Degrading Marine Bacteria: Isolation and Characterization

Degrading Marine Bacteria: Isolation and Characterization Isolation and Characterization of (PAH) Biodegrading Marine Bacteria Sulaiman Ali Alharbi1*, M.E.Zayed1, Arunachalam Chinnathambi1, Naiyf S. Alharbi1 and Milton Wainwright1,2 ABSTRACT Polyaromatic hydrocarbons (PAH) are considered to be important and dangerous pollutants which cause serious health problems and/or genetic defects in humans, as well as harming the flora and fauna of affected habitats. In this study, we interested in determining if bacteria can be successfully used to bioremediate PAH pollution as an alternative to physical and chemical methods. The bacteria used in this study were isolated from three PAH polluted sites of Mediterranean Sea, off Alexandria, Egypt. The study is devoted to the isolation of bacteria that can degrade three low molecular weight PAHs and to determine the effect of pH on this process. Nine phenanthrene-degrading, seven-naphthalene-degrading and eight anthracene-degrading bacteria were isolated, by enrichment, from the marine water sample. All of the isolates grew on the PAHs (phenanthrene, anthracene and naphthalene) at varying rates and utilized them as sole source of carbon and energy. Key words:   Polyaromatic hydrocarbons (PAH), Biodegrading marine bacteria, Environmental contamination, Marine water, Introduction Polycyclic aromatic hydrocarbons (PAHs) are contaminants of aquatic and terrestrial ecosystems which generated continuously by the inadvertently incomplete combustion of organic matter, for instance in forest fires, home heating, traffic, and waste incineration1. PAHs constitute a large and diverse class of organic compounds and are generally described as molecules which consist of three or more fused aromatic rings in various structural configurations2. Polycyclic aromatic hydrocarbons are composed of fused, aromatic rings whose biochemical persistence arises from dense clouds of Ï€-electrons on both sides of the ring structures, thereby making them resistant to nucleophilic attack3. Environments contaminated with PAHs are deemed hazardous because of their carcinogenic, mutagenic and teratogenic effects4,5 and low molecular weight PAHs such as naphthalene (the simplest, containing two benzene rings), anthracene and phenanthrene (both of which contain three benzene rings) are also known to possess potentially hazardous health effects6. A variety of techniques have been applied to the treatment of environments contaminated by PAH containing petroleum hydrocarbons, notably physical treatments using thermal or chemical processes7. However, these treatments are generally time consuming and expensive8,9. Microbial bioremediation however, provides a potentially cheap and effective means of bio-remediating PAH-contaminated environments10. The ability of microorganisms to degrade PAHs is well documented11, 12 and microbial degradation is a major environmental process affecting the fate of PAHs in both terrestrial and aquatic ecosystems13. Bioremediation using microbes converts toxic or persistent organic molecules into harmless end products, such as carbon dioxide and water 14. Unfortunately PAHs possess physical properties, such as low aqueous solubility and high solid water distribution ratios, which militate against their rapid microbial utilization resulting in their accumulation in the terrestrial and aquatic environments1. It is imperative therefore that the factors which influence the bioavailability and decomposition of PAHs in the environment be studied and optimized7. The aim of the work described here was to screening environmental samples for bacteria that are capable of degrading PAHs and use them a sole carbon and energy source. Materials and Methods Isolation of bacteria: Bacteria were isolated from three PAH-polluted sites of the Mediterranean Sea, Alexandria, Egypt, where PAH pollutants have been continually released. All isolates were preserved in 750 ÃŽ ¼l LB culture supplemented with 250 ÃŽ ¼l of 60% glycerol and maintained at -80oC. Media used: Bushnell-Haas (BH) medium, Luria-Bertani, Nutrient broth and Blood agar were used for the isolation of bacteria. All media were prepared using distilled water and sterilized by autoclaving at 120oC for 20 min. Medium- pH was adjusted as required before sterilization using   1N NaOH or 1N HCl. Hydrocarbon stock solutions: Stock solutions of each PAH (100 mg/ml) were prepared in ethyl acetate and sterilized by filtration. Isolation of phenanthrene, anthracene and naphthalene degrading bacteria: Water samples were collected from three PAH-polluted sites in the Mediterranean