Copyright © 2022 University of Bucharest Rom Biotechnol Lett.

2022; 27(1): 3309-3314

Printed in Romania. All rights reserved doi: 10.25083/rbl/27.1/3309-3314
ISSN print: 1224-5984
ISSN online: 2248-3942

Received for publication, December, 01, 2019

Accepted, January, 28, 2022

Original paper

The assessment of the soil mineralization processes along

the traffic corridors from urban and rural areas in Romania

I. PĂCEŞILĂ1,2, E. RADU1, C. C. BÎRSAN1, M. CONSTANTIN1,2

1
Institute of Biology Bucharest, Spl. Independentei, 296, 060031, Bucharest, Romania
2
The Research Institute of the University of Bucharest, ICUB, Bucharest, Romania

Abstract The soils of the green areas located near the traffic corridors accumulate different pollutants,
especially heavy metals, which have an important impact on the microbial decomposition
processes. The extracellular enzymes can be used as an indicator of the metabolic activity of
the microorganism communities. Four extracellular enzymes - α and β glucosidase, alkaline
phosphatase and alanine aminopeptidase - were assessed in green areas close to the traffic
zones in several urban, peri-urban and rural localities in Romania. The data showed that the
highest values of enzymatic activities were registered in rural areas. At the same time, the
spatial dynamics of the studied enzymes revealed an important variability between the three
types of sampled areas, without a clear pattern of their distribution.

Keywords soil mineralisation, extracellular enzymatic activity

To cite this article: PĂCEŞILĂ I, RADU E, BÎRSAN CC, CONSTANTIN M. The assessment of the
soil mineralization processes along the traffic corridors from urban and rural areas in Romania. Rom
Biotechnol Lett. 2022; 27(1): 3309-3314. DOI: 10.25083/rbl/27.1/3309-3314.

*Corresponding author: I. PĂCEŞILĂ, Institute of Biology Bucharest, Spl. Independentei, 296, 060031, Bucharest,
Romania.
E-mail: [email protected]
I. PĂCEŞILĂ et al

human population, as well as for the plant and animal

communities.
Introduction The purpose of the present study is to analyze and
highlight the dynamics of the extracellular enzyme
The extracellular enzymatic activity it is well intensity in vegetated soils from territories along different
acknowledged to play an important role in the traffic corridors in urban, peri-urban and rural localities in
mineralization of the detrital organic matter (DOM) and in Romania.
the nutrient recycling process in soils, contributing to the
ecosystems functioning. The extracellular enzymes are
released in soils by numerous microorganisms (especially
by bacteria and fungi), but also by the roots of some plants.
Materials and methods
They hydrolyze the large organic polymeric molecules (a The samples were taken from the first 5 cm below the
major component of DOM) to simpler molecules that can surface layer of the soil, between April and May 2016, from
be used by microorganisms or plants as a source of several urban and rural localities in the NE, E and SE part
nutrients and energy. Due to their action, these enzymes are of Romania. All the 60 sampled points were placed in
involved in the main biogeochemical cycles of the biogenic central or peripheral areas of the localities, in the proximity
elements like C, N, S, etc. Thus, the evaluation of the of trafficked roads, in parks or in the green areas of the
extracellular enzymes activity could conduct to a better blocks of flats. The sample size in different counties
understanding of the DOM composition, a major followed the density of the air quality monitoring stations
component in the soil formation. The quantitative and in the chosen locations, according to the National Network
qualitative alterations of DOM influence the composition of the Air Quality Monitoring in Romania (www.calitateaer.ro
and distribution of microorganism communities and, [9])), in order to asses possible pressures induced by the
therefore, the specific of the biochemical processes carried airborne pollution across the heterogeneity of our study
out at the soil level - processes of a great importance for area (ŞTEFĂNUŢ & al. [10]).
defining the characteristics of different types of soil
(IGALAVITHANA & al. [1]; DATTA & al. [2]; ZAHIR &
al. [3]; SHERENE [4]).
The intensity of the enzymatic activity in soils varies
with the pressure of the environmental - physical, chemical
and biological factors, of natural or anthropogenic origin.
In the urban and rural inhabited areas, the presence of
increasing quantities of heavy metals in soils (Ni, Cd, Cu
etc.) is often recognized as a major factor that influences
the activity of the extracellular enzymes. Another factor
that can impact the decomposition processes in soil is
represented by the pollution with fertilizers (especially of
organic nature) used in the agricultural treatments. Most
frequently, these fertilizers stimulate the enzymatic activity
in soil and, thus, modify the rates of the biogeochemical
processes at this level (KIM [5]; PETRISOR & LAZAR
[6]; IGALAVITHANA et al. [1]).
The composition of the vegetation that covers the soils
also influences the extracellular enzymatic processes in
respect to the quantity and quality of the detrital organic
matter produced. The enzymatic activity in soil is more
intense in its upper layer, often related to a higher microbial
biomass and detrital organic matter amount of this
substrate. In the rhizosphere, the intense activity of many
enzymes is sustained by a higher amount of the cellular
debris and root excretes released in the environment (DICK
& KANDELER [7]).
Regarding the urban and suburban areas, the soil Figure 1. The study area and the sampling stations
undergoes different biochemical changes due to
anthropogenic activities: agriculture, forestry, industrial The enzymatic activity was assessed using the substrate
activities, infrastructure and support activities etc. consumption method (OBST [11]). Four enzymes involved
(MOREL & al. [8]). Thus, the urban soils are characterized in soil mineralization processes (BANERJE et al. [12];
by a higher heterogeneity and are frequently contaminated ZHANG et al. [13]; NIEMI et al. [14]) have been
with important quantities of pollutants such as heavy metals investigated: α glucosidase (α GLC) and β glucosidase
and hydrocarbons, which represent a real thread for the (GLC) associated with the C cycle, the alkaline

