mechanisms controlling seasonal...

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MECHANISMS CONTROLLING SEASONAL HYDROCHEMISTRY OF VAMANAPURAMRIVER BASIN, KERALA, INDIA

Vol. 13, No. 2, June, 2018

ISSN: 0973-4155ECO CHRONICLE

PP: 43 - 50

RNI No. KERENG/2006/19177

ABSTRACT

The present study focuses on the hydrochemistry of Vamanapuram River Basin (VRB). Samples were collected fromtwenty five stations with respect to seasons. The data shows lowland and midland stations having exceedingly highvalues. The impact of anthropogenic pollution is severe during monsoon, compared to non-monsoon, owing to the influxfrom sub-basins with heavy precipitation. Na plays the dominant role in the river for all seasons. The dominance ofcations are in the order Na>Ca>Mg>K (monsoon), Na>Ca>Mg>K (post monsoon) and Na>K>Ca>Mg (pre monsoon)and anions in the order Cl>SO4>HCO3>Si2O3 for all the seasons. All nutrients studied were within affordable limits. ThePiper tri-linear diagram revealed that VRB carries mainly 17 types of waters during the study period; 7 water types areflowing through different terrains of the VRB and variations are prevalent according to seasons. The water types of theVRB are controlled mainly by rocks in nature and human interventions that are particularly evident on monsoon. It wasalso observed that during the flow, with regard to the changes in season, the water type changed in each point. TheGibbs diagram proclaims the soil matrix / aquifer-surface water interaction and it seems to be the major process thatcontrols the groundwater chemistry of this area. The water confining to the river is free from ions attributing corrosionand can be safely transported through pipelines, except T- 4 in post monsoon season.

INTRODUCTION

The hydrology of streams is extensively confined to thelithology of the upper reaches of basins. Rock weatheringin the watershed helps in understanding the chemicalweathering as water plays a major part in rock weathering.Deforestation gains importance as a cause for erosion;the mass erosion of fertile substrates not only turns farm-land into barren land but also adds nutrients to the ultimatesinks viz., rivers and lakes, thus virtually causing chemicalcontamination (Chapman, et al., 2001). The excessiveuse of organic fertilizers and manure used as foragricultural enrichment can cause eutrophication in themain streams, followed by ecosystem imbalance (Billen,et al., 2007; Corriveau, et al., 2010).

Eventhough, the spatial and temporal disparities in waterquality are highly prominent and the controlling factorssuch as climate (temperature, humidity, wind andprecipitation), rock types, vegetation, groundwatercontribution, rainwater and flow rate variations (Fritzsons,et al., 2003) add impetus to the thrilling enthusiasm ofresearchers, yet the interactions between the physical

and chemical environment of rivers and their ecologicalstatus are poorly understood (Hilton, et al., 2006). Hence,it is necessary to check the water quality of streamsperiodically, which can have long term engrossment withearth’s maternity.

Gibbs and McIntyre (1970) proposed a diagramwhich represents the ratio of (Na++ K+) /(Na++K++Ca2+) and Cl- / (Cl-+ HCO3-) as a functionof TDS. Based on the Gibbs diagram, there are threemajor mechanisms that regulate the chemistry ofsurface water, viz., evaporation dominance,precipitation dominance and rock dominance.

Study AreaThe Vamanapuram river originates from Chemmunjimottai (elevation:1717m. a.m.s.l.), in the high rangesof the Western Ghats in southern Kerala, one ofthe eight prime “hottest hotspots” of biologicaldiversity in the world, and traverses through thehighland and midland of Thiruvananthapuram and

Vinod Gopal, V1., Dhanya, G2. and Sabu Joseph3

Department of Environmental Sciences, University of Kerala, Trivandrum, [email protected] [email protected] [email protected]

44 ECO-CHRONICLE

Fig. 1 Location map- Vamanapuram River Basin

Kollam districts before debouching into the Anchuthengulake in the lowland region at Chiryankeezhu, Kerala(Fig.1). The Vamanapuram River Basin (VRB) liesbetween 835' to 850’ N latitudes and 7640’ to 7715' Elongitudes and is bounded by Nedumangadu taluk ofThiruvananthapuram district in the south, Kottarakkarataluk of Kollam district in the north, Tamil Nadu State inthe east and the Arabian Sea in the west. It nourishes 29panchayats and one municipality located within the basin.

MATERIALS AND METHODS

Prior to the selection of surface water sampling sites,the study area (i.e., Vamanapuram River Basin, VRB)was visited to identify the stressed locations on the river.Based on this, a total of 25 sampling sites covering themain stream (n= 20; S1 to S20), major tributaries (n= 4;T1 to T4) and river estuary (n=1; L) were selected withan interval of ~5 km. Surface water samples (Tot. No. =75) were collected (~1L each) ~5m away from the marginon the right bank of the river for three seasons (viz.,Monsoon (MON), August 2012; Post-monsoon (POM),December 2012 and Pre-monsoon (PRM), March 2013)for a period of one year. The portrait of sampling stationsis given in Fig.1.

Gibbs ratioGibbs ratios are calculated by the following formulae.Gibbs Ratio I Cation = [(Na+ + K+) / (Na+ + K+ + Ca2+)]Gibbs Ratio II Anion = [Cl- / (Cl- + HCO3

-)]Where all the ion concentrations are expressed in meq/l.

Hydrochemical FaciesThe facies are a function of lithology, solution kinetics,and flow patterns and to evaluate the hydrogeochemistry

(Raju et al., 2009) of surface waters in the VRB, thechemical data of important anions and cations such asCO3

-, HCO3-, SO4

2-, Cl-, Ca2+, Mg2+, Na+, and K+ are plottedon the Piper-trilinear diagram (Piper, 1994) with the helpof Aquachem Software to infer hydrochemical facies inorder to understand and identify the suitability of watercomposition in different classes. The diamond-shapedfield between the two triangles is used to represent thecomposition of water with respect to both cations andanions from which the relation between alkaline earths(Ca2++Mg2+) and alkali metals (Na++K+) can beunderstood. The points for both the cations and anionsare plotted on the appropriate triangle diagrams(Ramkumar et al., 2010). The positions of the points areprojected parallel to the magnesium and sulphate axes,respectively, until they intersect in the centre field(Wasim, et al., 2014).

RESULTS AND DISCUSSION

Major physico-chemical characteristics of surface watersamples (n=75) for three seasons, viz., monsoon (MON),post-monsoon (POM) and pre-monsoon (PRM) are givenin Table 1.

Gibbs Surface Water ChemistryFrom the plot (Table 2 & Fig. 3(a) – 5(b) it could berealized that, in PRM and POM seasons, most of thesamples (n= 19) of the study area fell in the category ofrock dominance and the remaining six samples were inthe evaporation dominance. So, the weathering of rockminerals and the soil - water interaction could be themain processes which contribute the major ions tosurface water of study area. During monsoon season,the majority of water samples (n= 19), the major cations

45ECO-CHRONICLE

Tabl

e 1.

Sea

sona

l flu

ctua

tions

of h

ydro

chem

ical

var

iabl

es i

n Va

man

apur

am R

iver

Bas

in

46 ECO-CHRONICLE

Loci

MON POM PRM

Gibbs

Cation

Gibbs

Anion

Gibbs

Cation

Gibbs

Anion

Gibbs

Cation

Gibbs

Anion

S 1 0.6 0.03 0.16 0.06 0.53 0.14

T 1 0.47 0.04 0.28 0.07 0.44 0.17

S 2 0.43 0.03 0.27 0.08 0.37 0.1

S 3 0.61 0.02 0.37 0.08 0.51 0.1

T 2 0.66 0.02 0.27 0.21 0.68 0.12

S 4 0.63 0.03 0.22 0.06 0.42 0.12

S 5 0.63 0.03 0.17 0.12 0.47 0.13

S 6 0.63 0.04 0.17 0.08 0.47 0.13

S 7 0.58 0.02 0.25 0.12 0.65 0.12

S 8 0.65 0.05 0.16 0.03 0.39 0.13

S 9 0.68 0.03 0.19 0.03 0.67 0.13

S 10 0.65 0.05 0.16 0.03 0.51 0.12

S 11 0.49 0.03 0.11 0.03 0.18 0.09

S 12 0.67 0.04 0.17 0.1 0.67 0.14

T 3 0.67 0.1 0.24 0.09 0.68 0.12

S 13 0.7 0.04 0.4 0.11 0.51 0.12

S 14 0.72 0.03 0.24 0.08 0.65 0.07

S 15 0.69 0.02 0.27 0.07 0.64 0.11

S 16 0.72 0.06 0.2 0.1 0.47 0.12

S 17 0.92 0.29 0.9 0.79 0.91 0.69

S 18 0.98 0.52 0.94 0.89 0.95 0.71

T 4 0.89 0.22 0.71 0.61 0.97 0.64

S 19 0.88 0.55 0.89 0.81 0.95 0.63

S 20 0.93 0.69 0.89 0.84 0.95 0.6

L 0.71 0.86 0.72 0.92 0.96 0.52

were contributed by precipitation dominance and for therest, evaporation dominance. At the same time, thesource of anions in majority of water samples (n=19)were met from precipitation dominance and the rest wasfrom rock dominance and evaporation dominance.

The Hydrochemical FaciesThe chemical data of important anions and cations suchas CO3

-, HCO3-, SO4

2-, Cl-, Ca2+, Mg2+, Na+, and K+ areplotted on the Piper-trilinear diagram (Piper, 1944) toinfer hydrochemical facies in order to understand andidentify the suitability of water composition in differentclasses. The surface water samples from the VRB have

Table 2. Mechanisms controlling surface waterchemistry (Gibbs ratio)

80 60 40 20 20 40 60 80

20

40

60

80

20

40

60

80

20

40

60

80

20

40

60

80

Ca Na+K HCO3+CO3 Cl

Mg SO4

<=Ca + Mg

Cl +

SO4

=>

S1

S2

S3

S4

S5

S6

S7

S8

S9

S10

S11

S12

S13

S14

S15

S16

S17

S18

S19

S20

T1

T2

T3

T4

L

S1

S2

S3

S4

S5

S6

S7

S8

S9

S10

S11

S12

S13

S14

S15

S16

S17

S18

S19

S20

T1

T2

T3

T4

L80 60 40 20 20 40 60 80

20

40

60

80

20

40

60

80

20

40

60

80

20

40

60

80

Ca Na+K HCO3+CO3 Cl

Mg SO4

<=Ca + Mg

Cl +

SO4

=>

S1

S2

S3

S4

S5

S6

S7

S8

S9

S10

S11

S12

S13

S14

S15

S16

S17

S18

S19

S20

T1

T2

T3

T4

L

S1

S2

S3

S4

S5

S6

S7

S8

S9

S10

S11

S12

S13

S14

S15

S16

S17

S18

S19

S20

T1

T2

T3

T4

L

80 60 40 20 20 40 60 80

20

40

60

80

20

40

60

80

20

40

60

80

20

40

60

80

Ca Na+K HCO3+CO3 Cl

Mg SO4

<=Ca + Mg

Cl +

SO4

=>

S1

S2

S3

S4

S5

S6

S7

S8

S9

S10

S11

S12

S13

S14

S15

S16

S17

S18

S19

S20

T1

T2

T3

T4

L

S1

S2

S3

S4

S5

S6

S7

S8

S9

S10

S11

S12

S13

S14

S15

S16

S17

S18

S19

S20

T1

T2

T3

T4

L

Fig. 2(a) Piper-trilinear diagram for Monsoon - VRB

Fig. 2(c) Piper-trilinear diagram for Pre-monsoon – VRB

Fig. 2(b) Piper-trilinear diagram for Post-monsoon – VRB

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Fig. 3 (a) Cations controlling mechanisms of water quality -MON

Fig. 3(b) Anions controlling mechanisms of water quality -MON

Fig. 4(a) Cations controlling mechanisms of water quality -POM

Fig. 4(b) Anions controlling mechanisms of water quality -POM

Fig. 5(a) Cations controlling mechanisms of water quality -PRM

Fig. 5(b) Anions controlling mechanisms of water quality -PRM

48 ECO-CHRONICLE

been plotted on Piper diagram (Fig. 2(a) to 2(c)) andthese ammonium sulphate fertilizers used in fields alsocontribute) and the remaining have non-carbonatehardness (Secondary salinity results from humanactivities like land development and agriculture).Common forms of secondary salinity are the result ofrising groundwater tables (from excessive irrigation) orthe use of poor quality water, clearing vegetationchanges in land use and sea water intrusion. In PRM,80% of the water is controlled by strong acids (naturaland anthropogenic). In POM, 52% of the water iscontrolled by alkaline earth metals (major source of ‘Ca’and ‘Mg’ are rock weathering by nature and by artificialas sewage and some industrial wastes), cause forhardness.

Loci

Water Type

Monsoon Post-monsoonPre-

monsoon

S1 Mg-Cl-HCO3 Mg-Ca-Cl Mg-Cl

T1 Ca-Cl-HCO3 Mg-Cl Mg-Cl

S2 Cl-HCO3 Mg-Cl Mg-Cl-SO4


T2 Mg-Cl-HCO3 Mg-Cl Mg-Cl

S4 Mg-Cl-HCO3 Mg-Ca-Cl-HCO3 Mg-Cl

S5 Cl-HCO3 Ca-Mg-Cl Mg-Cl

S6 Mg-Cl-HCO3 Ca-Cl-HCO3 Cl

S7 Cl-HCO3 Mg-Ca-Cl Mg-Cl

S8 Na-Cl-HCO3 Mg-Ca-Cl-HCO3 Mg-Ca-Cl

S9 Mg-Cl-HCO3 Ca-Mg-Cl-HCO3 Mg-Cl

S10 Mg-Cl-HCO3 Ca-Mg-Cl-HCO3 Mg-Cl

S11 Mg-Cl-HCO3 Ca-Cl-HCO3 Ca-Cl


T3 Na-Cl Ca-Cl-HCO3 Mg-Cl

S13 Mg-Cl-HCO3 Mg-Cl Mg-Cl


S15 Cl-HCO3 Mg-Cl Mg-Cl


S17 Na-Mg-Cl Na-Cl Na-Cl


T4 Na-Mg-Cl Na-Cl Na-Cl



L Na-Ca-Cl-SO4 Na-Ca-Cl Na-Cl-SO4

diagrams revealed the analogies, dissimilarities andtypes of different water in the study areas which wereidentified and summarized in the Table 3.

