ABSTRACT
This study was carried out to determine the effects of carbon source on the antifungal activities of some isolated species of Actinomycetes. Two (2) isolates of Actinomycetes and Streptomyces species were tested against two phytopathogens (Alternaria and Collectotrichum) On screening for its ability to enhance antifungal activity of actinomycetes, basal medium supplemented with glucose as a carbon source was found to be the best for antibiotic production by Actinomyces with the zone of inhibition of (17mm) against Alternaria and (21mm) against Collectrotrichum, this is followed by soluble starch with the zone of inhibition of (15mm) and (18mm) against Alternaria and Collectrotrichum, lactose (5mm and 4mm) and sucrose (2mm and 1mm) whereas fructose had repressed the production of the antimicrobial substance. Basal medium supplemented with soluble starch (12mm and 14mm) as a carbon source was found to be the best for antibiotic production by Streptomyces; this is followed by glucose (10mm and 11mm), lactose (5 and 6) and sucrose (4mm and 6mm) whereas no antimicrobial activity was detected using fructose. The results of this study showed that Actinomyces and Streptomyces species produces antifungal substances which are particularly active against the test organisms; Alternaria and Collectotrichum. Also, soluble starch and glucose were good carbon sources for optimal production of secondary metabolites.
TABLE OF CONTENTS
Title
Page i
Certification ii
Dedication iii
Acknowledgement iv
Table
of Contents v
List
of Tables vii
Abstract viii
CHAPTER ONE
1.0 Introduction 1
1.1 Aims and Objectives 3
CHAPTER TWO
2.0 Literature Review 4
2.1 The Actinomycetes 4
2.2 Streptomyces 6
2.3 Classification
of Streptomyces 7
2.4 The
Life Cycle of Streptomyces 7
2.5 Streptomyces Habitats 8
2.5.1 Hay
and other organic material 8
2.5.2 Fresh
water and marine habitats 9
2.6 Streptomyces Requirements 9
2.7 Streptomyces Metabolites 10
2.8 Isolation
of Streptomyces 10
2.9 importances
of streptomyces 11
2.10 Streptomyces
and antibiotics 12
2.11 Antibacterial
activity of Streptomyces 14
2.11.1 Sampling 14
2.12 Nutrient
Requirements for Antibiotic Production 14
2.13 Antibacterial
Activity Protocols 16
2.13.1 Cross
streak method 16
2.13.2 Agar
overlay method 16
2.13.3 Disc
Diffusion assay 16
2.13.4 Agar
well diffusion assay 17
2.14 Factors Affecting Antibiotic Production 17
2.14.1 Carbon Source 17
2.14.2 Nitrogen source 18
2.14.3 Inorganic phosphate 18
2.14.4 Trace
metals 19
CHAPTER THREE
3.0 Materials and Methods 20
3.1 Study Area 20
3.2 Sample Collection 20
3.3 Preparation of Culture Media 20
3.4 Sample Inoculation 21
3.5 Screening
for Antimicrobial Activity Of The Isolates 21
CHAPTER FOUR
4.0 Result 23
CHAPTER
FIVE
5.0 Discussion
and Conclusion 26
5.1 Discussion 26
5.2 Conclusion 27
References 28
Appendix 36
LIST
OF TABLES
TABLE
|
TITLE
|
PAGE
|
1
|
Effects of carbon source
on the antifungal substance production (activity) by Actinomycetes against Alternaria
and Collectotrichum
|
24
|
2
|
Effects of carbon source
on the antifungal substance production (activity) by Streptomyces against Alternaria
and Collectotrichum
|
25
|
CHAPTER ONE
1.0 INTRODUCTION
Streptomyces,
Greek adjective streptos (pliant or bent)
myces (fungus) therefore, Streptomyces means pliant or bent fungus (Ambarwati
et al., 2012). Streptomyces species
are gram-positive filamentous bacteria that belong to the Actinommycetales.