Sea, off Alexandria, Egypt, where PAH pollutants have been continually released into these aquatic environments without any control. Water samples (50ml) were collected from the contaminated sites under aseptic conditions. Phenanthrene, anthracene and naphthalene degrading bacteria were isolated from water samples by spreading onto solid medium; 100  µl of each water sample was spread over the surface Bushnell-Haas agar plates containing (100, 200, 300, 400, 500 and 600 mg/l) of either, phenanthrene, anthracene or naphthalene, as the sole carbon and energy source. The plates were then incubated at 30oC for 7 days. Assay of phenanthrene, anthracene and naphthalene degradation: Phenanthrene, anthracene and naphthalene degradation by the bacteria under a variety of concentrations was determined using 250 ml Erlenmeyer flasks containing 100 ml Bushnell-Haas broth supplemented with phenanthrene, anthracene and naphthalene in the concentration range,10, 30, 50, 70, 90, 130 to 150 mg/l.. The cultures were inoculated by transferring 1 ml of nutrient broth of pre culture medium of the strain under test. The cultures were then incubated at 30oC and 200 rpm; bacterial growth was daily evaluated by measuring the increase of OD600nm of the culture. Determination of naphthalene, phenanthrene and anthracene residues in the culture medium: The concentration of naphthalene and phenanthrene residues in the culture medium was determined by measuring the optical density at a wavelength of 254 nm and 275 nm respectively15. Extraction was carried out in a separator funnel, by mixing for two minutes an aliquot of the culture medium with an equal volume of hexane. The resulting organic phase was then used for the spectrophotometric readings. In some cases the sample was diluted with hexane in order to bring it out within the range of the calibration line (0.01-0.07 mg/ml for naphthalene and 0.001-0.1 mg/ml for phenanthrene). For the determination of anthracene residue, aliquots of the culture medium were mixed for two minutes with an equal volume of ethyl acetate and the optical density of the resulting organic phase was measured at a wavelength of 254 nm using ethyl acetate as a blank. For preparation of standard curve of anthracene, a stock solution of 10 ppm in ethyl acetate was prepared and aliquots in the range of 0.2 to 1 ppm were separately measured at 254 nm16. Utilization of carbon source: All purified isolates were tested for growth on 0.01%, of either, naphthalene, phenanthrene, anthracene or phenol which were added as sole carbon sources to BH liquid medium. Sterilized BH medium containing the desired amount of hydrocarbon source was inoculated with the test strain and incubated in an orbital shaker at 200 rpm and 30oC for 72 h. Growth was tested by measuring the increase of OD600nm of the cultures. Effect of pH on the degradation of naphthalene, phenanthrene and anthracene: In order to determine the effect of pH on naphthalene, phenanthrene and anthracene degradation, 50 ml of BH broth cultures were first prepared at the following pH; 2, 3, 4, 5, 6, 7, 8, 9, 10 and 11. After sterilization, 50 mg/l naphthalene, phenanthrene and anthracene was added to each flask, separately. 0.5 ml of Nutrient Broth overnight culture of bacterial strains (OD600 1.3-1.5) was added to each flask. The flasks were incubated at 30oC and 200 rpm for 72h. Bacterial growth was estimated using spectrophotometer at OD600nm of the cultures. Results and Discussion Isolation and selection of phenanthrene, naphthalene and anthracene biodegrading bacteria: Phenanthrene, naphthalene and anthracene degrading bacteria were isolated from water samples.   A range of bacterial colony types were observed on the different carbon source-containing plates, nine isolates were found to utilize phenanthrene, seven   grew on the naphthalene plates and eight isolates grew on anthracene amended media. Isolates Ph1, Ph2, Ph3, Ph4, Ph5 and Ph7 showed the ability to degrade phenanthrene as sole carbon sorce. The optical density (OD600)   for the bacterial isolates respective was was; 0.16, 0.512, 0.17, 0.482, 0.632 and 0.24. Isolates Ph6, Ph8 and Ph9 on the otherhand failed to utilize phenanthrene as sole carbon and energy source (Fig.1). Among the tested isolates only Ph5 isolate showed sustantial   growth rate on phenanthrene when compared with other tested isolates. According to the ioslates which showed no degrdation of the tetsed PAH, We assume that these isolates which did not degrade PAH may have lost this ability during preservation, or may have lost genes which control the utilization of this substrate; assumptions mirored in the work of   Zhao, et al., (2009)17, who reported that some phenanthrene degrading isolates lost their ability to degrade phenanthrene after a period of more than three weeks of preservation. Figure 1. Degradation of phenanthrene (20 mgl) by 9 bacterial isolates (named; Ph1-Ph9).       