Available online at http://www.urpjournals.

com

International Journal of Research in Pure and Applied Microbiology

Universal Research Publications. All rights reserved

ISSN 22773843

Original Article
Biological control of Calotropis gigantea leaf extracts against pathogenic fungus,
infecting Oryza sativa
Rajendhiran Viji1, Palanisamy Alaguraja2, Panagal Mani3. Velavan S4
1
Department of Biotechnology, Ponnaiyah Ramajayam Arts and science college, Thanjavur, Affiliated for Bharathidasan
University, Tiruchirappalli, Tamil Nadu, India, Pin: 620024.
2
Department of Geology, Bharathidasan University, Tiruchirappalli, Tamil Nadu, India, Pin: 620024.
3
The Sharmila Institute of Medicinal Products Research Academy (SIMPRA), Thanjavur, Affiliated for Bharathidasan
University, Tiruchirappalli, Tamil Nadu, India, Pin: 620024
4
Department of Biochemistry, Marudhupandiyar College, Thanjavur.
Corresponding author: master.maniji@gmail.com
Received 18 September 2013; accepted 03 October 2013
Abstract
The medicinal plant extracts biologically preventing fungal pathogens impacts of the cultivation rice field. These
approaches are traditionally several years used for plants various parts like leaf, stem, root, fruit and seeds are effectively
control pathogens. In this study, biologically control of rice crop affecting major fungal pathogen strains were tested for
their survival against leaf extracts of Calotropis gigantea plant commonly called Erukku in Tamilnadu, India. The infected
rice plant leaves were collected from various locations of paddy field the fungal strains infecting the plant were isolated
and identified as Aspergillus species, Penicillum species and Aspergillus niger. The medicinal plant leaves were collected
from the area surrounding the paddy field. The test plant leaves extracts were prepared by acetone and sterile distilled
water and examined for antifungal activity by agar well diffusion methods for minimum inhibition of fungal growth zones.
Results indicated that acetone extracts of Calotropis gigantea effectively controlled the growth of Aspergillus sp 7.1 mm,
Penicillum sp 7.4 mm, and A. niger 7.0 mm inhibition zones respectively, which were higher than aqueous extract
Aspergillus sp 10.5 mm, Penicillum sp 10.8 mm, and A. niger 10.7 mm of test plant. In future, these extracts may be used
as valuable agents for biological control and agro-eco-friendly pesticide various cultivation fields.
2013 Universal Research Publications. All rights reserved
Key words: Rice, Fungus, Antifungal activity, Calotropis gigantea, Thanjavur
INTRODUCTION
Rice is one of the important crops in the world, providing
more calories for the human beings who are consuming
around the world and its consumption has increased
significantly, because of the increasing population.
Currently, the growth of rice plants is highly affected by
pathogenic fungi, which leads to scarcity that is becoming a
global issue (FAO, 2009; Mahender et al., 2012). The
UN/FAO report stated that food production needs to be
increased by 40% in 2030 and 70% in 2050 (FAO, 2009).
In addition, because of food scarcity, more than 925 million
people are dying (FAO, 2010). Therefore, global level rice
production is immensely required for the survival of human
being. Especially, 96% of rice has been produced by Asian
countries (IRRI, 2011: http://irri.org). In the past few
decades, rice production is affected by various factors;
particularly, 20% to 30% of rice production is significantly
affected by microbial diseases worldwide (Savary et al.,
2012). Particularly, fungal pathogen infection is one of the

107

major problems in rice production, which is the most

naturally occurring abundant pathogenic fungal species in
paddy field. It can also grow in a little nutrient available
environment with potential to survive and tolerate
(Manandhar et al., 2012). Furthermore, 11% to 30% crop
lost per annum is because of the fungal pathogens impacts,
which represents a yield loss about 157 million tones as
reported by global scenarios (ICAR, 2012). The paddy
fields are vigorously affected by mycotoxins, such as
aflatoxins (Aspergillus), ochratoxins (Aspergillus and
Penicillium), and trichothecenes and fumonisins
(Fusarium) (Perrone et al., 2007). Because of the infection
of crops with these mycotoxins, million dollars of
economic losses in crop yield has been reported (Council
for Agricultural Science and Technology, 2003). These
impacts on the health of human beings estimated that 4.5
billion people population were exposed to aflatoxins in
worldwide (Williams et al., 2007). Fungal disease control
plays a crucial part in cultivation of paddy field in global

