International Journal of Environmental Health Research

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Kinetic study on biodegradation of glyphosate

with unacclimated activated sludge

Djaber Tazdaït, Rym Salah, Hocine Grib, Nadia Abdi & Nabil Mameri

To cite this article: Djaber Tazdaït, Rym Salah, Hocine Grib, Nadia Abdi & Nabil Mameri (2018):
Kinetic study on biodegradation of glyphosate with unacclimated activated sludge, International
Journal of Environmental Health Research, DOI: 10.1080/09603123.2018.1487043

To link to this article: https://doi.org/10.1080/09603123.2018.1487043

Published online: 22 Jun 2018.

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INTERNATIONAL JOURNAL OF ENVIRONMENTAL HEALTH RESEARCH
https://doi.org/10.1080/09603123.2018.1487043

ARTICLE

Kinetic study on biodegradation of glyphosate with

unacclimated activated sludge
Djaber Tazdaïta,b, Rym Salaha, Hocine Gribb, Nadia Abdib and Nabil Mamerib
a
Department of Biochemistry and Microbiology, Faculty of Biological and Agronomical Sciences, Mouloud
Mammeri University of Tizi-Ouzou, Tizi-Ouzou, Algeria; bLaboratory of Bioengineering and Process Engineering,
National Polytechnic School, Algiers, Algeria

ABSTRACT ARTICLE HISTORY

This article is concerned with the study of biodegradation of an organo- Received 20 March 2018
phosphorus herbicide (glyphosate) using unacclimated activated sludge. Accepted 6 June 2018
Glyphosate at different concentrations (0.1, 0.5, 1, 2 and 5 g/L) was tested KEYWORDS
for cellular growth. On the other hand, the effect of glyphosate on its Activated sludge;
own biodegradation was studied by evaluating the fittings of different biodegradation; glyphosate;
kinetic models (Andrews, Aiba et al., Han and Levenspiel, Luong, Tessier, inhibition; modelling
Webb, Tseng and Wayman, Yano and Koga). According to the obtained
results, the activated sludge was able to use glyphosate as the sole
carbon source; however, 2 and 5 g/L glyphosate seemed to inhibit
cellular growth. Moreover, glyphosate at initial concentrations of 0.1,
0.5 and 1 g/L was completely degraded within 4, 13 and 18 h of
incubation, respectively. Yano and Koga model was the best-fit model
(R2 = 0.999, F = 173,106 and P = 0.000006).

Introduction
Increasing demand pertaining to the pesticides has raised serious environmental concerns, which
should be considered more seriously by public opinion. Among the large number of marketed
pesticides, glyphosate is one of the most used in the world (Firth et al. 2007; Bento et al. 2017).
Glyphosate [N-(phosphonomethyl) glycine; CAS registry No 1071–83-6; C3H8NO5P] (Figure 1) is
commonly used as pesticide for weed control (Food and Agriculture Organization of the United
Nations (FAO) 2016), but it is toxic for human cells (Townsend et al. 2017) and animals (Bai and
Ogbourne 2016; Samanta et al. 2016), and its carcinogenicity potential is currently controversial
(Tarazona et al. 2017). Glyphosate is a systemic non-selective herbicide (Gomes et al. 2014), which
acts by inhibiting the activity of enolpyruvyl shikimate-3-phosphate synthase, an enzyme involved
in the aromatic amino acid biosynthesis pathway (Siehl et al. 1997; Schönbrunn et al. 2001; Duke
et al. 2012). Consequently, this effect impedes the biosynthesis of a number of essential metabo-
lites, and thus negatively impacts many vital plant physiological processes such as photosynthesis
(Gomes et al. 2014). Given the fact that glyphosate reaches aquatic media mainly via surface run-
off, it would therefore be appropriate to focus the investigations on its degradation processes in
such media. Biological process using microorganisms offers an important economic and societal
benefits, both because of its environment friendly character and because it consumes less energy
and fewer resources (Becker and Seagren 2010).

CONTACT Djaber Tazdaït djaber.tazdait@ummto.dz Department of Biochemistry and Microbiology, Faculty of

Biological and Agronomical Sciences, Mouloud Mammeri University of Tizi-Ouzou, P.O. Box 17 RP 15000 Hasnaoua, Tizi-Ouzou,
Algeria
© 2018 Informa UK Limited, trading as Taylor & Francis Group
2 D. TAZDAÏT ET AL.

