Rafiq et al., IJPSR, 2018; Vol. 9(3): 1002-1011.

E-ISSN: 0975-8232; P-ISSN: 2320-5148

IJPSR (2018), Volume 9, Issue 3 (Research Article)

Received on 30 May, 2017; received in revised form, 11 October, 2017; accepted, 20 October, 2017; published 01 March, 2018

ISOLATION AND IDENTIFICATION OF ANTIBIOTIC PRODUCING MICROORGANISMS

FROM SOIL
Adeel Rafiq 1, Shabir Ahmad Khan 1, Ali Akbar*1, Muhammad Shafi 2, Imran Ali 3, Fazal Ur Rehman 1,
Rozina Rashid 1, Gulmakai Shakoor 1 and Muhammad Anwar 1
Department of Microbiology 1, Faculty of Life Sciences, Center for Advanced Studies in Vaccinology and
Biotechnology 2, Institute of Biochemistry 3, University of Balochistan Quetta, 87300, Pakistan.
ABSTRACT: Keywords:
Antibiotics are most important commercially used secondary
metabolites, produced by many soil microorganisms i.e., bacteria and fungi
Secondary metabolites,
Soil bacteria, Soil fungi and employed in wide range. Most important antibiotics used today are of
Correspondence to Author: microbial origin. The emergence of the antibiotic resistance and need of
Dr. Ali Akbar broad spectrum antibiotics is in focus and in demand. In present study, soil
samples from different areas were collected i.e., the sampling is classified
Assistant Professor,
based on its micro and macro environment (waste polluted soil) (normal
Department of Microbiology,
Faculty of Life Science, University of street soil) and (agricultural soil), from a local soil and analyzed for the
Balochistan Quetta, 87300, Pakistan. antibiotic production. After primary screening, bacterial isolates were
identified as Micrococcus roseus, Brevibacterium sp., Bacillus subtilis,
E-mail: [email protected] Bacillus anthracis and Bacillus cerus, through biochemical characterization,
and fungal isolates were identified as Tricho-cladium opacum, Rhizocotania
sp., Epicoccum nipponicum, Aspergillus niger and Cladosporium
cladosporides through microscopic and macroscopic identification and
checked for antibiotic activity against some common gram positive and
negative bacteria namely, Staphylococcus aureus and Escherichia coli. The
antibiotic test indicates that Bacillus subtilis, Bacillus anthracis, Epicoccum
nipponicum, Aspergillus niger and Cladosporium cladosporides showed
antimicrobial activity against Saureus whereas against E. coli, Bacillus
anthracis, Bacillus cerus, Bacillus subtilis, Trichocladium opacum and
Cladosporium cladosporides produces zone of inhibition. This study
suggests that Bacillus species have the potential to produce antibiotics and
can be used to control the microbial growth in future. Strains of antibiotic
producing fungi could be harnessed by pharmaceutical industries and used in
medicinal purposes. This work may provide potential information on the
antibiotic production and further be used for the control of microbial strains.
INTRODUCTION: Antibiotics are a natural In 1928, the term antibiotic appeared as antibiosis
substance of biological, synthetic or semi-synthetic in French microbiological literature whereas as
origin 1. later in 1942 the term antibiotic was introduced by
QUICK RESPONSE CODE Waksman.
DOI:
10.13040/IJPSR.0975-8232.9(3).1002-11 The demand for new antibiotics growing day by
day due to the emergence of multiple pathogens
Article can be accessed online on: that are resistant to antibiotics cures for formerly
www.ijpsr.com life-threatening diseases. In recent years several
DOI link: http://dx.doi.org/10.13040/IJPSR.0975-8232.9(3).1002-11
microorganisms that are able to produce antibiotics
are grown on the artificial media for the intensive
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search for antibody producing microorganisms. The According to the International center of
most important antibiotics include Amino- information on antibiotics, 338 species of fungi are
glycosides, Penicillin, Macrolides, Glycopeptides, able to produce antibiotics. In soil habitat, fungi
Cephalosporins and Tetracyclines 2. Soil is a compete against different other microorganisms
complex and very diverse environment providing and may induce antimicrobial production 21.