In studies using naphthalene, only two isolates Na6 and Na7 showed dehradation   with the latter being the best naphthalene degrader (Fig.2). The other isolates, Na1, Na2, Na3, Na4 and Na5, in contrast, failed to use naphthalene as a sole carbon and energy source. Regarding the anthracene-degarding isolates, five- An1, An2, An3, An4 and An7 degraded the substrate with ODs respectively oft: 0.174, 0.614, 0.551, 0.482 and 0.164 (Fig.3). Isolate,   An2 isolate was the msot active anthracene degrader,while isolates An5, An6 and An8 were unable   to use anthracene as a sole source carbon and energy source. Figure 2.   Degradation of naphthalene (20 mgl) by 7 bacterial isolates (named; Na1-Na7). Figure 3. Degradation of anthracene (20 mgl) by the bacterial isolates (named; An1-An8). Effect of pH on hydrocarbons degradation by the different bacterial isolates: The effect of pH (2.0, 3.0, 4.0,   up to 11.0) on PAH degradation by the isolated bacteria Ph5, Na7 and An2 was investigated in BH medium containing 50 mgl phenanthrene and 20 mgl naphthalene or anthracene respectively. In the case of phenanthrene, the optimum pH for bacterial growth and phenanthrene degradation was pH 7. At pH 6 and pH 8, bacterial growth and substrate degradation was markedly reduced (Fig.4). Shin et al.,(2008)18 reported that, relatively high mineralization rates of phenanthrene are found over a pH range of 6-8, with maximum mineralization rate occurring at pH 6 in a mineral salt medium. In the present study we found that the pH 7 is the optimum for obtaining a high mineralization rate of phenanthrene in BH medium; in agreement with Simarro, et al., (2011)19, our results confirm that the optimal pH value for the degradation of this substrate,   in BH medium, is pH7. Figure 4. The ability of the selected Ph5 isolate to degrade 50 mg/l phenanthrene at different pH. Growth and biodegradation depends on the type of PAHs used and the optimum pH range is very variable20. Some acid resistant Gram-positive bacteria, such as Mycobacterium sp., show better PAH degradation capabilities under acid conditions, largely because low pH seems to render such Mycobacteria more permeable to hydrophobic substrates21. However, other microorganisms belonging to Pseudomonas genus tend to prefer neutral pH conditions. In agreement with previous works19, our results confirm that neutral pH is optimum for the biodegradation of PAHs. Our results are also in agreement with Bisht, et al.(2010)22, who reported the ability of D. radiodurans SBA6 and B. circulans SBA12 to degrade naphthalene and anthracene over in the pH range of 5.0 to 11 (Fig.5). Othman et al.(2009)23 observed that changes in media pH can alter the electrical charge on various chemicals groups in enzymes molecules, thereby probably altering the enzymes ability to bind its substrate and catalyze a given rea ction. Imbalance of the electrical charges in very acidic and alkali conditions can also disrupt hydrogen bonds and other weak forces that maintain enzyme structure. Such disruption of enzyme structure is called denaturation, a phenomenon which leads to poor rates of biodegradation. Figure 5. Determination of the ability of Na7 and An2 isolate to degrade 20 mg/l naphthalene and anthracene respectively, over a range of pH. Utilization of different hydrocarbons by the same bacterium: Phenol (a single ring compound) was also included in this experiment as an example of a low molecular weight PAHs to provide a comparison with the high molecular weights hydrocarbons e.g. naphthalene (two rings), anthracene, and phenanthrene (both three compounds). Isolate Ph5 was shown to use phenol, naphthalene and anthracene as sole carbon and energy source (Fig.6). These results are slightly different from those reported by Zhao, et al. (2009)17, who reported that a bacterial isolate (ZP2) was able to degrade phenanthrene and naphthalene but failed to degrade anthracene as sole carbon source. Figure 6. Assimilation of 0.01% of naphthalene, anthracene and phenol as a sole   carbon source in Bushnell-Haas medium by Ph5 isolate. The same response was seen in the other two isolates Na7 and An2.   