THE RELATIONSHIP BETWEEN THE NITRATES IN

IRRIGATION WATERS AND THEIR ACCUMULATION IN

LEGUMES FROM TWO BUZĂU AREAS

CIOFU ALEXANDRU I., APOSTOL MARIUS,LILIANA TUDOREANU

University of Agricultural Science and Veterinary
Medicine, 59 Mărăşti blvd. 011464, Bucharest, Romania

Abstact The paper presents the results of two experiments with plants
(lettuce, carrots and onions) cultivated upstream and downstream of Buzau
River. The differences between the nitrates concentration in the irrigation water
used in the studied areas were significant (p<0.0001). The mean concentration
of nitrates in the irrigation water from Sageata was 5.51 ± 0.29 mg/l while in
Vernesti 3.16 ± 0.29 mg/l. The 95% conficence interval for the nitrates
concentration in the irrigation water was 4.93 – 6.01ppm for Sageata area and
2.58 – 3.74 ppm for Verneşti area.
Irrigation water quality, which was influence by industrial pollution
from Buzau town, has influenced the nitrates concentration of legumes cultivated
in private farms which was increased in the farms downstream of river Buzau
compared with the upstream farms. There is a simple linear correlation between
the NO3- concentration in plants and NO3 concentration in the irrigation water.
The coefficient of determination was r2adj = 0.79 for carrots, followed by r2adj
= 0.78 for lettuce, and r2adj = 0.70 for onions.

Along quality demands such as flow, efficiency and safety during

exploatation, the water suplies for irrigations need to meet the quality standards
for chemical composition, toxicity and many other parameters (2, 7, 8,).
Some of the possible negative influences of the irrigation water on plants
growth are the reduction of plant water availability due to water high salt
concentration, complexation/reduction of available nutrients and toxic effects due
to high concentration of pollutants. The irrigation water may be an important
vector for pathogens transmission and an important factor of qualitative and
quantitative modification for agroecosystems (4, 5, 6).
A major risk is repesented by the high nitrates and nitrites water
concentration that may accumulate in plants leading to the formation of
nitrosamines in the organisms. The European and national regulation are
stipulating a 50 mg NO3- /l (1, 7, 8) upper concentration limit for nitrates
concentration in surface waters used for irrigation. The same regulations mention
that the maximum addissible limits for legumes is 200 – 4500 ppm according to
species, time of cultivation and harvest time.
Nitrates are considered an important parameter in food safety and human healts. The
objective of our researche was to study the nitrate accumulation in legumes cultivated in
various conditions and the corelation of nitrate concentration in legumes to nitrates
concentration in soils and in the irrigation water.

679
 MATERIALS AND METHODS
The experiment took place in two areas of Buzau County where the irrigation
water is colected from Buzau River. These two areas are situated uptream of Buzau
town – Vernesti - and downstream of Buzau town - Sageata. Three farms were chose
in each area forming the 6 experimental sites where 3 legume species (table1) were
cultivates.

Table 1
The experimental protocol

Experiment Location Variants

The influence of the - Verneşti (upstrewm from Buzău town) 3 legume cultures :
irrigation water on x three farms (G1-G3) - lettuce - Mona
legumes cultivated in - Săgeata (downstream from Buzău - onion - Buzău
different areas and town) x three farms (G1-G3) - carrots -
farms from Buzau Nantes
county

