N US A N T A RA B I O S C I E N C E ISSN: 2087-3948

Vol. 16, No. 2, pp. 284-291 E-ISSN: 2087-3956

November 2024 DOI: 10.13057/nusbiosci/n160215

Imidacloprid degradation by potential soil bacteria isolated from rice

fields in Grobogan, Central Java, Indonesia

MUHAMMAD ALFIYAN HERMAWAN, ARTINI PANGASTUTI, RATNA SETYANINGSIH

Faculty of Mathematics and Natural Sciences, Graduate Program of Bioscience, Universitas Sebelas Maret. Jl. Ir. Sutami 36A Surakarta 57126, Central
Java, Indonesia. Tel./fax.: +62-271-663375, email: artini_p@staff.uns.ac.id

Manuscript received: 31 July 2024. Revision accepted: 16 October 2024.

Abstract. Hermawan MA, Pangastuti A, Setyaningsih R. 2024. Imidacloprid degradation by potential ⠀soil bacteria isolated⠀from rice
fields in⠀Grobogan, Central Java, Indonesia. Nusantara Bioscience 16: 284-291. Imidacloprid a widely used pesticide is known for its
polar nature, resistance to evaporation, and persistence in soil. When concentrations exceed environmental thresholds, imidacloprid can
act as a pollutant, disrupting ecosystems, altering soil pH, and decreasing soil fertility. This study aimed to isolate and i dentify soil
bacteria from rice fields capable of degrading imidacloprid and to highlight their potential role in bioremediation. Isolated bacteria are
identified based on morphological characteristics, their ability to degrade imidacloprid and through molecular tests using 16S rRNA.
Four bacterial colonies were obtained from the isolation results with different morphological variations. The degradation test results
showed that the isolates were able to grow in media containing imidacloprid and were able to reduce imidacloprid by 26.66-31.75%.
Based on 16S rRNA gene analysis, isolate IT1 was identified as Enterobacterales, IT2 was identified as the Enterobacteriaceae, IT3 as
Pectobacterium aroidearum strain CCRMPA670, and IT4 was identified as Bacillus thuringiensis strain FDAARGOS_791.

Keywords: Bacillus thuringiensis, Bacteria, biodegradation, imidacloprid, Pectobacterium aroidearum, rice field

INTRODUCTION Grobogan District has a rice harvest area of 179,124

hectares, with the highest rice production in Central Java,
The effective use of pesticides in controlling pests is a Indonesia, with a production of 787,275 tons-GKG in
short-term solution. Pesticides create a dependency among 2022. Grobogan District is indeed the largest among other
farmers to consistently use them as a determinant factor for districts in Central Java, so it has become an essential
high yields and quality agricultural products (Putri et al. factor in Indonesia’s rice production. Considering that it is
2021). Imidacloprid is a type of pesticide that is widely the largest rice harvest area in Central Java, it is essential to
used and its effect lasts about 156 days (Zamule et al. maintain the soil condition of the area so as not to
2021). Previous studies have reported varying half-life experience an increase in soil acidity due to the residue of
values for imidacloprid in different soil types, namely 455- the imidacloprid pesticide which in terms affects the rice
518 days on sandy clay soils in Australia and 233-366 days crop productivity level. Imidacloprid can cause the
on muddy clay soil in India (Bhattacherjee et al. 2020). accumulation of pesticide residues in the soil, potentially
Imidacloprid exceeding the environmental threshold killing the diversity of soil fauna, increasing plant pest
becomes a pollutant and can disrupt the natural balance. Its resistance and reducing soil fertility (Bandeira et al. 2020).
uncontrolled use of imidacloprid can lead to various Imidacloprid can undergo natural environmental
problems (Erguven and Demirci 2021). Approximately processes, including hydrolysis, photodegradation and
20% of imidacloprid pesticides hit the target, while the biodegradation. Biodegradation is a promising process for
remaining 80% falls into the soil, causing soil acidification reducing residues due to its relatively easy, selective,
and reducing soil fertility (Sabourmoghaddam et al. 2015). effective, safe, and cost-efficient operation (Hu et al. 2013).
Continuous use of imidacloprid leads to environmental Cycoń and Seget (2015) demonstrated that various
accumulation, resulting in soil and water pollution, bacterial isolates can degrade imidacloprid residues as the
potentially accumulating in the food chain (Bhattacherjee sole carbon or nitrogen source or through metabolic
et al. 2020). Its residues in the soil affect the decline in the transformation. Bhattacherjee et al. (2020) reported that
diversity of soil fauna (Bandeira et al. 2020). This occurs Burkholderia cepacia from agricultural land can degrade
because imidacloprid can influence the growth and 50 µg/mL of imidacloprid by 69% within 20 days. Gupta et
reproduction of soil fauna, reducing the quantity and al. (2016) used Pseudomonas sp. RPT 52 with a 0.5 mM
variety of existing fauna. Additionally, imidacloprid can imidacloprid solution, achieving approximately 46.5%
disrupt interactions among soil fauna, disturbing the degradation within 40 hours.
balance of the soil ecosystem. The low quantity of soil This study aimed to isolate and identify soil bacteria
fauna will reduce their contribution to soil quality and from rice fields capable of degrading imidacloprid and to
productivity. highlight their potential role in bioremediation. The
urgency of research is getting bacterial isolates that have
 HERMAWAN et al. – Imidacloprid degradation by potential soil bacteria isolated 285

