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Energy, Environment, and Sustainability

Sunita J. Varjani
Avinash Kumar Agarwal
Edgard Gnansounou
Baskar Gurunathan Editors

Bioremediation:
Applications for
Environmental
Protection and
Management
Energy, Environment, and Sustainability

Series editors
Avinash Kumar Agarwal, Department of Mechanical Engineering, Indian Institute
of Technology, Kanpur, Uttar Pradesh, India
Ashok Pandey, Distinguished Scientist, CSIR-Indian Institute of Toxicology
Research, Lucknow, India
This books series publishes cutting edge monographs and professional books
focused on all aspects of energy and environmental sustainability, especially as it
relates to energy concerns. The Series is published in partnership with the
International Society for Energy, Environment, and Sustainability.
The books in these series are editor or authored by top researchers and professional
across the globe. The series aims at publishing state-of-the-art research and
development in areas including, but not limited to:
• Renewable Energy
• Alternative Fuels
• Engines and Locomotives
• Combustion and Propulsion
• Fossil Fuels
• Carbon Capture
• Control and Automation for Energy
• Environmental Pollution
• Waste Management
• Transportation Sustainability

More information about this series at http://www.springer.com/series/15901

Sunita J. Varjani ⋅ Avinash Kumar Agarwal
Edgard Gnansounou ⋅ Baskar Gurunathan
Editors

Bioremediation: Applications
for Environmental Protection
and Management

123
Editors
Sunita J. Varjani Edgard Gnansounou
Gujarat Pollution Control Board Bioenergy and Energy Planning Research
Gandhinagar, Gujarat Group (BPE)
India Swiss Federal Institute of Technology
Lausanne (EPFL)
Avinash Kumar Agarwal Lausanne
Department of Mechanical Engineering Switzerland
Indian Institute of Technology Kanpur
Kanpur, Uttar Pradesh Baskar Gurunathan
India Department of Biotechnology
St. Joseph’s College of Engineering
Chennai, Tamil Nadu
India

ISSN 2522-8366 ISSN 2522-8374 (electronic)

Energy, Environment, and Sustainability
ISBN 978-981-10-7484-4 ISBN 978-981-10-7485-1 (eBook)
https://doi.org/10.1007/978-981-10-7485-1
Library of Congress Control Number: 2017959893

© Springer Nature Singapore Pte Ltd. 2018

This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part
of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations,
recitation, broadcasting, reproduction on microfilms or in any other physical way, and transmission
or information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar
methodology now known or hereafter developed.
The use of general descriptive names, registered names, trademarks, service marks, etc. in this
publication does not imply, even in the absence of a specific statement, that such names are exempt from
the relevant protective laws and regulations and therefore free for general use.
The publisher, the authors and the editors are safe to assume that the advice and information in this
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The registered company address is: 152 Beach Road, #21-01/04 Gateway East, Singapore 189721, Singapore
Preface

Varieties of pollutants are released into the environment day by day due to increase
in population and urbanization which leads to environmental pollution, viz. air, land
and water. Pollutants may be organic or inorganic. Environmental Protection and
Management is a burning issue for different industrial sectors. Many conventional
engineering-based physicochemical decontamination methods for remediation
of these pollutants are expensive. The increasing costs and limited efficiency of
traditional physicochemical treatments have spurred the development of green
technologies which ultimately leads to the sustainable development of the
environment.
The first international conference on ‘Sustainable Energy and Environmental
Challenges’ (SEEC-2017) was organized under the auspices of ‘International
Society for Energy and Environmental Sustainability’ (ISEES) by the ‘Center of
Innovative and Applied Bioprocessing’ (CIAB), Mohali, held from 26–28 February
2017. ISEES was founded at IIT Kanpur in January 2014 with the aim of spreading
knowledge in the fields of energy, environment, sustainability and combustion. The
society’s goal is to contribute to the development of clean, affordable and secure
energy resource and a sustainable environment for the society and to spread
knowledge in the above-mentioned areas and awareness about the environmental
challenges, which the world is facing today. ISEES is involved in various activities
such as conducting workshops, seminars and conferences in the domains of its
interest. The society also recognizes the outstanding works done by the young
scientists and engineers for their contributions in these fields by conferring them
awards under various categories.
This conference provided a platform for discussions between eminent scientists
and engineers from various countries including India, the USA, South Korea,
Norway, Malaysia and Australia. In this conference, eminent speakers from all over
the world presented their views related to different aspects of energy, combustion,
emissions and alternative energy resource for sustainable development and cleaner
environment. The conference started with four mini-symposiums on very topical
themes, which included (i) New Fuels and Advanced Engine Combustion,
(ii) Sustainable Energy, (iii) Experimental and Numerical Combustion and

