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REVIEW
Toxicity, monitoring and biodegradation of the fungicide
carbendazim
Simranjeet Singh
1
•Nasib Singh
3
•Vijay Kumar
2
•Shivika Datta
1
•
Abdul Basit Wani
2
•Damnita Singh
1
•Karan Singh
4
•Joginder Singh
1
Received: 2 April 2016 / Accepted: 17 May 2016
ÓSpringer International Publishing Switzerland 2016
Abstract The increasing use of toxic pesticides is a major
environmental concern. Carbendazim is a systemic fungi-
cide having wide applications for controlling fungal diseases
in agriculture, forestry and veterinary medicines. Carben-
dazim is a major pollutant detectable in food, soil and water.
Carbendazim extensive and repeated use induces acute and
delayed toxic effects on humans, invertebrates, aquatic life
forms and soil microorganisms. Here, we review the pollu-
tion, non-target toxicity and microbial degradation of car-
bendazim for crop and veterinary purposes. We found that
carbendazim causes embryotoxicity, apoptosis, terato-
genicity, infertility, hepatocellular dysfunction, endocrine-
disrupting effects, disruption of haematological functions,
mitotic spindle abnormalities, mutagenic and aneugenic
effect. We also found that carbendazim disrupted the
microbial community structure in various ecosystems. The
detection of carbendazim in soil and reservoir sites is per-
formed by spectroscopic, chromatographic, voltammetric,
nanoparticles, carbon electrodes and mass spectrometry. A
review of the degradation of carbendazim shows that car-
bendazim undergoes partial to complete biodegradation in
the soil and water by Azospirillum, Aeromonas, Alternaria,
Bacillus, Brevibacillus, Nocardioides, Pseudomonas, Ral-
stonia, Rhodococcus, Sphingomonas, Streptomyces and
Trichoderma.
Keywords Carbendazim Fungicide Soil
microorganisms Biodegradation Monitoring
Introduction
Pesticides found extensive applications in agricultural and
veterinary practices worldwide since the last four decades.
The global annual consumption of pesticides is *two
million tonnes where Europe accounts for 45 %, USA
accounts for 25 % and other countries account for the
remaining 25 % usage. The total annual consumption of
pesticides in India is in the form of insecticides (80 %),
herbicides (15 %), fungicides (2 %) and other pesticides
(3 %) (De et al. 2014). Fungal pathogens cause significant
damages to the food crops every year resulting in poor
yields, deficient food contents and huge economic loss.
Hence, use of fungicides gains favour to circumvent these
losses (Karlsson et al. 2014; Patel et al. 2015). Carben-
dazim (methyl 1H-benzimidazol-2-ylcarbamate) is a sys-
temic broad-spectrum fungicide having chemical formula
C
9
H
9
N
3
O
2
and mol. wt. 191.19 (Table 1). It is also
obtained as degradation products of thiophanate-methyl
and benomyl fungicides (Mazellier et al. 2003; Fang et al.
2012). Carbendazim is used worldwide as pre- and post-
harvest treatment to control the Ascomycetes, Fungi
imperfecti and Basidiomycetes fungal diseases on various
vegetables, fruits and several other plants such as banana,
mango, strawberries, oranges, pineapples, pomes, cereals,
sugar beet, fodder beet, rape seed, ornamental plants,
&Joginder Singh
joginder.15005@lpu.co.in
1
Department of Biotechnology, Lovely Professional
University, Phagwara 144411, Punjab, India
2
Department of Chemistry, Lovely Professional University,
Phagwara 144411, Punjab, India
3
Department of Microbiology, Akal College of Basic
Sciences, Eternal University,
Baru Sahib 173101, Himachal Pradesh, India
4
Department of Chemistry, Akal College of Basic Sciences,
Eternal University, Baru Sahib 173101, Himachal Pradesh,
India
123
Environ Chem Lett
DOI 10.1007/s10311-016-0566-2
medicinal herbs and turf grasses (Tortella et al. 2013; Devi
et al. 2015). Further, carbendazim in combination with
mancozeb exhibited an effective control of sunflower leaf
blight, chilli rots and mango anthracnose (Devi et al.
2015). In addition, carbendazim is also used in paint,
textile, paper and leather industries (Selmanoglu et al.
2001). It is among the top five pesticides used in India with
annual consumption of 1992 metric tonnes (Bhushan et al.
2013). It stands second after mancozeb in terms of most
consumed carbamates in India. It is registered for 18 crops
by The Central Insecticides Board and Registration Com-
mittee (CIBRC) in India. These crops are paddy, wheat,
barley, tapioca, cotton, jute, groundnut, sugarbeet, peas
cluster, beans, cucurbits, brinjal, apples, grapes, walnut,
rose and ber mango (Bhushan et al. 2013).
Carbendazim is classified in the hazardous category of
chemicals by World Health Organization. Carbendazim
along with carbomyl are classified as possible human
carcinogens (Goodson et al. 2015). The European Com-
mission has categorized it in the priority list of endocrine-
disrupting chemicals (Ferreira et al. 2008). Due to its
severe toxicity and persistent nature, carbendazim has been
banned in Australia, most of European Union and USA
(Zhang et al. 2013; Huan et al. 2016). The orange juices
imported from Brazil to Australia and USA have also
undergone extensive screening before their availability to
the human consumption. However, UK, Portugal (Euro-
pean Union Pesticide Database 2015) and developing
countries such as China, Brazil and India are still permit-
ting the production and use of carbendazim in various
formulations. Its repeated applications lead to accumula-
tion and contamination of various ecosystems with long-
lasting impacts on soil sustainability and human and ani-
mal health. Here, we attempt to summarize the current
status and analyse the trends in in vivo toxicities, envi-
ronmental monitoring and microbial degradation of
carbendazim.
Toxicity of carbendazim against living organisms
Widespread usage of pesticides and fungicides in agricul-
ture, industrial and recreational activities resulted in daily
exposure of human, livestock and soil/aquatic animals to
these potentially fatal health hazards (Banyiova et al.
2016). The acceptable daily intake (ADI) of carbendazim
is 0.03 mg/kg/day in India (Sharma 2007). In a recent
study, the foliar use of 12 % carbendazim and 63 %
mancozeb combination on mango fruits was found to be
safe for both crop and consumer health (Devi et al. 2015).
However, the excessive and repeated uses of carbendazim
in agriculture have raised a growing concern for the health
and safety of human, livestock, aquatic animals and non-
target microorganisms. Depending on the nature of the
Table 1 Chemical and physical properties of carbendazim and its derivatives
General name IUPAC name Chemical
formula
Mol. wt.
(g/mol)
Solubility in
water
Log K
OW
(at
25 °C)
Density Henry’s law constant (atm.
m
3
/mol)
Carbendazim Methyl 1H-benzimidazol-2-ylcarbamate C
6
H
9
N
3
O
2
191.18 8 mg/L 1.49 270 kg/
m
3
1.02 910
-9
at 20 °C
2-Aminobenzimidazole 1H-benzimidazol-2-amine C
7
H
7
N
3
133.15 \1 mg/mL 0.9 *1.4 g/
cm
3
–
Benzimidazole 1H-benzimidazole C
7
H
6
N
2
118.13 2010 mg/L 1.32 1.23 g/
cm
3
3.7 910
-7
at 25 °C
2-Hydroxybenzimidazole 1,3-dihydrobenzimidazol-2-one C
7
H
6
N
2
O 134.13 – 1.1 – –
Benomyl (Parent moiety) Methyl N-[1-(butylcarbamoyl) benzimidazol-2-
yl]carbamate
C
14
H
18
N
4
O
3
290.32 3.8 mg/L 1.36 0.38 g/
cm
3
4.93 910
-12
at 25 °C
IUPAC: International Union of Pure and Applied Chemistry
Environ Chem Lett
123
soil, residual carbendazim has a half-life of about 3 days to
12 months (Torstensson and Wessen 1984; Jones et al.
2004; Pourreza et al. 2015). Its environmental persistence
is due to the benzimidazolic ring which resists degradation
(Barnett et al. 1996). Carbendazim degradation results in
the formation of 2-amino-benzimidazole, a highly toxic
component, which binds to the spindle microtubules
causing the nuclear division blockade (Yenjerla et al.
2009).
