soil dehydrogenase enzyme activity in natural and mine soil - a review

Middle-East Journal of Scientific Research 13 (7): 898-906, 2013ISSN 1990-9233© IDOSI Publications, 2013DOI: 10.5829/idosi.mejsr.2013.13.7.2801

Corresponding Author: S. Kumar, Department of Environmental Science & Engineering / Centre for Mining Environment,Indian School of Mines; Dhanbad 826004, India.

898

Soil Dehydrogenase Enzyme Activity in Natural and Mine Soil - A Review

S. Kumar, S. Chaudhuri and S.K. Maiti1 2 1

Department of Environmental Science & Engineering / Centre for Mining Environment,1

Indian School of Mines; Dhanbad 826004, IndiaDepartment of Mining Engineering, Indian School of Mines; Dhanbad -826004 India2

Abstract: Soil enzyme activities are very sensitive to both natural and anthropogenic disturbances and showa quick response to the induced changes. Soil dehydrogenase enzymes are one of the main components of soilenzymatic activities participating in and assuring the correct sequence of all the biochemical routes in soilbiogeochemical cycles. Dehydrogenase activity is measured by two methods using the TTC and INT substrate;however, various authors reported poor results when TTC is used as substrate. Different biotic and abioticfactors such as incubation time and temperature, pre-incubation, soil aeration and moisture content havesignificant effect on dehydrogenase activity in soil. Highest dehydrogenase activity is reported from forest soilin autumn seasons while the disturbed soil from coal mines soils containing lowest dehydrogenase activitiesalong the soil erosion gradient of experimental slopes. Least value of enzymes activity is reported from pollutedsites than restored and undisturbed sites. Dehydrogenase enzyme is often used as a measure of any disruptioncaused by pesticides, trace elements or management practices to the soil, as well as a direct measure of soilmicrobial activity.

Key words: Dehydrogenase Activity Substrate Mine Soil Natural Soil

INTRODUCTION and facilitate the development of plant cover. It is an

Soil quality and its degradation depend on a large insight into the relative changes in below-ground systemnumber of physical, chemical, biological, microbiological functioning as a plant community develops over time.and biochemical properties, the last two being the most Enzyme activity in soil results from the activity ofsensitive since, they respond rapidly to changes. The accumulated enzymes and from enzymatic activity ofmicrobiological activity of a soil directly influence proliferating microorganisms [3]. They are usuallyecosystem stability and fertility and it is widely accepted associated with viable proliferating cells, but enzymes canthat a good level of microbiological activity is essential for be excreted from a living cell or released into the soilmaintaining soil quality. The soil microbiological activity solution from dead cells. Study of soil enzymes givesviz., the enzymatic activities play a key role in soil nutrient information about the release of nutrients in soil bycycling, its activity is essential in both the mineralisation means of organic matter degradation and microbialand transformation of organic matters and plant nutrients activity as well as indicators of ecological change.in soil ecosystem [1]. Soil enzymes analysis helps to establish correlation

Soil enzyme activities are very sensitive to both with soil fertilization, microbial activity, biochemicalnatural and anthropogenic disturbances and show a quick cycling of various elements in soil, degree of pollutionresponse to the induced changes [2]. Therefore, enzyme (heavy metals) and to assess the succession stage of anactivities can be considered effective indicators of soil ecosystem. So, measurements of enzyme activity inquality changes resulting from environmental stress or degraded soils have useful in examining impacts ofmanagement practices. These soil enzymes play a environmental change or management on soil enzymefundamental role in establishing biogeochemical cycles activities. Several works have been reported the potential

important aspect of the below-ground processes and give

Middle-East J. Sci. Res., 13 (7): 898-906, 2013

899

use of enzyme activity as an index of soil productivity or activity thus serves as an indicator of the microbiologicalmicrobial activity [4, 5]. One of the general criteria used to redox systems and may be considered a good measure ofdetermine microbial activity and biomass in soil is soil microbial oxidative activities in soils [12]. H can bedehydrogenase activity (DHA). transferred to soluble tetrazolium salts (e.g., TTC, INT)