From the plots and tables (3 & 4), it is evident that seasonalfluctuations do occur in water samples. During MON, outof the 25 samples, majority (47%) fell in the Mg-Cl-HCO3

type, followed by 5 samples (20%) in Na-Mg-Cl facies, 4samples (16%) in Cl- HCO3 facies and the remaining arein Na-Cl-HCO3, Na-Cl, Ca-Cl-HCO3 and Na-Ca-Cl-HCO3

facies, respectively. In POM the water samples showed ascattering pattern, and a generalisation is not possible.Na-Cl facies was occupied by six water samples (24%)another six samples (24%) belonged to Mg-Ca-Cl facies.Four water samples (16%) were located in the Mg-Clcategory, while three (12%) were in Ca-Cl-HCO3 facies.At the same time, 2 samples (8%) were of Ca-Mg-Cl-HCO3

type and the other two were (8%) in the Mg-Ca-Cl-HCO3

facies. The remaining water samples fell in Na-Ca-Cl (4%)and Ca-Mg-Cl (4%) category. During PRM, major watertype was Mg-Cl and this occupied by 15 samples (60%),and another five (20%) belonged to Na-Cl facies. Theremaining water samples were belonged to Ca-Cl (4%),Mg-Cl-SO4 (4%), Na-Cl-SO4 (4%), Cl (4%), and Mg-Ca-Cl(4%) category, respectively.

Appelo and Postma (1996)suggested that the Ca-Clwater type indicated the active process of saline watermixing where, Na+ from sea water is exchanged for Ca2+

adsorbed on the clays. It is not considered that ion-exchange during the saline water incursion is the onlything responsible for this water type here, but it may alsobe from atmospheric dry deposition, wastewater fromseptic tanks and other sources of domestic and industrialwastewaters, fertilizers, etc.

The mixed Ca-Mg-Cl water type indicated that thesesamples occurred under the rock types of Ca-Mg-Cl andthe water type were referred to as base exchange waterions compared to available alkali metal ions (Na + K) inequivalent concentrations. Relatively high Cl - ionconcentrations in the water might be due to the alterationof biotite in the weathered and fracture zones of theunderlying biotite-rich basement rocks, viz., biotitegneisses, biotite schists, biotite granites, etc. (Samanta, etal., 2013). These excess HCO3

- ions then caused therelease of earth alkaline ions into the solution by exchangereaction with exchanger such as clay minerals (Drever,1997).

The water types of the VRB are controlled mainly byrocks in nature and human interventions particularlyevident in MON. About 52% of samples are neutral,having no cationic- anionic domination, and 28% ofsamples are acidic (acidity from SO4 in rocks and

Table 3 Water classification of Vamanapuram River Basin(Piper Diagram)

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The Piper tri-linear diagram reveals that the surface watersamples collected from the different points of the VRBcarry mainly 17 types of waters during the study period.On an average, 7 water types are flowing throughdifferent terrains of the VRB and variations are seenaccording to seasons. In MON and PRM season, 7 typesof waters were flowing along the VRB, while in POM itbecame 8 types. But during the flow, when the seasonchanged the water type in each point was also changed.In the river estuary the water type was Na-Ca-Cl-SO4

type during MON, and changed to Na-Ca-Cl facies inPOM and Na-Cl-SO4 in PRM season.

The water samples from S1, 3, 4, 6, 9, 10, 11, 12, 13,14, 16 and T2 were distributed in Mg-Cl-HCO3 facies inMON and in POM the S1, 3, 12, 14 and 16 were changedto Mg-Ca-Cl water type and in PRM these waters furtherchanged to Mg-Cl category.

The water from S4, S6 and S11 were changed to Mg-Ca-Cl-HCO3, Ca-Cl-HCO3 and Ca-Cl-HCO3 facies inPOM and in PRM it converted to Mg-Cl, Cl and Ca-Clrespectively. While the samples such as S9 and S10were changed to Ca-Mg-Cl-HCO3 and Ca-Mg-Cl-HCO3

type in POM, while in PRM, both these waters exhibitedMg-Cl type character. During POM and PRM season,the water distributed in S13 and T2 (Mg-Cl-HCO3) werechanged to Mg-Cl category. During MON, the watersamples from S2, S5, S7 and S15 shows Cl- HCO3 watertype and S2 and S15 were changed to Mg-Cl category,S5 changed to Ca-Mg-Cl category and S7 changed toMg-Ca-Cl category. But during PRM season S5, S7 andS15 were showed Mg-Cl type water and S2 showed Mg-

Subdivisionof the

diamondCharacteristics of corresponding subdivision of diamond shaped fields

No. of samples

MON POM PRM

1 Alkali earth (Ca2++Mg2+) exceeds alkalies (Na++K+) 13

2 Alkalies (Na++K+) exceeds alkaline earth (Ca2++Mg2+)

3 Weak acids (CO3-+HCO3

-) exceeds strong acids (SO42-+Cl-)

4 Strong acids (SO42-+Cl-) exceeds weak acids (CO3

-+HCO3-) 7 5 20

5Carbonate hardness (Secondary alkalinity) exceeds 50% (Chemical properties

are dominated by alkaline earth and weak acids)

6Non-carbonate hardness (Secondary salinity) exceeds 50% (Chemical

properties are dominated by alkaline earth and strong acids)5 7 5

7Carbonate alkalinity (Primary salinity) exceeds 50% (Chemical properties are

dominated by alkaline earth and weak acids)

8Carbonate alkalinity (Primary alkalinity) exceeds 50% (Chemical properties are

dominated by alkalies and weak acids)

9 Mixed types (No cation-anion pairs exceeds 50%) 13

Cl-SO4 water. The water samples from S17, S18, S19,S20 and T4 were located in Na-Mg-Cl facies during MONand were changed to Na-Cl facies in the POM and PRMseason. The water sample collected from S8 was Na-Cl-HCO3 type in MON and changed to Mg-Ca-Cl-HCO3

in POM and Mg-Ca-Cl in PRM season. The T3 and T4samples were belongs to Na-Cl and Na-Mg-Cl type inMON season, changed to Ca-Cl-HCO3 and Na-Cl typein POM and Mg-Cl and Na-Cl type in PRM season.

CONCLUSION

The quality analysis of water samples covering the mainstream, major tributaries and river estuary for MON, POMand PRM has revealed that physico-chemicalparameters do show both spatial and temporal variations.Spatially, the various parameters, in general, exhibit anincreasing trend towards downstream. Various cationslike Ca, Mg, Na, K are within permissible limits. However,Cl registers an increasing trend in the downstream.Seasonally, PRM shows higher values for most of thephysical parameters. Whereas, some parameters likeNa & Cl are dominating in POM. Na plays a dominantrole in the river for all seasons and the dominance ofcations are Na>Ca>Mg>K (MON), Na>Ca>Mg>K (POM)and Na>K>Ca>Mg (PRM). Again, dominance of anionsvaries Cl>SO4>HCO3>Si2O3 for all the seasons andnutrients are in affordable limit.

Gibbs diagram indicates that during PRM and POMseasons, rock weathering (soil - water interaction) andevaporation are the major process control the waterchemistry, while during MON precipitation and

Table 4. Seasonal variations of water types of Vamanapuram River Basin (Piper Diagram)

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evaporation are the major process controlling waterchemistry. In order to identify the hydrochemical faciesof water for different seasons using Piper diagram, ithas been observed that facies are changing withseasons. During MON 47% samples belong to Mg-Cl-HCO3 type. However, during POM, no facies type isdominating and 24% of the samples show each Mg-Ca-Cl and Mg-Cl category. During PRM 60% of samplebelonged to Mg-Cl type.

REFERENCES

Chapman, P. M., Birge, W. J., Adams, W. J., Barrick, R.,Bott, T. L., Burton, A., Collier, T. K., Cumberland, H. L.,Douglas, W. S., Johnson, L. L., Luther, G. W., O‘Connor,T., Page, D. S., Sibly, P., Standley, L. L. and Wenning,R. J., 2001. “Learned discourse: Sediment quality values(SQVs): Challenges and recommendations”, SETACGlobe, vol. 2, pp. 24–26.

Billen, G., Garnier, J., Nemery, J., Sebilo, M., Sferratore,A., Barles, S. and Benoît, M., 2007. “A long-term view ofnutrient transfers through the Seine river continuum”,Science of the total Environment, vol. 375(1), pp. 80-97.

Corriveau, J., van Bochove, E. and Cluis, D., 2010.“Sources of nitrite in streams of an intensively croppedwatershed”, Water Environment Research, vol. 82(7),pp. 622-632.

Fritzsons, E., Hindi, E. C., Mantovani, L. E. and Rizzi, N.E., 2003. “Conseqüências da alteração da vazão sobrealguns parâmetros de qualidade de água fluvial”, RevistaFloresta, vol. 33(2), pp. 201-214.

Hilton, J., O’Hare, M., Bowes, M. J. and Jones, J. I.,2006. “How green is my river? A new paradigm ofeutrophication in rivers”, Science of the TotalEnvironment, vol. 365(1), pp. 66-83.

Gibbs, J. and McIntyre, G. A., 1970. “The diagram, amethod for comparing sequences”, European Journalof Biochemistry, vol. 16(1), pp. 1-11.

Raju, N. J., Ram, P. and Dey, S., 2009. “GroundwaterQuality in the Lower Varuna River Basin, VaranasiDistrict, Uttar Pradesh, India”, Journal of the GeologicalSociety of India, vol. 7, pp. 178-192.

Piper, M., 1944. “A graphical procedure in the geochemicalinterpretation of water analysis”, Transactions AmericanGeophysical Union, vol. 25, pp. 914–928.

Ramkumar, T., Venkatramanan, S., Mary, A. I., Tamilselvi,M. and Ramesh, G., 2010. “Hydrogeochemical qualityof groundwater in Vedaraniyam town, Tamilnadu, India”,Research Journal of Environmental and Earth Sciences,vol. 2(1), pp. 44-48.

Wasim, S., Khurshid, S., Shah, Z. and Raghuvanshi, D., 2014.“Groundwater Quality in Parts of Central Ganga Basin, AligarhCity, Uttar Pradesh, India”, The Proceedings of the NationalAcademy of Sciences, India, vol. 80(1), pp. 123-142.

Appelo, C. A. J. and Postma, D., 1996. “Ion exchangeand sorption”, Geochemistry, Groundwater and Pollution,Balkema, pp. 142–201.

Samanta, P., Mukherjee, A. K., Pal, S., Senapati, T.,Mondal, S. and Ghosh, A. R., 2013. “Major ion chemistryand water quality assessment of waterbodies at Golapbagarea under Barddhaman Municipality of Burdwan District,West Bengal, India”, International Journal ofEnvironmental Sciences, vol. 3(6), pp. 1938-1956.

Drever, J. I., 1997. “Weathering processes”, In: Saether,O. and de Caricat, P. (Eds.), Geochemical Processes,Weathering and Groundwater Recharge in Catchments,Taylor & Francis publications, Rotterdam, Balkema,Chapter 1, pp. 3–19.

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EFFECT OF NITRATE AND PHOSPHATE LEVELS ON BIOCHEMICAL CONTENTS AND FATTYACID METHYL ESTERS PROFILE OF MONORAPHIDIUM CONTORTUM (THURET)

Ajayan, K.V*1., P.S. Manaswini2 and C.C. Harilal11Division of Environmental Science, Department of Botany, University of Calicut, Malappuram District, Kerala.

2Department of Environmental Science, University of Calicut, Malappuram District, Kerala.Corresponding Author: email: [email protected]*

Vol. 13, No. 2, June, 2018


PP: 51 - 59


ABSTRACTExploration of value added products from new algal species are the future assets for food, cosmetics and pharmaceuticalindustries. The present study was carried out to enhance the biochemical and fatty acid synthesis of Monoraphidiumcontortum, to get a hold of single cell protein production. We conducted experimental culture using various concentrationsof NaNO3 as nitrogen source and K2HPO4+KH2PO4 as source of phosphate for assessing the changes in single cellattributes. Maximum cell concentration of 47.2×105 and 14.4×105 cells mL-1 were observed in P1 (Phosphate) and N1(Nitrate) sets, than control. The highest specific growth (0.192 µd-1) and division rate (k=0.277) was observed in P1sets. The highest protein (48.45mg L-1) and carbohydrate contents (21.4mg L-1) were observed in P1 and N2 setsrespectively. In contrast, highest total lipid content 30.5% and 28% was obtained in P1 and N1 concentration respectively.The higher C/N ratio 7.91 (N1) and 7.54 (P1) showed under limited N and P culture conditions. Maximum TFA contentof control (70.5%), N1 (82.4%) and P1 (84.6%) sets were significantly varied. From these results it can be concludedthat biochemical contents and fatty acid profile of M. contortum is really promising for value addition.Key words: Monoraphidium; NP modulation; FAMEs; CHNS, aquafeeds

INTRODUCTION

Microalgae are the natural base for the entire aquaticfood chain. Microalgal species commonly used for foodand cosmetics include Arthrospira, Chlorella Sp.,Dunaliella Sp., and Haematococcus Sp. (Buono et al.,2014). Beneficial supplements derived from microalgaehave been the essential focal point of microalgalbiotechnology for a long time. Various studies conductedby researchers have demonstrated that the algal proteinsare of high quality and comparable to conventional crops.To date, a high algal biomass contents are marketed ashealth supplements, cosmetics or animal feed (Ghasemiet al., 2011). In the present market, Chlorella, Dunaliellaand Spirulina have dominated commercially due to theirhigh protein content and good nutritional value (Castro-puyana et al., 2013). Algal biomass are also a depositorysite for various amino acids, vitamins, minerals(potassium, sodium, magnesium, iron and calcium),carotene and other beneficial substances (Guil-Guerreroand Rodriguez- Garcia, 2008) and are considered as safe(GRAS) by the European Food Safety Authority (EFSA)(Chacon-Lee and Gozalez-Marino, 2010). Proteincontent of Arthrospira platensis (50-70%), Chlorella

vulgaris (38-58%); Nannochloropsis oculata (22-37%),Porphyridium cruentum (8-56%), and Haematococcuspluvialis (45- 50%) were noted to be high on a dry weightbasis (Safi et al., 2014). The product derived from algalbiomass mainly direct to pharmaceutical andnutraceutical market for more than hundreds of milliondollars (Apt and Behrens, 1999).

The wide interest in using microalgal biomass asfeedstock for biofuel (Chisti, 2007), pharmacology(Converti et al., 2009), coloring agent (Ajayan et al., 2012)etc. is mainly due to their mass production, higher growthrate and accumulation of valuable molecules. Similar toneutral lipids, glycolipids are a class of amphiphiliccompounds, which play an important role inpharmaceutical, food and cosmetic (Wang et al., 2016)industries. Astaxanthin pigments from H. pluvialis, arehighly used for salmonoids, shrimps, lobsters andcrayfish diets, to increase the pink colour in flesh, to getmaximum price in market (Muller-Feuga, 2000). Theincreased growth and resultant biochemical propertiesof microalgae can be improved by various concentration

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of nitrate (Pancha et al., 2014) and phosphate, pH,photoperiod (George et al., 2014) and light intensity(Ajayan and Selvaraju, 2011).