v
vi Preface

(iv) Environmental Remediation and Rail Road Transport. The conference had 14
technical sessions on topics related to energy and environmental sustainability and a
panel discussion on ‘Challenges, Opportunities and Directions of Technical Edu-
cation & Research in the Area of Energy, Environment and Sustainability’ to wrap
up the three-day technical extravaganza. The conference included 2 plenary talks,
12 keynote talks, 42 invited talks from prominent scientists, 49 contributed talks
and 120 posters. A total of 234 participants and speakers attended this three-day
conference, which hosted Dr. V. K. Saraswat, Member, NITI Aayog, India, as a
chief guest for the award ceremony of ISEES. This conference laid out the road
map for technology development, opportunities and challenges in this technology
domain. The technical sessions in the conference included Advances in IC Engines
and Fuels; Conversion of Biomass to Biofuels; Combustion Processes; Renewable
Energy: Prospects and Technologies; Waste to Wealth—Chemicals and Fuels;
Energy Conversion Systems; Numerical Simulation of Combustion Processes;
Alternate Fuels for IC Engines; Sprays and Heterogeneous Combustion of
Coal/Biomass; Biomass Conversion to Fuels and Chemicals—Thermochemical
Processes; Utilization of Biofuels; and Environmental Protection and Health. All
these topics are very relevant for the country and the world in the present context.
The society is grateful to Prof. Ashok Pandey for organizing and hosting this
conference, which led to the germination of this series of monographs, which
included 16 books related to different aspects of energy, environment and sus-
tainability. This is the first time that such a voluminous and high-quality outcome
has been achieved by any society in India from one conference.
The editors would like to express their sincere gratitude to the authors for
submitting their work in a timely manner and revising it appropriately at short
notice. We would like to express our special thanks to reviewers for reviewing
various chapters of this monograph and provided their valuable suggestions to
improve the manuscripts. We acknowledge the support received from various
funding agencies and organizations for successfully conducting the first ISEES
conference SEEC-2017, where these monographs germinated. These include
Department of Science and Technology, Government of India (special thanks to Dr.
Sanjay Bajpai); TSI, India (special thanks to Dr. Deepak Sharma); Tesscorn, India
(special thanks to Sh. Satyanarayana); AVL India; Horiba, India; Springer (special
thanks to Swati Mehershi); CIAB (special thanks to Dr. Sangwan).
Due to some limitations such as cost and efficiency, conventional remediation
methods of polluted sites, there is a need for the development of alternative tech-
nologies for in situ applications, particularly based on biological remediation
capabilities of plants and microorganisms. Green technologies for clean-up of
pollutants by biological means are used for Environmental Protection and Man-
agement. Hence, bioremediation technology is referred as an efficient, economic,
versatile and environmentally sound technique. The monograph is primarily
focused on every aspect of bioremediation technology practiced globally. It pro-
vides concise and updated literature on bioremediation technologies as a tool for
Environmental Protection and Management, which can be used by engineers,
scientists and academicians working in the field of bioremediation. Every chapter
Preface vii

of the book contains recent information and is clearly illustrated with tables, figures
and pictures in a more simple and scientific way.
The book shall include chapters on different aspects of recent advances in
bioremediation of different environmental pollutants. Some of the topics covered in
this book are: evaluation of next-generation sequencing technologies for environ-
mental monitoring in wastewater abatement; genetically modified organisms and its
impact on the enhancement of bioremediation; bioremediation of industrial
wastewater using bioelectrochemical treatment; phenol degradation from industrial
wastewater by engineered microbes; bioremediation of heavy metals; pesticides
bioremediation; mathematical modelling in bioremediation; application of microbes
in remediation of hazardous wastes: a review; phytoremediation of textile dyes and
effluents; biosorption strategies in the remediation of toxic pollutants from con-
taminated water bodies; phytoremediation technique for the removal of dye in
wastewater; role of nanofibers in bioremediation; bioremediation of volatile organic
compounds using biofilters; bioremediation of industrial and municipal wastewater
using microalgae; role of biosurfactants in enhancing the microbial degradation of
pyrene and so on.

Gandhinagar, India Sunita J. Varjani

Kanpur, India Avinash Kumar Agarwal
Lausanne, Switzerland Edgard Gnansounou
Chennai, India Baskar Gurunathan
Contents

1 Introduction to Environmental Protection and Management . . . . . 1

Sunita J. Varjani, Avinash Kumar Agarwal, Edgard Gnansounou
and Baskar Gurunathan
2 Mathematical Modeling in Bioremediation . . . . . . . . . . . . . . . . . . . 7
Parthasarthy Vijay and Margavelu Gopinath
3 Evaluation of Next-Generation Sequencing Technologies
for Environmental Monitoring in Wastewater Abatement . . . . . . . 29
P. Senthil Kumar, S. Suganya and Sunita J. Varjani
4 Genetically Modified Organisms and Its Impact on the
Enhancement of Bioremediation . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
Narasimhan Manoj Kumar, Chandrasekaran Muthukumaran,
Govindasamy Sharmila and Baskar Gurunathan
5 Integration of Lignin Removal from Black Liquor and
Biotransformation Process . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77
Krishnamurthi Tamilarasan, Periyar Selvam Sellamuthu
and Baskar Gurunathan
6 Role of Nanofibers in Bioremediation . . . . . . . . . . . . . . . . . . . . . . . 99
Sekar Aiswarya Devi, Muthukumar Harshiny
and Manickam Matheswaran
7 Bioremediation of Industrial Wastewater Using
Bioelectrochemical Treatment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 115
N. Samsudeen and Manickam Matheswaran
8 Biosorption Strategies in the Remediation of Toxic Pollutants
from Contaminated Water Bodies . . . . . . . . . . . . . . . . . . . . . . . . . . 127
P. Senthil Kumar and K. Grace Pavithra