Dehydrogenase Enzymatic Activity (DHA): Soil DHA determined calorimetrically after extraction with a solvent.determination in soils was first initiated by [6], since thenit has been widely used because of its simplicity as Different Method Used for Measurement ofcompared to other quantitative methods. The method was Dehydrogenase Activitylater on modified by [7, 8]. Soil dehydrogenase enzymes Use of Tetrazolium Salts: Dehydrogenase assays basedare one of the main components of soil enzymatic on the reduction of 2, 3, 5- triphenyltetrazolium chlorideactivities participating in and assuring the correct (TTC) to the creaming red-colored formazan (TPF),sequence of all the biochemical routes in soil have been used to determine microbial activity in soil.biogeochemical cycles [9]. It has also found that Water-soluble tetrazolium salts are the preferred oxidantsmeasurement of changes in soil enzyme activities may because they form water-insoluble colored formazansprovide a useful index of changes in soil quality [10]. which can be measured spectrophotometrically. Soil wasThe soil dehydrogenase activity in soils provides Prepared with CaCO and dispensed in three tubes as 6gcorrelative information on the biological activity and each. The total volume of fluids added to the soil wasmicrobial populations in soil. The basic idea of using soil 3.5 ml; this included any fluids added duringenzymes activity as a measure of microbial indicators for preincubation of the soil. Most substrate solutionsoil fertility was introduced and established by Waksman concentrations were 1% dextrose solutions. All of these[11]. Soil dehydrogenase activity is considered to exist in were sterilized by autoclaving and 1 ml was added to thesoils as integral parts of intact cells. They are not residing soil at the time of TTC addition. However, the substratesextracellular in the soil. Measurement of dehydrogenase were added as 0.5 ml of a double-strength solution if thereactivity represents immediate metabolic activities of soil was danger of exceeding the 3.5-ml fluid volume limit.microorganism at the time of the test. Soil dehydrogenase TTC (Calbiochem, San Diego, Calif.) was prepared as a 3%activity is an oxidative degradation process .i.e., aqueous solution and was sterilized by passage throughdehydrogenation of organic matter by transferring a 0.30-, um membrane filter (Millipore Corp., Bedford,hydrogen and electrons from substrate to acceptors. Mass.). Each tube received 1 ml of TTC, except for the soilDehydrogenase enzymes play a significant role in the blank, which received water instead. After addition, thebiological oxidation of soil organic matter by transferring water, substrate and TTC were simultaneously mixedprotons and electrons from substrates to accepters. through the soil with a sterile glass rod; then rubberWater-soluble tetrazolium salts are the preferred oxidants stoppers were inserted and the tubes were incubated atbecause they form water-insoluble colored formazans 37°C at different incubation period of 6hrs, 12hrs, &24hrs.which can be measured spectrophotometrically. Upon completion of incubation, the tubes could be

The intracellular dehydrogenase enzymes belong to extracted immediately. Each soil sample was transferredthe oxidoreductases and catalyse the oxidation of with methanol to a funnel containing Whatman no. 5 filterorganic compounds by separating two–H atoms. paper (W. and R. Balston, Ltd.) placed on a 100-mlMany specific dehydrogenases act as to transfer the graduated cylinder. Additional portions of methanol wereseparated H to either nicotinamide adenine dinucleotide passed through the soil until 50 ml of methanol,or nicotinamide adenine dinucleotide phosphate. containing the formazan, had been collected in theThrough these co-enzymes the H atoms take part in the graduated cylinder. If the filtrate passing from the funnelreductive processes of biosynthesis. Therefore, the still had a red color, additional methanol was passedoverall dehydrogenase activity of a soil depends on the through the soil until all formazan had been extracted andactivities of various dehydrogenases, which are a corrections in calculations were made for the additionalfundamental part of the enzyme system of all methanol. The red methanolic solutions of the formazanmicroorganisms (enzymes of the respiratory metabolism, were read at 485 nm against the extract from the non-TTCthe citrate cycle and N metabolism). Dehydrogenase soil blank by using a Spectronic 20 colorimeter

with the formation of red formazans, which can be

3


900

(Bausch and Lomb, Rochester, N.Y.). The values obtained at 30°C without added substrate. The 24 hrs incubationwere compared against a formazan (Calbiochem) standard periods seems to be the part of the dehydrogenasecurve prepared with methanol and they are reported as technique most likely to cause problems in themilligrams of formazan per gram of soil. interpretation of results. The dehydrogenase technique

Use of P-Iodonitrotetrazolium Violet, or INT salts: soil was modified to use a 6 hr, 37°C incubation with eitherVarious authors have used INT 2-(p-iodophenyl)-3- glucose or yeast extract as the electron donating(p-nitrophenyl)- 5-phenyltetrazolium chloride substrate by Casida [21]. He reported that the rate of(p-iodonitrotetrazolium violet, or INT) which act as an formazan production remained constant during this timeartificial electron acceptor as a substrate to determined interval and cellular multiplication did not occur. TheDHA in soil [13]. Later, Trevors et al. [14] used INT for technique was used to follow changes in the overallthe determination of dehydrogenase activity in soils, metabolic activities of microorganism in soil undergoingfinding it to be more suitable than TTC for this purpose. incubation with a limiting concentration of addedBenefield et al. [15], Grifiths [16] and Von Mersi and nutrients. Ross [22] studied soil dehydrogenaseSchinner [17] have reported that INT affords a more determinations conducted at 30°C. He too concluded thataccurate estimation of dehydrogenase activity than TTC, 6 hr incubation was more valid than longer incubations;which they attributed to its higher electron affinity however, he recommended a 1 hr anaerobic incubation.compared to TTC. As a consequence of this high electronaffinity, INT competes more effectively with oxygen for Pre-Incubation of Soil: Pre-incubation of soil withfree electrons and so it is not necessary to carry out the yeast extract or glucose increase the DHA of the soil.determinations under anaerobic conditions. Furthermore, Pre-incubation of a dry soil with moisture, however, alsounder both aerobic and anaerobic conditions, more INT caused an increase in DHA. A phenomenon somewhatthan TTC is reduced and so sensitivity is also improved similar to the latter was studied by Stevenson [23],[18]. They observed that the reduction from INT to INTF who pointed out that nutrients become more readily(iodonitrotetrazolium formazan) gives a more exact available when dry soil is remoistened. In addition,measurement of DHA in soil extracts, as O does not however, there probably is some breaking of dormancy for2