Figure 1. Chemical structure of glyphosate.

Several studies have dealt with microbial degradation of glyphosate (Araújo et al. 2003; Dick
et al. 1995; Forlani et al. 1999; Bazot and Lebeau 2008; Shushkova et al. 2012; Kryuchkova et al.
2014). Most of these studies were performed using pure microbial culture, and a very few reports
are available on the use of mixed cultures such as activated sludge culture. On the other hand,
although glyphosate biodegradation has been extensively studied, information on its kinetic
degradation aspects and especially on the inhibitory effect of the herbicide on its own biodegrada-
tion remained scanty. This is particularly important to investigate since the assessment of
substrate inhibition of enzymatic reactions is becoming increasingly crucial in the treatment of
toxic compounds in general, and pesticides degradation in particular (Hao et al. 2002). The
evaluation of such inhibitory effect can be achieved by using mathematical modelling approach.
In fact, several mathematical models have been elaborated and successfully applied to assess the
inhibitory effect of hazardous substrates on their own biodegradation. These models have been
derived from those describing the inhibitory effect of substrates on enzymatic activities and
involving a common substrate inhibition parameter called Ki (Tziotzios et al. 2008).
To the extent of our knowledge, no single study has attempted to evaluate different kinetic
models of the inhibitory effect of glyphosate on its own biodegradation. The aim of this work was
to investigate the biodegradation of glyphosate by indigenous activated sludge and to determine
statistically the most relevant kinetic model for glyphosate degradation inhibition.

Materials and methods

Microorganisms and cultivation media
In this study, a mixed culture of activated sludge, obtained from the aeration basin of urban
wastewater treatment plant in Mascara (Algeria), was used to study the biodegradation of
glyphosate. Before use, the raw activated sludge was first washed with warm tap water (34°C)
three times to remove suspended and/or adsorbed organic matters (higher temperatures tend to
favour desorption phenomenon), then washed with distilled water, and finally grown in
Erlenmeyer flasks using the following mineral salt medium (MSM): KH2PO4 (0.038 g),
MgSO4 (0.05 g), CaCl2 (0.05 g), urea (0.2 g), 1 L of deionized water (Kargi and Konya 2007), in
the presence of different concentrations of glyphosate ranging from 0.1 to 5 g/L.
Technical grade glyphosate (95% purity, water solubility 15.7 g/L at 20°C and pH 7) was
purchased from Dow AgroSciences (France).

Glyphosate biodegradation
Biomass growth and glyphosate biodegradation kinetic studies
The cultures were conducted at room temperature (23°C) at 100 rpm in 2000 mL Erlenmeyer
flasks containing 1000 mL of MSM inoculated with activated sludge (final biomass concentration
of about 300 mg MLSS/L). Glyphosate was added to the cultures to the required concentrations
(0.1–5 g/L). The final pH of the media was adjusted to 7.0 by using sodium hydroxide and
hydrochloric acid solutions. Samples were removed (22 ml) at regular time intervals (1 h) during
 INTERNATIONAL JOURNAL OF ENVIRONMENTAL HEALTH RESEARCH 3

22 h and were checked for biomass and glyphosate concentrations. The determination of residual
glyphosate concentrations was carried out on supernatants obtained by allowing samples to settle
under static conditions.
Specific growth rate (μ) values were estimated using the following equation (Equation 1):
lnðX=X0 Þ ¼ μt (1)
where X0 and X are biomass values at times t0 and t, respectively.
The following equation (Equation 2) was used for the calculation of substrate degradation
rates, r (g/L h):
r ¼ dS=dt (2)
where S is the substrate concentrations in g/L at time t, in h.

Model equations used for glyphosate biodegradation

In this study, different empirical mathematical models were tested.
Due to its mathematical simplicity and relevance, Andrews model (Edwards 1970) is one of the
most widely used model for describing substrate inhibition kinetics. Its equation is as follows
(Equation 3):



r ¼ rmax  S= Ks þ S þ S2 =Ki (3)

where Ks, Ki, r, rmax and S are half saturation constant (g/L) (substrate-affinity constant),
inhibition constant (g/l), substrate consumption rate (g/L h), maximum substrate consumption
rate (g/L h) and substrate concentration (mg/L), respectively.
The model elaborated by Tessier (Edwards 1970) aimed to describe substrate inhibition at
higher substrate concentration (Equation 4):
 S 
r ¼ rmax  eKi  eKs
S
(4)

