MICROBIOLOGICAL DETERIORATION OF PAPER

ABSTRACT

The aim of this study is microbial deteriorations of paper, with the following objectives to isolate and identify the bacteria present on each paper sample, to isolate and identify the fungal growth present on each paper sample, to ascertain the percentage occurrence of the various microorganisms in the samples evaluated, to also determine some compounds and enzymes that get degraded as a result of the growth of microorganisms on paper. The samples were cultured on MacConkey agar, Nutrient agar and Sabouraud Dextrose agar. The results obtained from the microbial analysis of the total numbers of Heterotrophic count ranges from 3.6 x 105to 9.7× 105, total coliform plate count ranges from 3.3× 105 to 6.4× 105 and total  fungi plate count 2.0 × 105 to 5.1× 105 in the used  paper samples. While for unused paper samples has Total heterotrophic counts ranging from 4.1 × 105to 7.0× 105 total coliform plate count 5.1 × 105 to 6.4 × 105cfu/g and total fungi plate count 3.3× 105 to 5.6× 105 A total of 8 bacteria and fungi isolate which comprised of 5 different species bacteria which are Escherichia coli, bacillus sp, micrococcus sp, pseudomonas sp and klebsiella sp and 3 species of fungi which are Aspergillus flavus, Aspergillus niger, Mucor alternaria. it was observed that bacteria isolates Escherichia coli, Micrococcus sp and Bacillus sp are the most frequently occurring isolates from deteriorated paper material sample used and unused with the percentage occurrence of (18.1%, 13.64% and 13.64%) and (25.0%, 12.5%, and 12.5%) respectively, while the lowest bacteria isolates were recorded in Klebsillaspp and Pseudomonas sp (9.09% and 9.09%) and Klebsiella sp for unused sample (6.25%). And for the fungi isolates Mucor alternaria and Aspergillus flavus were the most frequently occurring isolates from deteriorated paper sample with the percentage occurrence of (18.18% and 13.64%) respectively while the lowest of all is the Aspergillus niger which is (4.54%). The occurrence of fungi and bacteria on used and unused paper material suggests biological agent and chemical process that take place on these materials.

TABLE OF CONTENTS

Title page                                                                                                                                i

Certification                                                                                                                           ii

Declaration                                                                                                                             iii

Dedication                                                                                                                              iv

Acknowledgements                                                                                                                v

Table of content                                                                                                                      vi

List of tables                                                                                                                           x   

List of figures                                                                                                                         xi

Abstract                                                                                                                                  xii