interfere with the INT reduction. More formazan is cells and depending on the duration of incubation of theproduced from the INT reduction than TTC, both with remoistened soil, there can be cellular multiplication.anaerobic and aerobic incubation [18]. Substrate added to the soil for incubation trails usually

Different Factors Affecting the Dehydrogenase response is a component of the response to the substrate.Enzymatic Activities: Dehydrogenase activity can be Perhaps dry soils remoistened and incubated beforeconsidered to be a good measure of microbial oxidative substrates are added or dehydrogenase determinationsactivity in soils. It is usually determined by measuring the are made. Regardless of whether a soil has beenamount of an artificial electron acceptor reduced by pre-incubated, the dehydrogenase curve (formazanmicrobial activity, such as a soluble tetrazolium salt with production) does not remain linear during 24 hr ofa red colored reduced form (a formazan) that can be incubation but, instead, show a change to a decreaseddetermined colorimetrically following extraction with a rate at about 9 to 12 hr.suitable solvent. Studies of dehydrogenase enzymaticactivities in soil are important as they indicate the Soil Aeration: Soil aeration parameters such as oxygenpotential of the soil to support biochemical processes diffusion rate (ODR), redox potential (Eh), concentrationwhich are essential for the maintenance of soil fertility. of (Fe ), water content and bulk density have beenSoil dehydrogenase activity is often used as a measure of significantly effects on dehydrogenase activity in soilany disruption caused by pesticides, trace elements or (Brzezinska et al., [24]. They have found positivemanagement. correlation with Fe , water content and bulk density,

Incubation Time and Temperature: The most used Maximum DHA was observed at soil pH of 6.6-7.2.incubation periods is 24 hrs or longer at 37°C to study The soil physical properties and the content ofDHA. Klein et al, [19] incubated soil for 96 hr with decomposable organic matter are the main factorsglucose at 27°C and Skujins [20] incubated soil for 24 hrs determining the soil aeration status.

for measuring the metabolic activity of microorganism in

are added as an aqueous response is a component of the

2+

2+

while negative correlation in case of ODR and Eh.


901

Soil Moisture Content: The soil moisture influences both soil of aged less than 10 years, which containing aboutthe microbial activity and DHA in the soil. Since moisture 50.1±6.2 mg INTF g of soil h , another 10-20 years oldfound to be an important factor in microbial activity, after agricultural soil contains about 16.2±5.2 mg INTF g ofthe initial addition of water to the soils, 10 day were soil h and greater than 20 years abandoned agriculturalnecessary to reactivate the microbial population [25]. soil containing the same dehydrogenase activity as 10-20In succeeding weeks, the reduction in the soil moisture years old abandoned agricultural soil. The decrease incontent caused a corresponding reduction in the active activity they have reported may be with the passing timebiomass and thus also in DHA, which continued until may be due to progressive erosion of the abandoned soilsafter the 6 week of incubation, when water was again as a consequence of the low levels of plant cover and theth

added to the soil, once again allowing an increase in low levels of organic matter.enzyme activity. In soil having low moisture level the An experiment was conducted by Simek et al. [32]DHA was reported close to zero [26]. The highest DHA to study the evidence of rich microbial communities in thewas reported in rainy season and lowest in winter. Soil subsoil of a boreal acid sulphat soil conducive toDHA was positively and significantly correlated with soil greenhouse gas emissions from research farm at thepH, Ca, Mg, K and water content. Mersi and Schinner [27] University of Helsinki, Finland. They have found thatalso suggested that samples should be analysed dehydrogenase activity (DHA) was the highest in theimmediately if possible; where an immediate analysis is Ap horizon of both pedons 12.3±0.1µg TPF g h in thenot possible a maximum of 1 week and all analyses be non-acid sulphate soil and 14.3±0.1µg TPF g h in thecarried out with field moist soils. acid sulphate (AS) soil. In the B horizons, the DHA