Isolate, Na7 rapidly mineralized phenol, phenanthrene and anthracene when added as sole carbon sources (Fig.7). Isolate An2 degraded phenol, naphthalene and phenanthrene as sole sources of energy (Fig.8). An isolate used in a study by Dean-Ross, et al. (2001)24 in contrasts was able to mineralize anthracene and phenanthrene but not naphthalene when grown under identical conditions. Figure 7. Utilization of 0.01% of phenanthrene, anthracene and phenol as a sole   carbon source in Bushnell-Haas medium by the isolate Na7. Figure 8. Consumption of 0.01% of phenanthrene, naphthalene and phenol as a sole   carbon source in Bushnell-Haas medium by An2 isolate. Phenanthrene, naphthalene and anthracene are the main components of crude oil and ubiquitous in contaminated water and soil. These carbon-sources could be utilized by a range of living in these polluted environments. Hydrocarbon mineralization occurs in a variety of ways depending on the species of bacterium isolated and it use of a preferred pathway. In this study, the wide range carbon-source utilization of tested isolates confirms their ability to use potentially different degradation pathways. Al-Thani, et al.(2009)25, similarly reported that the acclimation of a microbial community to one substrate frequently results in the simultaneous acclimation to some, but not all structurally related molecules. As a result, individual microbial species have the ability to act on several structurally similar substrates and therefore more easily act on their analogues following initial exposure26,27. Conclusion: In this investigation, we fascinated in determining if bacteria can be successfully used to bioremediate PAH pollution as a substitute to physical and chemical methods.In conclusion, some of our isolates grew well on the tested 4 low molecular weight organic aromatic compounds and as also reported28, individual bacterial strains could degrade several PAHs, but tended to prefer a single one and also we establish that the pH 7 is the optimum for obtaining a high mineralization rate of phenanthrene in BH medium. Acknowledgements Authors extend their appreciation to the Deanship of Scientific Research at King Saud University for funding the work through the research group project No. RGP-VPP-332. References 1Johnsen, A. R., Wick, L. 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Degradation of phenanthrene by bacterial strain isolated from soil in oil refinery fields in Shanghai, China. J. Hazard. Mater. 164: 863-869. 18Shin, K., Kim, J. and Kim, K. 2008.Effect of biosurfactant addition on the biodegradation of phenanthrene in soil-water system. Environ. Eng. Res. 13: 8-13. 19Simarro, R., Gonzalez, N., Bautista, L. F., Sanz, R. and Molina, M. C. 2011. Optimization of key abiotic factors of PAH (naphthalene, phenanthrene and anthracene) biodegradation process by a bacterial consortium. Water Air Soil Pollut. 217: 365-374. 20Dibble, J.R. and Bartha, R. 1979.Effect of environmental parameters on the biodegradation of oil sludge. Appl. Environ. Microbiol. 37: 729-739. 21Kim, Y.H., Freeman, J.P., Moody, J.D., Engesse, K.H. and Cerniglia, C.E. 2005.Effects of pH on the degradation of phenanthrene and pyrene by Mycobacterium vanbaalenii PYR-1. Appl. Environ. Microbiol. 67: 275-285. 22Bisht, S., Pandey, P., Sood, A., Sharma, S. and Bisht, N. S. 2010.Biodegradation of naphthalene and anthracene by chemo-tactically active rhizobacteria of Populus deltoides. Braz. J. Microbiol. 41: 922-930. 23Othman, N., Hussain, N.H., Abd Karim, A.T. and Abdul-Talib, S. 2009.Isolation and optimization of napthalene degradative bacteria. International Conference on Sustainable Infrastructure and Built Environment in Developing Countries. Bandung, West Java, Indonesia.101-105. 24Dean-Ross, D., Moody, J. D., Freeman, J. P., Doerge, D. R. and Cerniglia, C. E. 2001.Metabolism of anthracene by Rhodococcus species. FEMS Microbiol. Lett. 204: 205-211. 25Al-Thani, R.F., Abd-El-Haleem, D.A.M. and Al-Shammri, M. 2009.Isolation and characterization of polyaromatic hydrocarbons-degrading bacteria from different Qatari soils. African J. of Microbiol. Resear. 3: 761-766. 26Bauer, J. and Capone, D. 1985. Degradation and mineralization of the polycyclic aromatic hydrocarbons anthracene and naphthalene in inter tidal marine sediments. Appl. Environ. Microbiol. 50: 81-90. 27Mitchell, J. and Cain, R. 1996. Rapid onset of the accelerated degradation of dicarboximide. Pesticides Sci. 48: 1-11. 28Tadros, M. G. and Hughes, J. B. 1997. Degradation of polycyclic hydrocarbons (PAHs) by indigenous mixed and pure cultures isolated from coastal sediments. Appl. Biochem. Biotechnol. 