The objective of this study was to establish the relationship between the quality
of the irrigation water (characterized by different nitrates and nitrites concentrations
befor and after crosing the Buzau town) and the level of nitrates concentration in the
soil and edable legume plant parts as well as the influence on the plants yield.
The experimental design was a field linear block design with four repetitions
with a total of 400 m2 for each specie, covering the area of 1200 m2 for the entire
experiment.
The environmental conditions for the experimental areas were:
- Mean temperature of 11.1 – 13.7 0 C; total precipitation of 539.4 – 372.2 mm;
relative air humidity 69.7 – 69.3 %.
- mollic-alluvial soils, clay-sandish texture, good permeability , pH 8.2, humus
2.57%, mean salts concentration under 0.05 %, low carbonated (4.50 % CaCO3), C :
N - 11, good nitrogen content (N total – 0.151 %) and very good Phosphorus
concentration (P total – 0.183 %; P mobile - > 144 ppm) and potassium (K mobile - >
268 ppm).
- According to the STAS 4706/88 and U.E regulations /1995, the quality
parameters for the Buzau river waters in the two tested sections had the following
characteristics:
upstream the Buzau town – quality category I, 73-75% degree of purity saprob
zone ß (light impurification);
downstream of Buzau town – general quality category II (inadequate values for
dissolved oxygen, mineralization, toxic substance concentrations) zona saprobă ß-α
(moderated pollution)
The agrotechniques used during the cultivation and vegetation period were
according to the specie needs. Fitosanitary treatments and specie adequate
fertilisation during vegetation with Complex III were applied as well as linear open
irrigation between plant beds in accordance with the studied cultures requirements.
Investigation methods and results interpretations
- The nitrate concentration was measured spectrofotometricaly (wavelength 410
nm and cuvette of 1cm)
- Soil nitrates - were measured using an ion-selective electrode for nitrate.
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 - The concentration of nitrates in plants from variant mean plant samples was
measured colorimetrically in water extract using fenodisulfonic acid.
- The yield was calculated by weighting the repetitions and variants.
- The ANOVA, Dunnet and Student tests as appropriate were used for the
statistical data analyses. The significance of correlation and regressions were
interpreted too. The data were statistically analysed using JMP 5.0 software.

RESULTS AND DISCUSSIONS

The results form the two experimental years (table 2) show the a significant
difference between the plants cultivation areas
Table 2
Results concerning the influence of water quality on some legume cultures

Water Soil Plant

(mg/l NO3-) (ppm NO3-) (ppm NO3-)
Area Farm Specie
2001 2002 2001 2002 2001 2002

Lettuce 269 486

G1 Onion 2.82 3.88 16.25 24.38 150 233
Carrot 148 156
Lettuce 254 470
G2 Onion 1.38 3.52 15.47 21.96 171 315
Carrot 123 128
Vernesti

Lettuce 407 483

G3 Onion 3.09 4.27 19.09 29.37 250 324
Carrot 194 247
G4 Lettuce 377 716
Onion 4.22 6.48 55.30 81.06 346 362
Carrot 243 379
G5 Lettuce 342 608
Onion 3.46 5.35 30.41 74.88 286 356
Carrot 295 250
Săgeata

G6 Lettuce 453 864

Onion 5.95 7.63 69.23 102.0 294 556
Carrot 348 640

In the three farms from Vernesti, during the vegetation period of 2001, the average
nitrates concentration in the irrigation water was between 1.38 and 3.09 mg/l, compared
with the average values of 3.46-5.95 mg/l from Sageata which is situated downstream the
Buzau town. There were also recorded differences between the soil Nitrate concentration
between the two areas (15.47-19.09 ppm compared with 30.41-69.23 ppm).
Thus the plants accumulated nitrates in concentrations which were proportional with
the NO3- concentration in the irrigation water. Although the NO3- concentration in the water
and soil from the 6th farm compared with the 2nd farm increased by 4.3 times in water and
4.5 times in soil, the concentrations of NO3- in plants increased only by 1.8 times for lettuce,
by 1.7 times for onion and by 2.8 times for carrots. It can be highlighted the high sensitivity
of the carrots plants compared with the other studied species.
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 Comparing with the previous year, in 2002, the nitrate concentration in the
irrigation waters increased during the vegetation period induceing an increase of
nitrate content in soil and plants.
In Vernesti village the nitrates concentration varied between 3.53 mg/l and 4.27
mg/l in the irrigation water, and between 21.96 – 29.37 ppm in the soil with the highest
values registered in the G3 farm. The irrigation water (pumped from the Buzau river
after the river collected the residual waters from the Buzau town) used in the Sageata
area had a nitrate concentration 1.5 – 1.8 times higher. The concentration in soil
increased too by 3.4 – 3.5 times, the highest values being recorded in the G6 farm.
In these condition the nitrates concentration in plants increased
dramatically: up to 556 ppm for onions, 864 ppm for lettuce and 640 ppm for
carrots cultivated in the G6 farm for which the MAL vas exceeded (400ppm),
showing once again the high sensitivity of this specie to nitrate pollution.
Comparing the results, for the nitrate concentrations in the irrigation water,
and plants over the two years, it can be observed the cumulative pollution effect
of the irrigation water for the 6 farms in the Buzau area.
All values recorded during 2002 were increased compared with the
previous year. Moreover in the Sageata area, which is located downstream from
the Buzau town, the reported NO3 concentrations were higher, compared with the
values from Vernesti area.
8
NO3 CONCENTRATION IN WATER (mg/l)