Dehydrogenase Activity in Natural Soil: Estimates of the In the C horizons of both pedons, it increased again todehydrogenase activity of soil are simple to carry out and relatively high values. High microbial activity in thesemay be used more frequently for studying various aspects horizons of the AS soil are due to presence of largeof the oxidation of organic compounds in soil or for carbon and nitrogen stocks as well as high substrateindicating possible levels of biological activity in soil. induced respiration. The potential impact ofQuilchano et al. [28] studied the seasonal effect on soil environmental change on ecosystem structure anddehydrogenase activity in upper 10 cm of Mediterranean function and to understand the relationships betweenforest soils in Los Alcornocales Natural Park (Southern enzyme activity and biotic and abiotic factors was studiedSpain). They have found that the DHA of forest soil in by Li, et al. [33] at the Department of Geography, Israel.autumn 527±165 nmol p-iodonitrotetrazolium formazan They observed that with increasing aridity, the soil(INTF) g h was almost double that in summer 289±95 dehydrogenase activities decreased significantly in both1 1

nmol INTF g h , for one of the studied plots. During 0–2- and 5–10-cm soils. They have also observed that1 1

the dry season, DHA of forest control soils 324±85 nmol when the field-moist soil which was air-dried,INTF g h was higher than in the thinned and dehydrogenase activity was increased significantly.1 1

shrub-cleared forest 253±93 nmol INTF g h . Dehydrogenase activities at two sites Giv’at Ye’arim (GIV)1 1

The values of DHA obtained are almost double those and MIS were least in winter, with no significantreported by Leirós et al. (2000) in an Atlantic forest of difference between spring and summer samples at bothQ. robur, exposed to a temperate climate in north-west soil depths. At Kalia (KAL), the value of soilSpain, in soils containing a higher amount of soil organic dehydrogenase activity at 0–2 cm was reported least inmatter than those of the Los Alcornocales forest. Other the summer samples.authors also attributed the increase in microbial activity in Bonanomi et al. [34] studied the soil quality underforest [29] and in grassland soils [30] to higher soil intensive cultivation and tree orchards in Southern Italy.moisture contents. They have selected 20 agricultural farms in five

The soil microbial activity act as a biomarker of geographical areas of Southern Italy with different soildegradation and remediation processes of natural soil types. In each farm, with different management regimeand abandoned agricultural soils were studied by Pascual which are classified as high-input (HIMR, intensiveet al. [31] at the department of Soil and Water cultivation under plastic tunnels) or low-inputConservation and Organic Waste Management, Spain. (LIMR, tree orchards). Results showed that there is effectThey have observed that natural soil contains the of management regimes on enzymatic activities.61.1±4.6 mg INTF g of soil h compared to agricultural Dehydrogenase activities were reduced by 84%1 1

1 1

1

1

1 1

1 1

decreased to about 3% of the values in the Ap horizons.


902

(0.89 µg TPF g h vs 5.41µg TPF g h ) in the HIMR Dehydrogenase Activity in Degraded Soil: Moreno-de las1 1 1 1

compared to the LIMR, respectively. This result is Heras [38] studied the development of soil physicalconsistent with the findings of many studies that structure and biological functionality in mining spoilsreported a decline of enzymatic activities in cultivated affected by soil erosion in a Mediterranean-Continentalsoils when compared to the corresponding uncultivated environment of Spain. The dehydrogenase activityor less-disturbed soils [35]. They have also showed that reported considerably low in the most eroded slopesenzymatic activities were positively correlated only with which are ranged from 0.34–0.56 µg INTF g soil h .total soil organic C content and with fungal mycelium. The maximum dehydrogenase activities were reported

The drying and rewetting effects on C and N from least eroded slopes which ranged from 0.87-0.98 µgmineralization and microbial activity in surface and INTF g soil h . Exponential decreases in enzymaticsubsurface of California grassland soils (USA) were hydrolytic activities were found along the soil erosionstudied by Xiang et al. [36]. They have reported that gradient represented by the five experimental slopes.microbial activity, as measured by dehydrogenase These decreasing trends are in accordance with theactivity, was roughly 4-fold higher in surface than in differences found in soil organic matter and vegetationsubsurface soil. In both soils, dehydrogenase decreased development between the slopes. Sinha et al. [39] studiedslightly with the initial equilibration and decreased further the microbial characteristic of rhizospheric tree specieswith extended dry incubation. In both soils, used for re-vegetation of degraded land in Jharia Coalfielddehydrogenase increased with initial wetting, but (India). They have selected three groups of differentdecreased with increasing dry/wet cycles. In surface soils, plants species used for revegetation. In the first group,dehydrogenase levels in the 12-cycle treatment fell to containing the highest DHA activity and were reported aslevels below those of the continuously moist treatment. >60 mg TPF g h in A. marmelos, M. alba, A. indica,In subsurface soils, all the cycling treatments increased D. sissoo of different plant species; intermediate valuetotal dehydrogenase activity over the course of the of DHA were reported as 20–40 mg TPF g h underincubation; in the 4- and 6-cycle treatments T. indica, M oleifera, F. religiosa, E. jambolana,dehydrogenase activities at the end of the incubation B. bauhinia plantation; and low values <20 mg TPFwere higher than in the continuously moist treatment. g h were recorded under Eucalyptus, T. grandis,The increases in dehydrogenase activity in cycling soils, B. monosperma plantation. The lowest dehydrogenasehowever, were much was the dominant mechanism driving activity, observed under Eucalyptus could be due to theoverall C and biomass dynamics through multiple dry/wet allelopathic effect of root exudates. The presence of manycycles in these soils, dramatically so in the subsurface aromatic organic pollutants in the coal mine spoil couldsoil. also be one of the limiting factors for the survival of the