The model of Webb (Edwards 1970) (Equation 5) has been proposed to kinetically describe the
substrate inhibition by integrating an allosteric effect of enzymes with an empirical term β
(dimensionless).
     2 
S S
r ¼ rmax  S 1 þ β  = Ks þ S þ (5)
Ki Ki
Yano and Koga (1969) proposed an equation to model the inhibitory effect of substrates in the
case of continuous fermentation. The model expression is given as follows (Equation 6):
0 1
S
r ¼ rmax  @  A (6)
Ks þ S þ K1 þ KS2 2
S2 3

where K1 and K2 (g/L) are positive constants, which describe the substrate inhibition effect.
Aiba et al. (1968) formulated an empirical correlation as follows (Equation 7):
 
r ¼ rmax  S  eKi =ðKs þ SÞ
S
(7)

The model developed by Luong (1987) describes butanol inhibition on yeast growth as
(Equation 8):
rs ¼ ðrs max  S=ðKs þ SÞÞ  ð1  ðS=Sm ÞÞn (8)
4 D. TAZDAÏT ET AL.

where Sm is substrate concentration above which the microbial growth stops (g/L) and n
(dimensionless) is empirical constant.
Han and Levenspiel (1988) proposed a generalized nonlinear model as given by Equation 9):

rs ¼ rs max  S  ð1  ðS=Sm ÞÞn =ðS þ Ks  ð1  ðS=Sm ÞÞm Þ (9)

where n and m (dimensionless) are empirical constants.

The model proposed by Tseng and Wayman (1975) (Equation 10) takes into account the
inhibitory effect of different substrates (1-butanol, ethyl acetate, acetic acid and ethyl alcohol)
concentrations on the growth rate of two Candida species and a Saccharomyces in batch mode:
 
S
r ¼ rmax   Ki  ðS  Sm Þ (10)
Ks þ S

Analytical methods
Glyphosate determination
The determination of residual glyphosate concentrations was carried out according to the colori-
metric method reported by Bhaskara and Nagaraja (2006). Absorbance at 570 nm was measured
using UV-Vis spectrophotometer (RAYLEIGH UV-9200, China). Samples (2 ml) were removed at
regular time intervals (1 h) and were centrifuged at 5000 rpm for 30 min to remove cells. The
obtained cell-free supernatant was used for glyphosate determination.
Percent glyphosate removal was calculated using the following Equation (11):

Percent glyphosate removal ¼ ½ðCi  Cf Þ=Ci   100 (11)

where Ci and Cf are the glyphosate concentrations values at times 0 and t, respectively.

Estimation of biomass concentration

Microbial growth was estimated by measuring the spectrophotometric absorption of culture
samples. The absorption values were then converted into mixed liquor suspended solids (MLSS)
concentrations using a calibration curve that relates absorbance red at a wavelength of 600 nm
with a UV-Vis spectrophotometer (Shimadzu, UV mini 1240) to MLSS.
MLSS measurement was carried out by filtering a sample (20 ml) of activated sludge through a
glass microfiber filter (Whatman®, GE Healthcare, Belgium) previously weighted. The residue
retained on the filter was then dried at 105°C until a constant weight was attained, and MLSS
concentration (mg/L) was calculated according to the following Equation (12):

MLSS ¼ ðW1  W0 Þ=V (12)

where W1 is the sample and filter weight (mg), W0 is the weight of the filter (mg) and V is the
volume in L.
All samples were measured in duplicate and average values were used.

Software used
The eight kinetic models tested in this work were fitted to the experimental data and were solved
by nonlinear regression method using Statistica Release 7.0 software (StatSoft Inc., USA), which
utilizes the nonlinear least squares model estimation (Levenberg–Marquardt and Gauss–Newton
methods) for minimizing the sum of square of residuals and estimating the kinetic parameter
values.
 INTERNATIONAL JOURNAL OF ENVIRONMENTAL HEALTH RESEARCH 5

