STUDY OF AQUATIC MICROBIAL DIVERSITY BASED ON DIFFERENTIATION AND STRUCTURAL STAINING

ABSTRACT

The microbial diversity of aquatic organisms based on differentiation and structural staining was investigated. Samples of aquatic organisms were collected from three different aquatic environments which are Ogutalake, Onu- Imo river and Olokorostream in UmuahiaAbia State. A total of three (3) samples, each of the sample was collected in a sterile bottle amounting to 0.75cl and were randomly sampled and cultured on Nutrient agar, Mannitol Salt, MacConkey agar, Salmonella- Shigella agar. Results indicated a high prevalence of Staphylococcus, Shigellaetc.The bacterial isolated from three aquatic sources includesStaphylococcus, Escherichia coil, Klebsiella, Salmonella, Pseudomonas, Bacillus. The Result of this study showed high counts with diversmicroflora which might be of public health importance.

TABLE OF CONTENTS

Title Page i

Certification ii

Dedication iii

Acknowledgment iv

Abstracts v

Table of Contents vi

List of Tables vii

Chapter One

1.0 Introduction: 1

1.1 Aims and Objectives 3

Chapter Two

2.0 Literature Review    4

2.2 Types of Aquatic Diversity 4

2.2.1 Marine Aquatic Diversity: 4

2.2.2 Marine Bacterial Diversity: 4

2.2.3 Marine Division: Ecosystem Division:   5

2.3 Mangrove Ecosystem: 5

2.3.1 Microorganisms found in Mangrove Ecosystem 5

2.4 Coral Reefs:        6

2.4.1 Microorganisms found in Coral Reefs Ecosystem 6

2.5 Deep Sea    6

2.5.1 Microorganisms found in Deep Sea Ecosystem 7

2.6 Extreme Environments:- 7

2.7 Marine Actinomycetes – a Boundary Microorganism 8

2.7.1 Marine Actinomycetes as a Source of Antibiotics   8

2.7.2 Role of Marine Actinomycetes in Marine Environment:    9

2.8 Freshwater Aquatic Diversity 9

2.8.1 Three Basic Types of Freshwater Ecosystem: 9

2.8.2 Lentic Ecosystem (ponds)   9

2.8.3 Lakes 10

2.9 Freshwater Wetlands 11

2.9.1 Streams    11

2.9.2 Microorganisms found in Freshwater Habitat and the likely Diseases Caused 13

2.9.3 Pseudomonas aeruginosa:   13

2.9.4 Aeromonas: 14

2.9.5 Legionella 14

2.9.6 Mycobacterium arium complex (mac) 14

2.9.7 Helicobacter pylori 15

2.10 Diseases Caused by Aquatic Microorganisms 15

2.10.1 Giardia Lambia 15

2.10.2 Cryptosporidium 16

2.10.3 Hepatitis A:         16

2.10.4 Helminthes:        16

2.10.5 Cholera:     17

2.10.6 Typhoid Fever:    17

Chapter Three

3.0 Materials and Methods 18

3.1 The Study Area: 18

3.2 Collection of Samples: 18

3.3. Isolation of Organisms from Three Aquatic Environment namely

Stream, Lake and River 19

3.4 Methodology used in Staining Procedures 20

3.4.2 Endspore Staining 20

3.4.3 Flagella Staining 21

3.5 Determination of Groups of Isolates Basedon Differentiation Techniques 21

3.6 Determination of Groups of Isolates Based on Structural Techniques. 22

Chapter Four  

4.0 Results 23

Chapter Five  

5.1 Discussion: 29

5.2 Conclusion: 29

5.3 Recommendation: 30

Reference

Appendix

LIST OF TABLES

TABLE TITLE PAGE

4.1 Total Concentration of Samples and its Grams Stain Reaction and Serial 25

Dilution

4.2 Groups of the Isolates Based on Differentiation Techniques   26

4.3 Groups of the Isolates Based on Structural Techniques   27

4.4 Biochemical Test carried out on the Three Aquatic Samples 28

4.5 Occurrence of General Bacteria Isolated from the Three 29

Different Aquatic sources  

CHAPTER ONE

1.0 INTRODUCTION:

Aquatic microbial diversity is well understood to be a key component of aquatic ecosystem functioning (Cotner & Biddanda, 2002; Gessner et al., 2010), and major advances toward linking microbial diversity with ecosystem function have made in aquatic system (Horner Devine et al., 2003; Smith, 2007; Sanger et al., 2010).