The effect of moisture and disaggregation on the plant species and also the rhizospheric microbiologicalmicrobial activity of soil was studied by Paradelo et al. properties.[37] at Departamento de Edafolox ae Qu mica Agr cola, The influence of mycorrhizal fungi on the growth ofSpain. The dehydrogenase activity (DHA) of a vineyard different tree species and their nutrient uptake insoil was measured at different moisture conditions, in gypsum mine spoil in India were studied by Rao et al.different aggregate size fractions and after breaking [40]. They have observed that dehydrogenase activitiesthe existing aggregates. The highest values of DHA in the rhizosphere soils were significantly enhanced to(144 mg TPF kg day ) were found for the undisturbed varying degrees upon inoculation with G. fasciculatum.1 1

soil at a suction pressure of 0.1 MPa and decreasing The dehydrogenase soil enzymatic activities reportedvalues were obtained at pressures above and below that under the A.lebbeck plantation at the control site is 6.1water potential. The disruption of the aggregates strongly pkatal and the DHA value reported under A.lebbeckreduced the biological activity at suction pressures under plantation inoculated with G. fasciculatum is 8.2 pkatal.0.1 MPa, but had no effect in drier conditions. Both in The DHA activities recorded under the A.indica, 4.4disturbed and undisturbed soil, an important part of the pkatal in control site and the DHA activities found inbiological activity remained at high suction pressures inoculated site is 6.1 pkatal. These observations are in(over 0.1 MPa), with DHA values around 50% of the conformity with many reports of higher microbial activitymaximum values. in the rhizosphere of plants inoculated with AMF [41, 42].

1 1

1 1

1 1

1 1

1 1


903

Table 1: Status of Dehydrogenase Activity (DHA) in different types of land useSite description Types of soil DHA activity ReferencesMediterranean forest soils in Los Forest soil in autumn. 527±165 nmol (INTF) g h Quilchano et al. (2002).1 1

Alcornocales Natural Park (Southern Spain) Forest soil in summer 289±95 nmol INTF g h1 1

Forest control soils. 324±85 nmol INTF g h1 1

Shrub-cleared forest. 253±93 nmol INTF g h1 1

Natural soil and abandoned Natural soil 61.1±4.6 mg INTF g of soil h Pascual et al. (2000).1 1

agricultural soils, Spain. Agricultural soil 50.1±6.2 mg INTF g of soil h1 1

10-20 years old agricultural soil. 16.2±5.2 mg INTF g of soil h1 1

Research farm at the University Ap horizon in non-acid sulphate soil. 12.3±0.1µg TPF g h Simek et al. (2011)1 1

of Helsinki, Finland. Ap horizon in acid sulphate (AS) soil. 14.3±0.1µg TPF g h1 1

Cultivation and tree orchards in Southern Italy. Intensive cultivation under 0.89 µg TPF g h Bonanomi et al. (2011).1 1

plastic tunnels. 5.41µg TPF g h1 1

Low-input (LIMR, tree orchards).Vineyard soil, Spain Undisturbed soil at a suction pressure 144 mg TPF kg day Paradelo et al. (2009).1 1

of 0.1 MPa.Mining spoils affected by soil erosion in Eroded slopes 0.34-0.56 µg INTF g soil h Moreno-de las Heras (2009)1 1

a Mediterranean-Continental, Spian Least eroded slopes 0.87-0.98 µg INTF g soil h1 1

Re-vegetation of degraded land Rhizospheric soils of A. marmelos, >60 mg TPF g h Sinha et al. (2009).1 1

in Jharia Coalfield (India). M. alba, A. indica, D. sissoo plantation.Under under T. indica, M oleifera, 20-40 mg TPF g h1 1

F. religiosa, E. jambolana, B. bauhiniaplantation.Under Eucalyptus, T. grandis, <20 mg TPF g h1 1

B. monosperma plantationGypsum mine spoil, (India) Rhizosphere soils of A.lebbeck 6.1 pkatal Rao et al. (2001)

plantation at the control site.Rhizosphere soils of A.lebbeck plantation 8.2 pkatalinoculated with G. fasciculatum.