63(65): 865-870 Can Liberty and Equality be Reconciled in Political Theory? Can Liberty and Equality be Reconciled in Political Theory? The word reconcile means that, to find a way in which two situations or beliefs that are opposed to each other can agree and exist together. It is sometimes difficult to reconcile for instance science and religion. When two people are reconciled they become friendly again after they have argued, (Cambridge advanced learners dictionary). In this essay I will dwell much on whether liberty and equality can be reconciled by defining these concepts. Liberty and freedom are often used interchangeably, however they essentially mean the ability to think or act as one wishes. Gerald McCallum: â€Å"Freedom is always of something (an agent or agents) to do or not to do, become or not become something†. Most political theorists assume that people ought to be free unless there are compelling reasons for restricting their freedom. For instance J.S.Mill believes that, to be human is to enjoy a sphere of in which one is able to think, express ideas and lead a lifestyle of one’s own choosing. He went on to say, â€Å"even if a person finds himself in an opinion, he should be free to express that opinion† and â€Å"if all mankind, minus one, were of one opinion, and only one person were of the contrary opinion, mankind would not be justified to silence that person, than he, if he had the power, would be justified in silencing mankind† By allowing individuals to be free, society can progress, hence the ideas of mi nimal government and protection of human rights and freedoms. There are two concepts of liberty, these includes negative and positive liberty (Isaiah Berlin, 958). Negative liberty involves response to the question: what is the area within which the subject a person or group of persons is or should be left to do what he is able to do or be, without interference by other people. It implies freedom of choice, therefore, Law and Government should only serve to enlarge and not restrict freedoms, and privacy of the individual must be respected by recognizing a clear distinction between what is public and what is private and finally it advocates individual rationality as the basis for choice. On the other hand, positive liberty is involved in the answer to the question: what or who is the source of control or interference that can determine someone to do or be, also interested in the question ‘By whom am I governed?’ However, as opposed to â€Å"How much am I governed? G.C. Mc Callum, (1972). According to T.H. Green it means personal gr owth and self-development. Liberalism is one of the dominant political ideologies of the contemporary world that favors liberty in terms of equality. It is founded on the traditional notions of individualism: this reflects the belief in the supreme importance of the human individual as opposed to any social group or collective body. The liberal goal is therefore to create a society within which individuals can flourish and prosper with everyone pursing what they consider as good in any way they choose to define it. Therefore, individual freedoms or liberties are given priority over notions of equality, justice and authority. It is thus based on the presumption that freedom is a good thing and that any limitations of the same should be justified. It is strongly against limitation on freedom of individuals to pursue self interest. It however, favors private property and free enterprise, for instance freedom to buy and sell anything. And finally it is against state control and seconds that role of the state limit ed to maintaining law and order, in this respect, it should provide defense and oversee enforcement of contracts. Equality refers to the belief that human beings are born having equal rights. The Declaration of Independence asserts some of these rights to be life, liberty and the pursuit of happiness. Equality is not about blanket uniformity but rather to establish the legal, political or social conditions in which people will be able to enjoy equally worthwhile and satisfying lives. The principle of equality sometimes contains the assertion that people must have equal access to wealth and property in order for them to more fully realize the principle of equality. According to Oxford English Dictionary, equality implies the condition of having equal dignity, rank or privilege with others, the condition of being equal in power, ability, achievement of excellence, fairness, impartiality, due proportion, and proportionateness. Therefore, this term is defined in the light of equal conditions guaranteed to each for making the best of him. In the words of Barker, it implies that â€Å"whatever conditions are guaranteed to me in the form of rights, shall also, and in the same measure, be guaranteed to others and that whatever rights are given to others shall also be given to me†. So says Laski: undoubtedly, it implies