Results and discussion

Effect of glyphosate concentration on biomass growth
To investigate the effect of glyphosate as the sole carbon source on the growth of activated sludge
microorganisms, different concentrations of the herbicide were tested. Figure 2 presents the
kinetic profiles of biomass growth expressed as mg MLSS/L, for different glyphosate concentra-
tions ranging from 0.1 to 5 g/L. It can be seen from the figure that no biomass growth (specific
growth rate (µ) = 0 h−1) was observed at 2 and 5 g/L of glyphosate, while the growth profiles at 0.5
and 1 g/L of glyphosate started with a short latency stage, which lasted around 1 h, immediately
followed by an exponential phase with µ values of 0.027 and 0.057 h−1, respectively. The stationary
stage, which started after 7 h culture, was observed only for the experiment with 1 g/L of
glyphosate. Finally, the growth profiles decreased dramatically beyond 9 and 12 h for 0.5 and
1 g/L, respectively. Besides, no latency phase was observed for the experiment with 0.1 g/L of
glyphosate, the growth started directly with a brief exponential phase (µ = 0.22 h−1), which lasted
about 2 h. This is possibly due to rapid consumption of the pesticide. The results observed in these
experiments clearly indicated that the microorganisms forming the activated sludge were able to
grow on glyphosate tested at 0.1, 0.5 and 1 g/L. However, glyphosate at 0.5 and 1 g/L seemed to
support biomass growth, indicating that the herbicide can serve as carbon and/or energy sources
for the microorganisms under study. On the other hand, the absence of growth in the experiments
with 2 and 5 g/L could be explained by an inhibitory effect exerted by the herbicide on microbial
growth at such concentrations. This inhibitory effect is very likely exerted on the enzymes
activities (glyphosate deshydrogenase, carbon–phosphorous lyase) involved in the initiation of
biodegradation of glyphosate. These key enzymes generate product(s) that are compatible with
central metabolic pathways such as Embden–Meyerhof–Paranas pathway.
The effects of pesticides, including glyphosate, on growth of different microbial species have
been extensively studied (Pal et al. 2006; Tazdaït et al. 2013a; Prashar and Shah 2016). As
examples: Zain et al. (2013) have reported that the application of the following herbicides at
different concentrations: glyphosate (0.88, 1.76 and 3.52 mg/mL), paraquat (0.44, 0.88 and
1.76 mg/mL), metsulfuron-methyl (0.015, 0.03 and 0.06 mg/mL) and glufosinate- ammonium
(0.44, 0.88 and 1.76 mg/mL) exhibited an inhibitory activity on the growth of soil fungi. In
another study, glyphosate at 20 or more mg/L, 2,4-D at 200 mg/L and paraquat at 2 or more mg/L
have been found to completely inhibit the growth, the photosynthesis and chlorophyll a synthesis

Figure 2. Growth kinetics of activated sludge with different concentrations of glyphosate used as the sole carbon source (initial
cell concentration was about 300 mg MLSS/L).
6 D. TAZDAÏT ET AL.

of green alga Scenedesmus quadricauda Berb 614 (Wong 2000). However, the author reported that
the presence of low concentrations of 2,4-D (0.02 or 0.2 mg/L) and glyphosate (0.02 mg/L) exerted
a stimulatory activity on the three algal parameters. The study conducted by Shushkova et al.
(2009) showed that glyphosate (0.5 g/L) tested as the sole source of phosphorus seemed to inhibit
the growth of indigenous soil microflora. However, according to Moneke et al. (2010), glyphosate
showed no inhibition of the growth of Pseudomonas fluorescens and Acetobacter sp. at concentra-
tion of 0.25 g/L. More recently, Ðorđević and Ðorđević-Pejčev (2016) reported that bifenthrin at
different concentrations (0.25–10.0 µg/mL) had a very low effect on the growth of two fermenta-
tive microorganisms (Lactobacillus plantarum and Saccharomyces cerevisiae). On the other hand,
glyphosate (360 g/L) applied at 1 L/ha has been reported as negatively affecting the diversity of
bacterial communities in an agricultural soil, while high dose of the herbicide (10 L/ha) seemed to
exert an opposite effect (Cherni et al. 2015). In the light of all these observations, it could be
concluded that the inhibitory effect exerted by glyphosate on microbial growth varies widely
depending on its dose (increasing with increased dose) and the microbial target.