ix
x Contents

9 Bioremediation of Heavy Metals . . . . . . . . . . . . . . . . . . . . . . . . . . . 165

P. Senthil Kumar and E. Gunasundari
10 Pesticides Bioremediation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 197
P. Senthil Kumar, C. Femina Carolin and Sunita J. Varjani
11 Application of Microbes in Remediation of Hazardous Wastes:
A Review . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 223
Moni Kumari, Pooja Ghosh and Indu Shekhar Thakur
12 Phytoremediation Techniques for the Removal of Dye in
Wastewater . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 243
B. Bharathiraja, J. Jayamuthunagai, R. Praveenkumar and J. Iyyappan
13 Phenol Degradation from Industrial Wastewater by Engineered
Microbes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 253
Ravichandran Rathna and Ekambaram Nakkeeran
14 Insect Gut Bacteria and Their Potential Application in
Degradation of Lignocellulosic Biomass: A Review . . . . . . . . . . . . . 277
Rajesh Kumar Prasad, Soumya Chatterjee, Sonika Sharma,
Pranab Behari Mazumder, Mohan G. Vairale and P. Srinavas Raju
15 Bioremediation of Volatile Organic Compounds in Biofilters . . . . . 301
Margavelu Gopinath, Rose Havilah Pulla, K. S. Rajmohan,
Parthasarthy Vijay, Chandrasekaran Muthukumaran
and Baskar Gurunathan
16 Bioremediation of Industrial and Municipal Wastewater
Using Microalgae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 331
Baskar Gurunathan, I. Aberna Ebenezer Selvakumari, R. Aiswarya
and S. Renganthan
17 Phytoremediation of Textile Dye Effluents . . . . . . . . . . . . . . . . . . . 359
Shanmugaprakash Muthusamy, Dhilipkumar Govindaraj
and Karthik Rajendran
18 Role of Biosurfactants in Enhancing the Microbial
Degradation of Pyrene . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 375
Gauri Gupta, Avantika Chandra, Sunita J. Varjani,
Chiranjib Banerjee and Vipin Kumar
19 Bioremediation of Nitrate-Contaminated Wastewater
and Soil . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 387
K. S. Rajmohan, Margavelu Gopinath and Raghuram Chetty
Author Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 411
Editors and Contributors

About the Editors

Dr. Sunita J. Varjani is Scientific Officer at Gujarat Pollution
Control Board, Gandhinagar, Gujarat, India. She holds M.Sc.
degree in Microbiology (2009) and Ph.D. in Biotechnology
(2014). Her major areas of research are Industrial and Environ-
mental Microbiology/Biotechnology and Molecular Biology.
She has authored 35 publications, including 1 book, 19 book
chapters/reviews and 15 original research papers. She has won
several awards and honours, including Young Scientist Award at
AFRO-ASIAN Congress on Microbes for Human and Environ-
mental Health, New Delhi (2014), and Best Paper Awards for
oral presentations in national and international conferences in
2008, 2012 and 2013. She is a member of the editorial board of
Journal of Energy and Environmental Sustainability.

Prof. Avinash Kumar Agarwal joined IIT Kanpur in 2001 and

is currently a Poonam and Prabhu Goyal Endowed Chair Pro-
fessor. He was at ERC, University of Wisconsin, Madison, USA,
as a Postdoctoral Fellow (1999–2001). His areas of interest are
IC engines, combustion, alternative fuels, hydrogen, conven-
tional fuels, lubricating oil tribology, optical diagnostics, laser
ignition, HCCI, emission and particulate control and large bore
engines. He has published more than 160 peer-reviewed inter-
national journals and conference papers. He is Associate Editor
of ASME Journal of Energy Resources Technology and Inter-
national Journal of Vehicle Systems Modelling and Testing. He
has edited ‘Handbook of Combustion’ (5 volumes; 3168 pages),
published by Wiley VCH, Germany. He is a Fellow of SAE
(2012), a Fellow of ASME (2013) and a Fellow of INAE (2015).
He is the recipient of several prestigious awards such as
NASI-Reliance Industries Platinum Jubilee Award-2012; INAE
Silver Jubilee Young Engineer Award-2012; Dr. C.V. Raman
Young Teachers Award-2011; SAE International’s Ralph R.
Teetor Educational Award-2008; INSA Young Scientist

xi
xii Editors and Contributors

Award-2007; UICT Young Scientist Award-2007; INAE Young

Engineer Award-2005. He is the recipient of prestigious Shanti
Swarup Bhatnagar Award-2016 in Engineering Sciences. He is
the first combustion/IC engine researcher to get this honour.