versatile source of antibiotic producing organisms Filamentous fungi contaminated with broad
3
. Each year nearly 500 antibiotics were found, in spectrum antibiotics, inhibited by antibiotic
which 60% of antibiotics are obtained from the soil resistant bacteria are able to produce the secondary
4
. Recent analyses have shown that screening of metabolites. 20% of isolated antibiotics have been
soil for antimicrobial activities have been carried discovered from soil fungi 22. Many species of
out in many parts of the world 5. A tea-spoon of fungi including Penicillium, Aspergillus,
soil contains hundred million to one billion bacteria Cladosporium and Yeasts which were reported only
active in each acre of the soil. to be the food 23, 24, 25 are tremendous sources of
Bacillus species are gram positive, rod shaped, industrially important enzymes and secondary
sporulating and aerobic or facultative anaerobic metabolite production 26. Antibiotics produced by
bacteria that were most abundant bacterial strains the fungal species are widely used inthe
found in the soil 6 which were capable of producing chemotherapy 27 especially Fusidic acid,
dozens of antibiotics 7. Genus of Bacillus was Cephalosporin and Penicillin which have both
interesting to investigate and were considered antifungal and antibacterial activity. Fungi
microbial factory for the production of biologically represent an important source of potentially
active secondary metabolites 8, 9. Various studies powerful new pharmaceutical products 28.
confirmed Bacillus species to produce Nowadays, Pharmaceutical industries are
antimicrobial compounds having pharmaceutical investigating an array of fungi to increase the
and biotechnological importance 10, 11. The main number of discoveries 21. For many years ago, in
antibiotic producer of Bacillus sp. were B. cereus many societies fungi have been recognized as
(Zwittermicin, Cerexin) B. brevis (Tyrothricin, palatable nutritious food and are now acceptable as
Gramicidin), B. circulans (Circulin), B. a valuable source in pharmaceuticals for
licheniformis (Bacitracin) B. laterosporus development of the medicines 28. Several thousand
(Laterosporin), B. polymyxa (Colistin, Polymyxin), antibiotics have been isolated from soil
B. subtilis (Bacitracin, Polymyxin, Subtilin, microorganisms since the discovery of penicillin,
Difficidin, Mycobacillin,) B. pumilus (Pumulin) are but unfortunately these have been limited only to
mainly polypeptides 12 were mostly active against fifty, most of them being too toxic to humans 29.
gram positive bacteria 13. According to the food and drug administration
An antibiotic activity of B. pumilus and B. subtilis approximately 80% of antibiotics produced from
has been reported against Staphylococcus aureus natural habitat are fed to animals and only 20% of
and Micrococcus luteus 14. Bacillus species isolated them are used to treat infections in the humans.
from Jordanian soils shows antibacterial activity The objective of this study was to find antibiotic
against the methicillin-resistant Staphylococcus producing microorganisms and to check their
aureus 15. Cellulases, Subtilisins, and Amylases ability to inhibit the growth of S. aureus and E.
produced by B. subtilis had several industrial coli.
importances and were consumed by the laundry
industries 16. Fungi were extremely diversified MATERIAL AND METHODS:
worldwide 17. On earth, there are approximately 1.5 Collection of Sample: Soil samples from different
million species of the fungi out of which 70 - areas were collected i.e., the sampling is classified
100,000 species were discovered and 95% of these based on its micro and macro environment (waste
have yet to be discovered 18. Fungi are best source polluted soil) (normal street soil) and (agricultural
of antibiotics 19 and to search antibiotics from them soil), within Quetta city. Approximately 1 kg of
is very promising. Twenty best-selling medical soil sample was collected for further processing.
drugs worldwide were fungal derived 20. Soil sample was collected in such way to get the