Effect of substrate concentration on glyphosate biodegradation

To assess the biodegradability of glyphosate by the indigenous activated sludge culture tested,
different initial concentrations of the herbicide, used as the sole carbon source, were tested and the
kinetic profiles of its biodegradation expressed as percent removal were established (Figure 3). It
can be seen from the figure that the mixed culture used was able to transform the herbicide for all
the concentrations tested. It should be noticed that it was possible to completely remove the
herbicide when tested at 0.1, 0.5 and 1 g/L, after 4, 13 and 18 h of culture, respectively, while the
percent removal values for initial concentrations of 2 and 5 g/L were found only to be 52 and 42%,
respectively, after 20 h incubation. These results indicate that the aerobic heterotrophic micro-
organisms used have the necessary enzymes for transforming glyphosate tested at different
concentrations; however, their ability to use it as their sole carbon source is concentration-
dependent (0.5 and 1 g/L) (Figure 2).
On the other hand, decreasing in percent removal observed for high glyphosate concentrations
(2 and 5 g/L) could be due to an inhibitory effect exerted by the herbicide on its own biodegrada-
tion. Several reports have studied the use of glyphosate as the sole carbon or phosphorous source
by different microbial species, such as Pseudomonas fluorescens and Acetobacter sp. (Moneke et al.
2010), Geobacillus caldoxylosilyticus T20 (Obojska et al. 2002) and Enterobacter cloacae K7

Figure 3. Effect of glyphosate concentration on biodegradation rates (initial cell concentration was about 300 mg MLSS/L).
 INTERNATIONAL JOURNAL OF ENVIRONMENTAL HEALTH RESEARCH 7

Figure 4. Experimental and predicted substrate degradation rates at different glyphosate concentrations due to different
models.

(Kryuchkova et al. 2014). The inhibitory effect of glyphosate on its own biodegradation observed
in this study could be due to the inhibition of the enzymes, which play a key role in glyphosate
degradation. Thus, the ability of microorganisms to utilize glyphosate as the sole carbon source is
due to the conversion of glyphosate to aminomethylphosphonate (AMPA). AMPA is
8 D. TAZDAÏT ET AL.

dephosphorylated to methylamine, which in turn is broken down to give formaldehyde and

ammonia. The resulting formaldehyde finally enters the carbon metabolic pathways of cellular
growth. Formaldehyde can also result from sarcosine (N-methylglycine), which originates from
C–P cleavage of glyphosate by C–P lyase activity (Kryuchkova et al. 2014; Malik et al. 2014).

Modelling study
In an attempt to confirm the inhibitory effect of glyphosate on its own degradation, glyphosate
degradation rate values obtained experimentally, and those predicted by the different kinetic models
tested were plotted against initial glyphosate concentrations as presented in Figure 4. The maximum
degradation rate of 0.02 g/L h was obtained for initial glyphosate concentration of 0.1 g/L. The
degradation rate then decreased drastically (from 0.0082 to 0.0005 g/L h) with increasing glyphosate
concentration from 0.5 to 5 g/L suggesting an inhibitory effect on the biodegradation of the herbicide
especially at 2 and 5 g/L, since the percent removal values observed for these two concentrations are
low (52 and 42%, respectively). On the other hand, the observed degradation rate values (experi-
mental data) are positioned close to those predicted by all model equations, except the one of Han
and Levenspiel, suggesting that the models adequately describe the biodegradation of glyphosate by
the microorganisms of the activated sludge used.
This is confirmed by the values of determination coefficient (R2), Fisher (F) (ratio of two variances)
and P (the probability of how much the independent variable can be used to predict the dependent
variable) shown in Table 1, which clearly highlight the adequacy of the models tested in this study.
These three statistic criteria were used to select the best biokinetic model for the description of
glyphosate biodegradation. P values less than 0.05 and larger F values suggest that the initial
glyphosate concentrations can be used to predict the biodegradation rates values. R2 values close to
1 is an indication that the model well describes the relation between initial glyphosate concentrations
of glyphosate and its biodegradation rates. As shown in Table 1, Yano and Koga model’s predictions of
glyphosate degradation rates are the best (R2 = 0.999, F = 173,106 and P = 0.000006).
The estimated values of biokinetic parameters for each model are presented in Table 2. The
biokinetic parameters of the best fitted model (Yano and Koga model) are: rmax = 0.006 g/
L h, Ks = 0.064 g/L, K1 = 2.4 g/L and K2 = 1.23 g/L. The terms K1 and K2 are direct indicators
of substrate inhibition. The values of K1 and K2, obtained with Yano and Koga equation, are
higher than those reported for others toxic compounds (Tazdaït et al., 2013b; Agarry et al. 2008).
This suggests that the mixed microbial culture tested in this work exhibited a high resistance
toward high glyphosate concentrations. On the other hand, the low value of Ks (0.006 g/L) clearly
indicated that there is a good affinity between the substrate (glyphosate) and the microbial
enzymes involved in its biodegradation.
There are few reports dealing with the application of kinetic models to describe the substrate
inhibition effect. According to Agarry et al. (2008), Yano and Koga equation was found to be the
best model for describing the kinetics of phenol degradation by a mixed culture of Pseudomonas

