulphur (S), exchangeable calcium (Ca), magnesium (Mg), sodium (Na), DTPA iron (Fe), manganese (Mn), copper (Cu) and zinc (Zn) were analyzed and tabulated (Table-3 and 4). It was found that the soil composition is fit for the cultivation of plants, because it fulfils all the requirements for the growth of plants. Sprouting and growth of Himalayan Balsam and Crossandra plant leaves, uptakes of all the parameters were very good in both 1:2 and 1:3 spent wash as compared to1:1, SW and raw water. In both 1:1, 1:2 and 1:3 spent wash irrigation, the uptake of the nutrients such as fat, calcium, zinc, copper and vitamins carotene and vitamin c were almost similar but the uptake of the nutrients and parameters such as protein, fiber, carbohydrate, energy, magnesium and phosphorous were much more in the case of 1:1, 1:2, spent wash irrigation than 1:3, and raw water irrigations (Table-5).
This is due to the more absorption of plant nutrients present in spentwash by plants at higher dilutions. It was also found that no negative impact of heavy metals like lead, cadmium and nickel on the leaves of Himalayan Balsam and Crossandra plant. The soil was tested after the harvest; found that there was no adverse effect on soil characteristics (Table-4)
Englishhttp://ijcrr.com/abstract.php?article_id=2278http://ijcrr.com/article_html.php?did=22781. Wanted Himalayan Balsam, British isles.
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35. S.Chandraju, Thejovathi.C, Chidan Kumar.C.S., Sprouting and Growth studies of Rose and Hibiscus flowering plants irrigated by distillery spentwash. Research in Plant Biology, 1(2), 06-13. 2011.
36. S. Chandraju, Thejovathi.C, Chidan Kumar C.S., Studies on the Germination and Growth of Zinnia and Vinca seeds irrigated by Distillery spentwash. SISSTA, 2011.
37. S.Chandraju, Thejovathi.C, Chidan Kumar.C.S., Impact of Distillery spentwash irrigation on sprouting and Growth of Gardenia flowering plant. J. Chem. Pharm. Res. (In press)
38. S.Chandraju, Thejovathi C., Chidan Kumar C.S., Impact of Distillery spentwash irrigation on sprouting and Growth of Tagetes flowering plant Asian J. Research Chem, 2011 (Accepted).