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soil of crust and depth of at least 6 inches with the Staining Characterization: Gram staining and
help of sterile spatula and placed in sterile plastic spore staining was performed of the isolated and
bags for transportation to laboratory. sub-cultured bacterial colonies. In gram staining a
Preparation of Soil Sample: Three sterile test clean glass slide was taken. Smear was prepared by
tubes were taken, marked and labeled for each soil heat fixing and air drying. Drop of crystal violet
sample and were filled with 10 ml of distilled was added on smear and allowed to stand for 60 sec
water. A soil suspension is made by adding 1 g of and then washed with the distilled water. Then
the soil sample in 10 ml of distilled water in a first gram iodine was added and stands for 30 sec. After
test tube for each soil sample and is vortexes. 1 ml though decolorizer was added to title the slide
of this solution is taken and transferred to the followed by safranine and stand for 60 sec and was
second test tube and is vortexes and from second to washed and dried and viewed under 100X
third. Hence each soil sample was serially diluted microscope lens whereas in spore staining a clean
in a laminar air flow. glass slide was taken. A drop of normal saline was
added on a slide then colonies of bacteria were
Isolation of Microorganisms: Media was prepared picked using sterile loop from isolated sub-cultured
for isolation of bacteria and the principle media colonies grown on nutrient agar plates in a laminar
used for this purpose was nutrient agar medium. An air flow and allowed to dry. Then Malachite green
amount 1000 ml of distilled water in a beaker was was added on smear and steam for 5 - 7 min.
taken and 28 g of Nutrient agar powder was Safranine was added which act as counter strain
dissolved in it followed by sterilization in an and was allowed to stand for 60 sec. Slide was then
autoclave at 121 °C for 15 min and allowed to cool. washed and dried and viewed under 100X
After that media was poured in Petri plates and microscope lens.
allowed to solidify and placed in an incubator at
37°C for 24 h in order to check its sterility. Biochemical Characterization: For the
Whereas for the isolation of fungi Sabouraud identification of bacterial isolates biochemical tests
dextrose agar (SDA) and Potato dextrose agar were performed as described by the Bergey’s
(PDA) was used. manual i.e., IMViC tests (indole test, citrate
utilization test, catalase test, MR-VP test), triple
Inoculation of Sample: The samples were sugar iron test (TSI), oxidase, urease, nitrate
inoculated on nutrient agar plates in duplicate for reduction and blood hemolysis.
bacterial species isolation. An amount 0.1 ml of
each soil sample from selected dilutions (usually1: Microscopic and Macroscopic and Examination
100 and 1:1000) were taken and poured using pour of Fungi Isolates: Microscopic examination of
plate method on labelled nutrient agar plates. The each fungal isolates was done by picking fungi
Petri dishes were then inverted followed by mycelia with the help of a sterilized needle and was
incubation for 24 h at 37 °C to obtain the isolated placed on a glass slide containing a drop of
colonies. For fungus growth, aliquots of 0.1 ml of lactophenol cotton blue stain, and then covered
the last two dilutions were inoculated on two SDA with the cover slip and was viewed under
and PDA plates followed by incubation at 28 °C for microscope using 40 X and I00X objective lens of
3 - 5 days. the microscope.

Sub Culturing of Microorganisms: Bacterial The macroscopic examination of fungi isolates was
colonies with clear margins was picked and sub- carried out by comparing the fungi isolate with the
cultured on fresh nutrient agar plates using sterile Pictorial atlas of soil and seed fungi. Some
loopusing streak plate method in laminar air flow to morphological features were observed that includes
purify the isolates followed by incubation for 24 h physical appearance, color and growth pattern of
at 37 °C. For fungi, each discrete colony on SDA each fungus colony on SDA and PDA medium.
and PDA plates were further inoculated on fresh Test Microorganisms for Antimicrobial Activity
SDA and PDA plates followed by incubation at Determination: Test organisms i.e., E. coli and S.
28°C for 3 - 5 days. aureus were obtained from local private clinical

