Dye Is A Visible Pollution Biology Essay

Dye is a seeable pollution. Even a little colour of H2O beginnings could do them unacceptable to consumers though it may non be toxic to the same grade. The beginning of such pollution prevarications in the rapid addition in the usage of man-made dyes. More than 10,000 chemically different dyes are being manufactured. The release of dyes into Waterss by industries is unwanted and causes serious environmental jobs.

It contains assorted organic compounds and toxic substances which are risky and harmful to aquatic beings. Most of the fabric, paper and surfacing industry usage dyes to colourise their merchandises. In Malaysia, dyes and pigments is normally used in batik printing industry. Other than that, the growing of fabric and dress industry is spread outing during 2nd Malaysia program ( 1971-1975 ) . The discharged of effluent from these industry can foul the H2O beginning with dyes.There are several methods of remotion of dyes from waste H2O. Some of them are, by flotation, precipitation, oxidization, filtration, curdling, ozonation, supported liquid membrane and besides biological procedure.

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Meanwhile, a new and more environmental friendly method, the biosorption procedure is proven to be a promising procedure to take dyes from wastewater. This procedure is similar to adsorbent procedure which it is cost-efficient, easy to run, merely designed and insensitiveness to toxic substances. Recently, there has been considerable involvement in the usage of microbic biomass including algae, bacteriums, Fungis, and agricultural byproducts or residues as adsorbents to take dyes from aqueous solution by surface assimilation, frequently called biosorption.

In this survey, Rhizopus Oligosporus is used as biosorbent for removal Methylene Blue dyes. Rhizopus Oligosporus comes from nutrient processing-derived ( production of tempeh ) . Tempeh is celebrated nutrient in Malaysia and Indonesia. Methylene Blue ( MB ) , known as strong surface assimilations into solid, will be used as the biosorbate. MB is an of import basic dye normally used for publishing cotton and tannic acid, and dyeing leather. Even though it is non strongly risky, MB can do harmful effects to human and other life beings. Excessively much exposed of MB can do oculus Burnss and causes annoyance to the tegument.

The aims of this survey were to analyze the consequence of biosorbent dose and the pH of dye solution on the per centum dye.


The reading of dyes into environment by different industries has been aesthetically unwanted and excessively much of it will eventually do serious environmental consequence, aquatic or non-aquatic. This is due to its belongingss which are largely toxic, mutagenic, and carcinogenic. Dyes are doing pollution to the environment, for illustration, dyes adsorb and reflect sunshine from come ining H2O and therefore interfere the aquatic ecosystem. Dyes when release can hold sensitive and or chronic effects.

It is evidently, hence, look intoing the remotion of dyes is important environmental, proficient, and commercially of import.Today activated C is normally used for biosorption in several intervention workss. But the bring forthing costs for activated C is really high, there is a demand of an alternate stuff is more cost efficient.

Rhizopus oligosporusA is a fungus of the household Mucoraceae that is a by and large usedA starting motor cultureA for the place production ofA tempeh. The spores produce downy, whiteA mycelia, adhering the beans together to make an comestible “ bar ” of partially fermented soya beans. In production of tempeh, Rhizopus oligopsporus comes out as food-derived which makes this stuff are highly low in cost. It is besides safe because it comes from nutrient and does non harmful.


To analyze the consequence of biosorbent dose on the per centum dye remotion.

To analyze the consequence of varied pH of dye solution on the per centum dye remotion.


Rational of the Research

Pollution caused by dyes had affected the society with serious environmental consequence and wellness jobs to human organic structure. For that ground, remotion of dyes from aqueous solutions is indispensable. Problems caused by the fresh of nutrient processing-derived, Rhizopus oligoporous can be solved by doing usage of the biomass as a biosorbent to take dyes. Nowadays, activated C is normally used for surface assimilation in many intervention workss.

Furthermore, bring forthing costs for activated C is really high ; therefore an alternate stuff is needed to do the cost more efficient. Biosorption procedure is an of import alternate method to replace conventional method, which is high cost and more complicated compared to biosorption.

Current Research

Kumari and Abraham, 2006

A survey made by Kumari and Abraham ( 2006 ) on the biosorption public presentation of nonviable biomass of four Fungis such as R. nigricans, R. arrhizus, A.