Prof. Edgard Gnansounou is a Professor of Modelling and

planning of Energy Systems at the Swiss Federal Institute of
Technology Lausanne (EPFL) where he is Director of the
Bioenergy and Energy Planning Research Group. His current
research works comprise techno-economic and environmental
assessment of bio-refinery schemes based on the conversion of
agricultural residues. He is leading research projects in that field
in several countries including Brazil, Colombia and South
Africa. He is credited with numerous papers in high-impact
scientific journals. He is a member of the editorial board of
Bioresource Technology Journal. He graduated with an M.S. in
Civil Engineering and Ph.D. in Energy Systems at the Swiss
Federal Institute of Technology Lausanne. He was Visiting
Researcher at the Thayer College, Dartmouth School of Engi-
neering, with Professor Charles Wyman, USA, at Polytech of
Clermont-Ferrand, University Blaise Pascal, France, and at the
Center of Biofuels, the National Institute for Interdisciplinary
Science and Technology, with Prof. Ashok Pandey, Trivandrum,
India. He was also Visiting Professor of the African University
of Science of Technology, Abuja, Nigeria. He is a citizen of
Benin, Africa, and Switzerland.

Prof. Baskar Gurunathan is currently working as Professor of

Biotechnology in St. Joseph’s College of Engineering, Chennai,
India. He has set an example for scholarly research work during
the past 11 years. Since his postgraduation, he has published his
research work in 92 reputed national and international journals.
To his credit, he has presented more than 100 papers in national
and international forum and published 5 chapters in books.
Currently, his research works have got h-index of 14 and i10
index of 19 in Google Scholar with a total citation of 483. His
current research expertise in biofuels, therapeutic proteins,
microbial enzymes nanomedicine and nanocatalysis clearly
indicates his thirst towards research and development in multi-
disciplinary areas. Currently, he is the Management Council
Member of Biotechnology Research Society, India (2017–19).
He is also the Honorary Secretary of Indian Institute of Chemical
Engineers, Chennai Regional Centre (2016–17). He is also an
active life member of various professional bodies to keep abreast
of latest trends and developments. He has organized various
training programmes to enhance the knowledge of students,
faculties and researchers. He has received two funded projects of
total worth Rs. 55 lakhs from DST (Food Processing) and DBT
Editors and Contributors xiii

(Cancer Biology), GOI, during the academic year 2015–16. He

has received Outstanding Reviewer Award from Bioresource
Technology in 2013, Materials science and Engineering-C in
2015 and Biochemical Engineering Journal and Energy Con-
version and Management in 2016. The International Biopro-
cessing Association (International Forum on Industrial
Bioprocesses) has conferred him with Young Scientist
Award-2015 in recognition of his contributions in the area of
Bioprocess Technology. The Indian Society for Technical Edu-
cation has conferred him with ISTE-Syed Sajid Ali National
Award-2016 for his outstanding research work in the area of
Renewable Energy.

Contributors
Avinash Kumar Agarwal Department of Mechanical Engineering, Indian Insti-
tute of Technology Kanpur, Kanpur, India
R. Aiswarya Department of Biotechnology, St. Joseph’s College of Engineering,
Chennai, India
Sekar Aiswarya Devi Department of Chemical Engineering, National Institute of
Technology, Tiruchirappalli, India
Chiranjib Banerjee Department of Environmental Science & Engineering, Indian
Institute of Technology (ISM), Dhanbad, Jharkhand, India
B. Bharathiraja Vel Tech High Tech Dr. Rangarajan Dr. Sakunthala Engineering
College, Chennai, Tamilnadu, India
Avantika Chandra Department of Environmental Science & Engineering, Indian
Institute of Technology (ISM), Dhanbad, Jharkhand, India
Soumya Chatterjee Defence Research Laboratory, DRDO, Tezpur, Assam, India
Raghuram Chetty Department of Chemical Engineering, Indian Institute of
Technology Madras, Chennai, Tamil Nadu, India
C. Femina Carolin Department of Chemical Engineering, SSN College of
Engineering, Chennai, India
Pooja Ghosh Centre for Rural Development and Technology, Indian Institute of
Technology, New Delhi, India
Edgard Gnansounou Bioenergy and Energy Planning Research Group (BPE),
Swiss Federal Institute of Technology Lausanne (EPFL), Lausanne, Switzerland
xiv Editors and Contributors