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laboratory and tested for antimicrobial activity of After incubation colonial morphology was
antibiotic producing isolates using agar well observed. A total five culture strains of bacteria and
diffusion method. five fungal isolates were selected from all of the
Screening of Antimicrobial Activity: For soil samples i.e., (waste polluted soil, normal street
antibiotic production, Mueller Hinton agar (MHA) soil and agriculture land soil). The selected culture
media was prepared by adding 17 g of MHA media strains of bacteria were then subjected to gram
in 500 ml of distilled water and autoclaved at staining, spore staining and biochemical
121°C for 20 minutes. After sterilization, the media characterization tests and their results are shown in
was cooled and poured in Petri dishes and kept in Table 1. The results of the biochemical tests were
incubator at 37 °C for 24 h to check its sterility. checked using Bergey’s manual of systematic
Two test tubes were taken containing 2 ml of bacteriology; as a result their morphological
sterilized nutrient broth. Test organisms i.e., E. coli features were clearly observed.
and S. aureus were inoculated in it and kept in The selected isolated culture strains of bacteria
shaker incubator for 24 h. After incubation, were identified as Micrococcus roseus,
sterilized cotton buds were dipped in it and Brevibacterium sp, Bacillus subtilis, Bacillus
swabbed on MHA plates. Wells were made on anthracis and Bacillus cerus whereas the
MHA plates using sterile borer. Isolated and sub- microscopic examination of fungal isolates was
cultured bacterial and fungal colonies were carried out using lactophenol cotton blue staining
inoculated in test tubes containing 5 ml of NB for and its macroscopic examination was done by
bacteria and PDB for fungi isolates were kept in comparing the fungal isolate with the Pictorial atlas
shaker incubator at 37 °C for 24 h. After of soil and seed fungi as shown in Table 2. The
incubation, were centrifuged at 6000 rpm for 10 fungal isolates were identified as Trichocladium
min and their supernatants were poured in the wells opacum, Rhizocotania sp., Epicoccum nipponicum,
and kept in incubator for 48 hr. Zones of inhibition Aspergillus niger and Cladosporium cladosporides.
were observed. The identified cultures of bacteria and fungi were
RESULTS: A soil samples was collected i.e., from then checked for antibiotic production activity
(waste polluted soil, normal street soil and using agar well diffusion method. The zones of
agriculture land soil). Colonies were observed in inhibition were observed against the test bacteria
the crowded plate. Colonies showing clear margins (S. aureus and E. coli) as shown in Fig. 1 and Fig.
were sub cultured on the fresh medium plates. 2 and Table 3 and Table 4.

TABLE 1: IDENTIFICATION OF BACTERIAL ISOLATES COLLECTED FROM VARIOUS SOIL

Colonial morphology Cellular Biochemical characterization
characteristic
Isolates Codes

Identified
organism
Gram stain

Spore stain

Hemolysis
utilization

reduction
Elevation

Catalase
Pigment

Sulphur

Oxidase
Motility

MR-VP
Surface

Citrate

Nitrate
texture

Urease
Whole
colony

Indole
Edge

TSI

+ - - - - - + +/[d] A/A [d] - [+] -

slightly convex

Bacillus subtilis Brevibacteriums Micrococcus

N1B
circular

smooth

roseus
entire

Pink

+ - - - - [d] + -/- K/A - + [-] +

irregular

smooth

convex
yellow
entire

N2A
dry or rough

+ + - - + + + -/+ A/NC - - + +
umbonate
undulate
irregular

(wavy)

white,
dull

N2B

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+ + - - - + + [d]/+ A/A - - + -

whitish to whitish to cream

matt or granular

anthracis
entire to
undulate
irregular

Bacillus
raised
N3A

+ + - - + + + -/+ A/NC [+] [-] + +

Bacillus cerus
undulate
irregular

cream
N3B dry

flat
Note: + 90% or greater positive,- 90% or greater negative, [d] 26 - 75% of strains are positive, [+] 76 - 89% positive, [-] 76 -
89% negative, K/A Glucose fermentation only, Peptone catabolized, A/A Glucose, lactose or Sucrose fermentation, A/NC
Ferment sugar but did not grow in anaerobic environment of butt