Recently M. neglactum (Bogen et al., 2013) and M.minutum are highly used as feedstock for biofuelproduction under nitrogen limiting conditions. Nitrogenis an essential factor for the structural characteristics ofproteins and also affects the relative content of fatty acids(Spolaore et al., 2006). Previous studies on growth,protein and lipid productivity from algae have mostly beenperformed on Chlorella, Scenedesmus and Spirulina.Most of the microalgae are playing an important role inaquatic ecosystem by stabilizing pH, reducing bacterialgrowth and improving the quality of rearing medium (Lio-Po, 2005) and act as primary food source for fish andzooplankton.

Unlike the studies undertaken by investigators toexplore the lipid accumulation and biofuel productionfrom microalgae, studies on single cell proteinproperties and fatty acid production by microalgae arelimited. Microalgae can also be mixed with animal andfish feed ingredients and can be given as pellets or pasteor powder to improve the diet quality. M. contortum is afast-growing non-toxic microalga commonly found infreshwater ponds and aquarium tank. The present studyfocused on the enhancement of growth, pigments andchanges in the single cell characteristics such asprotein, carbohydrate, lipid, elemental composition andfatty acid profile of M. contortum under various nitrateand phosphate concentration in Bold’s Basal Medium(BBM).MATERIALS AND METHODSIsolation of microalgae

The strain isolated from an aquarium tank was purifiedby agar plating method and sub-cultured under controlledlab conditions. The culture of M. contortum wasmaintained in Bold’s Basal Medium (BBM) at 28°C inlaboratory conditions, illuminated by white fluorescentlamps at an intensity of 2000 lux in a 12/12 hr light/darkcycle.

Experimental setupThe culturing was done in 500 ml flasks containing 200ml of BBM with various concentration of NaNO3 (N1-0.125,N2-0.5, N3-1.0 g L-1) and K2HPO4 + KH2PO4 (P1-0.0187+0.0437 P2-0.0375 +0.0875, P3-0.15+0.35 g L -1) intriplicates and BBM as control containing 0.25g of NaNO3

, 0.075g of K2HPO4 and 0.175g of KH2PO4 per litrerespectively. Phosphate concentration was kept constantin all varying nitrate concentrations of the medium, andvice versa. For each concentration, desired amount of

algal inoculums (cells×104 per liter) were added. Theexperiment was done in a controlled condition with a lightintensity of 2000 lux, at 28°C temperature.

Cell concentration and specific growth rateCell concentration was determined by placing an aliquotof well-mixed culture suspension on a hemocytometer.Each sample was measured twice and the average valuewas used. The growth rate of a microalgal population isa measure of the increase in biomass over time and it isdetermined from the exponential phase. Specific growthrate (µ) was calculated with the following equation:

12

12 /

tt

NInN

Where, N2 and N1 are the no. of cells at times t2 and t1.Division rate, and Doubling time (T2) can becalculated using following formula: .

Biochemical analysisThe chlorophyll and carotenoid content was extractedby 5 ml of 600C DMSO solution. The absorbance at 480nm, 649 nm and 665 nm determined using UV-Visspectrophotometer (A&E AE-S60-2UPC) (Wellburn,1994). Total protein content was determined by themethod of Lowry et al., (1951). Total lipid content wasdetermined by a modified version of the Bligh and Dyermethod (1959). The carbohydrate content wasdetermined using the method of National RenewableEnergy Laboratory (NREL), USA (Van Wychen andLaurens, 2013; Nielsen, 2017).

Transesterification and fatty acid analysisLipid extracts were converted to methyl esters withmethanolic H2SO4 and hexane (Christie, 2010). The fattyacid methyl esters (FAME) samples were analyzed usinggas chromatography (Shimadzu 2010 Plus). Methylesters prepared from a FAME (C14-22) standard mixturewere purchased from Sigma Aldrich.

Organic elemental analysisThe elemental (C, H, N, S) composition (%) of driedbiomass (105°C for 24 hr in oven) was analyzed by theCHNS organic elemental analyzer (FLASH 2000, Thermoscientific ) using sulphanilamide as a reference standard.The measured values of the standard had <0.03%variation as compared to theoretical value of its C, H, N,S composition in all analytical results. All samples wereanalyzed in triplicates.

Statistical analysisThe statistical significance was evaluated by one-wayanalysis of variance (ANOVA) at (p< 0.05) and valuesare expressed in mean ± SD using SPSS version 16(SPSS, Cary, NC, USA).

53ECO-CHRONICLE

Fig. 1. Effect of different nitrate and phosphate levels on cell

number of M. contortum.

Fig. 2. Effect of different nitrate and phosphate levels onspecific growth rate (SGR) division rate (k) [primary Y axis]and doubling time (T2) [Secondary Y axis] of M. contortum.

Fig. 3. Effect of different nitrate and phosphate levels onprotein content of M. contortum. The data represents the

mean±SD of the mean for eight samples (n=8).

Fig. 4. Effect of different nitrate and phosphate levels oncarbohydrate yield of M. contortum. The data represents the

mean±SD of the mean for eight samples (n=8).


In this study, the effect of various concentrationsof nitrate and phosphate were applied to enhancethe SCP characteristics and fatty acid productionof M. contortum. Biochemical changes weresignificantly affected by nitrate and phosphateconcentration. It is confirmed, a narrow alterationof medium composition can induce growth andgrowth attributes of M. contortum. Common BBMcontains 0.25 g L-1 NaNO3, 0.075 g L-1 K2HPO4 and0.175 g L-1 KH2PO4 and it is considered as the basicrecommended NP ratio for most of the freshwateralgal growth. However; the ratio of NP utilizationby various algae and their biochemicalaccumulation is highly varied according to variousdosages (Liang et al., 2013). To determine whatconcentration might be better to support the growthof M. contortum, thus; the organism was grown inmodified BBM with various concentrations ofnitrate and phosphate to get a hold of single cellprotein production. The parameters such as cellconcentration, specific growth rate, pigment,biochemical and fatty acid content were observedfor 25 days of culturing period.

Cell concentration and specific growth rateEffect of different nitrate and phosphateconcentrations on the growth of M. contortum isshown in Fig.1. The maximum cell concentration(47.2×105 cells mL-1) was observed in P1 on the20thd of cultivation. On the contrary, cell densityunder N1 concentration (14.4×105 cells mL-1)showed significant variation at 19thd. Surprisingly,three-fold increase in cell concentration wasobserved under low phosphate (P1) concentrationwhereas increased nitrogen concentration muchlowered the cell growth. Previous reports had alsofound higher concentration of nitrogen caused aninhibition of cell division in Chlorella vulgaris(Przytocka-Jusiak et al., 1977) and C. pyrenoidosa(Wang et al., 2016). Our result also confirmed thatM. contortum can grow under lower nitrogen (N1)and phosphorus (P1) concentrations thanrecommended doses (control).

The effects of different nitrate and phosphateconcentrations on the specific growth rate (SGR)and division rate of M. contortum are shown inFig.2. The average specific growth rate was 0.112to 0.192 µd-1 observed in entire experimental sets.SGR of M. contortum was higher in P1 (0.192µd-1)than control 0.165 µd-1. In the same way, thedoubling time for P1 was much lesser (3.6 d-1) thancontrol (4.2 d -1) and nitrogen sets (>5 d -1).

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Fig. 5. Effect of different nitrate and phosphate levelson lipid content of M. contortum. The data representsthe mean±SD of the mean for eight samples (n=8).

Biomass dryweight (g/l)

Experimental condition

Days Control N1 N2 N3 P1 P2 P3

5 0.233±0.02 0.228±0.02 0.236±0.02 0.233±0.01 0.254±0.01 0.254±0.02 0.244±0.01

10 0.320±0.02 0.283±0.03 0.272±0.04 0.259±0.01 0.443±0.03 0.385±0.01 0.338±0.01

15 0.474±0.01 0.393±0.06 0.367±0.07 0.364±0.04 0.728±0.05 0.597±0.05 0.503±0.03

20 0.687±0.04 0.500±0.08 0.492±0.05 0.475±0.06 1.160±0.09 0.841±0.03 0.660±0 .05

Table 1. Biomass content of M. contortum during different experimental conditions. The data represents themean±SD of the mean for 3 samples (n=3).

ResponseParameters(mg/l)


Days Control N1 N2 N3 P1 P2 P3

Chlorophylla

5 0.618±0.018 0.453±0.049 0.436±0.016 0.460±0.009 0.749±0.013 0.66±0.02 0.64±0.01

10 0.49±0.016 0.567±0.013 0.444±0.018 0.539±0.01 0.56±0.049 0.602±0.02 0.539±0.009

15 1.50±0.050 1.076±0.036 1.096±0.05 0.938±0.035 1.75±0.036 1.14±0.041 0.83±0.035

20 1.38±0.071 1.235±0.058 1.269±0.071 1.116±0.019 2.86±0.058 1.53±0.023 1.33±0.02

Chlorophyllb

5 1.068±0.038 0.85±0.036 0.816±0.044 0.87±0.02 1.115±0.03 1.09±0.02 1.055±0.01

10 0.25±0.036 0.94±0.046 0.76±0.038 0.926±0.046 0.37±0.07 0.46±0.0242 0.44±0.01

15 0.28±0.05 1.23±0.041 1.155±0.05 1.075±0.015 0.87±0.06 0.564±0.061 0.495±0.05

20 0.66±0.071 1.186±0.05 1.19±0.031 1.13±0.04 1.687±0.025 0.776±0.076 0.697±0.039

Carotenoids

5 0.067±0.04 0.030±0.003 0.020±0.004 0.029±0.005 0.064±0.03 0.066±0.02 0.088±0.02

10 0.157±0.036 0.096±0.006 0.11±0.006 0.093±0.004 0.165±0.07 0.133±0.02 0.176±0.02

15 0.368±0.02 0.203±0.005 0.203±0.005 0.228±0.002 0.47±0.06 0.35±0.03 0.219±0.05

20 0.420±0.04 0.289±0.008 0.301±0.004 0.262±0.003 0.77±0.025 0.447±0.03 0.395±0.01

Totalchlorophyll

5 1.75±0.068 1.33±0.063 1.272±0.054 1.36±0.065 1.93±0.05 1.817±0.06 1.78±0.06

10 0.897±0.036 1.603±0.06 1.315±0.068 1.558±0.046 1.09±0.07 1.19±0.07 1.15±0.07

15 2.15±0.07 2.50±0.055 2.45±0.05 2.24±0.025 3.09±0.06 2.05±0.03 1.54±0.08

20 2.58±0.04 2.71±0.08 2.760±0.041 2.5±0.032 5.31±0.04 2.75±0.03 2.42±0.09

Table 2. Photosynthetic pigments of M. contortum during different experimental conditions. The data representsthe mean±SD of the mean for eight samples (n=8).

Consequently the division rate for P1 was significantlyhigh (0.277 per day) when compared to other sets. Thissuggests that algal cells continued to divide usingminimum available nutrients up to a state of depletionand this reflected on a sudden drop in cell number attheir stationary phase (22thd). The various nitrogen and

phosphorus sources are essential nutrients for microalgalgrowth and also necessary for the synthesis of vivoprotein, nucleic acids and chlorophyll (Wu and Miao,2014).

Under different nitrate and phosphate levels, biomassvalues were markedly increased in M. contortum(Table.1). The maximum biomass concentration(1.16±0.09g L-1) was obtained at cells grown in theconcentration of P1 than other sets. Wang et al., (2016)reported that 0.02g L-1 of hydrophosphate concentrationwas used to achieve maximum biomass of Chlorella(1.44g L-1) and Synechococcus (1.83gL-1). These resultsindicate that M. contortum is also been able to grow inphosphorus limiting conditions and produce good amountof biomass. It also confirms that, phosphorus starvationhas a greater influence on biomass concentration of M.contortum than nitrogen starvation. Moreover,phosphorus is the main component of nucleic acids,proteins and phospholipids (Wu and Miao, 2014).

55ECO-CHRONICLE

Organicsubstrate/Experimentalcondition

Nitrate and phosphate concentrations

Control N1 N2 N3 P1 P2 P3

Carbon (%) 40.6 44.3 43.8 41.1 46.03 42.05 43.6

Hydrogen (%) 5.9 7.5 6.31 6.2 6.8 6.4 6.32

Nitrogen (%) 8.61 5.6 8.1 6.22 6.1 8.14 8

Sulfur (%) 0.01 0.01 0.01 0.04 0.01 0.02 0.02

C/N 4.71 7.91 5.40 6.61 7.54 5.16 5.45

Table 3. Mean CHNS composition of microalgal biomass from various NP sets.

Table 4. Fatty acid composition of M. contortum during different experimental conditions on the harvesting day.The data represents the mean±SD of the mean for three samples (n=3).

Fatty acidcompositions(%)

Nitrate and phosphate concentrations

C N1 N2 N3 P1 P2 P3

C16:0 18±0.5 22±1.1 8.1±0.4 3.1±0.2 23.8±1.1 20.2±1.0 14±0.82

C16:1 3.1±0.1 3.4±0.2 0.0 0.0 3.1±0.2 1.1±0.1 2.6±0.1

C18:0 8.5±0.4 13.8±0.9 6.6±0.7 4.5±0.22 4±0.2 4.3±0.1 6.9±0.2

C18:1 22±1.0 30.2±1.5 11.6±0.81 13.5±0.84 33.7±1.4 21.1±0.92 19.6±0.95

C18:2 16.4±0.7 8±0.9 12.5±1.1 2.65±0.1 12.9±0.9 7.9±0.6 5.3±0.2

C18:3 2.5±0.1 5±0.2 3.7±0.2 0.0 7.1±0.4 1.8±0.1 3.2±0.1

SFA 26.5 35.8 14.7 7.6 27.8 24.5 20.9

MUFA 25.1 33.6 11.6 13.5 36.8 22.2 22.2

SFA+MUFA 51.6 69.4 26.3 21.1 64.6 46.7 43.1

PUFA 18.9 13 16.2 2.65 20 9.7 8.5

TFA 70.5 82.4 42.5 23.75 84.6 56.4 51.6

Others 29.5 17.6 57.5 76.25 15.4 43.6 45.4

Effect of nitrate and phosphate concentration onPigments

Microalgal pigments are the rich source of antioxidantsand mineral contents which direct the growth throughphotosynthesis. Table.2 shows the effects of nitrate andphosphate concentrations on pigments of M. contortum.The changes on pigment composition were evident fromthe colour of cultures. During the experimental periodthe highest value for Chl-a content (2.86±0.058 mg L-1)was observed at P1 on the 20thd of culture, whereaslowest Chl-a was observed at P3 sets. When comparedto the values of P1 sets, 55% of reduction in Chl-a (1.269mg L-1) was observed, when cells was grown under N2concentration. The cell concentration and pigmentcontent of M. contortum were higher in P1 than control.This demonstrated that the higher phosphateconcentration or previously recommended control BBMreduced the pigment accumulation and biomassproduction of this organism. Based on the resultsdescribed above, to achieve enhanced pigmentproduction in M. contortum, P1 was sufficient than

normal recommended dose of both phosphateconcentration (0.075 gL -1, 0.175 gL-1) in BBM. Thechanges in pigment accumulation in cells are consideredto be an adaptation mechanism against variousconditions and the ratio of Chl a/b as well as carotenoid/total chlorophyll increase might be due to the fluctuationin nutrient composition (Sarah et al., 2016).