Margavelu Gopinath Department of Chemical Engineering, University of Pet-

roleum and Energy Studies, Dehradun, Uttarakhand, India
Dhilipkumar Govindaraj Downstream Processing Laboratory, Department of
Biotechnology, Kumaraguru College of Technology, Coimbatore, India
K. Grace Pavithra Department of Chemical Engineering, SSN College of Engi-
neering, Chennai, India
E. Gunasundari Department of Chemical Engineering, SSN College of Engi-
neering, Chennai, India
Gauri Gupta Laboratory of Applied Microbiology, Department of Environmental
Science & Engineering, Indian Institute of Technology (ISM), Dhanbad, Jharkhand,
India
Baskar Gurunathan Department of Biotechnology, St. Joseph’s College of
Engineering, Chennai, Tamil Nadu, India
Muthukumar Harshiny Department of Chemical Engineering, National Institute
of Technology, Tiruchirappalli, India
J. Iyyappan Vel Tech High Tech Dr. Rangarajan Dr. Sakunthala Engineering
College, Chennai, Tamilnadu, India
J. Jayamuthunagai Centre for Biotechnology, Anna University, Chennai,
Tamilnadu, India
Narasimhan Manoj Kumar Department of Genetic Engineering, School of
Bioengineering, SRM University, Kattankulathur, Tamilnadu, India
Vipin Kumar Laboratory of Applied Microbiology, Department of Environmental
Science & Engineering, Indian Institute of Technology (ISM), Dhanbad, Jharkhand,
India
Moni Kumari School of Environmental Sciences, Jawaharlal Nehru University,
New Delhi, India
Manickam Matheswaran Department of Chemical Engineering, National Insti-
tute of Technology, Tiruchirappalli, India
Pranab Behari Mazumder Department of Biotechnology, Assam University,
Silchar, Assam, India
Chandrasekaran Muthukumaran Department of Industrial Biotechnology,
Government College of Technology, Coimbatore, Tamilnadu, India
Shanmugaprakash Muthusamy Downstream Processing Laboratory, Depart-
ment of Biotechnology, Kumaraguru College of Technology, Coimbatore, India
Editors and Contributors xv

Ekambaram Nakkeeran Research Laboratory, Department of Biotechnology,

SriVenkateswara College of Engineering (Autonomous), Sriperumbudur, Tamil
Nadu, India
Rajesh Kumar Prasad Defence Research Laboratory, DRDO, Tezpur, Assam,
India; Department of Biotechnology, Assam University, Silchar, Assam, India
R. Praveenkumar Department of Biotechnology, Arunai Engineering College,
Tiruvannamalai, Tamilnadu, India
Rose Havilah Pulla Department of Chemical Engineering, College of Engineering
Studies, University of Petroleum and Energy Studies, Dehradun, India
Karthik Rajendran MaREI Center, Environmental Research Institute, University
College Cork, Cork, Ireland; School of Engineering, University College Cork,
Cork, Ireland
K. S. Rajmohan Department of Chemical Engineering, College of Engineering
Studies, University of Petroleum and Energy Studies, Dehradun, India; Department
of Chemical Engineering, Indian Institute of Technology Madras, Chennai, Tamil
Nadu, India
P. Srinavas Raju Defence Research Laboratory, DRDO, Tezpur, Assam, India
Ravichandran Rathna Research Laboratory, Department of Biotechnology, Sri
Venkateswara College of Engineering (Autonomous), Sriperumbudur, Tamil Nadu,
India
S. Renganthan Department of Biotechnology, A.C.Tech., Anna University,
Chennai, India
N. Samsudeen Department of Chemical Engineering, National Institute of Tech-
nology, Tiruchirappalli, India
Periyar Selvam Sellamuthu Department of Food Process Engineering, SRM
University, Chennai, Tamilnadu, India
I. Aberna Ebenezer Selvakumari Department of Biotechnology, St. Joseph’s
College of Engineering, Chennai, India
P. Senthil Kumar Department of Chemical Engineering, SSN College of Engi-
neering, Chennai, India
Sonika Sharma Defence Research Laboratory, DRDO, Tezpur, Assam, India
Govindasamy Sharmila Department of Industrial Biotechnology, Government
College of Technology, Coimbatore, Tamilnadu, India
S. Suganya Department of Chemical Engineering, SSN College of Engineering,
Chennai, India
xvi Editors and Contributors

Krishnamurthi Tamilarasan Department of Chemical Engineering, SRM

University, Chennai, Tamilnadu, India
Indu Shekhar Thakur School of Environmental Sciences, Jawaharlal Nehru
University, New Delhi, India
Mohan G. Vairale Defence Research Laboratory, DRDO, Tezpur, Assam, India
Sunita J. Varjani Gujarat Pollution Control Board, Gandhinagar, Gujarat, India
Parthasarthy Vijay Department of Chemical Engineering, College of Engineer-
ing Studies, University of Petroleum and Energy Studies, Dehradun, India
Chapter 1
Introduction to Environmental Protection
and Management