TABLE 2: IDENTIFICATION OF FUNGAL ISOLATES

Isolate Codes Colonial morphology Identified organism
S1A Short, simple branched conidiophores, bearing single conidia apically. Dark Trichocladium opacum
green, aleuriosporous, ellipsoidal, clavate Conidia, usually 2 to 3
transversely septate, rarely 4-septate, warty marginally, slightly curved,
with conspicuous pedicels
S1B Pale brown hyphae, branched, septated and constricted closely near the Rhizoctonia sp.
main hyphae. Conidia not formed. Monilioid cells usually formed.
Sclerotia pale brown to brown, discrete or aggregated, various
in shape and size
S1C Brown, porodochia discoid, composed of pseudoparenchymatous Epicoccum nipponicum
stromata and conidia. Stromata mostly brown, globose. Lacking
conidiophores. Conidiogenous cells short, hyaline or pale brown,
aggregated or simple on stromata, holoblastic, cylindrical. Conidia
also formed on simple conidiogenous independent cells are ellipsoidal
or
subglobose, slightly constricted at septum and occasionally formed in
chains
S1D Pale brown or hyaline conidiophores usually simple, erect, thick walled, Aspergillus niger
with basally foot cells, globose vesicles. Composed of catenulate conidia
uniseriate or biseriatephialides on vesicles and phialides acutely tapered
at
apex. Conidia are single celled, phialosporous, black in mass, minutely
echinulate
S3A Erect, pale brown, branched conidiophores bearing catenulate conidia in Cladosporium
each branch. Blastosporous, pale brown or hyaline, ellipsoidal, ovate, cladosporioides
cylindrical, subgloboseconidia, Irregular in shape, apiculate at one end,
often truncate at other end

TABLE 3: INHIBITION ZONE SHOWN BY DIFFERENT BACTERIAL ISOLATESAGAINST TEST ORGANISMS

Isolate Codes Test organisms
S. aureus E. coli
N1B - -
N2A - -
N2B + +
N3A + +
N3B - +
Note: + means active against the target bacteria, - means no activity

TABLE 4: INHIBITION ZONE SHOWN BY DIFFERENT FUNGAL ISOLATES AGAINST TEST ORGANISMS
Isolate Codes Test organisms
S. aureus E. coli
S1A - +
S1B - -
S1C + -
S1D + -
S3A + +
Note: + means active against the target bacteria, - means no activity

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FIG. 1: CLEAR ZONES OF BACTERIAL ISOLATES

FIG. 2: CLEAR ZONES OF FUNGAL ISOLATES

DISCUSSION: In searching for new antibiotics, bacterial and fungal isolates were subjected to test
screening of microorganisms through relatively microorganism (S. aureus and E. coli) in order to
rapid and simple methods has been done for check their ability to produce antibiotics using agar
antibiotic production. Antibiotic production is a well diffusion method.
main feature of several kinds of soil
microorganisms i.e., bacteria and fungi and may In our study results indicates that Bacillus species
thereof represent a survival mechanism. Variation with the highest number of isolates produces clear
in temperature may also affect the synthesis of zone of inhibition against test microorganisms.
antibiotic production. The bacteria isolated from Bacillus species were dominant to shows antibiotic
soil shows antibiotic activity under normal growth activity against S. aureus and E coli. This finding is
condition and were found to inhibit some gram also corroborating the findings of Ahmed et al.,
positive as well as some gram-negative organism. 2013 who screened soil microorganisms for
The isolates of bacteria were not organic acid antibiotic production and reveals that only Bacillus
producer but they are antibiotic producers 30, 31. species exhibited antibacterial activity of all
bacteria isolated 32.
In present study bacteria and fungi were isolated
from soil samples (waste polluted soil, normal In an average of about 108 cells per gram
street soil and agriculture land soil). The selected rhizobacteria are found in the soil 33. Isolation of
culture strains of bacteria were then identified by similar species from soil samples has been reported
techniques, like gram staining, spore staining and by other workers elsewhere 34, 35. The antimicrobial
biochemical characterization tests and microscopic activity from a sediment habitat and resistance to
examination of fungal isolates was carried out antimicrobials of bacteria can easily explain the
using lactophenol cotton blue staining and its persistence and selection of such strains in this
macroscopic examination was done and both particular ecology 36.