The highest value of Chl-b (1.687±0.025 mg L-1) wasrecorded in lower phosphate concentration (P1) at 20 thdof culturing than that of control medium. A significantreduction in chl-b was noted under nitrate concentrations.At the same time, higher nitrate concentration (>control)significantly reduced the pigment production in thisorganism. The lowest pigment concentration might bedue to the stress imparted lower photosynthetic rate andlesser cell concentration (George et al., 2014).

The highest carotenoid content (0.77±0.025 mgL-1) wasobserved in P1on the 20thd than control (0.420±0.04 mgL-

1). From the above results suggest that more than P1concentration of phosphate significantly reduced the



56 ECO-CHRONICLE

synthesis of carotenoid content in the tested algae.Kilham et al., (1997), studied nutrient limitation onAnkistrodesmus falcatus and reported that phosphorusstarvation reduces chlorophyll content. In our study, thehighest carotenoid content (0.301±0.004mgL -1) wasobserved at N2 concentration, i .e. one fold ofrecommended BBM or control. Previously researchersexplained highest carotenoid content is indicative ofactive stress conditions due to the decrease in lightharvesting complex (George et al., 2014) and increaseof nitrate concentration. Przytocka-Jusiak et al., (1977)observed, rich nitrogen concentration causes aninhibitory effect on the cell division of C. vulgaris. Fromthe above results, it is confirmed that increasing of nitrateconcentration and decreasing of phosphateconcentration stimulate to accumulate more pigmentcontent in M. contortum.

Effect of nitrate and phosphate on biochemicalcontentsThe effect of various nitrate and phosphateconcentrations on the protein content of M. contortum isshown in Fig.3. In this study, nitrate and phosphatemodulation significantly influenced the protein content.Researchers reported various optimization approachesto maximize the biochemical accumulation, nutrientuptake, chlorophyll and biomass production formicroalgae cultivation (Abdelaziz et al., 2014). The typicalcomposition of the biomass of M. contortum have 19-35% of l ipid 28-45% of protein and 17-25% ofcarbohydrates contents in relation to their dry weightbasis (Jaruwan et al., 2014).

Microalgal biomass considered as a complete protein,directly it can be used as feed in the form of algal biomasspellet or as single cell proteins. The highest protein valueof M. contortum (48.45±2.5 mg L-1) was observed at P1concentration on the 15thd of culture. Lowest value ofprotein was observed at higher (P3) phosphateconcentration, which indicates that, in the P-limited state,excess N accumulates in the protein fraction. While underrich (0.5 g L-1 and 1.0 gL-1) nitrate concentration, themaximum protein content (15.47±0.71 mg L -1 and13.45±0.58 mgL-1) was observed on 20th d than controland other nitrate sets. Ajayan and Selvaraju (2011)reported NaNO3 and KNO3 increases the protein contentin S. platensis under different conditions.

Apart from their protein characteristics, microalgae arealso a rich source of useful carbohydrates existed in theform of starch, sugars, cellulose and otherpolysaccharides. The effect of different nitrate andphosphate concentrations on the carbohydrate contentof M. contortum is shown in Fig.4. The highestcarbohydrate content (21.42±0.41 mg L-1) was observed

at N2 concentration on 15 thd of culture. The resultsobtained from P2 concentration, the highest carbohydratecontent of 19.86±0.77 mg L-1 was achieved on the 10thd.The lowest protein content of M. contortum was obtainedat control culture. Differences in nutrient fluctuation maycause significantly on physiological and biochemicalsynthesis and resultant yield of lipid, protein andcarbohydrate content in cells (Hulatt et al., 2012). Majordecreases in total carbohydrate with the concomitantincrease in total protein with supplementation of nutrientsare reported in various algae (Young et al., 2009).Moreover, the first response to nutrient deprivation iscarbon skeletons channeled to lipid synthesis, in the laterstage of starvation where carbohydrates are degradedto support fatty acid and lipid synthesis (Risman-Yazdiet al., 2011).

The effect of different nitrate and phosphateconcentrations on the lipid percentage of M. contortumis shown in Fig.5. Upon the reduction of nitrateconcentration from recommended dosage of 0.25 to0.125 g L-1, the lipid content of M. contortum increasedsignificantly. The maximum lipid content of M. contortumobserved at low nitrate (N1) level was 28%, which wasapproximately one-fold increase than that of nitrate rich(<17%) culture. In the case of different phosphate levels,it is interesting to note that the maximum lipid content ofM. contortum (30.5±1.3%) was obtained in cells grownin P1 concentration, which was 1 times than that obtainedin phosphate rich (20±0.57%) culture. This confirmed thatvariations in NP ratio have different influence on M.contortum for total lipid accumulation. Li et al., (2010)reported, total lipid content of Scenedesmus increasedfrom 23% to 53% with a reduction in initial totalphosphorus (as phosphate) concentration of 0.1 from2.0 g L”1. Similarly, lipid synthesis lags behind that ofstarch in N-starved Chlamydomonas reinhardtii andemphasized that lipid synthesis occurs only when thecarbon supply exceeds the capacity for starch synthesis(Fan et al., 2012). Previous studies also showed thatnitrate limitation stimulated an increase in lipid contentof about 6.7% in single cell protein algae Haematococcuspluvialis (Recht et al., 2012), 40.2% in Chlorella sp. NC-MKM and 37.3 % in Scenedesmus acutus NC-M2. Toachieve best combinations of protein, carbohydrate andlipid in single cell nutrient supply to algae must beconsidered. In present SCP targeting study on M.contortum, the total lipid content was increased to 10%by phosphate limitation. These results suggested thatM. contortum has higher content of lipid and it can bemade suitable for various supplemented feeds. Thepossible reason towards the increase in lipid content innitrogen and phosphorus limitation is that lipid maintainsnormal metabolic function during exponential stage. Theamount of nitrogen and phophorus present in any type

57ECO-CHRONICLE

of water are considered as essential factors and hasdirect influence on microalgal lipid accumulation (Li etal., 2010). The elemental composition (% C, H, N and S)of Monoraphidium biomass differed under variousexperimental sets is presented in Table 3. In this studyC, H, N and S elemental composition of biomass varied40.6-46.03, 5.9-7.5, 5.6-8.61 and 0.01-0.04, respectively.C/N composition varied from 4.71to 7.91 and it wasobserved that C/N ratio was higher than control (4.71)under various NP concentrations. Higher C/N inmicroalgal biomass in N1 (7.91) and P1 (7.54) grownbiomass confirmed that lipid accumulation was due tonitrogen limitation in the cell.

The fatty acid profile of algal oils present in cells cultivatedunder various concentrations of nitrate and phosphateare presented in table 4. Major fatty acids detected inthe lipid fractions were C16:0, C16:1, C18:0, C18:1,C18:2 and C18:3. The dominant fatty acids C18:1 was33.7±1.4% and 30.2±1.5% noted at P1and N1respectively. Whereas, the amount in C16:0 wassignificantly varied among the sets. The highest saturatedfatty acid (35.8%) was observed under N1 set. Although,the amount of PUFA content of this organism wasobserved under P1 (20%) set. It was significantly higherthan that of control and nitrogen less cultures. The TFAcontent of M. contortum under control, N1 and P1 setswere 70.5, 82.4 and 84.6% respectively, which make thisorganism highly suitable for single cell protein productionand a detailed toxin analysis of this organism is requiredbefore its addition as a human supplemental diet.

CONCLUSIONS

The results targeting SCP production from M. contortumreveal that higher concentration of phosphate in BBMmight reduce the growth rate, biomass production andpigment composition. To achieve enhanced pigmentproduction in M. contortum, P1 concentration wassufficient than normal recommended dosage in BBM.The highest carbohydrate content of this species wasobtained at 0.5 g/L nitrate (N2) concentration and P2concentration respectively. Also, the highest lipidcontent was observed at P1 and N1 concentrationsrespectively. For the enhancement of protein production,P1 concentrat ion was suffic ient than normalrecommended dosage of BBM. The entire outcomes ofthe present study confirm that M. contortum can becultivated for various SCP products within certainlimitation of nitrogen and phosphorus concentrations.Moreover, lower nitrogen/phosphorus concentrations ornitrogen/phosphorus starvations were more likely toinduce a sharp change in pigment, protein,carbohydrate, lipid and fatty acid profile of thismicroalga.

ACKNOWLEDGEMENTS

The First author expresses his sincere thanks to DST ,SERB – Fast Track Young Scientist Scheme (FileNo.YSS/2014/000206), New Delhi for the financialsupport. The entire research team is thankful to CentralSophisticated Instrumentation Facility, University ofCalicut, for CHNS analysis.

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Vol. 13, No. 2, June, 2018


PP: 61 - 68


ASSESSMENT OF WATER QUALITY AND PLANKTON COMMUNITIES OF POND ECOSYSTEMSOF KULATHUPUZHA GRAMA PANCHAYAT, KOLLAM DISTRICT, KERALA

Vincy M. V. 1, Shimi. S. S.2, Brilliant, R.2, Anila, G.2 and Alexander, T.2

1Dept. of Microbiology & Biochemistry, St. Berchmans College, Changanacherry, Kerala.2Dept. of Environmental Sciences, St. John’s College, Anchal, Kerala.

Corresponding author: [email protected]

ABSTRACT

The present study has been carried out to evaluate the organic pollution level of six ponds of Kulathupuzha Gramapanchayat, Kollam District, Kerala. The water quality parameters of each sampling site were measured, which includetemperature, pH, dissolved oxygen, salinity, total hardness, sulphate, nitrate, phosphate, EC and TDS. The diversityand distribution of micro algal members were also worked out. Algal members from the pond were representingmembers of Chlorophyceae, Cyanophyceae, Desmidaceae and Bacillariophyceae. The Palmer’s algal index showedthat FP1 has high organic pollution. The data on physico-chemical analysis supported the Palmer’s organic pollutionindex.Key words: Plankton diversity, Pond ecosystems, Kulathupuzha grama panchayat

INTRODUCTION

Optimal water quality is obviously essential for asustainable environment. Water pollution is harmful notonly to aquatic animal breeding, but also to human health.Polluted or non-optimum levels of water qualityparameters and poor management practices can causea series of physiological alteration on the growth andeven survival of aquatic animals (Chang et al., 2017;Carbajal- Hernández et al., 2013). Moreover, unbalancednutrient inputs and assimilations will lead to thedeterioration of water quality (Tran et al., 2013). As animportant primary producer in aquatic environments, suchas ponds, lakes and rivers, phytoplanktons can be usedas biological indicators of water quality monitoring asthey are sensitive to changes in water quality (Jamshidet al., 2016). Thus, the relationship between water qualityand phytoplankton community have gained increasingattention in the assessment of natural and artificial waterbodies (Lv et al., 2014; Masmoudi et al., 2015).

The state of Kerala is blessed with a large number offreshwater resources, which serve as sources of waterfor drinking and recreational purposes. Many of thefreshwater resources are currently under threat due tohuman interference. The main reasons for this includeurbanization and industrialization. Land filling is yet

another problem leading to the destruction or impairmentof traditional ponds. In India many studies have beencarried out in lotic and lentic systems, but significant studyhas not been carried out in the ponds of Kerala, excepta few by Jose & Sreekumar (2005, 2006), Jose et al.(2008), Jose and Kumar (2011). The present study is anattempt to evaluate the pollution associated with the lenticsystems associated with a grama panchayat usingbiological and chemical methods.

Study areaKulathupuzha is a village in the eastern part of Kollamdistrict of Kerala, India. It covers an area of around 424.06m2 and comes under Anchal block in Punalur Taluk.Kulathupuzha forms a part of the southernmost reserveforests of the State. The forest linked with the presentlocation is predominantly of tropical wet - and semi -evergreen and moist deciduous types. Kulathupuzhaforests have the largest variety of exotic MyristicaSwamps in Kerala. Also the forest is the origin of Ithikaraand Kallada rivers.


Water samples were collected on a monthly basis fromthe selected sites of forested ponds and non-forested

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ponds of, Kulathupuzha Gramapanchayat, KollamDistrict, Kerala, for the period from December 2017 toMay 2018. For the present study, a total of six samplingpoints were selected; three each from forested pondsand non forested ponds.

Physico-chemical analysesSurface water samples were collected from all the sixsampling points. Collections were made using plasticcontainers of 2-L capacity. The plastic containers were

Site

Code

Site Name Nature of pond

FP1 Kalluvettam kuzhy

Research

Forest pond

FP2 Samnagar (Anakkuzhy) Forest pond

FP3 Samnagar (Pangalu

kadu)

Forest pond

NFP1 Kuzhivilakkarikkam Non forest pond

NFP2 Nellimoodu Non forest pond

NFP3 Kurishin moodu Non forest pond

Table 1. Sampling sites

Parameters FP1 FP2 FP3 NFP1 NFP2 NFP3 Max Min Mean±SD

Air Temp. (oC) 31 26.5 30 27.5 30.5 31 31 26.5 29.42±1.93

Water Temp. (oC) 28 25 25 25 25.5 29 29 25 26.25±1.78

PH 5.76 5.99 5.37 5.68 5.63 6.15 6.15 5.37 5.76±0.28

EC (µS) 29.15 57.95 67.5 74.47 73.83 75.66 75.66 29.15 63.09±17.89

TDS (mg/l) 23.07 46.41 54.14 59.72 59.17 60.59 60.59 23.07 50.52±14.45

Alkalinity (mg/l) 14 16 14 12 14 10 16 10 13.33±2.07

Acidity (mg/l) 82 18 80 46 46 12 82 12 47.33±29.6Total Hardness(mg/l) 8 8 10 12 10 12 12 8 10±1.79

DO (mg/l) 6.09 5.08 2.03 4.07 5.08 7.11 7.11 2.03 4.91±1.75

Salinity (ppt) 0.02 0.04 0.04 0.04 0.04 0.04 0.04 0.02 0.03±0.007

Sulphate (mg/l) 11 10 11 10 12 12 12 10 11±0.89

Phosphate (mg/l) 0.78 0.13 0.7 0.62 0.74 0.69 0.78 0.13 0.61±0.24

Nitrate (mg/l) 0.008 0.003 0.004 0.03 0.001 0.0002 0.03 0.0002 0.007±0.01

Chlorophyceae 1020000 580000 460000 402000 600000 220000 1020000 220000 547000±269499.54

Cynophyceae 160000 60000 100000 20000 20000 0 160000 0 60000±60663

Bacillariophyta 40000 100000 20000 0 80000 20000 100000 0 43333.33±38815.8

Desmidaceae 80000 320000 0 320000 400000 140000 400000 0 210000±158871.02

Zooplankton 400000 20000 60000 80000 20000 120000 400000 20000 116666.67±143898.11

Table 2: Physico-chemical analysis of forested and non forested ponds

Figure 1: Map showing study area

rinsed thoroughly with sampling water before use. Afterfilling the containers, they were sealed and transferredto the laboratory for physicochemical analysis. Fewphysico-chemical parameters like Temp and pH wereperformed on spot and other parameters like Electricconductivity, Total Alkalinity, Sulphate, Phosphate, andNitrate were analyzed in laboratory following APHA(2012), Trivedi and Goel (1998) and Wetzel and Likens(1991). Temperature was measured using a Celsiusthermometer. Conductivity and pH were measured usingConductivity meter and digital pH meter. DO wasestimated according to Winkler’s method (Grasshoff etal., 1999). Salinity was analyzed using standard methodsprescribed by Strickland and Parsons (1972).