International Journal of Research in Pure and Applied Microbiology 2013; 3(4): 107-112

level. Therefore, Worldwide there is increasing demand for

high-quality and safe food, free of chemical and physical
contaminants and pathogens. Chemical pesticides were
applied to the cultivation plants, but they are harmful.
Excessive use of fungicides resulted in accumulation of
toxic compounds affecting soil physicochemical, micro and
macronutrients, and soil fertility; and ingesting them causes
adverse effects to humans (WHO, 1990; Shetty et al.,
2012). The increasing demand of production and
regulations on the use of agrochemicals have lead to the
emergence of pathogens resistant to the products employed,
justifying the search for novel, active molecules and new
control strategies. Hence, to find an alternative method to
control the fungal pathogen is of the current need
(Dellavalle et al., 2011). Plant metabolites and plant-based
pesticides appear to be one of the better alternatives as they
are known to have minimal environmental impact and
danger to consumers in contrast to the synthetic pesticides
(Varma and Dubey, 1999; Mohana et al., 2011). On earth,
lakhs of medicinal plants are available (Schippmann et al.,
2002). Especially, India has medicinal flora counting more
than 7500 species. These medicinal plants offer low-cost
and safety health care solutions; these plants have potential
to control pathogenic microorganisms (Preethi et al., 2010;
Prakasha et al., 2010; Mazid et al., 2012). The C. gigantea
is a milkweed wasteland plant distributed in tropical and
subtropical regions of Asia and Africa, which is medicinal
plant it belongs to the family of Asclepiadaceous. The plant
growth range is maximum 3 meter long, It has oral, light
green leaves are very much succulent in nature and milky
colour stem and it not consumed by grazing animals. These
plant extracts have immense of value for agriculture plants,
by which pathogenic microbes bacteria, fungi, and
insecticidal are effectively controlled (Pathak and Argal,
2007) and human disease and infection control such as
milky sap is used to stop bleeding, and for treating
boils, scabies, burns, bruises, cuts, sores, wounds chest
colds and heart conditions. Tamilnadu plays a crucial part
in rice production of India-23.3% of region cultivates rice.
Especially, in Thanjavur district, rice cultivation is
important in southern Peninsula, because 60% of the area
cultivates paddy. Therefore, this study has been performed
to analyze the biological control of wasteland medicinal
plant, C. gigantea, over rice crop-affecting major fungal
pathogens.
MATERIAL AND METHODS
Description of study area
Thanjavur district is popularly known as rice bowl in
Tamilnadu, located on the eastern coast of Tamilnadu,
India (Fig 1), Latitude 10o 47 N and longitude 79o 10 E.
The district is world famous for location of the big temple
(Brihadishwara Temple) built by Rajarajacholan during the
11th century; the temple is one of UNESCO world heritage
sites. Total people population of district is 22.16 lakhs and
66% of people live in rural areas. Moreover, most of the
population here is depended upon on agricultures including
1, 44, 942 of total cultivators and 4, 10, 718 of agricultural
laborers. The specialty of this district is the large Cauvery
River flows through it, which drenches the Southern
Peninsula of India. This river water is immensely

108

supporting to the agriculture activities. Therefore, it has

been named as the Mother Cauvery and considered as
sacred by the people here. Out of 3, 39, 657 of total hectors
in this district, 2, 41, 292 hector areas are used for
agriculture purpose. Rice is the major cultivation crop in
this district, cultivated in three seasons such as Kuruvai,
Samba, and Thaladi. Kuruvai contributes 17.7% of
cultivation
with
duration
from
June/July
to
October/November (100115 days); Samba contributes
63% of cultivation from December/January to March/April;
and at end of the year, Thaladi contributes 15.6% of
cultivation with duration from October/November to
February/March (110125 days). In addition, support to
rice cultivation is through annual rainfall, 1053 mm
normally, and temperatures, maximum 36.6 oC and
minimum 22.8oC. More than 80% of lands in the district
are used for cultivation by the small farmers, in which less
than five hectares are used for paddy cultivation.