Table 1. Determination coefficients (R2), F values and P values for various kinetic models tested for the fitting of experimental
data of glyphosate degradation.
Model R2 F value P value*
Andrews (Edwards 1970) 0.994 327.72 0.00028
Aiba et al. (1968) 0.999 2645.74 0.000012
Han and Levenspiel (1988) 0.687 4.91 0.111
Luong (1987) 0.988 163 0.0008
Tessier (Edwards 1970) 0.959 44.47 0.0054
Webb (Edwards 1970) 0.998 837.17 0.0012
Tseng and Wayman (1975) 0.988 81.57 0.012
Yano and Koga (1969) 0.999 173,106 0.000006
*P value ˂ 0.05 was considered to be significant.
Table 2. Estimated values of kinetic parameters for various kinetic models tested for the fitting of experimental data of glyphosate biodegradation.
Estimated value of parameters
Model rmax (g/L·h) P value* Ks (g/L) P value* Ki (g/L) P value* K1 (g/L) P value* K2 (g/L) P value* Sm (g/L) P value* n m β P value*
Andrews (Edwards 1970) 0.018 0.158 0.34 0.313 0.412 0.301 - - - - - - - - - -
Aiba et al. (1968) 0.01 0.000 0.053 0.000 1.66 0.001 - - - - - - - - - -
Han and Levenspiel (1988) 0.000 0.91 0.085 0.472 - - - - - - 0.665 0.921 1 1 - -
Luong (1987) 0.007 0.002 0.066 0.000 - - - - - - 5 0.005 1 - - -
Tseng and Wayman (1975) 0.007 0.043 0.065 0.009 0.001 0.058 - - - - 0.038 0.96 - - - -
Webb (Edwards 1970) 0.011 0.026 0.053 0.012 1.54 0.19 - - - - - - - - 0.26 0.27
Tessier (Edwards 1970) 0.023 0.000 0.001 0.000 0.64 0.00 - - - - - - - - - -
Yano and Koga (1969) 0.006 0.000 0.064 0.000 - - 2.406 0.004 1.23 0.000 - - - - - -
*P value ˂ 0.05 was considered to be significant.
INTERNATIONAL JOURNAL OF ENVIRONMENTAL HEALTH RESEARCH
9
10 D. TAZDAÏT ET AL.

aeruginosa and Pseudomonas fluorescens. In another studies, it was found that Yano and Koga and
Andrews models (Tazdaït et al. 2013b), and Haldane model (Tazdaït et al. 2015) were the best
fitted for malathion biodestruction by indigenous activated sludge. Combarros et al. (2015) who
investigated the biodegradation of thiocyanate by a pure culture of Paracoccus thiocyanatus
concluded that Tessier model was the best fit for the experimental data. More recently,
Rajamanickam et al. (2017) investigated the potential of mixed microbial culture isolated from
cow dung compost to degrade toluene in batch mode. The authors showed that Levenspiel model
exhibited a good fit to the biodegradation kinetics data.

Conclusions
In this study, unacclimated activated sludge culture was tested for its potential to biodegrade
glyphosate under aerobic conditions. For this purpose, different concentrations of the herbicide
ranging from 0.1 to 5 g/L have been tested. Besides different kinetic models have been used to
assess the inhibitory effect caused by glyphosate on its own degradation. The obtained results
revealed that the mixed culture used was able to grow in the presence of 0.5 and 1 g/L glyphosate
and allowed a total removal of glyphosate at 0.1, 0.5 and 1 g/L, within 4, 13 and 18 h of culture,
respectively. Regarding the modelling study, it was found that, with the exception of Han and
Levenspiel model, all models tested presented a good fit to the biodegradation kinetic of glypho-
sate. Yano and Koga model generates the most accurate predictions regarding the biodegradation
rates of glyphosate. The microorganisms tested in this work can be a good candidate for treating
waters contaminated with high levels of glyphosate and should, in future studies, be assessed for
their ability to remove other organophosphate pesticides from water and soil.

Acknowledgements
The authors thank the anonymous reviewers and the Editor for valuable inputs to this article.

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