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Biological parametersSamples for microalgal analyses were attempted withthe help of plankton net made up of blotting silk no. 25(0.03-mm mesh) fitted to a wide-mouthed bottle. Formicro algal sampling, horizontal trawling was done at avery low speed and the net was allowed to sink to a depthof about 1 m below the water surface to prevent theformation of a bow wave. Approximately fifty liters of waterwas sieved through the net at one spot. Sample materialswere collected from each sampling points and fixed in

Plankton species FP1 FP2 FP3 NFP1 NFP2 NFP3 %Composition

Organisms /litre

CHLOROPHYCEAE

Oedogonium 80000 60000 20000 40000 20000 20000 7.31

Ulothrix 200000 40000 100000 10000 460000 60000 26.51

Spirogyra 280000 160000 220000 12000 80000 120000 26.51

Closterium 160000 0 0 160000 0 0 9.75

Volvox 0 0 0 160000 20000 0 5.48

Euastrum 80000 200000 80000 0 0 0 10.97

Cladophora 20000 0 0 0 0 0 0.61

Chlamydomonus 0 0 0 20000 0 0 0.61

Zygnema 20000 0 20000 0 0 20000 1.83

Chlorella 180000 120000 0 0 0 0 9.14

Actinastrum 0 0 20000 0 20000 0 1.22

CYNOPHYCEAE

Anabaena 20000 60000 0 0 20000 0 27.78

Nostoc 80000 0 40000 20000 0 0 38.89

Oscillatoria 0 0 40000 0 0 0 11.11

Spirulina 60000 0 20000 0 0 0 22.22

BACILLARIOPHYTA

Navicula 40000 0 20000 0 20000 20000 38.46

Pinnularia 0 80000 0 0 0 0 30.77

Fragilaria 0 20000 0 0 60000 0 30.77

DESMIDACEAE

Cosmarium 0 160000 0 300000 400000 60000 73.02

Desmidium 80000 140000 0 0 0 80000 23.81

Micrasterias 0 20000 0 20000 0 0 3.17

ZOOPLANKTON

Amoeba 20000 0 20000 0 0 40000 11.43

Paramecium 240000 0 40000 20000 20000 60000 54.29

Euglena 120000 0 0 20000 0 20000 22.86

Daphnia 20000 0 0 20000 0 0 5.71

Brachious 0 20000 0 20000 0 0 5.71

Table 3. Plankton species enumerated in forested and non-forested ponds

Lugol’s iodine and transported to the laboratory inpolythene bottles. Plankton enumeration was performedin the samples preserved in Lugol’s iodine, as describedby Rodhe et al., (1958) and Anderson (2005). Fifty-milliliter samples from each incubation were collectedand immediately distributed into amber bottles withpredispensed Lugol’s iodine, resulting in a finalconcentration of 2% (v/v) fixative. Fixed samples wereenumerated using a Sedgwick–Rafter counting slide ona light microscope. Counting of plankton was done with

64 ECO-CHRONICLE

Tabl

e 4:

Pea

rson

’s c

orre

latio

n co

-effi

cien

t (r)

for t

he p

hysi

co-c

hem

ical

var

iabl

es a

nd p

lank

ton

spec

ies

in fo

rest

ed a

nd n

on fo

rest

ed p

onds

65ECO-CHRONICLE

FP1 FP2 FP3 NFP1 NFP2 NFP3

CHLOROPHYCEAE

Oedogonium - - - - - -

Ulothrix - - - - - -

Spirogyra - - - - - -

Closterium 1 - - 1 - -

Volvox - - - - - -

Euastrum - - - - - -

Cladophora - - - - - -

Chlamydomonus - - - 4 - -

Zygnema - - - - -

Chlorella 3 3 - - - -

Actinastrum - - - - - -

CYNOPHYCEAE

Anabaena 1 1 - - 1 -

Nostoc - - - - - -

Oscillatoria 4 - 4 - - -

Spirulina - - - - - -

BACILLARIOPHYTA

Navicula 3 - 3 - 3 3

Pinnularia - - - - - -

Fragilaria - - - - - -

DESMIDACEAE

Cosmarium - - - - - -

Desmidium - - - - - -

Micrasterias - - - - - -

ZOOPLANKTON

Amoeba - - - - - -

Paramecium - - - - - -

Euglena 5 - - 5 - 5

Daphnia - - - - - -

Brachionus - 2 - 2 - -

Total 17 6 7 12 4 8

Table 5. Palmer’s algal pollution index values in six ponds

the help of “Sedgwick– Rafter counting cell” as per theprocedure given by Wetzel and Likens (2000). Sampleswere allowed to settle in the counting chamber for 3–5min. prior to enumeration. More than ten fields of view

were randomly selected across each slide and repeatedthree times. Identification of the planktons was donefollowing Smith (1950), Desikachary (1959), Kudo (1986)and Pennak (1989). The Karl Pearson’s correlation

66 ECO-CHRONICLE

coefficient was performed using Microsoft Excel 2007 todetermine the relationship among the various physico-chemical attributes and different phytoplanktonassemblages. For rating the water sample as high orlow organically polluted the Algal Generic Pollution Indexof Palmer (1969) was employed. According to Palmer’sAlgal Pollution Index, values between 0-10 indicate lackof organic pollution, 10-15 moderate pollution, 15-20probable high organic pollution and 20 and above asconfirmed to have high organic pollution.


The physical parameters of concern are temperature,pH, TDS and conductivity. Water temperature rangingbetween 13.5oC and 32oC is reported to be ideal for thedevelopment of planktonic organisms (Kamat, 2000). Theincrease in number of zooplanktons is also in accordancewith temperature of its habitat. Jeppesen et al. (2002)has stated that the abundance and diversity ofzooplanktons vary according to limnological features andthe trophic state of fresh water. The pH value rangedfrom as low as 5.37 in FP3 to 6.15 in NFP3 with averagevalue of 5.76. This falls below acceptable standard ofthe World Health Organization (WHO) indicating acidicpH of the ponds.

The TDS measurements varied from 23.07 to 60.59 mg/l with average values of 50.52 while the conductivitymeasurements varied from 29.15 μs/cm to 75.66 μs/cmwith average value of 63.09 μs/cm (Table 2). Theconcentration of nitrate was relatively very low in theentire pond samples. The present study shows that theconcentration of phosphate ranges from 0.13 to 0.78 mg/l. This condition enhances the growth of phytoplanktonpopulation. The highest value was reported at NFP1(0.026 mg/l). But the concentration of phosphate wasslightly higher than nitrate.

Dissolved oxygen (DO) is one of the most important andlimiting parameters of water quality assessment, whichmaintains aquatic life. It regulates the metabolicprocesses of aquatic organisms (Vaidya, 2011). In thepresent investigation, maximum DO value recorded inNFP3 was 7.17 mg/l. DO in FP3 was below the range ofWHO standard. This may be due to the process ofeutophication in the area, which enhanced the growth ofaquatic vegetation or phytoplankton. Algal blooms disruptnormal functioning of ecosystems, causing variety ofproblems, including lack of oxygen supply for fish andshell fish to survive.

Water hardness is commonly defined as the sum of thepolyvalent cations dissolved in water. The most commoncations are calcium and magnesium; although iron and

manganese may contribute. In the present investigation,maximum total hardness value recorded was 12 mg/l andminimum was 8 mg/l with a mean value 10 mg/l.

Phytoplankton abundance is closely associated withnutrient status, while zooplankton abundance is relatedto measures of primary production (Blouin, 1989). Thephytoplankton diversities also showed variations in eachpond ecosystems. The most abundant phytoplanktonspecies were coming under the class Chlorophyceae,and Cyanophyceae.

The statistical analysis of Pearson’s correlation coefficientis presented in Table 4. The surface water temperaturewas significantly positively correlated with pH, DO,phosphate, nitrate and zooplanktons. On the other hand,surface water temperature showed firm negativecorrelation with alkalinity. The pH showed significantpositive correlation with DO. The increase in electricalconductivity increases TDS, total hardness and salinity.TDS showed significant positive correlation with totalhardness and salinity and significant negative correlationwith acidity. Increasing phosphate in the pond alsoincreases total hardness, sulphate and salinity.Phosphate has negative correlation with pH and alkalinity.

In the present study, the algal flora was represented bymembers of four groups viz, Cyanophyceae,Chlorophyceae, Desmidaceae and Bacillariophyceae.Their presence is an indication of organic pollution ofthe water, as reported by Palmer (1969), Robert et al.(1974) and Hosmani and Bharati (1980).

The distribution pattern of phytoplankton in differentponds was summarized in Table 3. Twenty two algalgenera represented the micro algal flora. Among them,11 was representing Chlorophyceae, 4 to Cyanophyceae,3 each to Desmidaceae and Bacillariophyceae each. TheChlorococcales like Oedogonium, Ulothrix and Spirogyrawere well distributed in all the ponds studied. ThePalmer’s Algal Pollution Index was employed in all theponds and the results are given in Table 5. FP1 showedprobable high organic pollution and NFP1 showedmoderate organic pollution.

The occurrence and relative abundance of certainphytoplankton species can be related to water qualityand hence indicators of water pollution (Wan Maznahand Mansor, 2002; Unuoha et al., 2011). For example,Oscillatoria was rare in the pond ecosystems, but couldbe considered as potential toxic and polluted waterindicator (Kumari et al., 2008). Anabaena can beconsidered as having the potential of producingunfavorable odors and flavors (Palmer, 1980). Dý´az-Pardoet al. (1998) reported that a phytoplankton community

67ECO-CHRONICLE

dominated by Cchlorophyceans replaced byCyanophyceans, showed severe signs of eutrophicationin a subtropical lake in Mexico. Aquatic pollution can beobserved with fairly rapid and fairly marked reduction incertain species and possibly a proliferation of anotherspecies (Wu 1984; El-Sheekh et al., 2010). Navicula,Nitzschia and Euglena recorded during the study periodwere indicators of pollution (Palmer, 1969). Similarobservations were noted by Hosmani and Bharti, (1980);Trivedi, (1988); More and Nandan, (2000).

SUMMARY AND CONCLUSION

This paper summarizes the relationship of physico-chemical parameters and their influences onphytoplankton community of selected pond ecosystems.Water quality regulates biotic diversity and trophic levelof an ecosystem. Analysis of physico - chemicalparameters reflects the hydro geochemical and biologicalstatus of an ecosystem. Dominance of pollution tolerantgenera Euglena, Oscillatoria, Nitzschia and Naviculasupports the view to categorize the pond as eutrophic innature. The present investigation indicates phytoplanktonabundance closely associated with nutrient status, whilezooplankton abundance related to measures of primaryproduction, which leads to freshwater pollution. Hencenecessary steps should be taken to minimize the freshwater pollution by minimizing human activities such aswashing of clothes, bathing and other utilitarian needs.The consumption and utilization of polluted pond watermay cause health hazard to the local residence. It istherefore, advisable that the authorities should takeappropriate steps to check pond water contamination.This also helps in conserving these water resources foreffective planning and utilization.

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Smith, G. M. (1950). The freshwater algae of the UnitedStates (2nd ed.). New York: McGraw-Hill Book Company.Strickland, J. D. H. and Parsons, T. R. (1972). A PracticalHandbook of Sea Water Analysis, 2nd ed. FisheriesResearch Board, Canada.

Tran, L., Nunan, L., Redman, R. M., Mohney, L. L.,Pantoja, C. R. and Fitzsimmons, K. (2013).Determination of the infectious nature of the agent ofacute hepatopancreatic necrosis syndrome affectingpenaeid shrimp. Dis. Aquat. Org., 105(1): 45–55.

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Trivedy, R. K. and Goel, P. K. (1998). In: Chemical andbiological methods for water pollution studies.Environmental publication, Karad, Maharashtra, India,pp: 257.

Unuoha P. C., Nwankwo D. I., Chukwu L. O. andWim V.(2011). Spatiotemporal variations in phytoplanktonbiomass and diversity in a tropical eutrophic lagoon,Nigeria. J Am Sci., 7(8):33–46.

Vaidya, S. R. (2017). Biomonitoring of zooplankton toassess the quality of water in the nagpokhari ofKathmandu valley. International Journal of ZoologyStudies, 2(1): 61-65.

Wan Maznah, W. O. and Mansor, M. (2002). Aquaticpollution assessment based on attached diatomcommunities in the Pinang River Basin Malaysia.Hydrobiologia, 487: 229–241.

Wetzel, R. G. and Likens, G. E. (2000). Limnologicalanalyses. New York: Springer.

Wetzel, R. G. and Likens, G. E. (1991). Limnologicalanalyses. 2nd edition Springer-verlag New York, pp: 1-175.

Wu, J. T. (1984). Phytoplankton as bioindicator for waterquality in Taipei. Bot Bull Acad Sinica., 25: 205–214.

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PHYTOCHEMICAL AND ANTIBACTERIAL ANALYSIS OF AMORPHOPHALLUS HOHENACKERI(Schott) Engl. & Gehrm.

Raghavan Kavalan1, V. Abdul Jaleel2 and K.M. Gothandam3

1. Jawahar Navodaya Vidyalaya,Bagalur P.O, Bangalore, Karnataka.2. Department of Post Graduate Studies and Research in Botany, Sir Syed College, Taliparamba, Kerala.

3. School of Bio Sciences and Technology, VIT University, Vellore, Tamil Nadu.Corresponding author: [email protected]

Vol. 13, No. 2, June, 2018


PP: 69 - 74


ABSTRACTThe morphological features, phytochemical properties, total phenolic content, antioxidant and antibacterial propertiesof Amorphophallus hohenackeri (Schott) Engl. & Gehrm., a member of the family Araceae belongs to the Sect.Raphiophallus was investigated for its phytochemical constituents. Chemical extraction has been carried out usingsolvents like hexane, methanol and water. Phytochemical screening revealed the presence of phenols, flavonoids,alkaloids, saponins and carbohydrates. The antioxidant property of the tuber extract was also analyzed. Among thethree, methanol extract expressed maximum efficacy in DPPH radical scavenging assay. Hexane and aqueous extractexhibited relatively low radical scavenging property. Methanolic and aqueous extracts showed good antibacterialproperty against the five species of bacterial cultures studied. Hexane extract did not exhibit any antibacterial property.The present study suggests that the bioactive compounds present in the tubers of Amorphophallus hohenackeri havegreat potential as a natural source of antibacterial and antioxidant to replace chemical drugs in pharmaceuticalproducts with isolation and characterization of the bioactive compounds.Key Words: Araceae, Amorphophallus hohenackeri, Phytochemical, Antioxidant, Antibacterial activities.