Aquatic diversity means the variability among living organisms from all sources and the ecological complexes of which they are part and it includes diversity within species or between species and of ecosystem. Diversity generally means “variation” and “differentiation” and “diversification”. In contrast to uniformity” then denotes “irregularities,” variety and differences when assessing biological systems, diversity may also be seen as “richness” (Kratochwil, 1999).

The highest overall marine diversity occurs in the tropical Indo Western Pacific area, a region that includes waters off the coast of Asia, Southeast Africa, Northern Australia and the Pacific Islands. The higher levels of diversity appear directly related to ecosystem stress (Hoffeman and Persons 1991).

Aquatic microbes are essential in making our planet habitable. The term “aquatic microbe” covers a diversity of microorganisms, including Microalgae, Bacteria and Achaea, Protozoa, Fungi and Viruses. These organisms are exceeding small-only 1/8000^th the volume of a human cell and spanning about 1/100^th the diameter of a human cell and spanning about 1/00^th the diameter of a human hair, up to a million of them live in just one milliliter of seawater (Dudgeon et al.,; 2006)

However, a recent survey of microbial diversity studies in aquatic habitats showed that microbial diversity in lotic environments is less commonly studied than in marine and lake ecosystems, and that impacted systems are less commonly studied than unimpacted systems (Zinger et al., 2012). Streams and rivers are hotspots of microbially mediated carbon (C) and nutrients processing within landscapes (Fisher et al., 1998; McClain et al., 2003).

Microbial activity drives Organic Matter (OM) decomposition, whole stream respiration and carbon flow to higher trophic levels (Hieber and Gessner, 2002; Tank et al., 2010). Also, microbial nitrification, denitrification and heterotrophic nitrogen (N) uptake in small streams affects downstream water quality. (Peterson et al., 2001; Mulholland et al., 2008, Valett et al., 2008).      

Early stream bacterial diversity research consisted of culture for diversity assessments of bacterial loads in the water Column, resulting on predominately Pseudomonas specie isolates with variation in diversity correlated most strongly with water, temperature, storm events and sunshine (Bell et al., 1982). These studies relied upon microscopy for identification of fungi conidia following sporulation or substract  utilization profiling of bacterial isolates. Bacteria diversity was particularly difficult to define, beyond differentiating Gram-negative from Gram-positive cells (Gesey et al., 1984), before the availability of molecular tools. Because microbial processes in stream and river ecosystem are variable in space and time in response to differences in nutrient availability (Dodds et al., 2000), temperature (Boyero et al., 2011), quality or quantity (Gessner and Chauvet, 1994), hydrological factors or land use (Mulholland et al., 2008), a reasonable initial prediction is that microbial diversity might also respond to changes in these environmental variables.

Based on the differentiation and structural staining, this procedure divides bacteria into separate groups based in their staining properties and structures. Structural stains take advantage of differences between a particular structure and the rest of the cells. Gram staining which is differential staining technique is used to characterize bacteria as Gram positive and Gram negative, the fixed bacterial smear is subjected to crystal violet, iodine solution, Alcohol and Safranin respectively (Verstraelen et al, 2009).Another staining procedure used in staining aquatic microbes is Endospore stains which are used for the identification of endospores or spores in aquatic environment (McClelland, 2001).

Flagella stain are  also used to stain structures which cannot be seen under ordinary microscope, hence the surface is coated with a precipitate to form a colladal substance and this perceptible serves as a layer of stainable material filamentous fungi (Asperigillus nigri) have been frequently isolated from aquatic environments such as rivers, streams. (M Wurzbacher et al., 2010) and so flagella stain are used to identify them.

1.1 AIMS AND OBJECTIVES

v To isolate microorganisms from three aquatic environments; fresh water, flowing River and lake.

v To determine the groups of the isolates based on differentiation techniques.

v  To determine the groups of the isolates based on structural techniques.

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