Fig 1. Thanjavur located on the eastern coast of Tamilnadu,

India

Fig 2. Shows Inhibition zone of Aspergillus sp

Sample collection, Isolation, and Identification of fungal
strains
The infected fresh rice plant leaves sample were collected
from paddy field in Mariamman Kovil villages (Kothattai,
Kalakudi, and Kulichiappatu), Thanjavur district,
Tamilnadu, India. Collected sample were stored in ice box
at 4C and sample were processed within 3 h. Uninfected
fresh C. gigantea plant leaves were collected from natural

International Journal of Research in Pure and Applied Microbiology 2013; 3(4): 107-112

population growing in the paddy field near wasteland

areas. The C. gigantea plant voucher specimen, BDUT
1284, Herbarium was deposited in Department of Botany,
School of Plant Science, Bharathidasan University,
Tiruchirappalli, Tamilnadu, India. The infected leaves
weighted 1 g, and to which 1 ml sterile distilled water was
added in mortar pestle, the sample was well crushed, and
collected 1 ml of sample was dissolved in 9 ml of sterile
distilled water. The test tube was shaken thoroughly to get
the uniform distribution of the leaf sample. The samples
were serially diluted from 102 to 105 1-ml of the diluted
and plated onto sterilized Potato Dextrose Agar medium
(PDA) supplemented with 1% streptomycin (1 g of
streptomycin was mixed thoroughly in 100 ml of sterilized
distilled water). The plates were incubated at 35C for 72
h. Three replicates were maintained for each sample. Each
replication plates predominant colonies were selected and
transferred to nutrient broth; the cultures were streaked on
PDA medium. After the isolation of the micro fungi, their
pure cultures were made by single-spore culture method
and culture organisms were identified from Lacto phenol
cotton blue method (Opasols and Adewoye, 2010).
Preparation of extract
The fresh C. gigantea leaves were thoroughly cleaned
with sterile distilled water and weighed (100 g), cut into
pieces of small sizes, and aqueous extract was macerated
with 100 ml sterile distilled water at 50C for two days in
water bath. After completion, the extract was filtered with
sterile Whatman No.1 filter paper and sterilized at 120C
for 30 min. Method has been modified by Satish et al.,
2007; acetone extract, weighed 100 g, was cut into pieces
of small sizes by sterile knife. Leaf was extracted with 100
ml acetone solution, macerated for 2 days at 50C in water
bath. After completion, the extract was filtered with sterile
Whatman No. 1 filter paper and sterilized at 120C for 30
min. All the extracts were subjected to biological control
against the test fungal organisms.
Determination of antifungal activity
PDA medium with different concentrations of 2%, 4%,
6%, 8%, and 10% of the aqueous and acetone plant
extracts were prepared. About 15 ml of the medium was
poured into each petriplate and allowed to solidify. The
test cut organisms were inoculated to the PDA medium.
However, the simple streak nutrient agar plates serve as
well growth condition for test organisms, where they were
cut using the 6-mm diameter sterile steel cap mouth,
transferred in to the medium, and incubated at 35C for 72
h. Then, the zones of inhibition appearing around the well
were measured (Vadlapudi, 2012) and method was
modified.
RESULTS AND DISCUSSIONS
This investigation clearly revealed that C. gigantea plant
leaf acetone and aqueous extract showed respective
percentage control over the rice crop affecting pathogenic
fungal strains, Aspergillus sp, Penicillum sp, and A. niger.
The medicinal plant extraction methods have been proven
to be most effective method to control the pathogenic
microorganisms (Cowan, 1999). A number of previous
studies have demonstrated antimicrobial activity of C.
gigantea plant with different types of solvent extraction