Niger and A. japonica ) and one barm which is S. cerevisiae handling five different reactive dyes. These beings are all produced as by-products in agitation industries and other surveies show that they are good biosorbents of heavy metals and radioactive compounds. The consequence of Kumari and Abraham ‘s survey shows that most of these beings besides are good biosorbents of reactive dyes.

Merely A. japonica shows comparatively low values for the five dyes tested which is around 4-20 % surface assimilation. Aspergillus niger adsorbed between 58-68 % . S. cerevisiae and R. nigricans were the most first-class adsorbents ( 55-91 % ) and was for some dyes similar to the values given by surface assimilation with activated C. These two beings were so studied further. The pH of the dye solutions was around 5.

9-6.1, but there was besides an experiment done at pH 10. The surface assimilation was most efficient at pH 6 while at pH 10, S. cerevisiae can non absorb any dye at all. Furthermore, R. nigricans merely could adsorb less important sums of reactive green and reactive blue.

Khalaf, 2008

A survey made by Khalaf ( 2008 ) focused on fabric effluent intervention by non-viable biomass of Aspergillus Niger and the alga Spirogyra.

The dye solution contained the commercial Synazol reactive dye, a mixture solution with one ruddy and one yellow dye. The biosorption experiments were performed at different initial pH between 1 to 8, different temperature ( 15-45 °C ) and different biomass burden which is 4 to 12g/L. The A.niger and Spirogyra biomass was inactivated by either gamma radiation or autoclaving. Autoclaving resulted in the highest biosorption values. Harmonizing to Fu and Viraraghavan ( 2000 and 2002 ) , autoclaved of A. Niger had higher biosorption capacity compared to populating biomass.

The surface features of the biomass are changed in a manner that recovers the dye biosorption capacity. A possible account to that is that the autoclaving disrupts the biomass construction and so exposes the surface assimilation sites even more. The consequence of the survey showed that the highest biosorption values were obtained at pH 3 and the lowest values occurred around pH 6 to 8. The optimum temperature and biomass burden for the biosorption was found to be at 30 °C severally at 8 g/L.

Advantages and Important of the Research

Normally, dyes are complicated to biodegrade ( Fu and Viraraghavan, 2000 ) . Application of biological procedures to handle coloured effluent is yet to be to the full established. Among other intervention engineerings, surface assimilation is quickly deriving important method of handling effluent.

Activated C is the most efficient and ever used as adsorbent ( El-Guendi, 1991 ) . However, its high cost has prevented its application, so it is indispensable to look for other alternate low-priced adsorbent which can be replace activated C.Comparing to the other available engineerings such as precipitation, ion exchange, reverse-osmosis and surface assimilation, biosorption gives tantamount public presentation at a really low cost. Apart from cost effectivity and competitory public presentation, other advantages are possible regeneration at low cost, handiness of known procedure equipment, sludge free operation and recovery of the sorbate ( Volesky, 1999 ) .

The usage of dead biomass is enhanced instead than unrecorded biomass because toxic or pollutants from nature can non consequence on the sorption procedure. Other than that, there is no demand for either holding any foods or keeping a growing environment.Furthermore, fungous biomass seems to be good stuff because it can be produced economically utilizing simple and economical growing media. Furthermore, fungous biomass is besides accessible as byproduct or waste stuff from assorted industrial procedures ( Fu and Viraraghavan, 2000 ) . It is merely and economically available anyplace, particularly in locations with hot and humid such as Malaysia.

4.4 Scope of the Study

This survey investigates the biosorption abilities of inactive Rhizopus oligosporus for the remotion of Methylene Blue ( MB ) dye from aqueous solutions.

There are two parametric quantities to be studied which are pH of the dye solution and effects of biosorbent dose.

4.4.1 Biosorption

Biosorption can be defined as the remotion of stuffs such as organic compounds, metal ions and dyes molecules by inactive, inanimate biomass ( stuffs of biological beginning ) as alternate low cost and eco-friendly engineering ( Farooq et al.

2010 ) . It is a belongings of certain types of inactive, dead, microbic biomass to adhere and concentrate heavy metals and other types of molecules or ions from dilute aqueous solution. Biomass exhibits this belongings, moving merely every bit chemical substance, as an ion exchange of biological beginning. It is peculiarly the cell wall construction of certain algae, Fungis and bacteriums which was found responsible for this phenomenon. In similar significance stated by D.J.