Reclamation of post-mining Loam sandy texture soil 72.7±26.9 µg INTF g h Chodak et al. (2010).1 1

barrens lands, Poland Sandy soil 52.2±11.2 µg INTF g h1 1

Under Birch plantation 77.3±26.8 µg INTF g h1 1

Mixed plantation 61.4±26.4 µg INTF g h1 1

Pine plantation. 48.7±14.8 µg INTF g h1 1

Heavy metal-contaminated and Non-polluted area 71.4±5.2 µg TPF g dw h Hinojosa et al. (2004).1 1

reclaimed soils, Spain. Reclaimed area 53.0±0.5 µg TPF g dw h1 1

Polluted area 2.9±12.1 µg TPF g dw h1 1

Chodak et al. [43] studied the re-establishment of soil The effect of soil moisture pre-treatment on statusmicrobial communities is a prerequisite for successful and health of soil microbiological enzymatic activities actreclamation of post-mining barrens to assess the effect of as indicators of heavy metal-contaminated andtexture of soil substrate and the planted tree species on reclaimed soils were studied by Hinojosa et al. [44], inmicrobial properties of mine soils reclaimed for forestry in Spain. The different soil samples were collected fromPoland. The dehydrogenase enzyme activity reported in polluted plot just underneath the pyrite mud layer, whichloam sandy texture soil is 72.7±26.9 µg INTF g h and was removed prior to sampling. The field moisture content1 1

in sandy soil it is 52.2±11.2 µg INTF g h . The effect of of soil samples reported was 4.0% (±2.8). It has been1 1

different plantation on DHA activities reported are under observed that DHA activities were significantly greaterBirch plantation is 77.3±26.8 µg INTF g h , in mixed for pre-treated rewetted and incubated soils. The greater1 1

plantation the activities is 61.4±26.4 µg INTF g h and amount of DHA activities were found in non-polluted area1 1

in Pine plantation it is 48.7±14.8 µg INTF g h . The is 71.4±5.2 µg TPF g dw h , average activities were1 1

afforested loamy sands contained signidficantly more reported in reclaimed area as 53.0±0.5 µg TPF g dw horganic matter and total nitrogen and had higher pH than and least value reported in polluted area as 2.9±12.1 µgthe sands. Consequently, they maintained larger and more TPF g dw h . Enzyme activities decreased significantlyactive microbial biomass and higher DHA activities. with increasing degree of pollution: non-polluted soil

1 1

1 1

1 1


904

showed the highest enzyme activity values, polluted increase in DHA because nutrients become morethe lowest and restored soil intermediate values. readily available when dry soil is remoistened.However, rewetting generally increased enzyme activities The DHA activity was greater in surface than sub-of non-polluted and reclaimed soils which improved surface soil which is due to accumulation of greaterdiscrimination from polluted treatments. Polluted soils had organic matter. It has been also found that the DHAmuch smaller increases or even decreases in enzyme activity was greater in natural soil than the degradedactivities with rewetting. soil.

Importance of Dehydrogenase Enzymes Activity in Soil: were significantly enhanced to varying degrees uponDehydrogenase enzyme is often used as a measure of any inoculation with G. fasciculatum as reported fromdisruption caused by pesticides, trace elements or the growth of different tree species and their nutrientmanagement practices to the soil, as well as a direct uptake in gypsum mine spoil.measure of soil microbial activity [45, 46]. It can also The DHA activity was found greater in forest soilsindicate the type and significance of pollution in soils. than grassland area. The DHA activity was alsoIt has been found that dehydrogenase enzyme is high found maximum in loamy sandy soil than sandy soil.in soils polluted with pulp and paper mill effluents [47] The afforested loamy sands contained significantlybut low in soils polluted with fly ash [48]. Similarly, higher more organic matter and total nitrogen and hadactivities of dehydrogenases have been reported at low higher pH than the sands.doses of pesticides and, lower activities of the enzyme athigher doses of pesticides [49]. As most areas of REFERENCESthe world are often polluted by different industrialbio-chemical products, better understanding of the role of 1. Dick, W.A. and M.A. Tabatabai, 1993. Significancethis enzyme in environmental science will open greater and potential uses of soil enzymes, pp: 95-127, In B.possibilities of using it as a diagnostic tool for better Metting (ed.), Soil Microbial Ecology, Marcel Dekker,ecosystem assessment and amelioration. New York.

CONCLUSIONS indicators of soil health. In: Pankhurst CE, Doube

The following are the significant findings emerging health. CAB International, New York, pp: 121-156.from the present study. 3. Kiss, S., M. Dragan-Bularda and D. Radulescu, 1975.