109

Fig 3 Shows Inhibition zone of Penicellum sp

methods (Hemalatha et al., 2011). Acetone extracts
exhibited to potentially suppress the growth of test fungal
organisms and the aqueous extracts showed minimum
inhibition zone value, which was less effective compared to
the acetone extract. The results showed that minimum
inhibition zone value was dependent on the concentration
of extract. Increasing the extract concentration level
enhanced the antifungal properties (Dellavalle et al., 2011).
The aqueous and acetone extract showed minimum
inhibition zone of Aspergillus sp at 10% concentration (Fig
2). The Aspergillus sp exhibited result is more important
part in paddy field. Because, worldwide 7,000 verities of
rice cultivation and its deterioration for several fungal
genera, particularly Aspergillus sp has leading to
mycotoxins production cause the complete lack of major
key nutrients such as carbohydrates, starch, glucose,
sucrose, dextrin, etc (Astoreca et al., 2012). This impacts
human consuming rice provides 80% of the calories level
decreased. (Stern, 2003). In India, most of the regions were
affected by the Aspergillus sp producing aflatoxins (Reddy
et al., 2008; Manafi, 2011). In addition, a number of
Aspergillus and other pathogenic fungal species vigorously
affect the paddy fields of Tamilnadu (Palaniswami et al.,
1989; Udagawa, 1976). However, best suitable conditions
for Aspergillus sp growth are temperatures between 30 and
35C (Shehu and Bello, 2011), humidity and climate
change etc, which are highly supported in tropical region of
Indian atmosphere (Sautour et al., 2001, 2002). Moreover,
Penicillum sp producing mycotoxins are potential to
modulate plant immune function (Richetti et al., 2003;
Keblys et al., 2004; Stec et al., 2008) which in turn causes
easier affecting and spread of other various diseases in
cultivation field. In Penicillum sp, minimum inhibition
zone values (Fig 3) were at 10% concentrations of acetone
and aqueous extracts. The ochratoxin A, fumonisin B2, and
aflatoxins producing A. niger mycotoxins significantly
caused huge economic loss of food production (Noonimabc
et al., 2009; Abdalall, 2009) and ailments to human beings,
such as problems in liver, kidney, nervous system, muscles,
skin, respiratory organs, digestive tract, genital organs etc
(Rai and Mehrotra, 2005; Truckesses and Scott, 2008).
A.niger causes the stack burn disease in rice; control of
disease is an inevitable necessity, as its sustainability factor
is very crucial in terms of protecting and preserving the
plants and human beings (Gonzales et al., 2002). In results
of A. niger, minimum inhibition zone values were 10.7 mm

International Journal of Research in Pure and Applied Microbiology 2013; 3(4): 107-112