Ju et Al. in his research, the consumption or accretion of chemicals by biomass is known as biosorption.


4.2 Biosorbent

There are a batch of attempts were made in many surveies for the remotion of dyes from either aqueous solution or effluents. This includes the usage of metal hydrated oxides, clays, sunflower chaffs, hardwood, fertilisers and steel wastes. The biosorbent used in the survey were prepared from autoclaving and drying Rhizhopus oligosporus ( biomass ) . The dye solution will be handling with inactivated Rhizopus oligosporus. This is because, if the cells of the Fungis are active they are easy affected by toxic compounds and chemicals in the waste H2O and they may so foul the environment by let go ofing toxins.Another trouble when covering with active biomass is that it could non be stored at room temperature for long clip periods before it may botch.

When the biomass is dead and dried it could be stored and transported easier. Based on the old survey, autoclaved of biomass has a higher biosorption capacity compared to populating biomass. ( Fu and Viraraghan, 2000 and 2002 ) . These facts are the grounds why inactivated biomass by autoclaving is chosen for this experiment. Rhizopous Oligosporous

Rhizopus oligosporous is a genus of casts that is found in dirt and works stuff. Normally, it is used as starting motor to do tempeh, a nutrient made of fermented soy beans. Rhizopus oligosporus is the dominant tempeh fungus ( Sharma and Sarbhoy,1984 ) although some other molds, such as R. Oryzae and Mucor spp, may besides lend to the spirit, texture or alimentary value ( Wiesel, Rehm and Bisping,1997 ) .

( Heseltine et al. ) isolated Indonesia and found that merely Rhizophus could do tempeh in pure civilization agitation. They besides found that the 40 strains of Rhizophus studied, 25 of them are R.oligosporus others are R.stolonifier, R.arrhizus.

R.oryzae, and R.formosaensis.This determination was besides confirmed by ( Saono et al ) , who isolated 118 civilizations from 81 tempeh samples collected from markets in assorted parts of Indonesia.

Figure 1: Rhizopous Oligosporous

4.4.3 Absorbate

Formula Weight for MB is 320 gmol-1, with the chemical expression, C16H18N3SCl, and wavelength figure is 662nm. MB is non by and large considered as acutely toxic, but it can hold assorted harmful effects.

On inspiration, it can increase to short periods of rapid or hard external respiration, while by consumption through the oral cavity it produces a combustion esthesis and may do sickness, purging, diarrhoea, and gastritis. MB is one of the commonly used commercial dyes and compared to the other dyes, it has high glare and colour strength, ensuing in high seeable colour, even at really low dye concentration. ( Fu and Viraraghavan, 2000 ) .

4.4.4 Design of Experiment ( DOE )

DOE is used to construct set of experiments of minimal figure with optimal status. This current research consists of two parametric quantities, which is pH of dye solution and dose of biosorbent. The surface charge on each fungous biomass is preponderantly negative at pH 3.

0 until 10.0 due to the presence of ionised groups such as carboxyl, phosphate, and amino groups ( Zumriye et al. 2008 ) . However at pH values below the isoelectric point ( & lt ; 3.0 ) ( Wu and Yu, 2006 ) , the overall surface charge on fungous cells becomes positive due to protonation of nitrogen-containing functional groups such as aminoalkanes which are the major biosorption sites for dye removal.While the scope for the dose of biosorbent is between 0.

1 until 0.6g. Other than that, DOE is besides used to optimise the optimal status for both parametric quantities in order to do the dye clearer.


Figure 2: Flow Chart of the ExperimentRhizopus OligosporousCultivation on PDABiosorbent Preparation( Autoclaving at 121A° C for 30 proceedingss )( Drying at 80 A°C at 24 hours )Biosorbtion Optimization ( Using DOE )pHDoseNoPercentage Dye RemovalBiosorbent Analysis:UV-vis Mass spectrometerYesDatas AnalysisThe experiment could be divided into two different parts.The first portion is the biosorbent readying and the 2nd portion is the biosorption optimisation.

5.1 Biosorbent Preparation

The solid agar, Potato Dextrose Agar ( PDA ) , is prepared by blending a prepared pulverization with de-ionized H2O. The composing of the agar is 5.

0g/L murphy, 20.0g/L dextroglucose, 13.0g/L agar and 0.1g/L Chloromycetin.