Carbendazim is known to manifest embryotoxicity,
germ cell apoptosis, teratogenesis, infertility and develop-
mental toxicity in different mammalian species (Table 4,
Carter et al. 1987; Mantovani et al. 1998; Minta et al. 2004;
Yenjerla et al. 2009; Rama et al. 2014). Microtubules
which are formed by non-covalent bindings of a- and b-
tubulin are responsible for chromosome segregation during
mitosis and meiosis (Rama et al. 2014). Tubulin exists
in vivo as a/bheterodimer, and carbendazim is known to
disrupt the dynamics of assembly and disassembly of
microtubule. Carbendazim inhibits the microtubule poly-
merization in fungal and mammalian cells by acting with
b-tubulin causing disruption of microtubule assembly,
which leads to impaired segregation of chromosomes
during cell division (Lacey and Watson 1985; Davidse
1986; Yenjerla et al. 2009; Pacheco et al. 2012; Rama et al.
2014). Experimental studies in mice and rats revealed that
carbendazim causes hepatocellular dysfunction (Janardhan
et al. 1987: Muthuviveganandavel et al. 2008; Dikic et al.
2012; Salihu et al. 2015), affects hematopoiesis (Zubrod
et al. 2014), increased the oestrogen levels (Morinaga et al.
2004), increased the androgen receptor mRNA levels (Hsu
et al. 2011), exerted endocrine-disrupting effects (Lu et al.
2004; Yunlong et al. 2009; Banyiova et al. 2016), exerted
oxidative stress in Leydig cells and testes (Correa et al.
2002; Rajeswari and Kanmani 2009; Adedara et al. 2013;
Sakr and Shalaby 2014; Rama et al. 2014) and decreased
the total WBC and platelets counts (Salihu et al. 2015).
Carbendazim also affects the reproductive functions in
hamsters (Gray et al. 1990) and Japanese quails (Aire
2005). Chronic or sub-chronic exposure of carbendazim
has been found to cause testicular damage (Prashantkumar
et al. 2012) and disrupt the biochemical, hepatic, renal and
haematological functions in male goats (Prashantkumar
et al. 2012; Daundkar and Rampal 2014).
Humans are exposed to carbendazim either directly (oral
ingestion, inhalation and dermal contact) or indirectly
(ingestion of contaminated water, food items and occupa-
tional exposure) (Bakirci et al. 2014; Salihu et al. 2015;
Olayemi 2015). Carbendazim is reported to inhibit the
proliferation of murine and human cancer cell lines
(Hammond et al. 2001; Yenjerla et al. 2009) and tumour
xenografts (Hao 2002). It is also reported to inhibit the
proliferation of human breast cancer cells (MCF7) by
inducing mitotic spindle abnormalities, metaphase arrest
and suppressing tubulin dynamic instability (Yenjerla et al.
2009). Carbendazim decreased the viability, altered the
gene expression and induced apoptosis in the human tro-
phoblast cell line (Adedara et al. 2013). It is a non-heri-
table gene mutagen (Kosasa et al. 1999) and exerts potent
aneugenic effect on human lymphocytes (Bentley et al.
2000). Lethal dose, 50 % (LD
50
) values of carbendazim
against different aquatic invertebrates, fishes, rodents and
mammals are shown in Table 2. Industrial production
process and extensive use in agriculture and forestry
facilitate the entry of carbendazim in aquatic systems. Its
concentration in such environments was found to be
4.5 mg/L (Chatupote and Panapitukkul 2005; Silva et al.
2015). It exhibited toxicity to the eggs of prussian carp
Carassius gibelio at a dose of 0.036 mg/L (Ludwikowska
et al. 2013). Its deleterious effects on biochemical param-
eters, embryonic development and gene expression are
known against earthworm Eisenia fetida (Rico et al. 2016;
Huan et al. 2016), milkfish Chanos chanos (Palanikumar
et al. 2014), African clawed frog Xenopus laevis (Yoon
et al. 2008) and zebrafish Danio rerio (Jiang et al. 2015;
Andrade et al. 2016). The research on the mechanistic
aspects and long-term ecological consequences of carben-
dazim on aquatic animals, however, is still in infancy.
Carbendazim when applied to agricultural crops, forest
areas or turf grasses finally reaches the soil where it is
responsible for alteration in the soil microbial balance via
different pathways. The effect of pesticides on resident
microflora of soil is affected by chemical nature of the
pesticide, dose levels, frequency of use, environmental
persistence, bioavailability and the mechanism of action
(John and Shaike 2015). Soil microflora is directly or
indirectly exposed to lethal doses of carbendazim. Studies
have indicated the negative influence of residual carben-
dazim on soil microbial community composition, microbial
counts and microbial-mediated ecosystem functions
(Yunlong et al. 2009; Wang et al. 2009a,b,2016).
Niewiadomska (2004) found that in the presence of car-
bendazim 20 % and thiram 45 % mixture, the nitrogenase
activity of Rhizobium leguminosarum bv trifolii KGL was
dropped by 80 %. However, 100 % increase was witnessed
in Azotobacter counts. In another study, deleterious effects
on the populations of nitrogen fixing, nitrifying and cel-
lulolytic bacteria were observed following treatment with
carbendazim-mancozeb mixture (Fawole et al. 2010).
Similarly, Wang et al. (2009a,b,2012) reported the dose-
dependent negative influence on the microbial diversity in
carbendazim-treated soils. Further, there was a decrease in
soil enzymatic (cellulose and pectinase) activities. Car-
bendazim either alone or in combination with chloram-
phenicol exhibited inhibitory effect of the fungal:bacterial
ratios and a variable response on soil enzymatic activities
Environ Chem Lett
123
Table 2 Toxicity profiles of carbendazim against different categories of living organisms
Living organism Common name Category Exposure
time (h)
EC
50
LD
50
References
Anas platyrhynchos Common mallard Aves (Anatidae) – – 615 mg/Kg Parsons et al. (2010)
Aphidius matricariae Aphid parasite Hexapodia (Braconidae) – – [30 LR50 g/ha Jansen (1999)
Aphidius rhopalosiphi Braconid parasitic wasp Hexapodia (Braconidae) – – [3000 LR50 g/ha Bernard et al. (2010)
Apis mellifera Bee Insecta (Apidae) – – [50 lg/Bee Mullin et al. (2010)
Bithynia tentaculata Mud bithynia Mollusca (Bithyniidae) 96 – – Daam et al. (2009)
Canis lupus familiaris Domestic dog Mammalia (Canidae) – – [10,000 mg/Kg Gupta and Aggarwal (2007)
Capra aegagrus Goat Mammalia (Bovidae) 90 days – Oxidative stress at 50 mg/Kg Daundkar and Rampal (2014)
Chanos chanos Milk fish Pisces (Chanidae) 96 – 11.5 lg/L Palanikumar et al. (2014)
Chaoborus obscuriceps Systema Dipterorum Insecta (Chaoboridae) 96 [3435 lg/L – Daam et al. (2009)
Chlorella pyrenoidosa Chlorella algae Chlorophyta (Chlorellaceae) 96 340 lg/L – Canton (1976)
Danio rerio Zebrafish Actinopterygii (Cyprinidae) 8 days – Apoptosis and immunotoxicity Jiang et al. (2015)
Danio rerio Zebrafish Actinopterygii (Cyprinidae) 96 h 0.85–1.6 mg/L 1.75 mg/L Andrade et al. (2016)
Daphnia Magna Water flea Arthropoda (Daphniidae) 96 87 91 lg/L Solomon et al. (2008)
Daphnia magna Water flea Arthopoda (Daphniidae) 48 – 2.89–23.12 lg/L Silva et al. (2015)
Dero digitata Naiadid worm Annelida (Naididae) 96 980 lg/L – Van Wijngaarden et al.
(1998)
Dugesia lugubris Planarian Platyheminthes (Dugesiidae) 96 25 lg/L 134 lg/L Solomon et al. (2008)
Dugesia lugubris Planarian Platyhelminthes (Planariidae) 96 178 lg/L – Van Wijngaarden et al.