6
Means comparison
5 Student’s test
4
Village Mean
3
SAGEATA A 5.515
2
VERNESTI B 3.160
1
SAGEATA VERNESTI Each Pair
Student's t
Levels not connected by
0.05
same letter are significantly
Fig.1. Comparison of NO3- (mg/l) concentration in water; different
F= 33.8943; p= 0.0001 (ANOVA)

The statistical analyses (using the JMP software) concerning the irrigation
waters nitrates concentrations for the three legume species in the 6 farms from the
2 areas are presented in figure 1. There is a significant difference (p < 0.0001)
between the two areas.
The mean value for the nitrates concentration in the irrigation water was of
5.51 ± 0.29 mg/l in Sageata area while for the Vernesti area the mean value was
3.16 ± 0.29 mg/l, due to a 1.74 time increase of nitrates in the Buzau river after
flowing through Buzau town.
The 95% confidence interval for the nitrates concentration in the irrigation
water was 4.93 – 6.01ppm NO3 for Sageata compared with 2.58 – 3.74 ppm for
Verneşti, having specific influences over the cultures.
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 Figure 2 presents representation of the statistical data analysis for nitrate
concentration in the cultures’ soil, which were irrigated with water having
different nitrate concentrations.
There is a significant difference (p < 0.0001) between the nitrates concentrations in
the soil cultivated with lettuce, onion and carrot plants in the 6 farms
110
100
NO3 Concentration IN SOIL (ppm)

90
80
70
Means comparison
60
Student’s test
50
40
30
Localitate Media
20 SAGEATA A 68.813
10 VERNESTI B 21.086
SAGEATA VERNESTI Each Pair
Student's t
For 2001 and 2002 0.05 Levels not connected by same
- letter are significantly different
Fig. 2. Comparison of NO3 (ppm) concentrations in
soil (F= 75. 3152 p= 0.0001 (ANOVA)

The 95% confidence for nitrates concentration was of 60.91 – 76.71 ppm NO3 in
Sageata compared to 13.18 – 28.99 ppmNO3 inVerneşti.
The mean comparison shows that in the Sageata village’s soils, the nitrates
concentration was of 68.81± 3.89 ppm while in the Vernesti village the nitrates
concentration in soil was of 21.09 ± 3.89ppm. The 3.26 times increase of the nitrate
concentration in the soil of the downstream village shows the great influence of nitrate
concentration increase in the irrigation water.

600
NO3 CONCENTRATION IN ONIONS (ppm)

500

400
Means comparison Student
300 test

200 Level Mean

SAGEATA A 366.666
100
SAGEATA VERNESTI Each Pair
VERNESTI B 240.500
Student's t
For 2001 and 2002 0.05
Levels not connected by
Fig. 3. Comparison of the concentrations of same letter are significantly
-
NO3 (ppm) in onions; F= 6.4631; p= 0.0292 (ANOVA) different

There are very significant differences between the experimental results (fig.1 and fig. 2).