a high ability to grow in rice fields that are applied by Isolation and purification of potential pesticide
imidacloprid and the ability to degrade the residue of the imidacloprid degrading bacterial isolates
imidacloprid pesticide and know the level of efficiency of The soil was dried and ground with a mortar and then
the isolates selected to re-feminist the imidacloprid residue sieved through a 0.2 mM mesh to remove physical
in the soil. impurities. Each soil sample was weighed and 5 grams
were taken using an analytical balance. Vortex was used to
homogenize the soil samples after they were put in bottles
MATERIALS AND METHODS with 45 mL of distilled water. Subsequently, centrifugation
was carried out at 10,000 x g for 20 minutes (Irfan et al.
Study area 2021), referred to as a 10-1 dilution. One millilitre of the
The sampling was conducted in the Godong, Wirosari, liquid was pipetted from the 10-1 dilution and added to a
and Ngaringan Sub-districts of Grobogan, Central Java, reaction tube holding nine millilitres of distilled water to
Indonesia (Figure 1). Grobogan is located at an altitude of make a 10-2 dilution. This process was repeated
100-500 meters above sea level with coordinates 7° 1' sequentially up to a 10-7 dilution using the serial dilution
18.188" S 110° 57' 45.306" E. The land in Grobogan is technique. Serial dilution was performed to reduce the
mostly used for the agricultural sector, such as rice fields density of microorganisms in the soil samples, facilitating
and plantations. the isolation of purer bacterial colonies on culture media
(Bhattacherjee et al. 2020).
Sample collection All the dilutions (10-1 to 10-7) were spread using the
The samples used were collected from the Grobogan pour plate method on 15 mL of MSM agar media
region, consisting of soil exposed to imidacloprid supplemented with 2 ppm of imidacloprid as the sole
pesticides based on a long history of using the pest- carbon source in petri dishes. The minimal salts medium
repellent pesticide for brown planthopper, namely Avidor (MSM, g/L) consisted of K2HPO4 2.27 g, KH2PO4 0.95 g,
25 WP brand (imidacloprid 25%). The sampling locations and (NH4)2SO4 0.67 g per 1 L of deionized water, adjusted
were at three points in the Sub-districts of Godong, to pH 7.0 (Coleman 2002). The inoculated plates were then
Wirosari, and Ngaringan, with soil samples taken from the incubated for 48 hours at 28°C (Yadav et al. 2021).
central area due to the likelihood of containing a significant Bacteria obtained from the mixed culture were purified
amount of pesticide residues and being the main rice using the quadrant streaking method with four streaks until
cultivation area. Using a scoop, 500 grams of soil samples no other bacterial mixtures were present. Pure isolates were
were taken from the top layer of soil (depth of 0–15 cm) also inoculated into glycerol stocks and stored in the
(Gautam and Dubey 2022). The collected soil was then freezer.
placed in an ice box (filled with ice bags to maintain a
temperature of ±4°C) to preserve the soil conditions (Alwi
et al. 2023).