After that, the PDA is autoclaved at 121°C for 30 proceedingss. The Petri dish incorporating the PDA is so laid down at room temperature until the medium is turned into solid mass. When it is solid the dish are incubated for 1 twenty-four hours. After incubation one could be excluded that other biomass is turning at the agar. The rhizopus oligoporus turn on PDA is stored in a electric refrigerator at 4°C. By inoculating the civilizations further onto fresh PDA, the biomass is deactivated. After vaccination, the dish is incubated for at least 5 yearss.

Following, the biosorbent is inactivated by being autoclaved at 121°C for 30 proceedingss. The mixture is so filtered to acquire rid of the supernatant and to acquire merely biomass. After filtration, the biosorbent is dried in an oven at 80°C for 24 hours. When the biomass is dry it is powdered and eventually it is ready for the biosorption experiment.

5.2 Biosorption Optimization

Table 1: Optimization of Biosorption

Optimization Design

Design of Experiment ( DOE )

ParameterpHBiosorbent DoseUnit of measurement

gSymbolX1X2Low degree-120.1Medium Level050.

35High Degree+180.6Table 2: Design of Experiment

Design Of Experiment ( DOE )




( Coded )

Biosorbent Dose

( Coded )


( non-coded )

Biosorbent Dosage ( g )

( non-coded )






352620050.35Two different biosorption experiments is performed harmonizing to two different parametric quantities ; biosorbent dose and pH. Each experiment will be done in different intervals and in extra samples. The dye solutions used is prepared by blending the methylene bluish stock solution with a H2O solution at a NaCl concentration of 30g/L. The concentration of the methylene bluish stock solution is 1g/L.

The biosorption will take topographic point in an brooder at temperature 30+/-1 °C, a shaking of 150 revolutions per minute and during 16 hours. After the biosorption the samples will be filtrated to acquire the supernatants. The optical density of that solution is measured by a UV/Vis Spectrophotometer and the optical density units are so converted into concentration units by utilizing a standard curve.

5.2.1 Biosorption dose

The biosorption experiments with different biosorbent dose are performed at natural pH and an initial dye concentration of 50mg/L. The dye solution is prepared by taking 50ml of the stock solution and adding de-ionized H2O until the volume is 1 litre.

Different biosorbent doses is used ; 0 until 0.6g. The biosorbent is weighted and put into a shingle flask. Then 50 milliliter of the dye solution is added to every conelike flask.


2.2 pH

Now the biosorption experiments will be performed at different pH. As in the experiments above the initial dye concentration will be 50mg/L. The proper biosorbent dose to utilize in this experiment is got in the old experiment. Different initial pH will be tested ; 2, 4, 6,8, 10 and 12. The pH of the dye solutions is prepared by adding 1M HCl or NaOH solutions.


3 Data Analysis

Design of experiment ( DOE ) is use to pull out the information from experimental informations or consequences.The standardization criterion on the Absorbance versus concentration is plotted. The sum of dye absorb by biosorbent is calculated by the undermentioned expression ; ( mg dye / mg biosorbent )Where ; qt is the surface assimilation capacity,C0 is the initial dye concentration,Ct is the dye concentration at clip, T,V is the volume of the solution, andW is the mass of biosorbent used,The dye remotion per centum can be calculated by the undermentioned expression ;Removal per centum ( decolourization ) ( % )Where ; C0 is the initial dye concentration, andCt is the dye concentration at clip, T.

6.0 Gantt Chart



Semester 1

Semester 2































Searching / Gathering informationProposal composingProposal entry / proposal defense mechanismProposal rectificationBiosorbent developmentBiosorbent provingProcedure surveyOptimization surveyBiosorbent word pictureAnalyze informations and consequences obtainedThesis composingConcluding study entry and VIVATable 3: Gantt chart for the Final Year Undertaking




Gathering inside informations information about Methylene BlueOctober 2012Collect biosorbent and developed itNovember 2013Prepared all stuff and setup sing biosorption procedureNovember 2013Biosorbent developmentDecember 2013Optimization by DOEJanuary 2013Biosorbent word pictureFebruary 2013Table 4: Milestones


For this current research, the cost can be reduced by operating in reactors of moderate size and by minimising heat and power demands. This is due to the temperature used in this survey is in a room temperature.

This is because, the increase of heat and power generated are affected the cost.


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