CHAPTER ONE

1.1       Introduction

1.2       Types of Paper

1.2.1    Some Paper Other Types Include

1.3       Uses of Paper

1.4       Factors of Deterioration of Paper

1.4.1    Environmental Factors:

1.5       Aim and Objectives

1.5.1    Objectives

CHAPTER TWO

2.1       Literature Review

2.2       Foxing: A Bio deterioration of Paper

2.3       Mechanisms Underlying Foxing Spot Formation

2.3.1    Foxing as A Consequence of Fungal Colonization of Paper

2.4       Methods for the Identification and Assessment of Foxing On Paper

2.4.1    Culturing

2.4.2    Visible and Ultra-Violet Light Examination

2.4.3    Scanning Electron Microscopy and Energy Dispersive X-Ray Spectral Analysis

2.5       Biodeterioration Control and Sterilization Treatments for Paper

2.5.1    Freeze-Drying

2.5.2    Fumigation with Ethylene Oxide

2.6       Microorganism Involved in the Detoriation of Paper

2.6.1    Chaetomium Globosum

2.6.2    Geomyces Pannorum

2.6.3    Davidiella Tassiana

2.6.4    Basidiomycetes

2.7       The Use of Antifungals on Paper Based Collections

2.7.1    Chemical Methods

2.7.1.1 Alcohols

2.7.1.2 Ethanol

2.7.1.3 Essential Oils

2.7.1.4 Phenol Derivatives.

2.7.2    Physical Methods

2.7.2.1 Dehydration

2.7.2.2 High Frequency Current

2.7.2.3 Ultraviolet Radiation

2.7.2.4 Temperature Extremes

CHAPTER THREE

3.1       Material and Methods

3.2       Sample Collection

3.3       Materials

3.4       Isolation of Bacteria

3.4.1    Test Paper-Based Materials

3.5       Isolation and Identification of Bacteria

3.5.1    Examination of the Total Number of Bacteria in Paper-Based Materials

3.6       1solation and Identification of Fungi

3.7       Determination of Fungal Load

3.8       Identification of Isolated Bacteria and Fungi

3.9       Microscopic Examination

3.9.1    Cultural Characteristics

3.9.2    Morphological Characteristics

3.10     Biochemical Characteristics

3.10.1  Motility Test

3.10.2  Catalase Test

3.10.3  Oxidase Test

3.10.4  Indole Production

3.10.5  Methyl Red Reaction

3.10.6  Vogesproskauer (Vp) Reaction

3.10.7 Citrate Utilization

3.11 Carbohydrate Fermentation

3.12 Statistical Analysis

CHAPTER FOUR 

4.1       Results                                                                                                                       

CHAPTER FIVE

5.1       Discussion, Conclusion and Recommendation

5.2       Discussion

5.3       Conclusion

5.4       Recommendations

LIST OF TABLES

CHAPTER ONE

1.1      INTRODUCTION

Paper is a thin material produced by pressing together moist fibres of cellulose pulp derived from wood, rags or grasses, and drying them into flexible sheets. It is a versatile material with many uses, including writing, printing, packaging, cleaning, and a number of industrial and construction processes. Paper is mainly made of cellulose and secondary substances such as lignin, hemicellulose, pectin, waxes, tannins, proteins and mineral constituents. In time, paper undergoes processes of deterioration, which cause the scission of macromolecular chains, an irreversible process called paper aging. The chemical, physical or microbial deteriorations can affect the cellulose fibers of the chemical composition of paper, depending on the raw material and the manufacturing procedure used (Pasquariello et al., 2005).

Microbialbiodeterioration is the result of either the combined or separate activity of several organisms on different substrates and under varied ecological conditions, the understanding and stoppage of its effects require the concurrence of the different branches of biology, biochemistry, chemistry, physics-chemistry, etc. (Michaelsenet al., 2010). The chemical destructions are principally caused by the oxidation of the cellulose chains. Generation of free carbonyl groups and redox reactions can involve all paper constituents. The ink, glue and impurities as well as other organic or inorganic matters can negatively alter the process of paper ageing. Most often, the physical deterioration caused by light, temperature, humidity can enhance the biodegradation processes. The chemical hydrolysis of cellulose can stimulate the attack of microorganisms (Michaelsenet al., 2010).

Paper microbial deterioration leads to different types of losses/destructions depending on the organism involved (Michaelsenet al., 2010). Of the varied range of biological agents, three main categories can be distinguished: filamentous fungi, cellulolytic bacteria and insects. The literature estimates that there are over 200 species of fungi and bacteria causing paper degradation. Some of these are found in the raw materials, while others contaminate paper when coming into contact with air.

Degradation caused by microbial growth occurs in general as: chromatic alterations in the form of stains in a wide variety of colors (e.g. purple, yellow, brown, black, red etc.), shape and size due to the presence of a pigmented mycelium, of spores or, in certain circumstances, pigments produced by bacteria or fungi; structural alterations of the main components caused by enzymes (e.g. cellulases, proteases etc.) produced by different types of microorganisms, which ultimately lead to paper embrittlement or even its partial destruction; alterations to the essential added components (e.g. adhesives, plasticisers, antioxidants etc.) due to the attack of microorganisms (Pasquariello et al., 2005).

The presence of bacteria is indicated by appearance of isolated spots of different colors of low intensity at first. Once the attack begins, such spots acquire bigger sizes and deep hues: red, brown, black etc., depending on the species, pH value and so on. Fungi can colonize and degrade an extensive range of heritage assets. They secrete destructive acids and are able to depolymerize cellulose fibers. Many widely-spread species, such as Aspergillus sp., Alternaria sp., Chaetomium sp., Penicillium sp. can degrade cellulose and are often associated with the holdings of libraries (Konkol et al., 2009). The appearance of foxing (an extensively studied phenomenon lately) also was attributed to the species belonging to the Bacillus genera, as the causing agent (De Paolis et al., Lippincott; 2008), even though this degradation process is described as being triggered by the fungal attack as well. Species of actinomycetes and bacteria secreting strong, staining pigments and organic acids causing foxing (Strzelczyk et al., 2004) were identified. Still from foxing stains, (Michaelsen et al.,2010) isolated cellulolytic bacteria, namely Bacillus sp., Acinetobacter spp., Kochuria spp., Stenotrophomanasmaltophilia, Clostridium colinum. (Rakotonirainy et al., 2007) managed to isolate from foxing stains species of fungi belonging to the following genera: Aspergillus, Bjerkandera, Chaetomium, Gloeotinia, Penicillium, Polyporus, Saccharicola, Trichoderma and Ulocladium. Some species were detected only in 1-2 spots, while others (i.e. Penicilliumminioluteum, Gloeotiniatenulenta) were very frequently encountered.