INTRODUCTION

Plants of the genus Amorphophallus belongs to the familyAraceae, have a long history of use in tropical andsubtropical Asia as a food source and as a traditionalmedicine. About 200 species of Amorphophallus aredistributed throughout the world and 20 species arereported from India (Abdul Jaleel et.al., 2011). All thewild relatives of Amorphophallus except A. paeoniifoliusare rare. A detailed study of this plant will help inidentifying the presence of various bioactive compoundsresponsible for its therapeutic uses. It is estimated thatonly less than 10 percent of plants have been studiedfor their medicinal properties (Widjaja and Lester, 1978).A few species such as Elephant foot yam(Amorphophallus paeoniifolius var. campanulatus) arewidely cultivated and used as vegetable. Many wildrelatives of the genus Amorphophallus are traditionallyused as medicine (Hettershield and Ittenbach, 1996).The tuberous corms of Amorphophallus are reported tobe used for treatment of piles, cysts and tumors(Ravikumar and Ved, 2004), cure for snake bite by tribal’sin some villages of Rajasthan, India (Jain et al., 2005,

Kavitha et al., 2011), in piles, acute rheumatism,abdominal tumors, boils, asthma and enlargement ofspleen (Yusuf et al, 1994). The therapeutic effect ofmedicinal plants is due to its biologically activecompounds. Most of the Phytochemical and other studiesconducted so far in the genus are confined to only someof the widely available species like A. paeoniifolius(Elephant Foot Yam - both cultivated and wild) and A.commutatus. Other species of the genus are unexploredperhaps due to its rare nature and non-availability.Morphological, phytochemical and antibacterialproperties of wild and indigenous plant A. commutatusvar. wayanadensis have been carried out ( Arjun etal.,2012); hepatoprotective effect of polyphenols of A.commutatus var. wayanadensis (Sreena Raj andGothandam, 2014); Phytochemical and anti-bacterialanalysis of A. commutatus (Sagarika Damle and AtulKotian, 2015); radical scavenging activity of A.commutatus (Kavita Krishna et al., 2012). Phenoliccontent and antioxidant capacity of A. commutatus andA. paeoniifolius (Shete, et al., 2015); Pharmacognostic

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evaluation and phytochemical analysis of A. paeoniifloiuswas investigated earlier by Yadu Nandan Dey and AjoyKumar Ghosh, 2010; Yadu Nandan Dey et al., 2012;Madhurima et al., 2012; Jyoti D Vora et al. 2015; ManjuMadhavan and Regi Raphel, 2012; Firdouse and Alam,2011). The Morphology and taxonomy of Amorphophallushohenackeri was carried out in the revision of IndianAraceae (Abdul Jaleel et al., 2011). But i t ’sPhytochemical and other properties have not beenstudied till now. In the present study, tuber extracts ofthe corm of Amorphophallus hohenackeri areinvestigated for i ts phytochemical constituents,antioxidant and ant- bacterial properties.


The tuber of the plant Amorphophallus hohenackeriwere collected from the natural locality of Calicut

University campus, Thenjipalam, Malappuram Districtand from Ponniam, Kannur District of Kerala. The plantspecies were identified and authenticated by one ofthe author Dr. V. Abdul Jaleel, Department of PostGraduate Studies and Research in Botany, Sir SyedCollege, Taliparamba, Kerala and was deposited inSir Syed College Herbarium and maintained in theAroid Home, Sir Syed college, Taliparamba, KannurDistrict, Kerala

MorphologyThe species of Amorphophallus exhibit variation inshape and size of tuber, petiole, spathe, spadix,appendix and the individual female flowers.Taxonomically it is one of the most difficult genera ofthe family due to various reasons including the timingof emergence of inflorescences and their relatively shortactive period of existence.

Figure1. Amorphophallus hohenackeri (Schott) Engl.& Gerhm.

a.Tuber; b. A portion of petiole showing lamina; c. A portion of petiole showing mottling; d. Inflorescence;e. A portion of spadix enlarged shoing female, male and neutral flowers; f. Infructescence.

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Tubers depressed sub-globose, 2 - 3.5cm diam. Petiole35-68 cm long, 8-14 mm diam. at the base, smooth, paleyellowish green with dark green mottling and minutecream speckles. Lamina 30-45 cm diam., acuminate atapex. Peduncle 18-50 cm long, 6-10 mm diam. at base,identical with petiole in colour and pattern of mottling.Spathe light pinkish yellow with purplish black mottlingand a pale purplish streak along the median outside,yellowish green within, 10.5-15.5 cm long and 4.0-5.5cm broad. Spadix more or less equaling the length ofthe spathe and it consists of a basal female zone(ca.1cm) followed by neutral zone(ca.4mm) arranged in1-2 rows with round to elongate neutral flowers, malezone(ca.1.5cm) and appendix (ca.10cm). Fruits oblong,green when young and turning scarlet-red when ripe.Phenology: Flowering: March-May; fruiting: May-August.Distribution: Very limited in distribution, known only fromKerala and at limited places in Karnataka.

Preparation of extracts:The tuber corms were properly washed, followed bysurface sterilization using 1% of Sodium hypochlorite(Maina et al., 2010). The tubers were chopped intopieces; sun dried, and powered using an electric blender.Extraction was done by Soxhlet apparatus (Gennaro etal., 2008) using solvents viz: Hexane, methanol andwater in the increasing order of their polarity. Theobtained solvent extracts were concentrated usingvacuum distillation process and the extracts were storedin a refrigerator at 4°C (Ayvaz et al., 2008; Sultana etal., 2009).

Bacterial Strains:Five species of human pathogenic bacteria wereobtained from the National Collection of IndustrialMicroorganisms (NCIM) Pune, India, available withSchool of Biosciences and Technology, VIT University.The gram-negative bacterial species used forexperimentation were Escherichia coli, Proteus mirabilisand Salmonella typhi. The gram-positive strains usedwere Bacillus cereus and Listeria monocytogenes.These strains were preserved at 4°C in the nutrient brothas stock cultures and were sub-cultured for 24 h at 37°Cprior to use.

Phytochemical analysis:Phytochemical tests were carried out on the Hexane,methanol and aqueous extract of plant materials usingstandard procedures (Trease and Evans, 1978; Edeogaet al., 2005). Phytochemical screening tests were carriedout on hexane, methane and aqueous extracts of tuberusing standard procedures. The analysis was done totest the presence of phytochemicals such ascarbohydrates, (Molisch’s test), reducing sugars(Fehling’s test), tannins (Ferric Chloride test), flavonoids

(Shinoda test), steroids (Liebermann’s –Buchard’s test),alkaloids (Wagner’s Test), anthraquinones (Borntrager’stest), glycosides (Killer-Kilian test), Phyto tannins,terpenoids, saponins (Frothing test) and phenols (FerricChloride test).

Total Phenolic content:The amounts of phenolics in plant extracts weredetermined with Folin-Ciocalteu reagent. To 1ml of eachsample, 0.5ml of 10% dilution Folin-Ciocalteu reagentand 1ml of Na2CO3 (20% w/v) were added and theresulting mixture was incubated at room temperature andin darkness for 2 hours, with intermediate shaking. Theabsorbance of all samples was measured at 760nm.Results were expressed as milligrams of Gallic acidequivalent per gram of dry weight.

DPPH assay:The DPPH assay was performed to determine the freeradical scavenging potential of the extract. 1ml of 0.2mMDPPH(2,2-diphenyl-1-picrylhydrazyl) in methanol wasmixed with 4ml of different concentration of extracts andstandards and it was incubated in darkness for 30min.The free radical (1,1-diphenyl-2-picryhydrazyl), which isabsorbing UV-light at 517 nm will be reduced in thepresence of antioxidant compound contained in theextract and the reaction will form a yellow molecule whichwill not absorb at the working wavelength. The morepotential the extract is, higher the free radicalscavenging.

Nitric oxide radical scavenging assay:Nitric oxide scavenging activity of the extracts wasdetermined. 1ml of 10 mM Sodium Nitroprusside wasmixed with 1ml of different concentrations of extractsand standards and the mixture was incubated at 37for 150 min. After incubation, 1ml of the mixture wastaken out to which 1ml of Griess’ reagent (1%sulphanilamide and 0.1% naphthyl ethylenediaminedihydrochloride in 2% o-phosphoric acid) wasadded and the absorbance were measured at 546nm.The procedure is based on the principle that, sodiumnitroprusside (SNP) in aqueous solution at physiologicalpH spontaneously generates nitric oxide which interactswith oxygen to produce nitrite ions. These nitrite ionscan react with Griess’ reagent and to form a chromophoreabsorbing at 546 nm. Scavengers of nitric oxide competewith oxygen, leading to a reduction in the production ofnitrite ions. The absorbance of the chromophore formedwill be measured at 546 nm. The percentage ofscavenging was calculated as per standard procedure.

Anti-bacterial assayThe anti-bacterial assay was performed on both grampositive and gram-negative bacterial species by well

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diffusion method (Onkar and Dhingra, 1995). The Petriplates were poured with approximately 25 ml autoclavednutrient agar media (HIMEDIA). Using a micropipette,standardized inoculums (0.1 ml) of 0.5McFarlandturbidity standards, equivalent to 5 × 10 8cfu/ml(McFarland, 1907) was asceptically spread on thesurface of nutrient agar plate. After drying, four wellswere punched on each plate using a sterile cork borerof 8 mm diameter (Bradshaw, 1992). 0.1 ml of eachextract (concentration of 100mg/ml each) was pipettedinto respective wells (Ayfer and Turgay, 2003), 10%Dimethyl Sulfoxide was used a vehicle control.Kanamycin (0.5 mg/ml) a broad-spectrum antibiotic isused as positive drug control. The Petri plates wereincubated overnight at 37°C and the anti-bacterial activitywas measured after 18 hours of incubation. The diameterof ZOI was also measured.


Phytochemical analysis:Total Phenolic contentThe total phenolic content of the three extracts viz.hexane, methanol and water were determined using thelinear regression equation of the Gallic acid calibrationcurve (Y=0.0307X-0.14; R2=0.9903) and then the totalphenol content is expressed as mg equivalent of Gallicacid per gram of extract. The Total phenolic content inthree extract has been shown in Table 2. Total phenoliccontent was estimated using Folin- Ciocalteu reagent.Total phenolic content of the different fractions ofAmorphophallus hohenackeri were solvent dependentand expressed as mg equivalent of Gallic acid per gramof extract. Highest phenolic content has been shown inwater.

DPPH assayDPPH radical scavenging activity is one of the mostwidely used method for screening the antioxidant activity

Chemical constituent Name of test Hexane Methanol Water

Carbohydrates Molisch’s Test + - -Reducing sugar Fehling’s test - - -Tannins Ferric chloride test - - -Flavonoids Shinoda test - - +Alkaloids Wagner’s test - + -Anthraquinones Borntrager’s test - - -Glycosides Keller-Killian test - - -Phytotanins Ferric chlorides test - - -Terpenoids Salkowski test - - -Fats and Oil - + - -Saponins Frothing test - - +Phenolic Ferric chlorides test + +

+ indicate presence, - indicate absence.

Table 1: Qualitative phytochemical analysis of Amorphophallus hohenackeri tuber extracts viz. hexane,methanol and water.

of the extract. Table 3 shows the antioxidant activities ofhexane, methanol and aqueous extracts ofAmorphophyllus hohenakeri. The highest DPPH activitywas found in methanolic extract, which is 86.76. Hexaneand water extract showed lesser activity. Upon comparingwith standard reading, ascorbic acid also showed similaractivity with maximum of 95.33 %. From this, it can bestated that the extract showed scavenging activity nearerto ascorbic acid standard.

Nitric Oxide Radical ScavengingNitric Oxide radical inhibition study proved that extractis a potent scavenger of nitric oxide generated fromsodium nitroprusside which reacts with oxygen to formnitrite. The extract inhibits nitrite formation by competingwith oxygen, leading to reduced production of nitric oxide.Table- 4 shows the antioxidant activity of hexane,methanol and water extracts of Amorphophyllushohenakeri. Moderate to high nitric oxide radicalscavenging was observed in 50-250µg/ml of methanolextract. The percentage of scavenging has beenincreased with the increasing concentration of extract.Table 4 shows that methanolic extract shows highestpercentage of inhibition which is 93.51% at concentrationof 250 µg/ml and the lowest inhibition expressed inhexane extract.

Antibacterial assayAgar well diffusion method is the widely accepted methodfor the evolution of antibacterial activity of samples.Preliminary screening for the antibacterial activity of theextract was performed with various pathogenic bacterialstrains. Among the three extract, water extract ofAmorphophallus hohenackeri showed potentialinhibition. Antibacterial activity of all the three extractstested on five bacterial strains are shown in Table 5.The results showed that methanol extract ofAmorphophallus hohenackeri has highest antibacterial

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Solvents Total Phenol content(in mg eq.of GA/gm of extract)

Methanol 79.05Hexane 68.72Water 93.51

Table 2: Total phenolic content in Amorphophallushohenackeri with different solvents

activity of (16mm) inhibit ion zone against L.monocytogenes and Bacillus cereus, (14mm) inhibitionzone against E-coli, followed by water extract whichshowed zone of inhibition (14mm) against B.cereus,12mm against E-coli, S. typhi and L. monocytegenes.

CONCLUSION

The phytochemical screening revealed the presenceof chemical constituents like carbohydrates, phenols,flavonoids, alkaloids and saponins in different extractsthat form the foundation of their pharmacological activity.Phytochemical investigation conducted previously inAmorphophallus paeoniifolius and Amorphophallus

Table 3: DPPH radical scavenging activity ofAmorphophallus hohenackeri’s extracts in differentsolvents.

Conc.(µg/ml)

Hexane Methanol Water

50 45.10 69.37 52.01100 47.37 74.14 52.09150 48.80 81.30 53.30200 50.01 84.07 54.08250 51.76 86.76 54.09

Conc.(µg/ml)

Hexane Methanol Water

50 10.96 78.29 38.59100 13.85 87.57 40.61150 15.78 90.91 43.17200 18.01 92.39 45.89250 19.90 93.51 46.29

Table 4: Nitric Oxide Radical Scavenging ofAmorphophallus hohenackeri’s extracts.