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On agar media cultural characteristics displayed by antibiotics with bactericidal activity. Among
bacteria, were used to identify bacteria because of identified fungal isolates Epicoccum nipponicum,
their specific and different growth patterns 37. It has Aspergillus niger, Cladosporium cladosporides and
been reported that Bacillus species and other spore Trichocladium opacum produces clear zone of
forming bacteria carry genes for the production of inhibition against test microorganisms.
antibiotics and breakdown of diverse carbon source
38
. It has been reported that Bacitracin produced by Similar studies on distinctive fungal species were
Bacillus species inhibits both E. coli and S. aureus carried out by different scientists. Makut and
38
. Hassan et al., 2014 identified fourteen isolates Owolewa, 2011 screened the fungal isolates
of antibiotic producing Bacillus species from soil isolated from soil for antibiotic production. Results
11
. For the synthesis of secondary metabolites revealed that Candida albicans was inhibited by all
Bacillus species are well known with remarkable fungal isolates whereas E. coli were inhibited by
diversity both in its function and structure 39. There Rhizopus stolonifera and Aspergillus fumigatus.
is an argument with Aslim et al., 2002 who Trichoderma viride and Alternaria alternata not
documented that strains of Bacillus had greater inhibit Staphylococcus aureus whereas
effects on gram positive bacteria as compared to Pseudomonas aeruginosa was not inhibited by
gram negative bacteria 40. Curvularia lunata, Aspergillus flavus and
Cladosporium herbarum 5.
B. subtilis has the potential to produce antibiotics
and has been recognized for past 50 years. B. Svahn et al., 2012 recognized sixty one strains of
subtilis is an endospore forming rhizobacterium 41. filamentous fungi predominantly various
Sonenshein et al., 2002 collected several wild type Aspergillus species were identified. Majority of
B. subtilis, having the potential to produce more Aspergillus strains shows antibiotic activity against
than two dozen of antibiotics 41. B. subtilis C126 beta lactamase producing E. coli, methicillin-
strain from sugar cane fermentation have the resistant S. aureus, Enterococcus faecalis and
potential to produce polypeptide antibiotic, Candida albicans 45. Miyake et al., 2009 reported
Bacitracin. Production of Bacitracin by B. subtilis that many Aspergillus species have been able to
is a pH dependent which gave maximum produce antioxidants 46. Gugnani 2003 reported
production at pH of 7.8 - 8. Strains of B. cereus that some Aspergillus species are utilized
from a soil sample have the ability to produce industrially for various enzymes production 47.
Bacteriocin and was active against most gram
positive but not against gram negative bacteria. Ifediora 2011 conducted a study to determine the
M15 strain of B cereus possesses inhibitory effect presence of fungi that has the ability to produce
against both gram positive and gram-negative antibiotics in rhizosphere and sewage and check
bacteria. their antibiosis potency on test microorganisms
(Bacillus subtilis, Escherichia coli, Pseudomonas
Bacilli are predominant soil bacteria widely used in aeruginosa and Staphylococcus aureus). Six
industrial applications, particularly antibiotics species of fungi isolated from the soil samples were
production having medically, agriculturally and examined for their antimicrobial activities which
veterinary importance 42, 43. Bacillus species includes; Mucor sp., Curvularia sp., Aspergillus
preferred hosts for the production of many sp., Penicillium sp., Drechslera sp., Rhizopus sp.
improved and new products used in genomic and Two species; Aurebasidium sp. and
proteomics 44. To enhance the yield of Bacitracin it Cephalosporium sp. were isolated from the sewage.
is possible to clone and amplify the gene coding for All the fungal isolates were found to inhibit the
some key enzymes in the biosynthetic pathways of growth of at least one of the test organisms.
Bacitracin. Bacillus subtilis was inhibited by the Rhizopus sp.,
Mucor sp., Drechslera sp., Aurebasidium sp. and
In the present study, we also identified that Curvularia sp. Pseudomonas aeruginosa was
filamentous fungi contaminated with extraordinary inhibited by all the strains except Cephalosporium
levels of broad spectrum antibiotics inhibited by sp. and Curvularia sp. Only Curvularia sp. and
antibiotic resistant bacteria are able to produce Aurebasidium sp. inhibit E. coli whereas