Figure 1. Locations of soil sampling sites: Wirosari, Godong, and Ngaringan Sub-districts of Grobogan, Central Java, Indonesia (7° 1'
18.188" S 110° 57' 45.306" E)
286 N U SA NTA RA B I OSC IE NC E 16 (2): 284-291, November 2024

Growth of bacterial isolates to measure Optical Density (OD) at 600 nm. The bacterial
Bacterial isolates were cultured in erlenmeyer flasks growth curve was generated based on the OD values
containing 250 mL of Minimal Salts Medium (MSM) obtained over time. Following the construction of the
supplemented with two ppm imidacloprid. Subsequently, bacterial growth curve, a test for bacterial resistance to
the cultures were placed on a shaker incubator at 28°C and imidacloprid was conducted by observing the OD values at
150 rpm for 24 hours, allowing them to reach the 600 nm under various concentrations of imidacloprid.
exponential growth phase. A 10 mL sample was extracted Subsequently, a test of imidacloprid pesticide degradation
and centrifuged at 8000 x g for 15 minutes, the supernatant by bacteria was performed using High-Performance Liquid
was then discarded and replaced with 10 mL of sterile Chromatography (HPLC). The data obtained consisted of
distilled water, followed by vortex. The Optical Density residual imidacloprid concentrations after incubation with
(OD) values were measured using a UV-Vis bacteria. The acquired data were then analyzed
spectrophotometer at a wavelength of 600 nm (Mishra et al. descriptively and qualitatively. The sequences obtained
2014). Absorbance values were recorded every three hours from the sequencing process underwent a similarity test
until the bacterial culture entered the stationary phase. using BLASTn features on NCBI, utilizing the 'nr/nt' or
'Bacteria' database.
Imidacloprid pesticide degradation test
The isolates cultured were transferred in a volume of 10
mL into a reaction tube containing 250 mL of liquid MSM RESULTS AND DISCUSSION
with two ppm imidacloprid. The reaction tube was
incubated at 170 rpm and 28°C. Samples were detected and Imidacloprid residue
measured on days 0, 3, 5, 7, 10, 12, 14, 17, 19, and 21. The soil sample from Wirosari exhibited the highest
Pesticide imidacloprid degradation was assessed using residue levels compared to the soil samples from Godong
High-Performance Liquid Chromatography (HPLC) (Hu et and Ngaringan (Table 1). According to the Indonesian
al. 2013). Minister of Health and Minister of Agriculture Decree No.
881/MENKES/SKB/VIII/1996 and No.
Identification of selected bacterial isolates 711/Kpts/TP.270/8/1996 Regarding the Maximum Residue
The bacterial DNA genome from the isolate was Limits of Pesticides in Agricultural Products, the maximum
extracted using the Quick-DNATM Kit from Zymo residue limit for imidacloprid in soil should be 0.5 ppm.
Research, following the manufacturer's protocol. DNA The soil in Ngaringan and Godong has imidacloprid
obtained was then used as a template for the Amplification residues approaching the maximum allowable limit, while
of DNA 16S rRNA. Amplification using a pair of 67F (5′- the Wirosari soil exceeds the imidacloprid residue limit
CCTACGGGNGGCWGCAG-3′) and Primer 1387R (5′- permitted by the Department of Agriculture.
ACTACHV GGGTATCTAATCC-3′) and for sequencing
using a primary 785F (5′-GGATTAGATACCCTGGTA-3′) Isolation of indigenous bacteria
and 907R (5′-CCGTCAATTCMTTTRAGTTT-3′) to target The isolates capable of growing on MSM +
the partial region of the 16S rRNA gene. PCR reactions imidacloprid media were coded as IT1, IT2, IT3, and IT4.
were performed in a thermocycler with 7.5 μL My Taq Red Color dissimilarity was the most noticeable aspect of
Mix buffer (2x), 0.5 μL forward primer, 0.5 μL reverse isolates characterization. According to Table 2 and Figure
primer, 0.5 µL DNA template, and 19.8 μL ddH2O. The 2, The color of isolates IT1, IT2, IT3, and IT4 were light
denaturation, amplification, and annealing processes each pink color, brick-red color, whitish-yellow color, and white
had 30 cycles. The cycle parameters were as follows: initial color, respectively.
primer denaturation at 95°C for 5 minutes, 30 cycles of
denaturation at 95°C for 45 seconds, annealing at 62°C for Bacterial growth
45 seconds, extension at 72°C for 2 minutes, and a final All bacterial isolates experienced an exponential growth
extension at 72 °C for 72 minutes (McCabe et al. 1999). phase from hour 3 to 51, except for isolate IT1, which that it
The PCR products were then kept at 4°C for subsequent ended its exponential phase at hour 54 (Figure 3). The
analysis by electrophoresis. The amplified products were bacterial growth results from the unique capabilities of each
electrophoresed on a 1% agarose gel. Electrophoresis was bacterial isolate in utilizing nutrients present in the media,
carried out for 45 minutes at an electric voltage of 84 V in ultimately leading to variations in metabolic efficiency.
1x TAE buffer. The DNA amplicons in the agarose gel
were stained with gel red dye. DNA visualization was
performed using a UV transilluminator (Gao et al. 2021).
The amplified DNA was then subjected to DNA Table 1. The content of imidacloprid residue Grobogan, Central
sequencing. The 16S rDNA amplicon was subsequently Java, Indonesia, soil sample
sent to a third party for sequencing processing.
Soil samples Residue (ppm)
Data analysis Ngaringan 0.47
The growth curve of imidacloprid degrading bacteria Godong 0.45
was constructed, and the findings were obtained by Wirosari 0.50
incubating the bacteria with a UV-Vis spectrophotometer
 HERMAWAN et al. – Imidacloprid degradation by potential soil bacteria isolated 287