(1998)
Eisenia fetida Earth worm Annelida (Lumbricidae) 336 – 3.0–35.2 mg/Kg Liu et al. (2012)
Eisenia foetida Earth worm Annelida (Lumbricidae) 14 days – 8.6 mg/Kg Huan et al. (2016)
Gammarus pulex Cancer pulex Arthropoda (Gammaridae) 96 55 lg/L 177 lg/L Solomon et al. (2008)
Mus musculus CD-1 mice Mammalia (Rodenta) – – Toxicity at 300 mg/Kg/day dose Farag et al. (2011)
Rattus norvegicus Brown rat Mammalia (Murids) – – [2000 mg/Kg Kumar (2001)
Rattus norvegicus Wistar rat Mammalia (Rodents) 14 days – Renal and hepatic dysfunction at
50 mg/Kg
Salihu et al. (2015)
Salmo gairdneri Rainbow trout Pisces (Salmonidae) 96 1800 lg/L – Canton (1976)
Scenedemus subspicatus Chodat Chlorophyceae
(Scenedesmaceae)
72 – [7.7 mg/L Dang and Smit (2008)
Scenedesmus
capricornutum
Green algae Chlorophyceae
(Scenedesmaceae)
72 – 300 lg/L Ma et al. (2002)
Stylaria lacustris Aquatic oligochaete worm Annelida (Tubificidae) 96 1060 lg/L – Daam et al. (2009)
Xenopus laevis African clawed frog Amphibia (Pipidae) – – 5.606 lM Yoon et al. (2008)
EC
50
=effective concentration, 50 %; LD
50
=lethal dose, 50 %; LR
50
=lethal rate, 50 %
Environ Chem Lett
123
(Yan et al. 2011). Enrichment of carbendazim-adapting
strains in the soil was found to alter the microbial com-
munity balance. However, its repeated applications led to
adaptation of soil microorganisms as revealed by compa-
rable Simpson and Shannon indexes of their community
structure from carbendazim-treated and untreated soils
(Yunlong et al. 2009). Considering the uncultivable nature
of many soil microbes, advance molecular techniques are
required to decipher the multifaceted effects of carben-
dazim on soil microbial health. The studies discussed
above indicate that soil and aquatic life forms are highly
vulnerable to carbendazim. Higher organisms are equally
susceptible to it, and the exact toxicological manifestations
depend on exposure levels and environmental persistence.
Accurate and reliable detection of carbendazim is thus
necessary for safety of human health and environment.
Environmental detection of carbendazim
Due to slow degradation, carbendazim persists in bare soils
for a long time (half-life of 6–12 months). In water, how-
ever, it dissipates rather rapidly as indicated by short half-
life of about 2-25 days (FAO 1998). Several techniques
such as gas chromatography, UV-visible spectroscopy,
capillary electrophoresis, high-performance liquid chro-
matography (HPLC) (Bicchi et al. 1989), ultraperformance
liquid chromatography, liquid chromatography/time-of
flight mass spectrometry, tandem mass spectrometry and
voltammetric methods have been utilized for reliable and
accurate quantitative estimation of carbendazim in various
commodities, soil samples and aquatic reservoirs (Alvarez
et al. 1997) (Table 3). Micellar electrokinetic chromatog-
raphy in conjunction with solid-phase extraction was used
to detect carbendazim residues in rice and wheat grains
with a detection limit of 0.05 ppm (Wu et al. 1997). (Patel
et al. 2015) used a colorimetric sensor based on
4-aminobenzenethiol functionalized silver nanoparticles
for detection of carbendazim in water, fruits and vegeta-
bles samples. Other techniques such as ELISA (Itak et al.
1993; Mountfort et al. 1994), cross section fluorimetry
(Zhu et al. 2008) and surface-enhanced Raman scattering
(Strickland and Batt 2009) have also been utilized for
carbendazim determination. Similarly, Furini et al. (2015)
assayed the carbendazim by adsorption onto the surface of
nanoparticles by surface-enhanced Raman scattering. UV-
visible spectroscopy was useful in detecting the complex
formation of carbendazim with 2, 2-Bipyridyl-Fe(III) or
potassium ferricyanide (Naidu et al. 2011). Zebrafish
(Danio rerio) and fresh water flea (Daphnia magna) have
Table 3 Analytical techniques applied for the detection and quantitative estimation of carbendazim or its derivatives in different type of samples
S.
no.
Technique Resource/medium References
1. UV-visible spectroscopy Soil, water Naidu et al. (2011)
2. High-performance liquid chromatography-mass spectrometry (HPLC/MS) Culture medium Fang et al. (2012)
3. Cross section fluorimetry Soil, water Zamora et al. (2000)
4. Enzyme-linked immunosorbent assay (ELISA) Fruit and vegetables Mountfort et al. (1994)
5. Micellar electrokinetic chromatography Grains Wu et al. (1997)
6. Gas chromatography mass spectrometry (supercritical fluid extraction method) Fruit and vegetables Anastassiades and
Schwack (1998)
7. Poly-pyrrole modified glassy carbon glass Soil, water Manisankar et al. (2005a;
2005b)
8. Sodium montmorillonite clay-modified glassy carbon electrodes – Manisankar et al. 2005b
9. Surface-enhanced Raman scattering Soil, water Strickland and Batt
(2009)
10. Colorimetric technique using 4-aminobenzenethiol-functionalized silver
nanoparticles (ABT-Ag NPs)
Fruits, vegetables, soil
and water
Patel et al. (2015)
11. Nanoparticles with surface-enhanced Raman scattering – Furini et al. (2015)
12. Voltammetric determination with carbon electrode modified with multi-walled
carbon nanotubes
Orange juice Petroni et al. (2016)
13. Square wave voltammetric determination with carbon electrode containing
fullerene/multiwalled carbon nanotubes
Soil Teadoum et al. (2016)
14. Liquid chromatography coupled with Orbitrap high-resolution mass spectrometry
(UHPLC-HRMS)
Air, soil and water Lopez et al. (2016)
Environ Chem Lett
123
also received significant attention as an environmental
monitoring model for studying the deleterious effects of
carbendazim on various groups of living organisms
(Schaack 2008; Segner 2009; Stollewerk 2010; Jiang et al.
2015). In a recent study, carbendazim was detected by
liquid chromatography coupled with Orbitrap high-
Table 4 Adverse and toxic
effects of fungicide
carbendazim on the metabolic
and physiological processes of
vertebrates, invertebrates and
microorganisms
Category Adverse and toxic effects
Vertebrates and invertebrates including human Birth deformities- hydrocephalus and lack of eyes
Foetotoxicity
Testicular atrophy
Potent aneugen
Inhibition of polymerization of tubulin
Hormone disruption
Mitotic spindle abnormalities
Reduces metaphase and inter-centromere distance
Structural changes in liver
Deformation in kidney
Malformed testis
Microorganisms Distortion of the cellular structure
Reduction in enzymatic activities
Disruption of microbial community structure
Table 5 Microorganisms
involved in the biodegradation
of carbendazim under in situ
and experimental conditions
Microbial species Geographical location/region References
Aeromonas hydrophila Poland Kalwasinska et al. (2008a)
Alternaria alternata Israel Yarden et al. (1990)
Alternaria alternata Israel Yarden et al. (1985)
Azospirillum brasilense XJ-H China Lin et al. (2011a,b)
Bacillus pumilus NY97-1 China Zhang et al. (2009)
Bacillus subtilis TL-171 India Salunkhe et al. (2014)
Bacillus subtilis TS-204 India Salunkhe et al. (2014)
Bipolaris tetramera Israel Yarden et al. (1985)
Brevibacillus borstelensis India Arya and Sharma (2015)
Burkholderia cepacia Poland Kalwasinska et al. (2008a)
Nocardioides soli China Sun et al. (2014)
Nocardioides sp. strain SG-4G Australia Pandey et al. (2010)
Pseudomonas fluorescens Poland Kalwasinska et al. (2008b)
Pseudomonas luteola Poland Kalwasinska et al. (2008a)
Pseudomonas sp. CBW China Fang et al. (2012)
Pseudomonas sp. China Xiao et al. (2013)
Ralstonia sp. 1-1 China Zhang et al. (2005)
Rhodococcus erythropolis CB11 USA Holtman and Kobayashi (1997)
Rhodococcus erythropolis djl-11 China Zhang et al. (2013)
Rhodococcus jialingiae djl-6-2 China Wang et al. (2010a,b)
Rhodococcus qingshengii China Xu et al. (2007)
Rhodococcus qingshengii djl-6 China Xu et al. (2007)
Rhodococcus sp. China Xiao et al. (2013)
Rhodococcus sp. djl-6 China Jing-Liang et al. (2006)
Sphingomonas sp. China Xiao et al. (2013)
Sphingomonas paucimobilis Poland Kalwasinska et al. (2008b)
Streptomyces albogriseolus India Arya and Sharma (2015)
Trichoderma sp. T2-2China Tian and Chen (2009)
Environ Chem Lett
123
resolution mass spectrometry (Lopez et al. 2016). The
analysis of available reports indicates significant advance-
ment in the detection of carbendazim in environmental
samples. However, there is still an immense need to
develop ultrasensitive, real-time and robust detection
methods (Table 4).