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 The assessment of the soil mineralization processes along the traffic corridors

phosphatase (AP) involved in the P cycle and the alanine The data suggest the presence of some less favorable
aminopeptidase (AMP) active in the N cycle (Tab. 1). The conditions for the MOD mineralization processes in the
intensity of the enzymatic activity was expressed in units of urban, highly trafficked areas. The accumulation of the
nmol substrate per gram per hour or per day (24 h). All the polluting substances, especially heavy metals from the traffic
processed data used in the cluster multivariate analysis emissions, can induce a significant impact in the
were log transformed. environment on the microorganism communities, mainly
The statistical analyzes were performed using the PAST decreasing their extracellular enzymatic activity (KHAN et
program ver. 2.17c (HAMMER & al. [15]) and following al. [16]). However, this was not a tendency for all the
PAST Reference manual methodology. analyzed sites; many of them in the urban area presented a
higher extracellular enzymatic activity than in the rural areas.
Table 1. The enzymatic activities assessed in the urban, The dynamic of mineralization processes is influenced by
peri-urban and rural localities from the NE, E and SE part many local factors, but the quantity and quality of the
of Romania substrate are playing a major role (BURNS [17] DICK &
KANDELER [7]). In our study, we aimed to sample a high
Enzymatic Measurement
Substrate variability of soil quality across the selected locations. On the
activity unit (nmol g-1h-1)
α Glucosidase p - nitrophenyl - α - p-nitrophenol
other hand, some categories of microorganisms are known to
(EC 3.2.1.20) D - glucopyranoside develop different mechanisms of resistance to the heavy
β Glucosidase p - nitrophenyl - β - p-nitrophenol metals stress (AZARBAD & al [18], HOOSTAL & al. [19]).
(EC 3.2.1.21) D - glucopyranoside As well, the rate of absorption of the heavy metals by the
Alkaline 4 - nitrophenyl - p-nitrophenol vegetal layer modulate their impact on soils (TANGAHU et
phosphatase phosphate al. [20]), and subsequently on the bacterial extracellular
(EC 3.1.3.1) enzymatic activity.
Alanine L - alanine - 4 - p-nitroanilline α GLC, registered the most intense activity in an urban
aminopeptidase nitroanilide - park from Bucharest (751 nmol p-nitrophenol g-1d-1, Titan
(EC 3.2.1) hydrochloride Park) and the lowest in a different type of urban site, in Iași,
when we sampled the green area located in the proximity of
Table 2. The intensity ranges of the assessed enzymatic the trafficked N. Iorga street (12 nmol p-nitrofenol g-1d-1).
activities across the study sites β GLC activity was the most intense in Bosia village
from Iași county (2031 nmol p-nitrophenol g-1d-1), at a
Enzymatic activity nmol substrate g-1d-1
sampling site placed near the main road, while its lowest
α Glucosidase (α GLC) 469,5 ± 457,5 value was assessed in an urban location, in Brăila - Brăila
β Glucosidase (β GLC) 1033,5 ± 997,5 county (32 nmol p-nitrophenol g-1d-1), at the green area
Alkaline phosphatase (ALP) 1100 ± 1037 close to the central Independenţei Square.
Alanine aminopeptidase 1574 511 AP phosphatase enzymatic activity varied between
(AMP) 2137 nmol p-nitrophenol g-1d-1 in Măgurele - Ilfov county
(at a station located in the garden of a block of flats located
to the outskirts of the city) and 63 nmol p-nitrophenol g-1d-
Results and discussion 1
, in Botoşani - Botoşani county (for a sample taken as well
from the garden of a block of flats, from M. Eminescu
By analyzing the spatial distribution of the
street).
extracellular enzymatic activity in the considered study
AMP registered its highest value in Iași (3085
areas, the highest intensity was mainly recorded for alanine
p-nitrofenol g-1d-1), in the rural area from the outskirts of
aminopeptidase and the lowest, for α-amylase, with large
the city, near M. Sadoveanu alley. Its minimum intensity
variations for each enzyme (Tab.2).
(63 nmol p-nitrophenol g-1d-1) was assessed at the same
In the rural areas, the enzymatic activity often
station where AP recorded its minimum intensity, namely
appeared to be higher than in the urban areas (Fig. 2). The
in Botoşani, in the garden of a block of flats.
exception was observed for AMP that registered a higher
In addition, because the intensity of soil
average intensity for the peri-urban area.
level enzymatic processes can be influenced by
soil physico-chemical characteristics
(BŁOŃSKA et al [21]), it is also important to
consider the types of soil identified in the
studied areas. According to the data from the
literature (STANILA & DUMITRU [22]) the ones
from the category chernisols (predominantly
chernozem), cambisols (predominantly
distriambosols) and luvisols (predominantly
luvosols) are prevalent. From table 3 it can be
Figure 2. The average activity of the extracellular enzymes
observed that, in general, α and β GLC were
studied in the urban, periurban and rural areas
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I. PĂCEŞILĂ et al