They are characterized by the ability to form reproductive mycelium from
vegetative mycelium in soil culture. The filamentous growth and the branching
of Streptomyces mycelia differentiate these organisms from the true
bacteria. Commonly, the genus Streptomyces has slender coenocytic
hyphae, the aerial mycelium at maturity forms chains of three to many spores (Bais
et al., 2012). Members of the genus
are soil inhabitants. They are common in wet than in dry areas having a pH of
about 6.5-8.0, and with the exception of few species that cause mycetoma; Streptomyces
are saprophytes. They may be found on vegetation, food products, manures,
peat, water basins, composts, silage, fresh water and river bottoms, dust and
plant residues.
For
many years, members of Actinomycetales were classified with fungi, with which
they share parallel evolution, but to which they are completely unrelated (Baniasadi,
2009). Various keys for the identification of Streptomyces have
been suggested and the most common of them employs four criteria: colour of
aerial mycelium, spore chain morphology, structure of spore surface, and
melanin formation. Generally, new bioactive products
from microbes continue to be discovered at an amazing pace: 500 per year (Baskaran et al., 2011). As a result of the increasing
prevalence of antibiotic-resistant pathogens and the pharmacological limitation
of antibiotics, there is exigency for new antimicrobial substances. In fact,
many of the known antibiotics produced by members of the family Bacillaceae are
polypeptides, which have proven generally to be somewhat unstable and difficult
to purify. Antibiotics produced by fungi, with a few notable exceptions, are
generally found to be too toxic for treatment of eukaryotes including plants (Dehnad
et al., 2010).
However, the antibiotics produced by Streptomyces are
comparatively recognized as generally safe and stable. For this reason Streptomyces
screening for the production of new antibiotics has been intensively
pursued for many years by a number of scientists. This resulted in the
characterization and purification of about 6000 distinct antibiotic substances
from Streptomyces species (Hassan et al., 2011).
Among the different types of drugs prevailing in the market,
antifungal antibiotics are very few but significant and have an important role
in the control of mycotic plant and animal diseases (Ilić et al., 2010). The search for new, safer, broad-
spectrum antifungal agents with greater potency has been progressing. The
reason for this is that when compared to antibacterial, fungi, like plant
cells, are eukaryotes and therefore agents that inhibit protein, RNA or DNA
biosynthesis in fungi have greater potential for toxicity on plant as well (Kariminik
and Baniasadi, 2010).
Recent reports have shown that Streptomyces continue to
remain an important source of antifungals examples included:
24-Demethylbafilomycin C1, Phenyl - 1- napthyl- phenyl acetamide and DPTB16,
and (6S,8aS,9S,11S,12aR)-6-hydroxy-9,10-dimethyldecahydrobenzo [d] azecine- 2,4,12(3H)-
trione (Kavitha et al., 2010). About 80% of
plant diseases can be traced to fungi. In Sudan; fungi infect some important
crops and cause serious diseases that lead to great losses in the production of
these crops. Examples include: Alternaria early blight on tomato, Alternaria
leaf spot on sesame, Macrophomina charcoal rot on sesame, Drechslera
leaf spot on sorghum, Colletotrichum tissue necrosis in beans, D.
maydis leaf blight on corn, and Fusarium wilt caused by Fusarium spp.
(Kharat et al., 2009). Continuous
screening of Streptomyces for secondary metabolites production can
possibly reveal a novel antifungal agent which can be used to treat one or more
of such plant diseases. Hence the main objective of this study has been
screen locally isolated Streptomyces for production of potent antifungal agents that can be used to control some selected
important fungal plant pathogens.
1.1 AIMS AND OBJECTIVES
The
aim of this study was to determine the effect of different carbon sources on
the antifungal activities of some isolated species of Actinomycetes against some test organisms.
The
objectives are;
1. To
determine the antifungal activities of Actinomycetes species against some test
organisms.
2. To
determine how carbon source could be manipulated to enhance the antifungal substances
produced by Actinomycetes species.
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