Photolytic degradation of carbendazim
Carbendazim applied on the crops and fields reaches the
soil ecosystem and subsequently is met with different
disposal or transfer fates. It is usually transferred to the
whole plant through roots, stem or leaves. As repeated
applications of carbendazim at high dose are usually
required to control fungal diseases, a serious environmental
risk for human, livestock, aquatic animals and soil micro-
flora is being faced, which needs immediate attention.
Photolytic degradation is one of the several pathways
responsible for carbendazim derivatization. The hydroxyl
radicals generated by UV photolysis of H
2
O
2
are quenched
by hydrogeno-carbonate and carbonate ions leading to
formation of carbonate (CO
3
2-
) ions which are highly effi-
cient in carbendazim degradation (Mazellier et al. 2003).
The degradation of carbendazim is also feasible via com-
bination of TiO
2
-based photo-catalysis, ozonation and UV
light action (Rajeswari and Kanmani 2009). In another
study, two different types of titanium dioxide were studied
where aeroxide P25 showed better efficiency than LR.
Addition of catalyst resulted in complete mineralization
within 60 min (Kaur et al. 2015). Iron (Fe)-doped TiO2
nanoparticles (size 25–34 nm) showed better photocat-
alytic degradation of carbendazim due to the presence of Fe
as measured by UV-vis spectrophotometer (Kaur et al.
2016).
Microbial biodegradation of carbendazim
Several research reports indicate that accelerated efforts are
being taken to develop convenient, environmental friendly
and economically feasible methods for pesticides detoxi-
fication such as bioaugmentation by utilizing resident
microflora or supplementation with specific microbial
culture or consortia (Javorekova et al. 2010). Microbial
degradation of carbendazim is affected by several biotic
factors (presence of the plants, competing microbes) and
abiotic factors (pH, salts, type of soil, humidity, etc.). Only
few microbial strains are capable to tolerate and degrade
carbendazim in situ or under experimental conditions as
depicted in Table 5. Bacteria and fungi usually cleaved the
methyl carbamate side chain of carbendazim parent struc-
ture leading to the generation of 2-amino-benzimidazole,
benzimidazole and 2-hydroxybenzimidazole derivatives
(Table 1, Figs. 1and 2).
Genus Rhodococcus is endowed with extraordinary
abilities to utilize numerous recalcitrant and toxic com-
pounds for its growth and metabolism. Several strains of
Fig. 1 Common biodegradation pathway of carbendazim (with added
3D property). RMS (root mean square; line), exclusion sphere (range
in A
˚), centroid (average position) drawn with Chemdraw Ultra ver.
8.0. MW represents molecular weight
Environ Chem Lett
123
Rhodococcus viz. R. erythropolis CB11 (Holtman and
Kobayashi 1997), R. erythropolis djl-11 (Zhang et al.
2013), R. jialingiae djl-6-2 (Wang et al. 2010a,b), R.
qingshengii djl-6 (Xu et al. 2006,2007) and Rhodococcus
sp. (Jing-Liang et al. 2006; Xiao et al. 2013) showed
degradation of carbendazim in contaminated soils, in lab-
oratory pot experiments and in culture media. Pseu-
domonas is another bacterium having magnificent genetic
ability to utilize diverse carbon sources for its survival, and
it has already been exploited for bioremediation purposes.
P. luteola (Kalwasinska et al. 2008a) and P. fluorescens
(Kalwasinska et al. 2008b) are reported for carbendazim
mineralization activity. In another study, Pseudomonas sp.
strain CBW utilizing carbendazim as sole carbon and
nitrogen source was identified on the basis of 16 s rRNA
sequence homology and biological assays (Fang et al.
2012). Several species of Nocardioides, a filamentous
actinomycete, have been implicated in biodegradation of
carbendazim.
Nocardioides sp. strain SG-4G isolated from carbendazim
contaminated area exhibited hydrolysis of carbendazim into
2-amino-benzimidazole (Pandey et al. 2010). Further, N. soli
Fig. 2 Microorganism-mediated degradation of fungicide carben-
dazim in in situ and under experimental conditions. The initial
cleavage of methyl carbamate side chain of parent moiety by
microbes leads to the generation of 2-aminobenzimidazole,
2-hydroxybenzimidazole and benzimidazole derivatives which are
then further degraded to through a series of intermediate steps
depending on microbial species involved
Environ Chem Lett
123
strain mbc-2(T) was isolated from the soil in China which has
undergone repeated applications of carbendazim (Sun et al.
2014). Apart from these, Ralstonia sp. 1-1 (Zhang et al. 2005),
Bacillus pumilus NY97-1 (Zhang et al. 2009), B. subtilis
(Salunkhe et al. (2014), Aeromonas hydrophila,Burkholderia
cepacia (Kalwasinska et al. 2008a), Sphingomonas pauci-
mobilis (Kalwasinska et al. 2008b), Azospirillum brasilense
XJ-H (Lin et al. 2011a,b)andSphingomonas sp. (Xiao et al.
2013) were also found to be potent carbendazim metabolizing
organisms from the soils of China, India and Poland (Table 5).
Some fungal strains, Alternaria alternata,Bipolaris tetramera
(Yarden et al. 1985;1990)andTrichoderma sp. T2-2 (Tian
and Chen 2009) have also been associated with degradation of
carbendazim in experimental studies. In a recent study, Sedum
alfredii (a Cd hyperaccumulator plant) reportedly enhanced
the carbendazim-degrading action of bacterial strains
belonging to B. subtilis, Paracoccus sp., Flavobacterium and
Pseudomonas sp. in pot experiments. This enhanced degra-
dation was attributed to the stimulation of microbial activity
and community structure by S. alfredii in rhizosphere soil
(Xiao et al. 2013). Another study revealed that Brevibacillus
borstelensis and Streptomyces albogriseolus were degrading
98 and 91 % carbendazim, respectively, in 24 h. Their com-
bined inoculation was, however, even more effective as
indicated by 97 % degradation in just 12 h (Arya and Sharma
2015). It is evident from the above that only limited microbial
strains have the potential to utilize and hence dispose car-
bendazim from the environment. Carbendazim exhibits long-
term environmental persistence;therefore, highly efficient,
competent and ecologically competitive microbes are needed
to bioremediate the severely contaminated soil and water
reservoirs. Encouragingly, new strains are being increasingly
isolated from diverse resources endowed with superior
bioremediation characteristics. Future strategies based on
genetically engineered microorganisms, immobilized micro-
bial carriers, bioaugmentation and the use of activated soil are
expected to offer eco-friendly and sustainable solutions for
carbendazim bioremediation.
Conclusion
As the demand for pesticides is rapidly increasing world-
wide, environmental accumulation, contamination and life-
threatening effects on living organisms are inevitable. The
increasing use of carbendazim and its serious toxic effects
are considered ever-increasing threat to the ecological
balance of the environment. There is an urgent need for
concerted efforts to minimize the worldwide use of
fungicides, and at the same time, it is equally important to
recognize their impact of the living organisms, soil
ecosystem and microorganisms. The laboratory-scale and
on-field monitoring of carbendazim and its derivatives is
facilitated mainly by spectroscopic, chromatographic and
voltammetric techniques. Recently, colorimetric and
nanoparticles-based methodologies have been recognized
to offer increased accuracy and sensitivity. With the rapid
advancements in ‘omics’ technologies, further insights into
microbial community structure and intra- or inter-microbial
interactions in microenvironments of polluted soil and
water are expected to be revealed. Another promising
factor in carbendazim degradation will be to generate
transgenic organisms having improved enzymatic reper-
toire, broad substrate range and superior adaptability in
toxic environments. Nanotechnology is poised to give
greater contribution towards sensitive, rapid and repeat-
able monitoring and degradation of carbendazim and its
metabolites. The future availability of biofungicides and
biopesticides will certainly add to our efforts for curtailing
environmental pollution by minimizing the reliance on
chemical pesticides, herbicides and fungicides including
carbendazim.