1.2       TYPES OF PAPER

Categories of paper

Paper may be classified into seven categories (Pasquariello et al., 2005).

·       Printing papers of wide variety.

·       Wrapping papers for the protection of goods and merchandise. This includes wax and kraft papers.

·       Writing paper suitable for stationery requirements. This includes ledger, bank, and bond paper.

·       Blotting papers containing little or no size.

·       Drawing papers usually with rough surfaces used by artists and designers, including cartridge paper.

·       Handmade papers including most decorative papers, Ingres papers, Japanese paper and tissues, all characterized by lack of grain direction.

·       Specialty papers including cigarette paper, toilet tissue, and other industrial papers

1.2.1 Some other paper types include

·       Bank paper

·       Banana paper

·       Bond paper

·       Book paper

·       Coated paper: glossy and matte surface

·       Construction paper/sugar paper

·       Cotton paper

·       Fish paper (vulcanized fibres for electrical insulation)

·       Inkjet paper

·       Kraft paper

·       Laid paper

·       Leather paper

·       Mummy paper

·       Oak tag paper

·       Sandpaper

·       Tyvek paper

·       Wallpaper

·       Waterproof paper

·       Wax paper

·       Wove paper

·       Xuan paper

1.3   USES OF PAPER

Paper can be produced with a wide variety of properties, depending on its intended use:

·       For representing value: paper money, bank note, cheque, security (see security paper), voucher and ticket.

·       For storing information: book, notebook, graph paper, magazine, newspaper, art, zine, letter

·       For personal use: diary, note to remind oneself, etc.; for temporary personal use: scratch paper

·       For communication: between individuals and/or groups of people.

·       For packaging: corrugated box, paper bag, envelope, Packing & Wrapping Paper, Paper string, Charta emporetica and wallpaper.

·       For cleaning: toilet paper, handkerchiefs, paper towels, facial tissue and cat litter

·       For construction: papier-mâché, origami, paper planes, quilling, paper honeycomb, used as a core material in composite materials, paper engineering, construction paper and paper clothing

·       For other uses: emery paper, sandpaper, blotting paper, litmus paper, universal indicator paper, paper chromatography, electrical insulation paper (see also dielectric and permittivity) and filter paper

1.4       FACTORS OF DETERIORATION OF PAPER

Deterioration is a change of original state of any material by interaction between the object and the factors of destruction. The different types of deterioration of the paper based materials are reflected in wear and tear, shrinkage, cracks, brittleness, warping, bio-infestation, discoloration, abrasion, hole, dust and dirt accumulation etc. Generally, library materials are susceptible to deterioration by the following factors:

1. Environmental (climatic factors) factors like light, heat, humidity and moisture, dust and dirt, water.

2. Biological factors:  Microorganisms, insects and rodents.

3. Chemical factors

4. Human factors and

5. Disasters

1.4.1    ENVIROMENTAL FACTORS:

(a) Light: Whether natural light or artificial light paper gets deteriorated when it isexposed to light. Especially sun light, has a serious damaging effect on written or printed paper materials. The ultraviolet radiation of light is mainly responsible for photochemical degradation of paper which takes place rapidly when paper is exposed to sun light in presence of air (oxygen). When some portion of cellulose is oxidized to oxycellulose, the long cellulose chains are broken and the paper becomes weak and brittle. Fading of ink and dye of the colored paper and yellowing of white paper also takes place due to the formation of oxycellulose. Artificial light like fluorescent tube light also radiates a high percentage of ultraviolet rays which cause deterioration by yellowing the paper. However, the amount of damage by light depends upon the following factors.

i) Intensity of light: as the intensity of light increases the rate of deterioration of the paper also increases.

ii) Duration of exposure: the duration of exposure of paper to light is directly proportional to its deterioration.

iii) Distance from the source of light: more the distance, less the damage.