Sunita J. Varjani, Avinash Kumar Agarwal, Edgard Gnansounou

and Baskar Gurunathan

Abstract Man’s environment consists of natural resources like air, land, water,
plants, and animals. With the progress of industrialization and civilization, man has
interacted with his surroundings and disturbed the nature. It leads to environmental
pollution, which cannot be eradicated by nature’s self-acting process, i.e., various
biogeochemical cycles. Environmental problems stem from two main categories of
human activities: (a) resources utilization at unsustainable levels and contamination
of the environment through pollution and (b) discharge of wastes at levels beyond
the earth’s and environment’s capacity to absorb them or render them harmless
which results in ecological damage and degradation of the environment. Environ-
mental damage around includes pollution of water and air and consequent health
problems, biodiversity loss, deterioration of buildings and monuments, soil fertility
loss, desertification, ozone depletion, and many more. Environmental protection
and management has become one of the foremost concerns of the world commu-
nity. International concern for environmental protection and management has
gained momentum with Stockholm Declaration in 1972. It is considered as Magna
Carta of environmental protection and sustainable development. Then a series of
global efforts have been undertaken internationally for protection of the environ-

S. J. Varjani (✉)
Gujarat Pollution Control Board, Paryavaran Bhavan, Gandhinagar, India
e-mail: drsvs18@gmail.com
A. K. Agarwal
Department of Mechanical Engineering, Indian Institute of Technology Kanpur,
Kanpur, India
e-mail: akag@iitk.ac.in
E. Gnansounou
Bioenergy and Energy Planning Research Group (BPE), Swiss Federal Institute
of Technology Lausanne (EPFL), Lausanne, Switzerland
e-mail: edgard.gnansounou@epfl.ch
B. Gurunathan
Department of Biotechnology, St. Joseph’s College of Engineering,
Chennai, Tamil Nadu, India
e-mail: baskarg@stjosephs.ac.in

© Springer Nature Singapore Pte Ltd. 2018 1

S. J. Varjani et al. (eds.), Bioremediation: Applications for Environmental
Protection and Management, Energy, Environment, and Sustainability,
https://doi.org/10.1007/978-981-10-7485-1_1
2 S. J. Varjani et al.

ment. Hence, environmental protection has become not only local, regional, or
national importance but also a global concern. Over the past several decades,
growing public awareness regarding threats to the environment, informed by
warnings from scientists, has led to demands that law protects the natural sur-
roundings on which human well-being depends. Under growing pressure from
national and international public opinion, governments began to demonstrate
concern over the general state of the environment introduced legislation to combat
pollution of inland waters, ocean, and air.

Keywords Biodegradation ⋅
Bioleaching ⋅
Genetically modified microorgan-
isms ⋅ Microalgae ⋅
Molecular tools ⋅
Nanofibers ⋅
Next-generation
sequencing

The waste generation and their disposal in natural water bodies become a serious
topic of concern. Consequently, there is a demand for new strategies and tech-
nologies to address wastewater treatment and subsequent recycle and reuse. Bio-
logical treatment process is advantageous and constitutes tools to biodegrade
organic matter, transfer toxic compounds into harmless products, and nutrient
removal in wastewater microbiology. Biomonitoring employs sentinel or indicator
species in water bodies to infer water quality, ecosystem health status, and to
protect public health from waterborne risks. Next-generation sequencing is one of
the most leveraging studies focussed on the ecology of microbial-mediated pro-
cesses that influence freshwater quality such as algal blooms, contaminant
biodegradation, and pathogen dissemination. One of the chapters in this book
discusses next-generation sequencing technologies for environmental monitoring in
wastewater abatement. However, essential hypothesis and utilization of contami-
nant transport demonstrated by numerical methods have been discussed in other
chapter.
Bioremediation is a process to degrade environmental pollutants, which are
introduced accidentally or deliberately, and causes a hazardous effect on the earth
and harms the normal life process. The conversion of these pollutants into less toxic
forms is the goal of bioremediation process that can be achieved by the use of
microorganisms. The bioremediation approaches have more advantages when
compared with traditional methods as it can be directly implemented at the targeted
pollutant site. Sometimes, bacteria and fungus when employed to decompose
chemical compounds showed limited ratio to metabolize the pollutants on their
own. The genetically modified organisms as well as immobilized microbes/their
products are applied nowadays in bioremediation process for effective removal of
pollutants, where the indigenous microbes cannot work efficiently. Genetically
modified microorganisms (GMOs) play an important role in remediating the
industrial waste, reducing the toxicity of some hazardous compounds, and helping
in removal of pollutants efficiently. A variety of molecular tools used for con-
struction of GMOs, pros and cons, ethical issues, and laws governing the appli-
cation of GMOs are explained.
1 Introduction to Environmental Protection and Management 3