Compliance with ethical standards
Conflicts of interest Authors declare that no conflicts of interest
exist.
References
Adedara IA, Vaithinathan S, Jubendradass R, Mathur PP, Farombi EO
(2013) Kolaviron prevents carbendazim-induced steroidogenic
dysfunction and apoptosis in testes of rats. Environ Toxicol
Pharmacol 35:444–453. doi:10.1016/j.etap.2013.01.010
Aire TA (2005) Short-term effects of carbendazim on the gross and
microscopic features of the testes of Japanese quails (Coturnix
coturnix japonica). Anat Embryol 210:43–49. doi:10.1007/
s00429-005-0001-0
Alvarez JLM, Calzo
´n JAG, Fonseca JML (1997) Catalytic polaro-
graphic prewave of cobalt (II) induced by carbendazim. Appli-
cation to the voltammetric determination of benomyl.
Electroanal 9:500–502. doi:10.1002/elan.1140090614
Anastassiades M, Schwack W (1998) Analysis of carbendazim,
benomyl, thiophanate methyl and 2,4-dichlorophenoxyacetic
acid in fruits and vegetables after supercritical fluid extraction.
J Chromatogr A 825:45–54. doi:10.1016/S0021-9673(98)00
691-8
Andrade TS, Henriques JF, Almeida AR, Machado AL, Koba O,
Giang PT, Soares AM, Domingues I (2016) Carbendazim
exposure induces developmental, biochemical and behavioural
disturbance in zebrafish embryos. Aquat Toxicol 170:390–399.
doi:10.1016/j.aquatox.2015.11.017
Arya R, Sharma AK (2015) Bioremediation of carbendazim, a
benzimidazole fungicide using Brevibacillus borstelensis and
Streptomyces albogriseolus together. Curr Pharm Biotechnol
17:185–189. doi:10.2174/1389201016666150930115737
Bakirci GT, Acay DB, Bakirci F, Otles S (2014) Pesticide residues in
fruits and vegetables from the Aegean region, Turkey. Food
Chem 160:379–392. doi:10.1016/j.foodchem.2014.02.051
Banyiova K, Necasova A, Kohoutek J, Justan I, C
ˇupr P (2016) New
experimental data on the human dermal absorption of simazine
and carbendazim help to refine the assessment of human
Environ Chem Lett
123
exposure. Chemosphere 145:148–156. doi:10.1016/j.chemo
sphere.2015.11.018
Barnett RM, Carone CD, Groom DE, Trippe TG, Wohl CG,
Armstrong B, Gee PS, Wagman GS, James F, Mangano M,
Monig K (1996) Review of particle physics. Phys Rev D 54:1.
doi:10.1103/PhysRevD.54.1
Bentley KS, Kirkland D, Murphy M, Marshall R (2000) Evaluation of
thresholds for benomyl-and carbendazim-induced aneuploidy in
cultured human lymphocytes using fluorescence in situ hybridiza-
tion. Mutat Res 464:41–51. doi:10.1016/S1383-5718(99)00165-5
Bernard MB, Cole P, Kobelt A, Horne PA, Altmann J, Wratten SD,
Yen AL (2010) Reducing the impact of pesticides on biological
control in Australian vineyards: pesticide mortality and fecun-
dity effects on an indicator species, the predatory mite Euseius
victoriensis (Acari: Phytoseiidae). J Eco Entomol 103:2061–
2071. doi:10.1603/EC09357
Bhushan C, Bhardwaj A, Misra SS (2013) State of pesticide
regulations in India. Report of Centre for Science and Environ-
ment, New Delhi
Bicchi C, Belliardo F, Cantamessa L, Gasparini G, Icardi M, Sesia E
(1989) Simultaneous determination of benzimidazole fungicides
by HPLC on apples, pears and their pulps. Pest Sci 25:355–360.
doi:10.1002/ps.2780250405
Canton JH (1976) The toxicity of benomyl, thiophanate-methyl, and
BCM to four freshwater organisms. Environ Contam Toxicol
16:214–218. doi:10.1007/BF01685230
Carter SD, Hess RA, Laskey JW (1987) The fungicide methyl
2-benzimidazole carbamate causes infertility in male Sprague-
Dawley rats. Biol Reprod 37:709–717. doi:10.1095/biolreprod37.
3.709
Chatupote W, Panapitukkul N (2005) Regional assessment of nutrient
and pesticide leaching in the vegetable production area of
Rattaphum catchment, Thailand. Water Air Soil Pollut Focus
5:165–173. doi:10.1007/s11267-005-7411-0
Correa LM, Nakai M, Strandgaard CS, Hess RA, Miller MG (2002)
Microtubules of the mouse testis exhibit differential sensitivity
to the microtubule disruptors carbendazim and colchicine.
Toxicol Sci 69:175–182. doi:10.1093/toxsci/69.1.175
Daam MA, Satapornvanit K, Van den BPJ, Nogueira AJ (2009)
Sensitivity of macroinvertebrates to carbendazim under semi-
field conditions in Thailand: implications for the use of
temperate toxicity data in a tropical risk assessment of fungi-
cides. Chemosphere 74:1187–1194. doi:10.1016/j.chemosphere.
2008.11.040
Dang Z, Smit CE (2008) Environmental risk limits for carbendazim.
Bilthoven, The Netherlands: National Institute for Public Health
and the Environment (RIVM). Letter Report (601716014)
EU Pesticide Database (2015) European Commission, (http://ec.
europa.eu/food/plant/pesticides/eu-pesticides-database/public/?ev
ent=homepageandlanguage=EN)
Daundkar PS, Rampal S (2014) Evaluation of ameliorative potential
of selenium on carbendazim induced oxidative stress in male
goats. Environ Toxicol Pharmacol 38:711–719. doi:10.1016/j.
etap.2014.09.007
Davidse LC (1986) Benzimidazole fungicides: mechanism of action
and biological impact. Ann Rev Phytopathol 24:43–65. doi:10.
1146/annurev.py.24.090186.000355
De A, Bose R, Kumar A, Mozumdar S (2014) Worldwide pesticide
use. In: Targeted Delivery of Pesticides Using Biodegradable
Polymeric Nanoparticles; Springer: New Delhi, India, pp 5–6
Devi PA, Paramasivam M, Prakasam V (2015) Degradation pattern and
risk assessment of carbendazim and mancozeb in mango fruits.
Environ Monit Assess 187:1–6. doi:10.1007/s10661-014-4142-6
Dikic D, Landeka I, Knez
ˇevic
´F, Mojsovic
´-C
´uic
´A, Benkovic
´V,
Horvat-Knez
ˇevic
´A, Rogic
´D (2012) Carbendazim impends
hepatic necrosis when combined with imazalil or cypermethrin.
Basic Clin Pharmacol Toxicol 110:433–440. doi:10.1111/j.1742-
7843.2011.00831.x
Fang H, Wang Y, Gao C, Yan H, Dong B, Yu Y (2012) Isolation and
characterization of Pseudomonas sp. CBW capable of degrading
carbendazim. Biodegradation 21:939–946. doi:10.1007/s10532-
010-9353-0
FAO (1998) Pesticides residues in food-1997 evaluations. Part I
Residues, FAO Plant production and protection paper, Rome,
p 146
Farag A, Ebrahim H, ElMazoudy R, Kadous E (2011) Developmental
toxicity of fungicide carbendazim in female mice. Birth Defe
Res B: Develop Reprod Toxicol 92:122–130. doi:10.1002/bdrb.