Imidacloprid pesticide degradation test CCRMPA670 and Bacillus thuringiensis strain

Bacterial isolates can grow in MSM + imidacloprid FDAARGOS_791. With this percentage, IT1 and IT2 were
media. Isolates IT1, IT2, IT3, and IT4 can reduce the identified into Enterobacterales and Enterobacteriaceae,
concentration of imidacloprid in MSM+imidacloprid 2 ppm respectively.
media. The highest (31.75%) percentage of decreased
imidacloprid was recorded in the IT1 isolate (Figure 4), and
the percentage of decreased imidacloprid IT3 isolates was
lower than the other three isolates. The percentage of
reduced imidacloprid was consistently increased from day 0
to the 21st day; this shows the potential for the isolate's
degradation or reduction of imidacloprid.

Identification of bacterial isolates

The results of BLAST-n analysis (Table 3) showed that
identified bacterial isolates had a similarity of 16S rRNA
gene sequences with bacteria from the genus
Pectobacterium and Bacillus. IT 1 and IT 2 had a
percentage of similarity <95% with a low category after
comparing species data in Genbank. Based on the 16S
rRNA enclosure gene, the results revealed that isolates IT1
and IT2 were not identified because the similarity was only
86.83% and 89.51% with Serratia nevei and S. marcescens,
respectively. Isolates IT3 and IT4 showed 99.83% and Figure 3. Growth of bacterial isolates on MSM with imidacloprid
98.57% similarities with Pectobacterium aroidearum strain media

A B

C D

Figure 2. Bacterial colonies. A. IT1, B. IT2, C. IT3, D. IT4

288 N U SA NTA RA B I OSC IE NC E 16 (2): 284-291, November 2024

Table 2. The characteristics of bacterial isolates from the rice fields in Grobogan, Central Java, Indonesia

Isolates code Elevation Margin Colony color Shape of colony Shape of cell
IT 1 Convex Entire Light pink Circular Bacilli
IT 2 Convex Entire Brick red Circular Bacilli
IT 3 Convex Entire Whitish yellow Circular Bacilli
IT 4 Convex Entire White Circular Bacilli

Table 3. Similarity of 16S rRNA gene sequences of bacterial isolates using the BLAST-n program

Isolates code Related species Query cover (%) Similarity (%) ACC Numbers
IT 1 Serratia nevei strain 2017-45-174 76 86.83 CP109739.1
IT 2 Serratia marcescens strain JW-CZ2 100 89.51 CP055161.1
IT 3 Pectobacterium aroidearum strain CCRMPA670 90 99.83 MN883868.1
IT 4 Bacillus thuringiensis strain FDAARGOS_791 100 98.57 CP054568.1

A C

B D

Figure 4. Growth of bacterial isolates on MSM + imidacloprid media: A. IT1, B. IT2, C. IT3, D. IT4