683
 These figures show the pollutant effect of industrial waters spilled in the
Buzau river on the agricultural soils. These influences are reflected by the results
obtained for the nitrates concentration from the legumes grown in the above
mentioned conditions. The ANOVA results for are presented in figure 3 – 5.
700
NO3 CONCENTRATION IN CARROTS (ppm)
600

500
Means comparison
400 Student test
300
Area Mean
200
SAGEATA A 359.166
100
Each Pair
SAGEATA VERNESTI
Student's t
VERNESTI B 166.000
For 2001 and 2002 0.05
Levels not connected by
Fig. 4. Comparison of concentrations of NO3- in same letter are significantly
carrots (ppm); F= 9.3301 p= 0.0122 (ANOVA ) different

The difference for the concentration of nitrates in onion bulbs (fig. 3) are
significant (p = 0.0292). The same response was observed for carrot plants for
which the nitrates concentration in roots harvested in Sageata are higher than in
Vernesti (fig. 4). The p (0.0122) value and the graphical representation from fig 3
and 4 show a higher influence of the cultivation environment on the carrot plants
compared to the onion plants.

1000
Continutul in NO3 din plantele de salata (ppm)

800
Means comparison
Student test
600

Level Mean
400
SAGEATA A 560.000
200 VERNESTI A 394.833
SAGEATA VERNESTI Each Pair
Student's t Levels not connected by
LOCALITATEA – cumulat 2001 şi 2002 0.05 same letter are
Fig. 5. Concentration of NO3 in lettuce plants; significantly different
F= 3. 0416 p= 0. 1118 (ANOVA)

For the lettuce plants (fig. 5), the p = 0.1118 value and the graphical
representation of the experimental results shows that there are no significant
diference in nitrates accumulation in the rosette due to nitrates from the irrigation
water and soils in the two studied areas
Linear simple correlations were obtained for NO3- concentration in plants
and NO3- in the irrigation water for all studied legumes (fig. 6 - 8).

684
 600
The regression equation is:

NO3 CONCENTRATION IN ONIONS (ppm)

CONC. NO3 IN ONIONS (ppm) =
500 -63.82963 + 75.253709 CONC.NO3 IN
WATER (mg/l)
400
2
R 0.813
2
300
R Adj 0.795
Observations (or 12
Sum Wgts)
200
Parameter Estimates
Term Estimat Er. Std t p|
100 e
0 2 4 6 8
Intercept -63.829 52.887 - <.0001
NO3 CONCENTRATION IN WATER (mg/l) 1.21
NO3 75.253 11.381 6.61
CONC. IN
-
Fig.6. The correlation between NO3 WATER
concentration in onions and the NO3- (mg/l)
concentration in the irrigation water
(p<0.0004)

The coefficient of determination is different for each specie showing the affinity
of each specie for nitrates accumulation from the irrigation water. Thus the most
sensitive are the carrot plants (r2adj = 0.79), followed by the lettuce plants (r2adj =
0.78), and the least sensitive being the onion plants (r2adj = 0.70).

The regression equation is:

600 CONC NO3 IN CARROTS (ppm)=
-63.82963 + 75.253709 CONC NO3 IN
NO3 CONC. IN CARROTS (ppm)

500 WATER (mg/l)

400
R2 0.813
300 R2 Adj 0.795
200 Observations 12

100
Parameters estimate
Term Estimate Er. Std t p
2 4 6 8 Intercept -63.829 52.887 -1.21 <.0001
NO3 CONC. IN WATER (mg/l) CONC. 75.253 11.381 6.61
NO3 IN
Fig.7. Correlation between the NO3- WATER
concentration in carrots and the NO3- (mg/l)
concentration in the irrigation water (p<0.0001)

Linear correlations were also obtained for the NO3- concentration in plants and
soil (fig.9 -11).

685
 1000

NO3 CONCENTRATION IN LETTUCE (ppm)

The regression equation is:
NO3 COC.IN LETUCE (ppm) = 77.33 +
800 92.23 NO3 CONC IN WATER (mg/l)

600 R2 0.808
R2 Adj 0.788
400 Nr de 12
observaţii
Parameter Estimates
200
Term Estimate Er. Std t p
0 2 4 6 8
NO3 CONCENTRATION IN WATER (mg/l)
Intercept 77.336 66.060 1.17 <.0001
Fig. 8. Corelation between the NO3- NO3 CONC.IN 92.237 14.216 6.49
concentration from lettuce plants and the NO3- WATER (mg/l)
concentration in irrigation waters (p<0.0001)