Soil dehydrogenase enzymes are one of the main Biological significance of enzymes in soil. Adv.components of soil enzymatic activities participating Agron., 27: 25-91.in and assuring the correct sequence of all the 4. Alef, K., P. Nannipieri and C. Trazar-Cepada, 1995.biochemical routes in soil biogeochemical cycles. Phosphatase activity. In Methods in applied soilIt has also found that measurement of changes in soil microbiology and biochemistry. Ed K. Alef and P.enzyme activities may provide a useful index of Nannipieri. Academic Press, London, pp: 335-344.changes in soil quality. 5. Dick, R.P., D. Breakwill and R. Turco, 1996. SoilDehydrogenase enzymatic activities is measured by enzymes activities and biodiversity measurementstwo methods, namely use TTC and INT as substrate, as integrating biological indicators. In Handbookhowever, various reported the poor results of DHA of methods for assessment of soil quality. Ed J.Wwhen TTC is used as substrate. Doran and A.J. Jones. Soil Sci. Soc. Am., Madison,Several biotic and abiotic factors affected soil pp: 247-272.dehydrogenase activity such as the incubation time 6. Lenhard, G., 1956. The dehydrogenase activity in soiland temperature, soil aeration status and soil as a measure of the activity of soil microorganisms. Z.moisture content and is also often used as a measure Pflanzenernaehr. Dueng. Bodenkd., 73: 1-11.of any disruption caused by pesticides, trace 7. Casida, L.E., Jr., D.A. Klein and T. Santoro, 1964. Soilelements or management. dehydrogenase activity. Soil Sci., 98: 371-376.Several workers have used incubation periods of 24 8. Von Mersi, W. and F. Schinner, 1991. An improvedhrs or longer at 37°C to study DHA. Some and accurate method for determining theresearchers modified with either glucose or yeast dehydrogenase activity of soils withextract as the electron donating substrate. Pre- iodonitrotetrazolium chloride. Biol. Fertil. Soils,incubation of dry soil with moisture, also caused an 11: 216-220.

The dehydrogenase activities in the rhizosphere soils

2. Dick, R.P., 1997. Soil enzyme activities as integrative

BM, Gupta VVSR (eds) Biological indicators of soil


905

9. Ladd, J.N., 1985. Soil enzymes. In: Soil Organic 23. Stevenson, L.L., 1956. Some observations on theMatter and Biological Activity. Nijhoff, Dordecht. microbial activity in remoistened air-dried soils. Plant

10. Visser, S. and D. Parkinson, 1992. Soil biological Soil., 8: 170-182.criteria as indicators of soil quality: Soil 24. Brzezinska1, M., Z. St pniewska and W.microorganisms. American Journal of Alternative St pniewski, 2001. Dehydrogenase and CatalaseAgriculture, 7: 3-37. Activity of Soil Irrigated with Municipal Wastewater.

11. Waksman, S.A., 1992. Microbiological analysis of soil Polish Journal of Environmental Studies,as an index of soil fertility. III. Influence of 10(5): 307-311.fertilization upon numbers of microorganisms in soil. 25. Marzadori, C., C. Ciavatta, D. Montecchio andSoil Sci., 14: 321-346. C. Gessa, 1969. Effect of lead pollution on different

12. Tabatabai, M.A., 1982. Soil enzymes. In: Page, A.L., soil enzymes activities. Biology Fertility Soils,Miller, R.H., Keeney, D.R. (Eds.), Methods of Soil 22: 235-241.Analysis. Part 2. Chemical and Microbiological 26. Marzadori, C., C. Ciavatta, C. MontecchioProperties. Agronomy No. 9. ASA and SSSA, and C. Gessa, 1996. Effects of lead pollution onMadison, WI, pp: 903-948. different soil enzyme activities. Biol. Fertil. Soil,

13. Curl and J. Sandberg, 1961. The measurement of 22: 53-58.dehydrogenase activity in marine organisms. Journal 27. Mersi, von, W. and F. Schinner, 1991. An improvedof Marine Research, 19: 1234-1238. and accurate method for determining the

14. Trevors, J.T., C.I. Mayfield and W.E. Inniss, 1982. dehydrogenase activity of soils withMeasurement of electron transport system (ETS) iodonitrotetrazolium chloride. Biol. Fertil. Soils,activity in soil. Microbial Ecology, 8: 163-168. 11: 216-22.

15. Benefield, C.B., P.J.A. Howard and D.M. 28. Quilchano, C. and T. Marañón, 2002. DehydrogenaseHoward, 1977. The estimation of dehydrogenase activity in Mediterranean forest soils. Biol Fertil Soils,activity in soil. Soil Biol Biochem., 6: 67-70. 35: 102-107.

16. Griffiths, B.S., 1989. Improved extraction of 29. Görres, J.H., M.J. Dichiaro, J.B. Lyons andiodonitrotetrazolium-formazan from soil with J.A. Amador, 1998. Spatial and temporal patterns ofdimethylformamide. Soil Biol. Biochem., soil biological activity in a forest and an old field. Soil21: 179-180. Biol Biochem., 30: 219-230.