20290
Fawole OB, Aluko M, Olowonihi TE (2010) Effects of a Carben-
dazim-Mancozeb fungicidal mixture on soil microbial popula-
tions and some enzyme activities in soil. Agrosearch 10:1–2.
doi:10.4314/agrosh.v10i1-2.69831
Ferreira AL, Loureiro S, Soares AM (2008) Toxicity prediction of
binary combinations of cadmium, carbendazim and low dis-
solved oxygen on Daphnia magna. Aqua Toxicol 89:28–39.
doi:10.1016/j.aquatox.2008.05.012
Furini LN, Sanchez-Cortes S, Lo
´pez-Toco
´n I, Otero JC, Aroca RF,
Constantino CJ (2015) Detection and quantitative analysis of
carbendazim herbicide on Ag nanoparticles via surface-enhanced
Raman scattering. J Raman Spectr 46:1095–1101. doi:10.1002/
jrs.4737
Goodson WH, Lowe L, Carpenter DO, Gilbertson M, Ali AM, de
Cerain Salsamendi AL et al (2015) Assessing the carcinogenic
potential of low-dose exposures to chemical mixtures in the
environment: the challenge ahead. Carcinogenesis 36:254–296.
doi:10.1093/carcin/bgv039
Gray LE, Ostby J, Linder R, Goldman J, Rehnberg G, Cooper R
(1990) Carbendazim-induced alterations of reproductive devel-
opment and function in the rat and hamster. Fund Appl Toxicol
15:281–297. doi:10.1016/0272-0590(90)90055-O
Gupta RC, Aggarwal M (2007) Toxicity of fungicides. Veterinary
Toxicology, New York
Hammond LA, Davidson K, Lawrence R, Camden JB, Von Hoff DD,
Weitman S, Izbicka E (2001) Exploring the mechanisms of
action of FB642 at the cellular level. J Cancer Res Clin Oncol
127:301–313
Hao LQ (2002) Advances on the pesticide residue analysis of Chinese
herbal medicine. J Huazhong Agric 2:20–24
Holtman MA, Kobayashi DY (1997) Identification of Rhodococcus
erythropolis isolates capable of degrading the fungicide carben-
dazim. Appl Microbiol Biotechnol 47:578–582. doi:10.1007/
s002530050976
Hsu YH, Chang CW, Chen MC, Yuan CY (2011) Carbendazim-
induced androgen receptor expression antagonized by flutamide
in male rats. J Food Drug Anal 19:4
Huan Z, Luo J, Xu Z, Xie D (2016) Acute toxicity and genotoxicity of
carbendazim, main impurities and metabolite to earthworms
(Eisenia foetida). Bull Environ Contam Toxicol 96:62–69.
doi:10.1007/s00128-015-1653-y
Itak JA, Selisker MY, Jourdan SW, Fleeker JR, Herzog DP (1993)
Determination of benomyl (as carbendazim) and carbendazim in
water, soil, and fruit juice by a magnetic particle-based
immunoassay. J Agric Food Chem 41:2329–2332. doi:10.1021/
jf00036a021
Janardhan A, Rao AB, Sisodia P (1987) Sub-chronic toxicity of
methyl benzimidazole carbamate in rats. Bull Environ Contam
Toxicol 38:890–898. doi:10.1007/BF01616717
Jansen JP (1999) Side effects of pesticides on adults of Aphidius
rhopalosiphi De Stefani-Perez (Hym.: Aphidiidae) in the labo-
ratory: results of the 8th Joint Pesticide Testing Programme.
Versailles, France 27–29 October 23:65–72
Environ Chem Lett
123
Javorekova SN, Svrcbreve ekova I, Makova J (2010) Influence of
benomyl and prometryn on the soil microbial activities and
community structures in pasture grasslands of Slovakia. J Envi-
ron Sci Health B 45:702–709. doi:10.1080/03601234.2010.
502463
Jiang J, Wu S, Wang Y, An X, Cai L, Zhao X, Wu C (2015)
Carbendazim has the potential to induce oxidative stress,
apoptosis, immunotoxicity and endocrine disruption during
zebrafish larvae development. Toxicol In Vitro 29:1473–1481.
doi:10.1016/j.tiv.2015.06.003
Jing-Liang X, Xiang-Yang G, Biao S, Zhi-Chun W, Kun W, Shun-
Peng L (2006) Isolation and characterization of a carbendazim-
degrading Rhodococcus sp. djl-6. Curr Microbiol 53:72–76.
doi:10.1007/s00284-005-0474-3
John EM, Shaike JM (2015) Chlorpyrifos: pollution and remediation.
Environ Chem Lett 13:269–291. doi:10.1007/s10311-015-
0513-7
Jones SE, Williams DJ, Holliman PJ, Taylor N, Baumann J, Forster B,
Van Gestel CAM, Rodrigues JML (2004) Ring-testing and field-
validation of a terrestrial model ecosystem (TME)-an instrument
for testing potentially harmful substances: fate of the model
chemical carbendazim. Ecotoxicol 3:29–42. doi:10.1023/B:
ECTX.0000012403.90709.c9
Kalwasin
´ska A, Ke˛sy J, Donderski W, Lalke-Porczyk E (2008a)
Biodegradation of carbendazim by planktonic and benthic
bacteria of eutrophic lake Chełm_
zyn
´skie. Pol J Environ Stud
17:515–523
Kalwasin
´ska A, Ke˛sy J, Donderski W (2008b) Biodegradation of
carbendazim by epiphytic and neustonic bacteria of eutrophic
chełm_
zyn
´skie lake. Pol J Microbiol 57:221–230
Karlsson I, Friberg H, Steinberg C, Persson P (2014) Fungicide
effects on fungal community composition in the wheat phyllo-
sphere. PLoS One 9:e111786. doi:10.1371/journal.pone.0111786
Kaur T, Toor AP, Wanchoo RK (2015) Parametric study on
degradation of fungicide carbendazim in dilute aqueous solutions
using nano TiO
2
. Desalination Water Treat 54:122–131. doi:10.
1080/19443994.2013.879081
Kaur T, Sraw A, Toor AP, Wanchoo RK (2016) Utilization of solar
energy for the degradation of carbendazim and propiconazole by
Fe doped TiO
2
. Sol Energy 125:65–66. doi:10.1016/j.solener.
2015.12.001
Kosasa T, Kuriya Y, Matsui K, Yamanishi Y (1999) Effect of
donepezil hydrochloride (E2020) on basal concentration of
extracellular acetylcholine in the hippocampus of rats. Eur J
Pharmacol 380:101–107. doi:10.1016/S0014-2999(99)00545-2
Kumar M (2001) Acute oral toxicity study with carbendazim
technical 98% in Wistar rats. Toxicology Department, Rallis
Research Centre, Bangalore, India. Report number: 3179/1.
Report date: 16 August 2001
Lacey E, Watson TR (1985) Structure-activity relationships of
benzimidazole carbamates as inhibitors of mammalian tubulin
in vitro. Biochem Pharmacol 34:1073–1077. doi:10.1016/0006-
2952(85)90611-2
Lin X, Za Hou, Zhao SF, Xie L, Li HJ, Li MZ (2011a) A high
efficient carbendazim-degrading bacterial strain: its isolation and
identification. Chin J Ecol 7:031
Lin X, Hou Z, Feng Y, Zhao S, Ye J (2011b) Isolation and
characteristics of efficient carbendazim degradation bacterium.
In: International Conference on Agricultural and Biosystems
Engineering, Advances in Biomedical Engineering. 1:384–388
Liu K, Pan X, Han Y, Tang F, Yu Y (2012) Estimating the toxicity of
the weak base carbendazim to the earthworm (Eisenia fetida)
using in situ pore water concentrations in different soils. Sci
Total Environ 438:26–32. doi:10.1016/j.talanta.2015.11.068
Lopez A, Yusa
`V, Millet M, Coscolla
`C (2016) Retrospective
screening of pesticide metabolites in ambient air using liquid
chromatography coupled to high-resolution mass spectrometry.
Talanta 150:27–36. doi:10.1016/j.talanta.2015.11.068
Lu SY, Liao JW, Kuo ML, Wang SC, Hwang JS, Ueng TH (2004)
Endocrine-disrupting activity in carbendazim-induced reproduc-
tive and developmental toxicity in rats. J Toxicol Env Heal A
67:1501–1515. doi:10.1080/15287390490486833
Ludwikowska A, Bojarski B, Socha M, Lutnicka H, Trzeciak KB
(2013) The effect of carbendazim on embryonic Prussian carp
(Carassius gibelio) development and hatching. Arch Polish
Fisheries 21:367–371. doi:10.2478/aopf-2013-0038
Ma J, Zheng R, Xu L, Wang S (2002) Differential sensitivity of two
green algae, Scenedesmus obliqnus and Chlorella pyrenoidosa to
12 pesticides. Ecotoxicol Environ Saf 52:57–61. doi:10.1006/
eesa.2002.2146
Manisankar P, Selvanathan G, Vedhi C (2005a) Utilisation of
polypyrrole modified electrode for the determination of pesti-
cides. Int J Environ Anal Chem 85:409–422. doi:10.1080/
03067310500050726
Manisankar P, Selvanathan G, Vedhi C (2005b) Utilization of sodium
montmorillonite clay-modified electrode for the determination of
isoproturon and carbendazim in soil and water samples. Appl
Clay Sci 29:249–257. doi:10.1016/j.clay.2005.01.006
Mantovani A, Maranghi F, Ricciardi C, Macrı
`C, Stazi AV, Attias L,
Zapponi GA (1998) Developmental toxicity of carbendazim:
comparison of no-observed-adverse-effect level and benchmark
dose approach. Food Chem Toxicol 36:37–45. doi:10.1016/
S0278-6915(97)00116-6
Mazellier P, Leroy E, De Laat J, Legube B (2003) Degradation of
carbendazim by UV/H
2
O
2
investigated by kinetic modelling.