600
NO3 CONCENTRATION IN ONIONS (ppm)

500
The regression equation is:
400 CONC. NO3 IN ONIONS (ppm) = 171.63906 +
2. 935 C ONC NO3 IN SOIL (ppm)
300
2
R 0.702
2
200 R Adj 0.672
Observations 12
100
0 25 50 75 100 125
Parameter Estimates
NO3 CONCENTRATION IN SOIL (ppm)
Term Estimate Std Er t
Fig.9. Corelation between NO3- Intercept 171.639 32.226 5.33 0.000
concentration in onions and the CONC.OF 2.935 0.603 4.86
- NO3 IN
NO3 concentration
in soil (p<0.0007) SOIL (ppm)

The strongest correlation was obtained for carrot plants (r2adj= 0.80), followed by
the correlation for onion plants (r2adj = 0.67) and the weakest correlation being
for the lettuce plants (r2adj = 0.66)

686
 700
The regression equation is:
600 NO3 CONC. IN CARROTS (ppm) =

NO3 CONC. IN CARROTS (ppm)

65.595438 + 4.3823781 CONC. NO3 IN
500
SOIL (ppm)
400
2
R 0.821
300 2
R Adj 0.803
200 Nr de observaţii 12

100 Parameter estimate

0 25 50 75 100 125 Term Estimate Er. Std t p|
NO3 CONC. IN SOIL (ppm) Intercept 65.595 34.485 1.90 0.086
NO3 CONC 4.382 0.646 6.78 <.0001
Fig. 10. Correlation between the NO3-
IN SOIL
concentration in carrots and the NO3- (ppm)
concentration in soil (p<0.0001)

1000
The regression equation is:
NO3 CONC. IN LETTUCE (ppm)

NO3 CONC. IN LETTUCE (ppm) = 254.39406

800 + 4.9615707 NO3 CONC. IN SOIL (ppm)
2
R 0.695
600 2
R Adj 0.665
Observations 12
400
Parameter Estimates
Term Estimate Er. Std t p
200
0 25 50 75 100 125 Intercept 254.394 55.354 4.60 0.0007
NO3 CONC. IN SOIL (ppm) NO3 CONC. 4.96157 1.037 4.78
IN SOIL
Fig.11. Correlation between NO3 (ppm)
concentration in lettuce plants and NO3
concentration in soil (p<0.0007)
AA A

CONCLUSION
- In the water- soil- plant relationship, water quality pays an important role,
in soil properties modification and plant growth processes, as well as for the yield
quality.
- Following the run of residual industrial waters in the Buzau river the nitrates
concentration had a very significant increase in the farms from downstream Buzau town
compared with farms situated upstream Buzau town, increasing by 1.5 -1.8 times in the
irrigation water and by 3.4 - 3.5 times in the soil.
- The pollution effect on irrigation water, soil and plants was cumulative;
all the values obtained during the second year being higher that in the first year.

687
 - The legumes had distinct responses to the nitrates concentration increase of the
irrigation water. A significant increase was observed for the carrot roots (2.8 times over
the MAL), and the onion bulbs (1.7 times), while for the lettuce plants the nitrate
concentration was not significantly influenced by the irrigation water.
- Linear correlations were obtained between NO3- concentration in the
irrigation water and nitrates concentration in plants.
- The strongest correlation between the analysed quality indexes are for
carrots, a sensitive specie to root nitrates accumulation influenced by increasing
nitrates concentration in irrigation water and soil. Onions are less sensitive to
increasing nitrates concentration in the irrigation water and the lettuce plants to
the nitrates increasing concentration in soils. Comparing the two species, onions
are less sensitive to the nitrate NO3- increase in water and the lettuce plants to the
increase of Nitrates concentration in soil.
- Analysing the results of this experiment and the coefficients of
determination it ca be concluded that, the irrigation water, due to its high nitrate
concentration, increases nitrates concentration in the soil and has a significant
influence on legume quality.
- The experimental results suggests that for reducing the risk of legumes
pollution with nitrates and for promoting sustainable agriculture the cultivation
area should be carefully chosen including an adequate choice of plant species and
testing the nitrates concentration in the irrigation water

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