more intense in the stations located in regions enzymatic activity across the sampling sites of
where chernisols are dominant, while AP and ALP different soil types did not show a high
recorded higher values in the regions where variation, suggesting that, most likely, the
cambisols predominate. In general, chernisols metabolic activity of the heterotrophic
are recognized to have a high content of organic microorganism communities involved in the
matter (VYSLOUŽILOVÁ et al [23]), but the geochemical circuits at C, N and P, was not
concentration and quality of organic matter may significantly influenced by the type of soil
widely vary in the case of all soil types, present in the sampling area.
depending on the specificity of the area .
(ZARNEA [24]). However, the extracellular

Table 3. The intensity of enzymatic activity assessed on different soil types

Enzymatic activity average value
(nmol substrate h-1 mL-1) α GLC β GLC ALP AMP
chernisols 215,16 284,67 355,16 641,25
cambisols 206,53 269,61 351,87 646,46
luvisols 205,98 277,95 351,62 623,12

For a better understanding of the spatial variability of (u – urban, p – peri-urban, r – rural) (Ward’s method, Coph.
each bacterial extracellular enzymatic activity in soil across Corr.: 0,696)
the study zones - urban, peri-urban, rural areas, we further
performed the cluster analysis following Ward’s method APL and α GLC enzymatic activity are the most similar
(Fig. 3). in rural and peri-urban locations. Instead, the variability of
Regarding the intensity of the substrate consumption β GLC enzymatic activities was the lowest between urban
performed by the four studied enzymes, we observed a and peri-urban areas, and AMP, between the urban and
similarity between α GLC and β GLC activities (Fig. 3). rural sites.
This could be the result of the amount of plant debris in the The differences between the intensity of the
analyzed soils, the two enzymes being involved in the extracellular enzymatic activities evaluated at different
degradation of the vegetal glucidic substrates such as locations could be explained by the variation of the
cellulose and starch. The activities of these two enzymes pressures exerted by the natural or anthropic environmental
also showed the smallest spatial variability in the three factors (KIM [4], DICK & KANDELER [8]).
types of studied areas. At the same time, we should mention The extracellular enzymes are considered to be very
that each enzyme revealed a specific dynamic per site type. sensitive to changes in the ecosystems state, including
pollution. Consequently, they can be used as early
indicators of soil quality and health (LI & al. [25]) based on
their property of quick response to changes in the
environment, in comparison to other parameters (RAO &
al. [26]). Due to this property, the study of the extracellular
enzymatic activity of the bacterial communities can be used
to improve the understanding of the dynamic and role of
soils in the evolution of different types of ecosystems.

Conclusions
The intensity of the assessed enzymatic activities
registered important variations across the sampling stations
in the surveyed area, a fact that could be explained by
considering the high variability of soil quality between the
selected locations. The highest intensity values were
recorded in most of the cases for the alanine
aminopeptidase and the lowest for the α-glucosidase
Figure 3. The cluster analysis of similarity of the enzymatic enzymatic activity. Generally, considering the entire data
activities evaluated across the tree types of sampled locations set, the enzymes analyzed from the rural soil substrate often
showed a higher intensity in comparison with the ones
analyzed from the urban areas samples. The pollution from

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 The assessment of the soil mineralization processes along the traffic corridors

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