fundamentally a leveling process. It means that no man shall be so placed in society that he can over-reach his neighbor to the extent which constitutes a denial of the latter’s citizenship†, J.J. Rousseau [(1762) 1969]. There are two forms of equality positive and negative. Positive form implies the adequate provision of opportunities for all. Here, the term ‘adequate opportunities’ does not mean ‘equal opportunities’. Since people differ in their needs and capacities and also in their efforts, they need different opportunities for their individual self-development, Friedrich Hayek and Keith Joseph (1979). Negative equality implies the absence of undue privileges. That is to say there should be no artificial grounds of discrimination, such as of religion, color, race, caste, wealth, sex, and many more. The underlying principle being that no talent should suffer from frustration for want of encouragement. It is still a challenge to provide a coherent defense of equality. This however requires separating out the various principles, and explaining what it is that is being equalized or equality is in danger of degenerating into a mere political slogan unless it is possible to a nswer the question ‘equality of what’?: is it income, or well being, the capacity to acquire certain goods, or something else? Socialism is also one of the dominant ideologies of the modern world that favors equality than liberty. Socialists believe in a Common Ownership: this relates to the ownership of property in the community as opposed to individual ownership. The argument here is that by placing property in joint community ownership, the common good is promoted rather than the good of individuals. To them a positive view of freedom must be examined in a social context and therefore in the context of resources of a material kind. The right to read and write, for example, requires the provision of schooling if such a right is to be meaningful. In the Socialist World Order all classes and private property are abolished. The means of production for instance, that is all land, machinery, factories and other productive resources must be under the democratic control of the people. Wage labor is unknown. All worked for the collective good and products are distributed according to the principle of â€Å"From all according to their ability to all according to their need. They however, call this a good society. Liberty and Equality are political concepts that strongly oppose each other, to be specific; between them there is what we call war of principles. For instance, Libertarians believes that Libertaria is the place where humankind should seek its future. This is because it is a society where people are truly free. They have the economic freedom to pursue their own ends in a market society. Indeed Libertaria is run exclusively on market lines. They do not believe in the good society as such, as market choices constantly fluctuate. Individuals must choose their own good life and the market is where they make their key decisions. On the other hand, Egalitarians believes that Egalitaria is the ideal place to which humankind should commit itself. It is the benchmark by which other societies can measure whether they are serving the aims which any good society should fulfill. Individuals believe that market decisions produce what people want. In Libertaria all make market choices and all receive from the market that which is their due. They are free from both the stifling collectivity of Communitarian and from the fear of having to make sacrifices, a fear which haunts Utilitarian. While Egalitaria would be the good society because it would be one in which all citizens are treated as being of equal worth. It would have basic political liberties, equality of opportunity and the rule of law, all of which are constitutionally guaranteed. It would also be dedicated to ensuring that these basic rights are of equal worth to their possessors (Lukes, 1993: 34). Libertaria has freedom of speech, of movement, of thought and of association. The rule of law prevails and there is no torture. They have a minimal state which oversees the system of free exchange, the courts, contractual disputes, the armed forces and the police. They realize that the state can build up its own interests which can conflict with those of private individuals. They therefore, keep its role to an absolute minimum. Freedom is their watchword and freedom to choose is prized above all else. On the other hand, the citizens of Egalitaria would be filled with a basic abhorrence of injustice and would strive to overcome any involuntary disadvantage whether such disadvantages are