17. Von Mersi, W. and F. Schinner, 1991. An improved 30. Banerjee, M.R., D.L. Burton, W.P.P. McCaugheryand accurate method for determining the and C.A. Grant, 2000. Influence of pasturedehydrogenase activity of soils with management on soil biological quality. J. Range.iodonitrotetrazolium chloride. Biol. Fertil. Soils, Manage., 53: 127-133.11: 216-220. 31. Pascual, J.A., C. Garcia, T. Hernandez,

18. Trevors, J.T., 1984. Dehydrogenase activity in J.L. Moreno and M. Ros, 2000. Soil microbialsoil: A comparison between the INT and TTC activity as a biomarker of degradation andassay, Soil Biology & Biochemistry, 16: 673-674. remediation processes. Soil Biology & Biochemistry,

19. Klein, D.A., T.C. Loh and R.L. Goulding, 1971. A 32: 1877-1883.rapid procedure to evaluate dehydrogenase activity 32. Simek, M., S. Virtanena, V. Kristufekb,of soils low in organic matter, Soil Biology & A. Simojoki and M. Yli-Halla, 2011. Evidence ofBiochemistry, 3: 385-387. rich microbial communities in the subsoil of a

20. Skujins, J., 1973. Dehydrogenase: an indicator of boreal acid sulphate soil conducive to greenhousebiological activities in arid soils. Bull. Ecol. Res., gas emissions. Agriculture, Ecosystems andCommun [Stockh] 17: 235-241. Environment, 140: 113-122.

21. Casida, L.E., 1977. Microbial Metabolic activity in 33. Li. Xiangzhen, and P. Saraha, 2003. Enzyme activitiessoil as measured by dehydrogenase determinations. along a climatic transect in the Judean Desert. Catena, Appl. Environ. Microbial., 34: 630-636. 53: 349-363.

22. Ross, D.J., 1971. Some factors influencing the 34. Bonanomi, et al., 2011. Assessing soil quality underestimation of dehydrogenase activities of some soils intensive cultivation and tree orchards in Southernunder pasture. Soil Biol. Biochem., 3: 97-110. Italy. Applied Soil Ecology, 47: 184-194.


906

35. Acosta-Martinez, V., D. Acosta-Mercado, 43. Chodak, M. and M. Niklinska, 2010. Effect of textureD. Sotomayor-Ramirez and L. CruzRodriguez, 2008. and tree species on microbial properties of mine soils.Microbial communities and enzymatic activities under Applied Soil Ecology, 46: 268-275.different management in semiarid soils. Appl. Soil 44. Hinojosa, et al., 2008. Effects of pyrite sludgeEcol., 38: 249-260. pollution on soil enzyme activities: Ecological dose-

36. Xiang Shu-Rong A. Doyle, P.A. Holden and response model. Science o f the total Environment,J.P. Schimel, 2008. Drying and rewetting effects on C 396: 89-99.and N mineralization and microbial activity in 45. McCarthy, G.W., R. Siddaramappa, R.J. Reight,surface and subsurface California grassland soils. E.E. Coddling and G. Gao, 1994. Evaluation of coalSoil Biology & Biochemistry, 40: 2281-2289. combustion by products as soil liming materials: their

37. Paradelo, R. and Mar a T. Barral, 2009. Effect of influence on soil pH and enzyme activities. Biol. Fert.moisture and disaggregation on the microbial activity Soils, 17: 167-172.of soil. Soil & Tillage Research, 104: 317-319. 46. Pitchel, J.R. and J.M. Hayes, 1990. Influence of fly

38. Moreno-de las Heras, M., 2009. Development of soil ash on soil microbial activity and populations. J.physical structure and biological functionality in Environ. Qual., 19: 593-597.mining spoils affected by soil erosion in a 47. Baruah, M. and R.R. Mishra, 1986. Effect ofMediterranean-Continental environment. Geoderma, herbicides butachlor, 2,4-d and oxyfluorfen on149: 249-256. enzyme activities and CO2 evolution in submerged

39. Sinha, et al., 2009. Rhizosphere soil microbial index of paddy field soil. Plant Soil, 96: 287-291.tree species in a coal mining ecosystem. Soil Biology 48. Leirós, M.C., C. Trasar-Cepeda, S. Seoane and F. Gil-& Biochemistry, 41: 1824-1832. Sotres, 2000. Biochemical properties of acid soils

40. Rao, A.V. and R. Tak, 2001. Influence of mycorrhizal under climax vegetation (Atlantic oakwood) in anfungi on the growth of different tree species and area of the European temperate-humid zone (Galicia,their nutrient uptake in gypsum mine spoil in India. NW Spain): general parameters. Soil BiolApplied Soil Ecology, 17: 279-284. Biochem., 32: 733-745.

41. Rao, A.V. and J.C. Tarafdar, 1993. Role of VAM fungi 49. Baruah, M. and R.R. Mishra, 1986. Effect ofin nutrient uptake and growth of clusterbean in an herbicides butachlor, 2, 4-d and oxyfluorfen onarid soil. Arid Soil Research and Rehabilitation, enzyme activities and CO 2 evolution in submerged7: 275-280. paddy field soil. Plant Soil, 96: 287-291.

42. Rao, A.V. and J.C. Tarafdar, 1998. Significance ofmicrorganisms in afforestation programs. Ann. AridZone, 37: 337-346.

soil dehydrogenase enzyme activity in natural and mine soil - a review

Documents