and 7.0 mm 10% of aqueous and acetone concentration

(Fig 4). A number of studies reported that biotic factors
significantly decrease the rice production compared to the
abiotic factors (Strange and Scott, 2005). Pathogenic biotic
organism control has evitable necessity of cultivation
paddy field. In our result, it is observed that wasteland
medicinal plant, C. gigantea, extracts potentially controls
biotic fungal organism, which may be because of
antimicrobial compounds, such as usharin, gigantin, calotropeol, -amyrin, fatty acids, hydrocarbons, tetracyclic
triterpene etc (Habib et al., 2007; Tenpe et al., 2007).
These are compounds effectively controlled pathogenic
microorganism in agriculture filed and various fields. In
addition C. gigantea medicinally used to human affecting
disease and infection such as fevers, rheumatism,
indigestion, cough, cold, eczema, asthma elephantiasis,
nausea, vomiting, leprosy, diarrhea effectively controlled.
These types of biological control to crop plants disease is
necessary for cultivation of rice in many countries where
day-by-day, the consumption of chemical pesticide
contaminated rice has been increased as an alternative to
chemical pesticide control. Especially, this type of
biological control methods has immense values for large
amount of chemical pesticide consuming and producing
countries. Particularly, as a largest rice producing nation,
India needs to adapt the alternative biological pest control.
At a global level, India ranks 12th place as a pesticide
producer (WHO, 2009) and second place as a pesticide user
in Asia (Shetty et al., 2010). The estimation of chemical
pesticide consumption is approximately 43, 00085, 000
tonnes; 76% of pesticide is applied for rice cultivation
(Subramanyam et al., 2012) and 45% of pesticide applied
for paddy, cotton crop, and wheat cultivations. This type of
chemical pesticide usage in cultivation process may
become a dangerous issue in future generation by affecting
natural characteristics of soil and plants, and thereby
leading to toxic accumulation in human beings. Therefore,
the biological control of plant pathogens plays more
important role as replacement for the application of
chemical pesticide in agriculture field, immensely
supporting as one of the eco-friendly pesticide approaches
for farmers.

Fig 4 Shows Inhibition zone of Aspergillus niger

CONCLUSIONS
The analysis of study showed that the C. gigantea plant
extraction exhibited remarkable biological control against
the rice-affecting pathogenic fungal strains even at low

110

concentrations. The leaf extracts can be used as alternative

source for the various other diseases control in cultivation
of agriculture plants.
ACKNOWLEDGEMENTS
The Sharmila Institute of Medicinal Products Research
Academy (SIMPRA) Thanjavur, supported by providing
necessity facilities for this research work. We are grateful
to founder and chairman Mr. R.J.A. Raj and Mariamman
Kovil village farmers.
REFERENCES
1. Abdalall, A.H.A. (2009). Production of aflatoxins by
Aspergillus flavus and Aspergillus niger strains
isolated from seeds of pulses. Journal of Food
Agriculture and Environment 7:3339.
2. Astoreca, A., Vaamonde, G., Dalcero, A., Ramos, A.J.,
and Mar, S. 2012. Modelling the effect of temperature
and water activity ofAspergillus flavus isolates from
corn. International Journal of Food Microbiology 156:
6067.
3. CAST (Council for Agricultural Science and
Technology), 2003.Mycotoxins: risks in plant, animal
and human systems. Task Force Report 139. Ames,
Iowa: CAST 199. Available at: www.cast-science.org.
4. Cowan, M.M. 1999. Plant products as antimicrobial
agents. Clinical Microbiology Reviews 10:564582.
5. Dellavalle, P.D., A. Cabrera, D. Alem, P. Larraaga, F.
Ferreira, and M.D. Rizza. 2011. Antifungal activity of
medicinal plant extracts against phytopathogenic
fungus Alternaria spp. Chilean Journal of Agricultural
Research 7:12.
6. FAO, (2009). Rice market monitor. Trade Markets 12.
7. FAO, 2010. The state of food insecurity in the world
addressing food insecurity in protracted crises.
8. Gonzales, Mew, T.W., (2002). A Handbook of Rice
Seed-Borne Fungi. IRRI Science Publishers 83.
9. Habib, M.R., F. Nikkon, M. Rahman, M.E. Haque, and
M.R. Karim. 2007. Isolation of stigmasterol and sitosterol from methanolic extract of root bark of
Calotropis gigantea (Linn). Pakistan Journal of
Biological Sciences 10:41744176.
10. Hemalatha, M., B. Arirudran, A. Thenmozhi, and
U.S.M. Rao. 2011. Antimicrobial effect of separate
extract of acetone, ethyl acetate, methanol and
aqueous from leaf of milkweed (Calotropis gigantea
L.). Asian Journal of Pharmaceutical Research 1:102
107.
11. ICAR - Indian Council of Agricultural Research, 2012.
Agbiotech digest 3:3.
12. Keblys, M., A. Bernhoft, C.C. Hofer, E. Morrison,
H.J.S. Larsen, and A. Flaoyen. 2004. The effects of the
Penicillium mycotoxins citrinin, cyclopiazonic acid,
ochratoxin A, patulin, penicillic acid, and roquefortine
C on in vitro proliferation of porcine lymphocytes.
Mycopathologia 158:317324.
13. Mahender, A., D.M. Swain, D. Gitishree, H.N.
Subudhi, and G.J.N. Rao. 2012. Molecular analysis of
native Manipur rice accessions for resistance against
blast. African Journal of Biotechnology 11:13211329.
14. Manafi, M. 2011. Evaluation of different mycotoxin
binders on aflatoxin B1 (Aspergillus parasiticus)