Environ Chem Lett 1:68–72. doi:10.1007/s10311-002-0010-7
Minta MA, Wilk IW,
_
Zmudzki J (2004) Embryotoxicity of carben-
dazim in rat and hamster micromass cultures. Bull Vet Inst
Pulawy 48:481–484
Morinaga H, Yanase T, Nomura M, Okabe T, Goto K, Harada N,
Nawata H (2004) A benzimidazole fungicide, benomyl, and its
metabolite, carbendazim, induce aromatase activity in a human
ovarian granulose-like tumor cell line (KGN). Endocrinology
145:1860–1869. doi:10.1210/en.2003-1182
Mountfort KA, Reynolds SL, Thorpe SA, White SN (1994) Compar-
ison of ELISA and HPLC techniques for the analysis of
carbendazim and thiabendazole residues in fruit and vegetables.
Food Agric Immunol 6:17–22. doi:10.1080/09540109409354
808
Mullin CA, Frazier M, Frazier JL, Ashcraft S, Simonds R, Pettis JS
(2010) High levels of miticides and agrochemicals in North
American apiaries: implications for honey bee health. PLoS One
5(3):e9754. doi:10.1371/journal.pone.0009754
Muthuviveganandavel V, Muthuraman P, Muthu S, Srikumar K
(2008) Toxic effects of carbendazim at low dose levels in male
rats. J Toxicol Sci 33:25–30. doi:10.2131/jts.33.25
Naidu PK, Niranjan T, Naidu NVS (2011) Spectrophotometric
determination of carbendazim in its formulations and environ-
mental samples. Int J Chem Tech Res 3:1728–1733
Niewiadomska A (2004) Effect of carbendazim, imazetapir and
thiram on nitrogenase activity, the number of microorganisms in
soil and yield of red clover (Trifolium pratense L.). Polish J
Environ Studies 13:403–410
Olayemi OA (2015) Comparative toxicity of two different pesticides
on the skin of Japanese quail (Cortunix japonica). World Vet J
5:13–18
Pacheco SE, Anderson LM, Sandrof MA, Vantangoli MM, Hall SJ,
Boekelheide K (2012) Sperm mRNA transcripts are indicators of
sub-chronic low dose testicular injury in the Fischer rat. PLoS
One 7:e44280. doi:10.1371/journal.pone.0044280
Palanikumar L, Kumaraguru AK, Ramakritinan CM, Anand M (2014)
Toxicity, biochemical and clastogenic response of chlorpyrifos
Environ Chem Lett
123
and carbendazim in milkfish Chanos chanos. Int J Environ Sci
Technol 11:765–774. doi:10.1007/s13762-013-0264-6
Pandey G, Dorrian SJ, Russell RJ, Brearley C, Kotsonis S, Oakeshott
JG (2010) Cloning and biochemical characterization of a novel
carbendazim (Methyl-1H-Benzimidazol-2-ylcarbamate)-hy-
drolyzing esterase from the newly isolated Nocardioides sp.
strain SG-4G and its potential for use in enzymatic bioremedi-
ation. Appl Environ Microbiol 76:2940–2945. doi:10.1128/
AEM.02990-09
Parsons KC, Mineau P, Renfrew RB (2010) Effects of pesticide use in
rice fields on birds. Waterbirds 33:193–218. doi:10.1675/063.
033.s115
Patel GM, Rohit JV, Singhal RK, Kailasa SK (2015) Recognition of
carbendazim fungicide in environmental samples by using
4-aminobenzenethiol functionalized silver nanoparticles as a
colorimetric sensor. Sensor Actuat B-Chem 206:684–691.
doi:10.1016/j.snb.2014.09.095
Petroni JM, Lucca BG, Fogliato DK, Ferreira VS (2016) Sensitive
approach for voltammetric determination of carbendazim based
on the use of an anionic surfactant. Electroanalysis [published
online]. doi: 10.1002/elan.201501069
Pourreza N, Rastegarzadeh S, Larki A (2015) Determination of
fungicide carbendazim in water and soil samples using disper-
sive liquid-liquid microextraction and microvolume UV-vis
spectrophotometry. Talanta 134:24–29. doi:10.1016/j.talanta.
2014.10.056
Prashantkumar W, Sethi RS, Pathak D, Rampal S, Saini SP (2012)
Testicular damage after chronic exposure to carbendazim in
male goats. Toxicol Environ Chem 94:1433–1442. doi:10.1080/
02772248.2012.693493
Rajeswari R, Kanmani S (2009) TiO
2
-based heterogeneous photocat-
alytic treatment combined with ozonation for carbendazim
degradation. Iran J Environ Health Sci Eng 6:61–66
Rama EM, Bortolan S, Vieira ML, Gerardin DC, Moreira EG (2014)
Reproductive and possible hormonal effects of carbendazim.
Regul Toxicol Pharmacol 69:476–486. doi:10.1016/j.yrtph.2014.
05.016
Rico A, Sabater C, Castillo MA
´(2016) Lethal and sub-lethal effects
of five pesticides used in rice farming on the earthworm Eisenia
fetida. Ecotoxicol Environ Saf 127:222–229. doi:10.1016/j.
ecoenv.2016.02.004
Sakr S, Shalaby SY (2014) Carbendazim-induced testicular damage
and oxidative stress in albino rats: ameliorative effect of licorice
aqueous extract. Toxicol Ind Health 230:259–267. doi:10.1177/
0748233712456059
Salihu M, Ajayi BO, Adedara IA, Farombi EO (2015) 6-Gingerol-rich
fraction from Zingiber officinale prevents hematotoxicity and
oxidative damage in kidney and liver of rats exposed to
carbendazim. J Diet Suppl 16:1–16. doi:10.3109/19390211.
2015.1107802
Salunkhe VP, Sawant IS, Banerjee K, Wadkar PN, Sawant SD,
Hingmire SA (2014) Kinetics of degradation of carbendazim by
B. subtilis strains: possibility of in situ detoxification. Environ
Monitor Assess 186:8599–8610. doi:10.1007/s10661-014-4027-
8
Schaack S (2008) Daphnia comes of age: an ecological model in the
genomic era. Mol Ecol 17:1634–1635. doi:10.1111/j.1365-294X.
2008.03698.x
Segner H (2009) Zebrafish (Danio rerio) as a model organism for
investigating endocrine disruption. Comp Biochem Physiol Part
C: Toxicol Pharmacol 149:187–195. doi:10.1016/j.cbpc.2008.10.
099
Selmanoglu G, Barlas N, Songu
¨r S, KocSkaya EA (2001) Carben-
dazim-induced haematological, biochemical and histopatholog-
ical changes to the liver and kidney of male rats. Hum Exp
Toxicol 20:625–630. doi:10.1191/096032701718890603
Sharma KK (2007) Pesticide residue analysis manual. Directorate of
Information and Publications of Agriculture. Indian Council of
Agricultural Research, New Delhi
Silva AR, Cardoso DN, Cruz A, Lourenc¸o J, Mendo S, Soares AM,
Loureiro S (2015) Ecotoxicity and genotoxicity of a binary
combination of triclosan and carbendazim to Daphnia magna.
Ecotoxicol Environ Saf 115:279–290. doi:10.1016/j.ecoenv.