Bioremediation of pollutants released in environment from various industries,

viz. paper and pulp, dye and dyes intermediates, metal, pesticide manufacturing,
pharmaceuticals, chemical manufacturing, petroleum refineries, petrochemical, coal
gasification operations, and tannery by various biological means, viz. microor-
ganisms, plant, and microalgae is discussed in-depth by authors of different
chapters. Heavy metals, radioactive waste, hydrocarbons, pesticides, phenol, and
nitrate are some of the leading toxic pollutants in the environment. Challenges are
faced in decontamination of these types of pollutants in soil and water for a long
period of time. A number of methods such as membrane technology, electro-Fenton
reaction, advanced oxidation process, nanotechnology play a major role in
removing toxic pollutants, but difficulties are seen in degradation of toxic sludge,
additional side reactions, high cost in initial installment and in maintenance, etc.
Roles of microorganisms and plants to remediate the organic and inorganic are
discussed in chapters of this book. Recent bioremediation technologies and meth-
ods for pollutant removal are also discussed.
Phenol and its derivatives are the most pondered substantial pollutant generated
from various industrial processes. Accumulation of phenol even at a lower con-
centration may be fatal to all living beings in the ecosystem. Overview of phenol
pollution, deleterious effects of phenol in ecosystem, biodegradation of phenols,
and significance of engineered microbes for phenol degradation are discussed.
Pyrene is a high-molecular-weight polycyclic aromatic hydrocarbon (PAH) with a
symmetrical structure, commonly found as a pollutant of air, water, and soil. Being
one of the most abundant high-molecular-weight pericondensed PAH and having its
structure similar to several carcinogenic PAHs, it is being used as a model com-
pound to study the degradation of high-molecular-weight PAHs. Therefore, its
removal from the environment is a challenging task for scientists. Microbial
degradation of pyrene by pure microbial cultures and microbial consortium has
been discussed by authors, which simultaneously emphasizes the role of surfactants
in enhancing degradation process.
Microbes such as bacteria and fungi are involved in biodegradation of lignin
present in the effluent of paper and pulp industry. Degradation of lignin by white-rot
fungi may be helpful for the biotechnical applications like biopulping, biobleaching
and pulp mill effluents treatment, and soil bioremediation. The abundance and
renewability of lignin potentially converted to valuable bioproduct may eventually
replace the existing technology in manufacturing industries.
Scarcity of pure water is a threatening issue worldwide. Water is essential for
human survival and all activities on the earth. The effluent water from industries
containing recalcitrant pollutants causes dangerous impacts to the environment and
human health. Immobilized nanofibers possessing enhanced catalytic activity, high
stability, and very good reusability of novel nanobiocomposites have remarkable
potential for the treatment of water and wastewater. It also plays a major role in safe
preservation of bioremediating bacteria for potential wastewater treatment appli-
cations. Nanofibers have become a popular carrier matrix for immobilization of
specific microorganisms. Simple, versatile, and cost-effective properties of nano-
fibers made them a promising tool for microbial integration which enhances the
4 S. J. Varjani et al.

bioremediation by efficient removal of contaminants such as dyes and heavy metals

from wastewater. One chapter of this book describes immobilization of specific
bacteria on electrospinning nanofibers and its application in bioremediation process.
However, the other chapter summarizes the research on bioelectrochemical systems
for bioremediation of organic matter as well as recovery of heavy metal ions from
the wastewater.
Biosorption is a process involving solid and liquid phases in which dissolved
species need to be sorbed. Low cost, high efficiency, and reusability of biosorbent
are some of the advantages of biosorption. Biosorption involves removal of toxic
pollutants by biomass. Some microorganisms are targeted for the removal of single
pollutant alone. Algae, bacteria, fungi, yeast, waste materials from agricultural and
food industries, etc., are used as biosorbents. Different mechanisms like precipi-
tation, absorption, adsorption, ion exchange are combined with biosorption in order
to treat toxic pollutants. Collective ideas of various pollutant removal techniques in
combination with biosorption and their applications to remediate water streams are
discussed.
The applications of certain microorganisms have gained importance in applied
environmental microbiology. Amongst them, biomineral processing is a field that
deals with metal mining from ores, concentrates, industrial wastes, overburdens,
etc., under the impact of microorganisms and/or their metabolites. Metals from poor
quality ore and mineral compounds are removed by bioleaching process which is
simple and low cost valuable technology. Metal recovery technique is widely
practiced for the recovery of copper, gold, iron, manganese, and lead. Treatment of
mineral industry effluents by microorganisms with incidental recovery of some
metal values constitutes a similarly important area of biomineral processing.
Microbial metal-leaching processes offer a possibility to recover metal values from
mineral resources not accessible by conventional mining. Microbes act as biocat-
alysts to convert metal compounds into their water-soluble forms in leaching pro-
cesses. Therefore, bioleaching has possible effect on metal retrieval and
detoxification of waste products of industry, coal mine, sewage sludge, and heavy
metal-contaminated soil.
In the developing countries, the usage of pesticides in the production of crops,
fruits, and vegetables increases the economic status which establishes the major
success in this field. Although the pesticide is an important aspect of the agricultural
practices, the vast handling of harmful pesticides is an ultimate concern to the air,
water, soil, and public health. Due to high impacts on human health, their appli-
cation has been limited and different scenarios are developed to clean up the
stubborn pesticides at different contaminated sites. Properties of polluted sites,
temperature, pH, and nature of the pollutants are important factors which play a
major role in the bioremediation process. Bioremediation technologies for cleaning
up the pesticides at polluted sites additionally, their fundamentals, advantages,
limitations, and the pesticides treated are summarized.
One of the major reasons behind the growing environmental pollution is illegal
disposal of waste. Due to the toxicity of waste, establishing efficient and envi-
ronmentally friendly method to degrade and detoxify these wastes represents an
1 Introduction to Environmental Protection and Management 5