(b) Heat: Usually the source of heat is high atmospheric temperature. Heat is measured in terms temperature either in Centigrade scale or Fahrenheit scale. High heat with low humidity causes dehydration of cellulose fibers and the paper becomes brittle. It loses its flexibility to the extent that it tends to crumble on touch. On the other hand, high temperature with high humidity creates the condition for the growth of moulds. If electric bulbs are used for lighting purpose, they increase room temperature as high powerful bulbs generate more heat. Besides extreme variation in temperature (say 5^oC in winter and 45^oC in summer) affects the physical condition of the library materials.

(c) Humidity and Moisture: Humidity is the amount of moisture in the atmospheric air. The moisture is measured in terms of relative humidity. All organic objects absorb water to a greater or lower extent and the water goes inside the object through surrounding air. Because of this absorbency property, the paper absorbs more moisture when there is high humidity. Certain amount of humidity is necessary for the flexibility of paper but in prolonged high humid condition, paper becomes soggy and the moisture weakens the fibers of paper. Moisture is the root cause of various types of physical, chemical and biological deterioration of library materials. It weakens the adhesive and makes the book binding loose. It also weakens the sizing elements of paper and causes spreading of ink.

(d) Dust and Dirt: Fine dry particles of any matter present in the air are known as dust. Dust, which is highly dangerous for the library and archival collection, composed of soil, tar, metallic substances, fungus spores and moisture among other things. Since dust is air borne it settles down on any surface of the object. Dust is hygroscopic in nature and when it is mixed with high humidity, it is transformed into dirt and if this dirt sticks to the surface of the books, it becomes difficult to remove. Dust and dirt are sources of both physical and chemical degradation of the library collection.

(e) Water: Water occurs in all the normal state of matter- solid, liquid and gas. It acts as a physical agent of deterioration by causing hygroscopic materials to undergo dimensional changes. Water, which is harmful for the library collection may come from sources like natural calamities, human negligence, from leaking roofs, defective plumbing and through open windows at the time of raining. Excessive water brings about biological attack on paper, which is usually manifested as the growth of fungus or mildew. The effects of water are stained paper, rotted leather, and smeared ink, weaken adhesive, sustained fungi etc. Water also does injury to the steel furniture due to rusting.

2. BIOLOGICAL FACTORS:

The deterioration caused by biological agents such as microorganisms, insects and rodents is generally known as biodeterioration. Almost all book components, be it paper, leather, textiles or straw board used for binding are prone to attacks by these biological agents. The problem of biodeterioration is a matter of considerable significance of tropical hot and humid climate like India.

These biological agents can be subdivided into:

(a) Microorganisms- Fungus or moulds, bacteria etc.

(b) Insects

(c) Rodents

(i) Microorganisms

i. Fungus: Fungus are a large heterogenous group of plant organisms. The fungal spores are present in the earth, water and air and remain in a dormant state for long periods. These spores sprout and grow when they have the required moisture and heat. Generally, fungi grow in a relative humidity range of 63-100% and temperature range of 15-35⁰ c. In libraries fungal growth is known as mould or mildew and they appear as brown/black vegetative growth on paper, leather and textiles. Fungus consume cellulose and also thrive on nutrients in leather, glues, pastes, binding threads etc. they weaken and stain the paper and can cause discoloration.

ii. Bacteria: Besides fungus, bacteria also decompose cellulose in paper and binding textiles.

(ii) Insects Even though there are thousands of insects, only certain insects badly damage the

Archive-library materials. They are silverfish, cockroaches, booklice, bookworms’ and termites.

1.5       AIM AND OBJECTIVES

The aim of the present study is to determine microbiological deterioration of different types of paper.

The objectives are

·       To isolate and identify the bacteria present on each paper sample.

·       To isolate and identify the fungal growth present on each paper sample.

·       To ascertain the percentage occurrence of the various microorganisms in the samples evaluated.

·       To also determine some compounds and enzymes that get degraded as a result of the growth of microorganisms on paper.

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