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Staphylococcus aureus was inhibited by the Mucor Alernariaspecie and Penicillium species indicates
sp., Cephalosporium sp., Aurebasidium sp. and moderate activity while Fusarium species,
Drechslera sp. P. aeruginosa was inhibited by all Humicolas specie and Torula species showed low
strains except Penicillium sp., Curvularia sp., activity 54. Mukunda et al., 2012 isolated fungi
Cephalosporium sp. and Rhizopus sp. 48. from soil of Western Ghats and screened for the
Idris et al., 2013 investigated the antimicrobial production of important hydrolytic enzymes such
activity of endophytic fungi isolated from as Cellulose, Protease, Amylase, CMCase, Lipase
medicinal plant Kigelia africana. Aspergillus and pigment production. Results indicate that
sp., Aspergillus flavus, Cladosporium sp. and Trichoderma, Aspergillus, Penicillium and
55
Curvularia lunata, and three unknown species were Cladosporium species were predominated .
screened for antibacterial test against CONCLUSION: The present study was an attempt
Staphylococcus aureus, E. coli and Bacillus to identify and characterize versatile strains of
subtilis. The inhibition zones ranged from 14 - 37 bacteria and fungi and to check their ability for
mm 49. antibiotic production. A number of different
Muhsinand Mohammad, 2012 checked bacterial and fungal isolates were found producing
antibacterial activity of fungi against S. aureus and clear zone of inhibition against the test
E. coli. The inhibition zones by fungal extracts microorganisms i.e., S. aureus and E. coli. The
ranged from 22 - 28 mm in diameters. MIC test study revealed that bacterial and fungal species
indicates that extract of D. australiensn exhibit have potential of antibiotics production. The
minimal inhibition ranges from 12.5 - 6.25 ug/ml potential antibiotic producer bacterial species
against S. aureus and E. coli 50. identified include Bacillus strains i.e., Bacillus
subtilis, Bacillus anthracis and Bacillus cerus.
Species of Trichocadium were frequently isolated
from the soil. Pothiraj et al., 2006 isolated various Whereas among fungal species Trichocladium
species of fungi i.e., Aspergillus niger, Aspergillus opacum, Epicoccum nipponicum, Aspergillus niger
terreus and Rhizopus stolonifer from a and Cladosporium cladosporides were dominated.
contaminated cassava waste soil by primary This study may contribute in providing information
selection, serial dilution and pour plate technique on the antibiotic producing microorganisms in soil.
and reported the production of cellulase by using Further characterization, purification, and structural
solid state fermentation technique 51. Sathyaprabha elucidation are recommended to know the novelty,
et al., 2011 isolated fungi namely, Aspergillus quality and commercial value of these antibiotics.
niger, Aspergillus fumigatus, Aspergillus nidulans ACKNOWLEDGEMENT: The authors extend
and Aspergillus versicolor from crude petroleum warm gratitude to all those, who helped in this
oil contaminated soil 52. Damisa et al., 2011 study one way or the other including the sanitation
isolated strain of Aspergillus niger from soil and cleaning staff of the Department of
samples in zaria. The samples were collected from Microbiology and Center for Advanced Studies in
different soil environment i.e., compost soils, rice Vaccinology and Biotechnology, University of
growing field, street soil, flower beds, maize farm Balochistan, Quetta, Pakistan.
and fallow farm land. They were screened for
cellulolytic efficacy. CONFLICT OF INTEREST: The authors declare
that there is no conflict of interest regarding this
However, microorganisms with higher α amylase manuscript.
activities facilitate the discovery of novel amylase
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How to cite this article:

Rafiq A, Khan SA, Akbar A, Shafi M, Ali I, Rehman FU, Rashid R, Shakoor G and Anwar M: Isolation and identification of antibiotic
producing microorganisms from soil. Int J Pharm Sci & Res 2018; 9(3): 1002-11. doi: 10.13040/IJPSR.0975-8232.9(3).1002-11.
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