Discussion Coribacteraceae, Coxiellaceae, and Rhodospirillaceae in

Imidacloprid residue can affect the presence of soil the soil, but imidacloprid increases the abundance of
bacteria through various mechanisms, both directly and Nitrospirae bacteria in soil. Nitrospirae bacteria play a role
indirectly. The impact of imidacloprid on soil bacteria in the nitrogen cycle, which is the conversion of ammonia
depends on several factors such as bacterial types, into nitrate, a source of nitrogen that is important for plants
imidacloprid concentrations, environmental conditions, and (Yu et al. 2020).
degradation of microorganisms. Excessive use of All growth of bacterial isolates undergoes an
imidacloprid can interfere with the balance of soil microbes exponential phase at a vulnerable time of 3 to 51 hours,
and decrease soil ecological function. Several studies have except IT1 isolates, which ended the exponential phase at
shown that imidacloprid residues can affect the 54 hours. The growth of these bacteria arises due to the
composition of bacterial communities in the soil. Akter et unique capabilities of each bacterial isolate in taking
al. (2023) reported that high imidacloprid concentrations advantage of the nutrients contained in the media, which
could reduce the abundance of Actinobacteria, ultimately impacts variations in metabolic efficiency. The
Bacteroidetes, and Proteobacteria in the soil. Astaykina et death of bacteria sensitive to imidacloprid can provide an
al. (2020) noted that imidacloprid changed the relative opportunity for bacteria that are resistant to breed.
abundance of several eukaryotic and prokaryotic genera, Mohammed and Badawy (2017) reported that soil bacteria
such as Apiotrichum, Gamicola, Humicola, Kitasatospora, can quickly degrade imidacloprid through various
Solicoccozyma, Sphingomonas, Streptomyces and metabolic pathways. Gonzalez and Aranda (2023) reported
Terrabacter, respectively. He also stated that imidacloprid that growth in the exponential phase is influenced by the
also reduces the relative abundance of Methylophilaceae, nature and shape of microbes in the environment, using
 HERMAWAN et al. – Imidacloprid degradation by potential soil bacteria isolated 289

nutrients in the growth medium, temperature conditions, control and a plant growth promoter. In another study,
and media pH. Then, enter the stationary phase until 72 Tepidibacillus decaturensis strain ST1 was able to degrade
hours. The stationary phase occurs if the number of bacterial imidacloprid effectively in liquid media, slurry, and soil
cells stops increases (Jõers et al. 2020). Although there is no microcosms (Tiwari et al. 2023).
growth in the stationary phase, cells can still grow and The results of molecular analysis revealed that IT1 was
divide themselves. In this phase, the number of growing identified at the level of the order Enterobacterales and IT2
bacteria is balanced with the number of dead bacteria (Risna isolates identified at the level of the Enterobacteriaceae
et al. 2022). Bacterial isolates demonstrate the ability to family. Isolates IT3 and IT4 were identified as
thrive in MSM+imidaclopridmedia due to their utilization Pectobacterium proidearum strain CCRMPA670 and B.
of carbon and nitrogen sources in the media. This aligns thuringiensis strain FDAARGOS_791. The similarity of
with the findings of Zamule et al. (2021), who reported that the 16S rRNA gene is one of the characteristics of a close-
various bacterial strains, including Pseudomonas related bacterium. The 16S rRNA gene is very
fluorescens, Pseudomonas putida, Pseudomonas conservative, so changes that occur in this gene usually
aeruginosa, Alcaligenes faecalis, Escherichia coli and occur slowly and gradually. This causes closely related
Streptococcus lactis can flourish in imidacloprid-containing bacteria to have a similar similarity to the 16S rRNA gene
media. (Sharma et al. 2014). Determination of potential bacterial
The highest percentage of decreased imidacloprid in identity is based on the criteria for the percentage of
test was 31.75%, in IT1 isolates, while the percentage of similarities ≥99% shows the similarity of species, the
reduction in IT3 isolates was lower than in the other three percentage of similarity ≥95%-<99% shows the similarity
isolates. The percentage of decreased imidacloprid was of the genus, and the percentage of similarity <95% shows
increased from day 0 to 21st day, this shows the potential the similarity of the family (Collins et al. 1994). Church et
for degradation or reduction of imidacloprid by the isolates. al. (2020) reported that comparing sequences of the 16S
Bacteria that can grow in MSM+imidacloprid media are rRNA gene can help distinguish organisms at the genus
bacteria that have enzymes that can break the chemical level across key bacterial phyla and classify strains at
structure of imidacloprid into simpler molecules, which various levels. Pectobacterium is included in the gram-
bacteria can then use as a source of carbon and nitrogen. negative bacteria Enterobacteriaceae found in rice fields'
Other bacteria that are unable to grow in soil (Rossmann et al. 2018). Bacillus is generally used as a
MSM+imidacloprid media do not have the enzymes needed plant growth booster agent found in plantations and rice
to utilize the carbon imidacloprid content. These bacteria fields (Akinrinlola et al. 2018). Pang et al. (2020) state that
have enzymes that can break imidacloprid, but these Pectobacterium can grow and survive under high levels of
enzymes are not efficient enough to produce enough energy imidacloprid. According to Vu et al. (2022),
for bacterial growth. Cycoń and Seget (2015) reported that Pectobacterium can resist imidacloprid pesticides, and the
the activity of the enzyme dehydrogenase, bacteria have developed mechanisms to protect themselves.
hexaphosphatase, and urease in soil bacteria given Ferreira et al. (2016) research states that B. thuringiensis is
imidacloprid has decreased performance. Every pesticide able to degradate imidacloprid. Other members of the
application that affects the microbial community and its genus Bacillus who also showed the ability to degrade
biochemical activity in the soil can be estimated to produce imidacloprid compounds such as Bacillus cereus (Talpur et
changes in the level of soil enzyme activity. Akoijam and al. 2023), and Bacillus wehenstephanensis (Shetti et al.
Singh (2015) observed that Bacillus aerophilus and B. 2021). Bacillus striatum, which contains CYP353D1v2
alkalinitrilicus are capable of degrading over 90% of genes exhibits strong resistance to imidacloprid (Pang et al.
imidacloprid in clay loam within 56 days. The degradation 2020). Soil-dwelling bacteria from the genus Bacillus have
produces metabolites, such as 6-chloronicotinic acid, the ability to break down pesticides into simpler residues.
nitrosimine, and imidacloprid-NTG, which remain B. cereus was identified as an efficient catalyst for
unaffected by sterilization. In another study, B. cepacia degrading imidacloprid, metabolizing 92% of it within 11
strain CH 9 was able to degrade 69% of the 50 ppm days at a neutral pH. Through optimization using the Box-
imidacloprid in 20 days after inoculation in MSM media Behnken design, the bacteria transformed imidacloprid into
(Bhattacherjee et al. 2020). Ochrobacterium sp. strain 6-CNA via the intermediate’s guanidine and 5-hydroxy
BCL-1 can degrade 67.67% from 50 ppm imidacloprid in imidacloprid (Talpur et al. 2023). The B. cereus is
48 hours after the application as mentioned in the literature considered a promising tool for removing imidacloprid
(Hu et al. 2013). Akoijam and Singh (2015) have observed from contaminated water and soil (Gangola et al. 2021).
that the loss of imidacloprid follows the first pseudo-order Isolating the enzyme responsible for this degradation could
kinetics when applied at levels of 50, 100, and 150 ppm in provide a pathway for commercial use of purified enzymes.
sandy clay enriched with B. aerophilus with a part-time In conclusion, 4 colonies obtained with different
value of 14.33, 15, 15, 05, and 18.81 days. A strain of B. morphological variations namely IT1, IT2, IT3, and IT4.
thuringiensis isolated from polluted marine sediments has Analysis of degradation activity using the HPLC method
been shown to degrade 71% of imidacloprid within 11 days showed that all isolates have the ability to grow in MSM,
(Obayori et al. 2024). Trichoderma, one of the most which contains imidacloprid and succeed in reducing the
promising biological control agents, is found across various imidacloprid content by 26.66-31.75%. Based on 16S
agricultural climates and is prevalent in soil and root rRNA gene analysis, isolate IT1 was identified as
ecosystems, it has the ability to serve as both a biological Enterobacterales, IT2 was identified into the
290 N U SA NTA RA B I OSC IE NC E 16 (2): 284-291, November 2024