the result of religion, class, gender, ethnicity or whatever. Egalitaria would not tolerate the gross inequalities which characterize some of the communities within Communitarian. Rights in Egalitaria would be universal, not subject to the dictates of history, culture and tradition. Considering the above argument it seems like each ideology is giving no space to the enjoyment of the other ideology, or in simple terms we can say that it is hard for liberty and equality to go together, however, in practice these terms are mutually reinforcing each other. It is also very hard to find a society that is purely libertarian or that is purely Egalitarian, this means that in libertarian the possibility of finding some element of equality is there, same applies to the egalitarian, there are some sort of liberties though to the slightest degree. Therefore, there are some instances where liberty and equality move together with a common goal for instance the promotion of welfare of the people. It is also of great important that liberty and equality should work hand in hand, just because they act as watch dog to each other, for instance, the relationship that is between them is that like of three arms of government executive, parliamentary and judiciary. The problem with establishing the realm of liberty is that there are a bewildering number of grounds upon which freedom can be upheld. In much liberal political thought freedom is closely related to the notion of rights. However, as many political theorists employ a value-free or social-scientific understanding of such terms, they are quite prepared to accept that certain freedoms such as the freedom to murder should be constrained. In that sense, liberty or license distinction merely begs the question: which freedoms are we willing to approve, and which ones are we justified in curtailing? Alternative means of distinguishing between liberty and license was proposed by J.S. Mill. As libertarian who believed that individual freedom was the basis of moral self-development, Mill proposed that individuals should enjoy the greatest possible realm of liberty. However, Mill also recognized that unrestrained liberty could become oppressive, objectionable, morally corrupt and even tyrannical. In on Liberty (1859) 1972) Mill proposed a clear distinction between self-regarding actions and other-regarding actions, suggesting that each individual should exercise sovereign control over his or her own body or life. The only justification for constraining the individual, Mill argued, was in the event of ‘harm’ being done to others. Mill emphasizes that liberty becomes license not only when the rights of another are violated, or when harm is done to others, but when liberty is unequally shared out. In this sense equality is seen as standing block to the existence of license . Equality is a multi-dimensional concept. Diverse opinions are put forth in locating the exact relationship between these two concepts. Writers like De Tocqueville and Lord Acton hold the view that liberty and equality are opposed to each other as they are antagonistic. The desire to have equality destroys the possibility of having full liberty. Achievement of equality demands positive state action. Equality needs a positive state and liberty needs a Negative State. The Elite theory of Democracy is against the principle of equality. But on the other hand writers like Maitland, Rousseau, Barker, Laski and others, hold the view that they are complementary to each other, in the sense that they both play an important part in human life, fulfilling separate but complementary. Since the principles of liberty and equality are ideals capable of being mutually realizable, one answer to the continuing debate regarding each principles precedence would be the check of each principle on the excesses of the other. A just society might hold itself to the standard that, while difficult to reconcile, it is still most desirable to strive to strike a balance between upholding the belief in the inherent equality of all individuals, and the protecting the right to liberty for all its members. L. T. Hothouse opined that liberty without equality is a high-sounding phrase with squalid results. Liberty lies in equality. Liberty without equality degenerates into license and equality without liberty lapses into uniformity. To Prof. Barker Equality in all its forms, must always be subject and instrumental to the free development of capacity; but if it be pressed to the length of uniformity; if uniformity be made to thwart the free development of capacity, the subject becomes th e master, and the world is turned topsy-turvy. Liberty