International Journal of Research in Pure and Applied Microbiology 2013; 3(4): 107-112

15.

16.

17.

18.

19.

20.
21.

22.

23.

24.
25.

26.

27.

28.

produced on rice (Oryza sativa) on fertility,

hatchability, embryonic mortality, residues in egg and
semen quality. Advances in Environmental Biology
5:38183825.
Mazid, M., T.A. Khan, and F. Mohammad. 2012.
Medicinal plants of rural India: a review of use by
Indian folks. International Journal of Pharmaceutical
Sciences. 2:286304.
Mohana, C. Devihalli, P. Pravin, V. Veena, and R.K.
Anandrao. 2011. Plant extract effect on Seed-borne
pathogenic fungi from seeds of paddy grown in
southern India. Journal of plant Protection Research
51:101-106.
Noonimabc, P., W. Mahakarnchanakulb, K.F.
Nielsend, J.C. Frisvadd, and R.A. Samsona. (2009).
Fumonisin B2 production by Aspergillus niger in Thai
coffeebeans.Food Additives&Contaminants26:94100.
Opasols, A.O., and S.O. Adewoye. 2010. Assessment
of degradability potential of Penicillium oxalicumon
crude oil. Applied Scientific Research 1:182188.
Palaniswami, A., A. Manickam, and S. Neelakantan.
1989. Surveillance of toxigenic fungi and aflatoxins in
rice and paddy grains. Madras Agricultural Journal
76:481489.
Pathak, A.K, and A. Argal. 2007. Analgesic activity of
Calotropis gigantea flower. Fitoterapia 78:40-42.
Perrone. G., A. Susca, G. Cozzi, K. Ehrlich, J. Varga,
J.C.Frisvad,M.Meijer.P.Noonim,W.Mahakarnchanakul
, and R.A. Samson. 2007. Biodiversity of Aspergillus
species in some important agricultural products.
Studies in Mycology 59:5366.
Prakasha, H.M., M. Krishnappa, Y.L. Krishnamurthy,
and S.V. Poornima. 2010. Folk medicine of NR
Puratalukin Chikamaglur district of Karnatka. Indian
Journal of Traditional Knowledge. 9:55-60.
Preethi, R., D.V. Vimal, and M. Loganathan. 2010.
Antimicrobial and antioxidant efficacy of some
medicinal plants against food borne pathogens.
Advances in Biological Research 4:122125.
Rai, V., and S. Mehrotra. (2005). Toxic contaminants
in herbal drugs. Environment News Archives 11:13.
Reddy, K.R.N., C.S. Reddy, and K. Muralidharan.
2008. Potential of botanicals and biocontrol agents on
growth and aflatoxin production by Aspergillus flavus
infecting
rice
grains.
Food
Control.
doi:10.1016/j.foodcont.2008.03.009.
Richetti, A., A. Cavallaro, T. Ainis, and V. Fimiani.
2003. Effect of some mycotoxins on superoxide anion
production of isolated human neutrophils and in whole
blood. Immunopharmacol Immunotoxicol 25:441449.
Satish, S., D.C. Mohana, M.P. Raghavendra, and K.A.
Raveesha. 2007. Antifungal activity of some plant
extracts against important seed borne pathogens of
Aspergillus sp. Journal of Agricultural Technology
3:109119.
Sautour, M., C.M. Soares, C. Divies, M. Bensoussan,
and P. Dantigny. 2002. Comparison of the effects of
temperature and water activity on growth rate of food
spoilage moulds. Journal of Industrial Microbiology
and Biotechnology 28:311316.