2015.02.022
Solomon KR, Brock TC, De Zwart D, Dyer SD, Posthuma L,
Richards S, van den Brink PJ (2008) (eds). Extrapolation
practice for ecotoxicological effect characterization of chemi-
cals. CRC Press
Stollewerk A (2010) The water flea Daphnia—a ‘new’model system
for ecology and evolution. J Biol 9:21. doi:10.1186/jbiol212
Strickland AD, Batt CA (2009) Detection of carbendazim by surface-
enhanced Raman scattering using cyclodextrin inclusion com-
plexes on gold nanorods. Anal Chem 81:2895–2903. doi:10.
1021/ac801626x
Sun LN, Zhang J, Gong FF, Wang X, Hu G, Li SP, Hong Q (2014)
Nocardioides soli sp. nov., a carbendazim-degrading bacterium
isolated from soil under the long-term application of carben-
dazim. Int J Syst Evol Microbiol 64:2047–2052. doi:10.1099/ijs.
0.057935-0
Teadoum DN, Noumbo SK, Arnaud KT, Ranil TT, Ze
´ADM, Tonle
IK (2016) Square wave voltammetric determination of residues
of carbendazim using a Fullerene/Multiwalled carbon nanotubes/
Nafion
Ò
/coated glassy carbon electrode. Int J Electrochem.
doi:10.1155/2016/7839708
Tian L, Chen F (2009) Characterization of a carbendazim-degrading
Trichoderma sp. T2-2 and its application in bioremediation. Wei
Sheng Wu Xue Bao 49:925–930
Torstensson L, Wessen B (1984) Interactions between the fungicide
benomyl and soil microorganisms. Soil Biol Biochem
16:445–452. doi:10.1016/0038-0717(84)90050-6
Tortella GR, Mella-Herrera RA, Sousa DZ, Rubilar O, Bricen
˜oG,
Parra L, Diez MC (2013) Carbendazim dissipation in the
biomixture of on-farm biopurification systems and its effect on
microbial communities. Chemosphere 93:1084–1093
Van Wijngaarden RPA, Crum SJH, Decraene K, Hattink J, Van
Kammen A (1998) Toxicicity of derosal (active ingredient
carbendazim) to aquatic invertebrates. Chemosphere
37:673–683. doi:10.1016/S0045-6535(98)00083-6
Wang X, Song M, Gao C, Dong B, Zhang Q, Fang H, Yu Y (2009a)
Carbendazim induces a temporary change in soil bacterial
community structure. J Environ Sci (China) 21:1679–1683.
doi:10.1016/S1001-0742(08)62473-0
Wang YS, Huang YJ, Chen WC, Yen JH (2009b) Effect of
carbendazim and pencycuron on soil bacterial community.
J Hazard Mat 172:84–91. doi:10.1016/j.jhazmat.2009.06.142
Wang Z, Xu J, Li Y, Wang K, Wang Y, Hong Q, Li WJ, Li SP
(2010a) Rhodococcus jialingiae sp. nov., an actinobacterium
isolated from sludge of a carbendazim wastewater treatment
facility. Int J Syst Evol Microbiol 60:378–381. doi:10.1099/ijs.0.
013219-0
Wang Z, Wang Y, Gong F, Zhang J, Hong Q, Li S (2010b)
Biodegradation of carbendazim by a novel actinobacterium
Rhodococcus jialingiae djl-6-2. Chemosphere 81:639–644.
doi:10.1016/j.chemosphere.2010.08.040
Wang X, Song M, Wang Y, Gao C, Zhang Q, Chu X, Fang H, Yu Y
(2012) Response of soil bacterial community to repeated
applications of carbendazim. Ecotoxicol Environ Saf 75:33–39.
doi:10.1016/j.ecoenv.2011.08.014
Wang C, Wang F, Zhang Q, Liang W (2016) Individual and combined
effects of tebuconazole and carbendazim on soil microbial
activity. Eur J Soil Biol 72:6–13. doi:10.1016/j.ejsobi.2015.12.
005
Environ Chem Lett
123
Wu YS, Lee HK, Li SFY (1997) Determination of carbendazim
residues in grains by solid-phase extraction and micellar
electrokinetic chromatography with ultraviolet detection. J Chro-
matogr Sci 35(11):513–518. doi:10.1093/chromsci/35.11.513
Xiao W, Wang H, Li T, Zhu Z, Zhang J, He Z, Yang X (2013)
Bioremediation of Cd and carbendazim co-contaminated soil by
Cd-hyperaccumulator Sedum alfredii associated with carben-
dazim-degrading bacterial strains. Environ Sci Pollut Res
20:380–389. doi:10.1007/s11356-012-0902-4
Xu JL, Wang ZC, Wang K, Li SP (2006) The isolation identification
and degradation characters of an efficient carbendazim-degrad-
ing bacterium. China Environ Sci 26(3):307–310
Xu JL, He J, Wang ZC, Wang K, Li WJ, Tang SK, Li SP (2007)
Rhodococcus qingshengii sp. nov., a carbendazim-degrading
bacterium. Int J Syst Evol Microbiol 57:2754–2757. doi:10.
1099/ijs.0.65095-0
Yan H, Wang D, Dong B, Tang F, Wang B, Fang H, Yu Y (2011)
Dissipation of carbendazim and chloramphenicol alone and in
combination and their effects on soil fungal: bacterial ratios and
soil enzyme activities. Chemosphere 84:634–641. doi:10.1016/j.
chemosphere.2011.03.038
Yarden O, Katan J, Aharonson N, Ben-Yephet Y (1985) Delayed and
enhanced degradation of benomyl and carbendazim in disin-
fected and fungicide-treated soils. Phytopathol 75:763–777
Yarden O, Salomon R, Katan J, Aharonson N (1990) Involvement of
fungi and bacteria in enhanced and nonenhanced biodegradation
of carbendazim and other benzimidazole compounds in soil. Can
J Microbiol 36:15–23. doi:10.1139/m90-004
Yenjerla M, Cox C, Wilson L, Jordan MA (2009) Carbendazim
inhibits cancer cell proliferation by suppressing microtubule
dynamics. J Pharmacol Exp Ther 328:390–398. doi:10.1124/jpet.
108.143537
Yoon CS, Jin JH, Park JH, Yeo CY, Kim SJ, Hwang YG, Hong SJ,
Cheong SW (2008) Toxic effects of carbendazim and n-butyl
isocyanate, metabolites of the fungicide benomyl, on early
development in the African clawed frog, Xenopus laevis.
Environ Toxicol 23:131–144. doi:10.1002/tox.20338
Yunlong YU, Xiaoqiang CHU, Guohui PANG, Xiang Y, Hua FANG
(2009) Effects of repeated applications of fungicide carbendazim
on its persistence and microbial community in soil. J Environ Sci
21(2):179–185. doi:10.1016/S1001-0742(08)62248-2
Zamora DP, Galera MM, Frenich AG, Vidal JM (2000) Trace
determination of carbendazim, fuberidazole and thiabendazole in
water by application of multivariate calibration to cross-sections
of three-dimensional excitation–emission matrix fluorescence.
Analyst 125(6):1167–1174. doi:10.1039/A909886K
Zhang GS, Jia XM, Cheng TF, Ma XH, Zhao YH (2005) Isolation and
characterization of a new carbendazim-degrading Ralstonia sp.
strain. World J Microbiol Biotechnol 21:265–269. doi:10.1007/
s11274-004-3628-8
Zhang L, Qiao X, Ma L (2009) Influence of environmental factors on
degradation of carbendazim by Bacillus pumilus strain NY97-1.
Int J Environ Pollut 38:309–317. doi:10.1504/IJEP.2009.027231
Zhang X, Huang Y, Harvey PR, Li H, Ren Y (2013) Isolation and
characterization of carbendazim-degrading Rhodococcus ery-
thropolis djl-11. PLoS One 8:1–6. doi:10.1371/journal.pone.
0074810
Zhu SH, Wu HL, Li BR, Xia AL, Han QJ, Zhang Y, Yu RQ (2008)
Determination of pesticides in honey using excitation–emission
matrix fluorescence coupled with second-order calibration and
second-order standard addition methods. Anal Chim Acta
619:165–172. doi:10.1016/j.aca.2008.05.005
Zubrod JP, Baudy P, Schulz R, Bundschuh M (2014) Effects of
current-use fungicides and their mixtures on the feeding and
survival of the key shredder Gammarus fossarum. Aquatic
Toxicol 150:133–143. doi:10.1016/j.aquatox.2014.03.002
Environ Chem Lett
123