Bacterial strain Kana-mycin

Methanolextract

Hexaneextract

Waterextract

Escherichia coli, 18 14 - 12Salmonella typhi 12 - - 12Listeriamonocytogenes

14 16 - 12

Bacillus cereus 18 16 - 14Proteus mirabilis 15 - - -

Table 5: Antimicrobial properties of Amorphophallushohenackeri against various strains of bacteria.

commutatus revealed the presence of such secondarymetabolites. Phenolics that possess antioxidant activitywas found in Amorphophallus hohenackeri also. Thedifferent extract showed good radical scavengingproperties on par with standard in DPPH and NitricOxide radical scavenging assessment. ThereforeAmorphophallus hohenackeri can be considered as anatural source of antioxidants and may be consideredin future to be used as antioxidant additive or asnutri t ional supplement and in cosmetic andpharmaceutical products with further detailed studies.The present study revealed that the methanolic extractof Amorphophallus hohenackeri showed good efficacyagainst the different bacterial strains used. Bothmethanolic and aqueous extracts exhibited potentialantimicrobial activity. The result of the presentpreliminary investigation substantiates the claim of thetraditional use of this plant in ayurvedic medicinepreparation. Further studies are required to isolate andcharacterize the structural elucidation of the bioactivecompounds for its pharmaceutical applications. Thespecies also require conservation as the populationsof this threatened species, once prevalent in Malabarregion of Kerala, are becoming less due to urbanizationand human activities.

ACKNOWLEDGEMENTS

The authors are thankful to the management of VITUniversity, Vellore, Tamil Nadu, for extending the labfacilities of the School of Biosciences and Technologyfor undertaking this study. Authors are also thankful tothe Principal, Sir Syed College, Taliparamba, Kannur,Kerala for offering facilities for research. First authorexpresses his gratitude to the Commissioner, NavodayaVidyalaya Samiti, Noida for granting permission toundertake the present study.

REFERENCES

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Arjun, R. Krishna, Asis Singh, V., Abdul Jaleel, SreenaRaj, S., Karthikeyan, and K.M. Gothandam (2013):Morphological, Phytochemical and Antibacterialproperties of wild and indigenous plant (Amorphophalluscommutatus). Journal of Medicinal Plant Research Vol.7 (13) 744-748.

Ayfer, A.D., Turgay, Z.E. (2003). Antimicrobial activitiesof various medicinal and commercial plant extracts. Turk.J. Biol. 27:157-162.

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Ayvaz, A., Osman, S., Salih, K. and Ismet, O. (2008):Insecticidal activity of the essential oils from different plantsagainst three stored product. J. Insect Sci. 10:1-13.

Bradshaw, L.J. (1992): Laboratory Microbiology, 4 th ed.Saunders College Publishing. Fort Worth, USA. P 435.Edeoga, H.O., Okwu, D.E. and Mbaebie, B.O. (2005).Phytochemical constituents of some Nigerian medicinalplants. Afr. J. Biotechnol. 4(7): 685-688.

Firdouse, S. and Alam, P. (2011). Phytochemicalinvestigation of extract of Amorphophallus campanulatustubers. Int. J. Phytomed. 3(1):32-35.

Gennaro, L., Sukhdev, S.H., Suman Preet, S., Khanujaand Dev Dutt, R. (2008). Extraction Technologies forMedicinal and Aromatic Plants. United Nations IndustrialDevelopment Organization and the International Centrefor Science and High Technology, Trieste, Italy. P 23.

Hetterscheid, W. L. A. and Ittenbach, S. (1996):Everything you always wanted to know aboutAmorphophallus, but were afraid to stick your nose into.Aroideana 19:7-131.

Jain, A., Katewa, S.S., Galav, P.K. and Sharma, P. (2005):Medicinal plant diversity of Sitamata wild life sanctuary,Rajasthan, India. J. Ethnopharmacol. 102:143-157.

Jyothi, D. Vora, M. Aswathi Sarman and M. NonitaMadhrani (2015): Biochemical, Organoleptic andAntimicrobial assessment of Elephant Foot yam(Amorphophallus paeonifolius). Journal of EnvironmentalScience, Toxicology and Food Technology. Vol.9, Issue5, Ver.1: 7-10.

Kavitha, K.R., Karthikeyani, S. and Krishna, K.S. (2011):Anti-bacterial activity of Amorphophallus commutatus, anendemic plant of Western Ghats, South India. J. Pharm.Res. 4 (3):710-711.

Kavitha Krishna, R., S. Krishna Kumar, M. Vinodha, AnuSebastian and R. Gomathi (2012): Invitro free radicalscavenging activity of Amorphophallus commutatus anendemic aroid of Western Ghats, South India.International research journal of pharmacy. 3(2) 133-137.

Mac Farland, J. (1907). The nephelometer: an instrumentfor establishing the number of bacteria used forcalculating the opsonic index for vaccines. J. Am. Med.assoc. 49:1176.

Madhurima, P., I.J. Kuppsat and K.L. Mankani (2012). Areview on Amorphophallus paeoniifolius. InternationalJournal of Advanced scientific research and Technology.Issue 2, vol. 2: 99-111.

Manju Madhavan and Regi Raphel (2012).Pharmacognostic and preliminary PhytochemicalScreening of the corm Amorphophallus paeoniifolius

(Dennst) Nicols. var. paeoniifolius. Academic Sciencesvol.4, suppl. 5: 238-240.

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Ravikumar, K. and Ved, D.K. (2004): Illustrated FieldGuide - 100 Red listed medicinal plants of conservationconcern in South India. FRLHT, Bangalore. pp. 1- 467.

Maina, S.M., Emongor, Q., Sharma, K.K., Gichuki, S.T.,Gathaara, M. and deVilliers SM (2010). Surface sterilanteffect on the regeneration efficiency from cotyledonexplants of groundnut (Arachis hypogea L.) varietiesadapted to Eastern and Southern Africa. Afr. J.Biotechnol.9 (20): 2866-2871.

Sagarika Damle and Atul Kotian (2015). Elemental,Phytochemical and Antibacterial analysis ofAmorphophallus commuatatus Lnn. International Journalof Recent Scientific Research. Vol. 6, Issue 5: 4308-4311.

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INDOOR AIR POLLUTION: SOURCES, HEALTH IMPACTS AND CONTROL

Roy, M., F. Shamim and A. DasDepartment of Environmental Science, Directorate of Distance Education, Vidyasagar University,

West Bengal, IndiaCorresponding author: [email protected]

ABSTRACT

Indoor air pollution has nowadays become a severe environmental hazard affecting the well-being of humans. Theterm “building related illness” has been coined to describe various illness and problems related to specific airbornecontaminants in buildings, which results in indoor pollution. Modern homes and office buildings are now days soconstructed that they trap pollutants like benzene, formaldehyde, trichloroethylene (TCE), etc. According to the USEPA (Environmental Protection Agency), indoor pollutants levels are 100 times higher than outdoor levels. The way tocontrol the indoor pollution is by restricting the polluting sources. A little modification and change in our daily lifepattern can overcome these issues. In addition, adopting the practice of house plants will not only decorate our homesbut also filter the harmful and toxic chemicals from our indoor air. The present study strategically elucidates solutionsto these problems.

Key Words: Indoor air pollution, Environmental hazards, Pollutants, Indoor plants

Review Article

INTRODUCTION

Clean and pure air is the foremost requisite for healthyliving of human beings. Anthropogenic emissions ofvarious gaseous and particulate matters into theatmosphere, known as primary pollutants, have led tothe formation of new pollutants by various chemicalreactions, known as secondary pollutants. People areexposed to extreme health risks due to increasingparticulate matters and other hazardous airborne agentsin the atmosphere. Outdoor air quality is affecting indoorair quality also. People take thousands of breaths daily,leading to a total intake of about 10,000 litres of air perday (Phalen, 1996). Consequently, the lung receivessignificant doses of many air contaminants, even thosepresent at seemingly low and trivial concentrations bothin indoor and outdoor environments.

Indoor air pollution can be defined as the degradation inthe indoor air quality, The Indoor pollution can be up to10 times worse than outdoor air pollution. Over a millionpeople in India die every year because of indoor airpollution (Kankaria, et.al, 2014). People spend maximumtime while being indoor than outdoor (Yang, 2017).

People tend to spend their time in different kinds of placessuch as their homes, workplaces, public places and otherindoor environments known as microenvironments.

Effects of indoor air pollution on health

The term “building related illness” has been coined todescribe illnesses such as bronchitis, asthma, etc. thatcan be traced to specific airborne contaminants inbuildings. There are carcinogens from tobacco smoking,building materials and furnishings, as well as from thenaturally occurring carcinogen, radon. Besides, manyother hazardous substances generate indoor pollution.Advances in construction technology have caused amuch greater use of synthetic building materials and inmodern days, buildings are made airtight to improveenergy efficiency which in turn results in poor ventilation.The principal sources of indoor air pollution are smokefrom combustion, releases from building materials, bioaerosols, radon, asbestos, pesticides, heavy metals,volatile organic matter and even tobacco smoke. Thecombustion products of biomass fuels contribute mostly

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Pollutant Properties Major Sources Health Impacts

Asbestos A natural mineral fiber usedin various building materials.All homes more than about20 years old are likely to haveasbestos

Damaged or deterioratingceiling, wall, and pipe insulation,Vinyl-asbestos floor material,Fireproof gaskets in heatshields, wood stoves andfurnaces, Acoustical materials,Thermal insulation, Exteriorsiding

No immediate symptoms, chest,abdominal and lungcancers and asbestosis.Asbestos can cause lung cancerespecially among smokers

BiologicalContaminants

Molds, mildews and fungi,bacteria, viruses, dust, mites

House dust, Infected humans oranimals, poorly maintainedhumidifiers, dehumidifiers & airconditioners, carpets and homefurnishings

Allergies and asthma,headaches, eye, nose and throatirritation, colds, flu, and pneumonia

CarbonMonoxide (CO)

Colorless, odorless gasproduced by incompletecombustion of all carbonfuels.

Heating equipments (furnaces,water heaters, fuel-fired spaceheaters) natural gas, kerosene,wood or coal stoves, fireplaces,cook tops and ovens, charcoalgrills, engines (gasoline, diesel)

Headaches, drowsiness, dizziness,impairment of respiration, vision &brain functioning,nausea, mental confusion, very highconcentration can cause death

TobaccoSmoke

Smoke exhaled by smokers,also known as side streamsmoke.

Cigarettes, Cigars, Pipes Eye, nose and throat irritation,Respiratory irritation (wheezing,coughing), Bronchitis andpneumonia (particularly in children)Increased risk of emphysema,lung cancer and heart diseases

Formaldehyde Pungent gas released intoair.

Pressed wood products, ureaformaldehyde foam, wallinsulation, carpets, draperies,furniture, fabrics paper products,glues, adhesives etc.

Allergic reactions,eye, nose and throat irritation,headachesnausea, dizziness, coughing etc.

Lead Natural element once usedas a component in gasoline,house paint, solder and waterpipes.

Household dust from leadpaints, lead-based paint,water from lead or lead-solderedpipes or brass fixtures, soil nearhighways / lead industries,Lead-glazed ceramic ware.

Damage to brain, kidneys, andnervous system, behavioural andlearning problems, slowed growth,anaemia, hearing loss etc.

Nitrogen oxidesand Sulfurdioxide

Gases formed by incompletecombustion of all carbonfuels.

Same as for carbon monoxide Damage to respiratory tract andlungs (Nitrogen dioxide), Irritation ofeyes, nose and respiratory system(sulfur dioxide)

Radon Colorless, tasteless andodorless gas that comes fromthe radioactive decay ofuranium or radium.

Earth and rock under buildings.Some earth-derived buildingmaterials groundwater and well-water.

Lung cancer, smokers are at higherrisk of developing radon inducedlung cancer

RespirableSuspendedParticulates(RSP)

Particles small enough toinhale, which come in avariety of sizes, shapes, andlevels of toxicity.

Wood-burning stoves,fireplaces, unvented kerosenespace heaters, gas-fired ranges,furnaces, water heaters,vacuum cleaning and housedust, Tobacco smoke, soappowders, pollen, lint, dust,cleaning and cooking sprays

Eye, nose, and throat irritation,respiratory infections and bronchitis,emphysema, lung cancer

V o l a t i l eO r g a n i cChemicals(VOCs)

Airborne chemicals containedin many household products

Aerosol sprays, hair sprays,perfumes, solvents, glues,cleaning agents, fabricsofteners, pesticides, paints,moth repellents, deodorizersand other household products,

Eye, nose, throat irritation,HeadachesLoss of coordination,confusion, damage to liver, kidneys,and brain, various types of cancer

Table 1. Common indoor air pollutants and their sources

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to indoor air pollution in developing nations. Some ofthe common house-hold items which we use in our dailylives can act as a potent source of indoor pollutants(Table 2). Some examples are candles, air fresheners,cosmetics, perfumes etc. The indoor air pollutants havepotential health effects. The ill-effects of indoor airpollution results in about 2 million premature deaths peryear (Kankaria, 2014) of which 44% are due to

Items Pollution Recommendation

AirFresheners

Almost all air fresheners emit toxic chemicals. Many top-selling freshenerscontain significant amounts of ethylene-based glycol ethers, which are knownto cause neurological and blood effects, including fatigue, nausea, tremor,and anaemia. Many air fresheners also contain phthalates, which are provenendocrine disruptors. These can be specifically harmful to infants and childrenwhose endocrine systems have not yet fully developed. Phthalates affect thedeveloping male sex organs and are linked to abnormally develop malegenitalia, poor semen quality and low testosterone levels.

A safer choice for purifyingthe air would be to keephouseplants. For fragrance,use essential oils.

Candles Most candles will pollute with harmful gases and sediments. The candle ismade from paraffin, vegetable oil, soy or beeswax. While burning, candlesreleases soot carbon particles that can lead to respiratory problems. Anotherchemical, acrolein, added to the wax as a solidifying agent is linked to the riskof lung cancer.Most of the candles are made of Paraffin. Paraffin is a by-product ofpetroleum, coal or shale that has been whitened by bleaching. Burningparaffin candles emits large amounts of benzene and toluene, both are knowncarcinogens. Other toxins in candle include artificial dyes and syntheticfragrances, especially those used for aromatherapy. These ingredients oftencontain toxic plasticizers and solvents.

Buy candles made ofbeeswax or vegetable oils,and with natural dyes andperfumes.

Carpet That famous “new carpet smell” is actually the off-gassing of hazardousvolatile organic compounds (VOCs), including toluene, benzene, ethylbenzene, formaldehyde, bromine, styrene, and acetone. Regular exposure tothese chemicals is known to cause headaches, throat and eye irritation,allergies, confusion, and drowsiness. Synthetic carpets made from nylon andolefin fibers typically off-gas the most. Exposure to these toxins can createlong-term health problems, including learning and memory impairment, birthdefects, decreased fertility, and diseases of the liver, thyroid, ovaries, kidneys,and blood. Benzene is a well-known human carcinogen and formaldehyde is aprobable human carcinogen. Some new carpets also contain the moth-proofing chemical naphthalene, which is known to produce toxic reactions,especially in new-borns. Some carpets also contain P-Dichlorobenzene, acarcinogen also known to produce foetal abnormalities.While older carpets no longer off-gas toxins, dust mites will infiltrate carpetsover time. The droppings can cause severe allergic reactions in many people,and scientists are just beginning to correlate dust mite exposure to asthma.We also add toxins into our carpets when we track in contaminated dirt, heavymetals and pesticides from outside on our shoes. Almost any toxic substancewe use near or within the home can settle into carpet fibers and later spreadinto the air.