111

29. Savary, S., F. Horgan, L. Willocquet, and K.L. Heong.

(2012). A review of principles for sustainable pest
management in rice. Crop Protection 32:5463.
30. Schippmann, N., D.J. Leaman, and A.B. Cunningham.
2002. Impact of cultivation and gathering of medicinal
plants on biodiversity: Global Trends and Issues.
Published FAO 122.
31. Shehu, K., and M.T. Bello. 2011. Effect of
environmental factors on the growth of Aspergillus
species associated with stored millet grains in sokoto.
Nigerian Journal of Basic and Applied Sciences
19:218223.
32. Shetty, P. K., M. Murugan, M. B. Hiremath, and K. G.
Sreeja. 2010. Farmers education and perception on
pesticide use and crop economies in Indian agriculture.
Journal of Experimental Sciences 1:3-8.
33. Shetty, P.K., M.B. Hiremath, M. Murugan, and R.B.
Nerli. 2012. Farmers health externalities in pesticide
use predominant regions in India. World Journal of
Science and Technology 1:111.
34. Stec, J., J. Rachubik, M. Szczotka, and J. Kuzmak.
2008. Effects of Penicillium mycotoxins: citrinin,
ochratoxin A, and patulin on in vitro proliferation of
bovine lymphocytes. Bulletin of the Veterinary
Institute in Pulawy 52:16367.
35. Strange, N.R and Scott, R.P. 2005. Plant disease:a
threat to global food security. Annual Review of
Phytopathology. 43:83116
36. Subramanyam, D., V. Lokanatha, W. Rajendra, and B.
Vijayalakshmi. 2012. Biomanagement of pesticides for
sustainable environment: an Indian scenario.
International Journal of Biological & Pharmaceutical
Research 3:826834.
37. Tenpe, C.R., A.B. Upaganlawar, P.A. Dongre, and
P.G. Yeole. 2007. Screening of methanolic extract of
Calotropis gigantea leaves for hepatoprotective
activity. Indian Drugs 7:874875.
38. Truckesses, M. W., and P.M. Scott. (2008).
Mycotoxins in botanicals and dried fruits. a review.
Food Addition Contamination 25:181192.
39. Udagawa, S. 1976. Distribution of mycotoxinproducing fungi in foods and soil from New Guinea
and Southeast Asia. In: Proceedings of the Japanese
Association of Mycotoxicology 2:1015.
40. Vadlapudi, V. 2012. In vitro antimicrobial activity of
plant extracts of Avicennia alba against some
important pathogens. Asian Pacific Journal of Tropical
Disease 408411.
41. Varma, J., and N.K. Dubey. 1999. Prospectives of
botanical and microbial products as pesticides of
tomorrow. Current Science 76:172179.
42. WHO, (1990).World Health Organization: Public
health impact of pesticides used in agriculture
(Geneva: WHO-UNEP).
43. WHO, 2009. Health implications from monocrotophos
use: a review of the evidence in India. World Health
Organization, Regional Office for South-East Asia,
New Delhi. Available on: http://203.90.70.117/
PDS_DOCS/B4293.pdf. Retrieved on: 15 February
2912.

International Journal of Research in Pure and Applied Microbiology 2013; 3(4): 107-112

44. Williams, J.H., T.D. Phillips, P.E. Jolly, K.J. Stiles,

C.M. Jolly, and D. Aggarwal. (2004). Human
aflatoxicosis in developing countries: a review of

toxicology, exposure, potential health consequences,

and interventions. The American Journal of Clinical
Nutrition 80:11061122.

Source of support: Nil; Conflict of interest: None declared

112

International Journal of Research in Pure and Applied Microbiology 2013; 3(4): 107-112