Enterobacteriaceae, IT3 as P. aroidearum strain imidacloprid in soils. Ecotoxicol Environ Saf 225: 112785. DOI:
10.1016/j.ecoenv.2021.112785.
CCRMPA670 and IT4 were identified as B. thuringiensis Gautam P, Dubey KS. 2022. Biodegradation of imidacloprid: Molecular
strain FDAARGOS_791. and kinetic analysis. Bioresour Technol 350: 126915. DOI:
10.1016/j.biortech.2022.126915.
Gonzalez JM, Aranda B. 2023. Microbial growth under limiting
conditions-future perspectives. Microorganisms 11: 1641. DOI:
ACKNOWLEDGEMENTS 10.3390/microorganisms11071641.
Gupta M, Mathur S, Sharma TK, Rana M, Gairola A, Navani NK. 2016.
We want to express our gratitude to the Laboratory in A study on metabolic prowess of Pseudomonas sp. RPT 52 to
Universitas Sebelas Maret (UNS) Surakarta, Indonesia and degrade imidacloprid, endosulfan and coragen. J Hazard Mater 301:
250-258. DOI: 10.1016/j.jhazmat.2015.08.055.
the government of Grobogan District, Ministry of Research, Hu G, Zhao Y, Liu B, Song F, You M. 2013. Isolation of an indigenous
Technology, Indonesia. imidacloprid-degrading bacterium and imidacloprid bioremediation
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