Consider removing yourshoes and leaving them bythe door every time youcome into the house. Notonly will it reduce toxins inyour home, and your floorswill stay cleaner too.

CleaningProducts

Many conventional household cleaning products contain harmful chemicalslike alcohol, chlorine, ammonia or petroleum based solvents, all of whichcauses eye or throat irritation and headaches.Some cleaning products release dangerous Volatile Organic Compounds(VOCs) that can contribute to chronic respiratory problems and aggravateallergies, asthma and other respiratory illnesses. Products containing VOCsinclude aerosol sprays, chlorine bleach, upholstery cleaners, furniture andfloor polish, and oven cleaners.Chlorine bleach is particularly dangerous. Mixing bleach with any acidiccleaner like ammonia or vinegar can create chlorine gas which can causeimmediate health problems, even death, when inhaled.Even “green” or “organic” cleaning products may contain ingredients that cancause health problems. Natural citrus fragrances in particular can producedangerous indoor pollutants.

The safest is to use lesstoxic, less expensivecleaners such as hydrogenperoxide (for sanitizing,stain removal andbleaching), tea tree oilor Thieve’s oil and water(for mold removal and as adisinfectant), baking soda,and white vinegar

Pneumonia, 54% due to Chronic Obstructive PulmonaryDiseases (COPD) and 2% from Lung Cancer (Bhat,2012). The most affected groups are women and youngerchildren, as they spend maximum time at home. Thepollutants commonly occurring indoor, their sources andimpacts are summarized in Table 1. The commonhousehold items which we use frequently and can actas indoor pollutants are listed in Table 2. A very common

Table 2. Some common house hold Items which act as a potent indoor pollutants

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Dryer sheets Many people like the smell of warm laundry just pulled from the dryer. Thewaxy surfactant is made of a mixture of quaternary ammonium salt (which islinked to asthma), silicon oil, or stearic acid (derived from animal fat) whichmelts in the heat of the dryer to coat clothes. Dryer sheets also containfragrances which contain toxins that get into the air when released from dryervent emissions.Laundry detergents and dryer sheets contain more than 25 volatile organiccompounds, including seven hazardous air pollutants. Of those, twochemicals, acetaldehyde and benzene are carcinogens.

Use a less toxic dryer sheetthat contains no fragrancesor masking agents.

Furniture Chemical fire retardants are common in a wide variety of household itemssuch as furniture, electronics, appliances and even baby products. Thesechemicals were mandated by a 1975 law called TB 117, but they have sincebeen proven ineffective in preventing fires and are linked to numerous healthand environmental problems. In fact, these chemicals can make fires moretoxic by forming deadly gases and soot—which are the real killers in mostfires. Fire retardants are most commonly found in furniture containingpolyurethane foam, including couches and upholstered chairs, futons andcarpet padding. They can also be found in children’s car seats, changing tablepads, portable crib mattresses, nap mats and nursing pillows.

It is nearly impossible toavoid fire retardantscompletely. However someprecautions can minimizethe exposure. This includechoosing products madewithout fire retardants.,damping and dusting thefurniture on a regular basis.,checking the TB 117-2013label and verifying with thestore that the product doesnot contain flameretardants., Covering ofcushions since exposedfoam can allow fireretardant chemicals toescape more quickly. Itemssuch as car seats andmattress pads shouldalways be completelyencased in protectivefabrics.

KitchenStove

A poorly ventilated kitchen can cause a huge amount of air pollution. Gasstoves emit nitrogen dioxide, which is formed when fuel is burned at hightemperatures. Nitrogen dioxide mixes with the air to form nitric acid and toxicorganic nitrates. These can irritate the lungs and lower resistance torespiratory infections such as influenza. According to the EPA, frequentexposure to high concentrations of nitrates may cause acute respiratoryillnesses in children.

Make sure your kitchen iswell ventilated. Installing aventilation fan or rangehood can greatly improveair quality. In case ofabsence of ventilation fan,be sure to cook with nearbywindows open.

Paint Paint typically contains VOCs, and can off-gas for weeks, even months after aroom is painted. Paint fumes can cause headaches, dizziness, nausea,asthma, exacerbation, fatigue, skin allergies, confusion and memoryimpairment.

When buying paint, choosebrands that are Zero-VOC.

Mosquitorepellentcoils

Use of mosquito repellent is of the significant and consistent cause of indoorair pollution. Nowadays different types of mosquito repellents (solid form orliquid) are used. When the liquid repellent is sprayed in the room, the toxicchemicals present in it will mix with the air. These chemicals are toxic not onlyfor mosquitoes but also for the humans. These chemicals have great effect oneyes, skin and brain cells. Spray causes neuron to die in the regions of thebrain that control movement of muscles, learning, memory and concentration.The smoke can also cause various health problems. It can affect eyes,throats, nose, kidney and lungs. It is causing head ache, irritation, damage tobrain, memory loss and other respiratory problems.The burning of mosquito coil reportedly produces higher levels of indoor PM2.5 and CO, which were identified as potential causative factors of adverserespiratory health effects

Efforts should be made toavoid of those items/activities which attractsmosquitoes.1. Temperature. Mosquitoesgenerally get attracted tothe cooler space.2. Moisture. Perspirationdraws attraction ofmosquitoes. Even smallamounts of water in theroom will help in their visit

and reasonable method to combat indoor air pollutionin these days is the maintenance of indoor plants, alsoknown as house plants, which effectively filter the indoorair pollutants, while decorating the houses and requireslittle expenses for their maintenance.

Air filtering indoor plants

Plants improve indoor air quality by removing toxins; filterdust and increases moisture content. They also reducehuman stress. Indoor plants like ferns, palms, and

Table 2. Some common house hold Items which act as a potent indoor pollutants

can cause a huge amount of air pollution.

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Plants Common Name Properties Image

Aglaonema sp. ChineseEvergreen

Removes benzene and formaldehyde.

Aloe barbadensis Aloe Vera Aloe vera is a succulent plant species. Itcleans the air perfectly. A single Aloe Veraplant can refresh any small apartment. Itremoves formaldehyde effectively fromindoor air. It is also known for its healingproperties. It can treat burns and colds.

Anthuriumandraeanum

Flamingo lily Removes toulene, xylene, ammonia andhydrocarbon

Chlorophytumcomosum

Spider Plant Removes formaldehyde, xylene andtoluene. Removes carbon monoxide

Chrysalidocarpuslutescens

Bamboo Palm Adds moisture to dry air and removesbenzene, trichloroethylene andformaldehyde

Chrysanthemummorifolium

Chrysanthemums This plant was found by NASA to be a realair-purifying plant. It removes ammonia,benzene, formaldehyde and xylene. It ispopular and inexpensive and can beeffective in outside environments too.

Dieffenbachiahybrids

Leopard Lily Its large leaf surface area helps to quicklyremove air contaminants from indoorspaces. These are poisonous if come indirect contact.

orchids can do wondrous things (Girman et.al., 2009).They also help us feel at one with nature, while draggingharmful pollutants out of the air. Indoor plants have very

high rates of photosynthesis, which allow them to groweven in very diffuse light. The leaves, roots, soil andmicro-organisms work symbiotically to remove pollutants,

Table 3. List of indoor plants and their role in indoor air purification.

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Plants Common name Properties Image

Dracaena fragrans/Deremensis

Janet Craig,Warneckii,Massangeana

Remove benzene, formaldehyde, xylene andtoluene

DraceanaMassangeana

Cornplant

The Corn Plant absorbs trichloroethylene

Epipremnumaureum

Money Plant This plant acts as an excellent natural anti-pollutant against common pollutants likebenzene, formaldehyde and carbon monoxide.

Ficus benjamina &Ficus alii

Weeping FigorBenjamin's Fig

Insect resistant and known for its braided trunk.It is effective at filtering formaldehyde. It alsoworks well to remove chloroform, which is foundin dyes and pesticides

Ficus elastica Rubber Plant Removes formaldehyde, may be toxic ifswallowed

Gerbera jameson Daisy, Gulbahar NASA reports that this plant is fantastic atremoving benzene, a known cancer-causingchemical. It also absorbs carbon dioxide andgives oxygen overnight.

Hedera helix English Ivy Ivy is a good remover of formaldehyde, benzene,toluene, xylene and trichloroethylene from theatmosphere.

Nephrolepis sp. Boston fern Absorbs formaldehyde, xylene and toluene

Ocimum basilicum Tulsi or Holy basil Tulsi releases a special kind of essential oilwhich frees the air from bacteria and substances

purify indoor air by filtering out toxins, pollutants and thecarbon-di-oxide we exhale, replacing them with oxygen.

Air pollutants are removed from the air by being absorbedthrough tiny pores in their leaves (Wolverton, 2005; Star


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Plants Common name Properties Image

Ocimum basilicum Tulsi or Holy basil Tulsi releases a special kind of essential oilwhich frees the air from bacteria and substancesthat cause diseases. It also releases ozonewhich contains 3 atoms of Oxygen (O3)

Philodendron sp. Arrowhead vine Climber Pant that removes formaldehyde,toluene, trichloroethylene and xylene from thesurrounding atmosphere. Cuttings grow easily ina moist environment

Phoenix roebelinii Dwarf date palm Removes formaldehyde and xylene fromsurrounding indoor air.

Rhapsis excelsa Lady palm Resistant to plant insects and will tolerate a widerange of indoor environments – besidesabsorbing volatile organic compounds.

Sansevieriatrifasciata 'Laurentii’

Snake plant orMother in law’stongue

Removes benzene, formaldehyde, xylene andtoluene.

Scindapsus sp. Pothos It removes benzene and formaldehyde from theair.

Spathiphyllum sp. Peace lily It has long dark green leaves and a unique whiteflower. It thrives in low light. It removes acetone,benzene, trichloroethylene and xylene

and Cengage, 2012; Pessarakli, 2016). They are movedthrough the plant, to the root zone, where they are brokendown by soil microbes. For example plants absorbvolatile organic compounds (VOC) from the air into theirleaves and then translocate them to their root, wheremicrobes act on them and break them down.Microorganisms in the soil can use trace amounts of

pollutants as a food source. The right selection of indoorplants can purify the indoor air and can protect us fromharmful indoor pollutants. A study by NASA (Wolveryonet.al., 1989) found that there were specific plants thatwere most effective at removing benzene, formaldehyde,trichloroethylene, xylene and ammonia from the air(Table 3).


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However there is some ambiguity regarding thebeneficial aspect of the indoor plants as they releasecarbon dioxide (CO2) during night. Most indoor plantsrelease CO2 into the interior at night because of plantrespiration process. Therefore, the introduction of plantsindoors may be effective in improving indoor air qualityduring the day but may have a negative effect on healthdue to an increase in carbon dioxide concentration atnight. However the reports revealed that there are minorvariations in this process, depending upon time of dayand time of year, the amounts of gases released orabsorbed is small and will not appreciably alter thecomposition of indoor air, even in rooms with a lot ofplants. Even greenhouses and botanical gardens usuallydon’t contain enough plants to significantly affect the ratioof oxygen vs. CO2 in the air. Some studies has alsorevealed that use of use fleshy plants such as cactusand mixed planting of CAM plants and C4 plants iseffective in removing indoor pollutants and balancing theindoor CO2 concentration (Lee and Kang, 2015).

Control Measures

There is a need to adopt various strategies to improveindoor air quality. People should be made aware aboutthe indoor pollution and its impacts. The few measureswhich could be adopted to prevent indoor pollution are:

Public awareness: One of the most important stepsin the prevention of indoor air pollution is education,spreading awareness among people about the issueand the serious threat. The education should helppeople in finding different ways of reducing exposuressuch as better kitchen management practices andprotection of children at home. People should alsobe educated about the use of alternative cleanersources of energy.

Change in pattern of fuel use: Fuel use depends onones’ habit, availability and its affordability. Themajority of low income families rely solely on directcombustion of biomass fuels for their cooking needsas this is the cheapest and easiest option availableto them; however, this could be rectified by promotingthe use of cleaner energy sources such as gobargas which utilizes cow dung to produce gas forcooking.

Modification of design of cooking stove: The stovesshould be modified from traditional smoky and leakycooking stoves to the ones which are fuel efficient,smokeless and have an exit (e.g., chimney).

Improvement in ventilation: During construction ofhouses, importance should be given to adequateventilation; for poorly ventilated houses, measuressuch as a window above the cooking stove and crossventilation though doors should be instituted.

Wise selection of materials: The common house holditems which we use on a regular basis can act as a

source of indoor pollutant. Hence general awarenessand knowledge about the detrimental effects of thisitems and wise and proper selection of items areessential to combat indoor pollution. Maximizing theuse of organic materials and plant products in placeof synthetic chemicals can reduce the source ofpollutants.

House Plants: Try to plant as many as house plantsas possible. This will not only filters harmful pollutantsfrom your home and offices, but will return you withpure oxygen. House plants add aesthetic beauty toour homes and offices and require only soil and wateras inputs.

REFERENCES

Bhat, Y.R., Manjunath, N., Sanjay, D. and Dhanya, Y.(2012) Association of indoor air pollution with acute lowerrespiratory tract infections in children under 5 years ofage, Paediatr. Int. Child Health. 32(3), 32-135.

Girman, J., Phillips, T. and Levin, H. (2009). CriticalReview: How well do house plants perform as indoor aircleaners, Proceedings of Healthy Buildings, 23, 667-672.

Kankaria, A., Nongkynrih, B. and Gupta, S. K. (2014)Indoor Air Pollution in India: Implications on Health andits Control. Indian Journal of Community Medicine:Official Publication of Indian Association of Preventive& Social Medicine. 39(4), 203-207.

Lee, J. and Kang, H. (2015) The Effect of ImprovingIndoor Air Quality using some C3 Plants and CAM Plants,Indian Journal of Science and Technology, 8(26),1 -7.

Linda, A. (2000). Common indoor air pollutants: Sourcesand health impacts. IAQ Fact Sheet 2, 1-3.

Pessarakli, M. (2016). Handbook of Photosynthesis,CRC Press.

Phalen, R. F. (1996). Methods in Inhalation Toxicology,CRC Press.

Starr, C. and Cengage, C.E. (2012) Biology Today andTomorrow with Physiology-Science.

Wolverton, B. (2005) How to grow fresh air: fiftyHouseplants that purify your home or offices, Insight:The Consumer Magazine.

Wolverton, B.C., Douglas, W.L. and Bounds, K. (1989)Interior landscape plants for indoor air pollutionabatement - Final Report, September 15, NASA.

Yang, Y. (2017). A numerical study of the particlepenetration coefficient of multi-bended building crack.Aerosol Air Qual. Res. 17, 290–301.

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