unites men but equality criticizes the social hierarchy and contributes towards the stability of the community ((1931) 1969) R.H. Tawney. Therefore liberty would be hollow without some measure of equality and equality would be meaningless without liberty. In this age of democracy where voting is a powerful weapon in the hands of the electorate economic equality is most essential condition because the economically powerful person will use his economic resources to gain political power. Political equality will be a mockery in the absence of economic equality. As per above argument it is hard to say that liberty and equality can be reconciled or not, in the sense that these terms are supported by different ideologies but which are reinforcing each other. So to say that they can be reconciled is forgetting that the terms are founded on different ideologies and on the same point to say that they cannot be reconciled is also forgetting that these terms are mutually reinforcing each other, hence making this question very contested one. Who determine that excessive use of liberties leads into license? What yard stick do they use to determine harm to others? And what is harm? It is good to know that what is harm to me cannot be harm to others and vice-versa. For instance, a society that is comprised of well-off and worse-off, come together and agree that they should be following the principle of â€Å"to each according to ability† can another man from a distance society come and say that the well-off in this particular society are harming the worse-off? This cannot be possible since what he calls it harm cannot be harm to others. In this case the worse-off choose to do what they can manage to, according to the level of their economic status, therefore, it is not harm to them, because they have decided to follow it based on their will and rationality. And again, who determine that excessive use of equality leads to uniformity? What is uniformity? What yard stick do they use to measure uniformity? In every time we talk of equality we must also note that there are some forms of inequality. What is equality to a particular society cannot be equality to the other, since society perceives things in different way all together. The issue of inequality will not come to an end since they were there before, they are here today and possibly they will be there tomorrow. What a just society would do is to come up with measures or initiative to overcome them. For instance legitimate inequality, affirmative action and positive discrimination where the society comprised of well-off and worse-off come to a consensus that they should accept the status of inequality and that they will be following a principle of â€Å"to each according to needy†, but the end result should be greater to the worse-off so that they should little by little cover the gap that is there between them. On the other hand this makes sure that talents, skills and ability are not hindered. Therefore, one cannot come and say, this is uniformity since what is uniformity to him cannot be uniformity to others. It is good to reflect on these questions and acknowledge that what is liberty or equality to the state cannot be liberty or equality to individuals or the collectives group and vice-versa. What was liberty or equality yesterday cannot be liberty or equality today. These things keeps on changing based on circumstances that people as a nation are going through. Therefore, liberty and equality are continuously changing depending on time and who uses them. Finally, based on argument above its very tricky and challenging to say that liberty and equality can be reconciled or not. However, beauty lies in the hands of the beholder; even though, there are no general examples that can be sighted to show that liberty and equality can be reconciled, but there is a room for reconciliation since they are complementary. Therefore, liberty and equality have a common end, the promotion of the value of the personality and the free development of its capacities. No one of these can be enjoyed in isolation. Bibliography Heywood, A. (2004). Introduction to Political Theory. In A. Heywood, Introduction to Political Theory (p. 252 and 284). china: Palgrave Mc Millan. J.C, J. (2004). Principle of modern politcal science. In J. Johari, Principle of modern politcal science (p. 200). New Dehli: Sterling. Hoffman J. and Graham P (2009) introduction to Political Theory. Professor Wizeman Chijere Chirwa, Dr. Fidelis Edge Kanyongolo and Dr. Edrinnie Kayambazithu. (2004). Building an informed Nation. In Building an informed Nation (p. 58). Lilongwe: Democracy Consolidation Programme.