important research challenge. Various physiochemical methods are applied all over
the world for solid waste management. The application of microbes to degrade
waste is gaining attention due to its environmental and economic benefits. Appli-
cation of microbes and factors affecting the bioremediation of hazardous wastes are
discussed. Authors have also discussed in detail about the prospects of waste
valorization for production of biopolymers, biofuels, biocompost, and industrial
enzymes.
Phytoremediation attempts the application of plants and microbes associated
with plant root systems to protect the environment by removal of pollutants.
Phytoremediation is capable of treating pollutants of dyes waste, which are derived
from various sources. Adaptation in genetic levels is basic attitude behind plants
that are able to manage the contaminants from the polluted site. The phytoreme-
diation techniques are classified in detail. Treatment of textile dyes using plant
remains an unfamiliar area of research. Mechanisms of uptake of different dyes by
plants have also been proposed by authors. Selection criteria of plants for achieving
high efficiency for treatment of dye contaminant wastewater have been projected.
Lignocellulosic biomass is most abundant in the environment. Enzymatic
breakdown of lignocellulose, an important component of common waste materials,
can be an essential step toward mitigating the wastes and generating biofuel. The
diverse microbial community is maintained within the insect gut as per their food
habit and ecological niche. Certain insects have enzymatic potential as they feed on
lignocellulosic materials for their nutrition. In this context, scientific community has
become interested to explore different insect gut microbial diversity through advent
of new technologies. Potential role of insect gut bacteria, aspects of colonization,
and role in degradation of lignocellulosic biomass are discussed. Further, the sig-
nificance of potential bacteria for harnessing the enzymes and appropriateness of
application in lignocellulosic wastes degradation are also discussed in one chapter.
Industrial and municipal wastewater contains numerous ingredients, and inter-
estingly, some of the compounds in wastewater, like nitrogen and phosphorus, are
identified as beneficial ingredients for microalgae cultures. Therefore, algal biore-
mediation can be considered as a feasible alternate technology for treating the
wastewater in a cost-effective and assertable way compared to conventional water
treatment process. These microalgal cultures are autotrophs, and they play a notable
role in remediation of wastewater by their photosynthetic ability. A win-win situ-
ation of using microalgae in the bioremediation of wastewater provides tertiary
biotreatment of wastewater coupled with production of potentially valuable biomass
as bioresource for biofuel or high-value by-products.
Water utilization is on a steep hike due to the urbanization and population
increase. On the other hand, pollution of freshwater due to human activities is
increasingly a major concern as it affects economy and growth of a nation. Among
various water pollutants, nitrogen compounds form a significant role in wastewater
contamination due to the increase in anthropogenic sources like agriculture. Nitrate
contamination in water and soil has become a growing environmental concern.
According to USEPA standards, the maximum contamination level for nitrate is
45 mg L−1, and the same standard is adopted by the Bureau of Indian Standards
6 S. J. Varjani et al.

(BIS). Among various technologies employed for treating nitrate-contaminated

water, biological denitrification is one of the more versatile and promising methods
widely being employed. The treatment of NO3− using bacteria referred as deni-
trification or bioremediation of nitrate has high separation efficiency.
Bio-denitrification processes, immobilization of microorganism and different
reactors employed for removing the nitrate from wastewater as well as reactor
designs ranging from fixed-bed reactors to biological-aerated filters have been
demonstrated for effective denitrification.
Volatile organic compounds (VOCs) are major air pollutants which are released
into the environment. Stationary sources such as petrochemical and pharmaceutical
industries release VOCs like toluene in the environment. In addition to it, VOCs
pollute air, soil, and water which are a growing environmental concern. Several
biological methods ranging from biotrickling filters to biofilters have been
demonstrated, and they are found to be economical. The biofilters are simple to
construct, easy to operate, and cost-effective. Major advantage of this method is that
the pollutant is converted into biodegradable waste which can decompose within a
moderate time frame, thus producing no secondary pollutants. Authors have
focused on bioremediation of volatile organic compounds in biofilters.
Chapter 2
Mathematical Modeling in Bioremediation

Parthasarthy Vijay and Margavelu Gopinath

Abstract Roughly 98% of the available freshwater is represented by the ground-

water on the planet. Protecting and re-establishing groundwater quality is of great
importance. A major threat to the resources of groundwater is soil and aquifer pollu-
tion by hazardous wastes. This pervasive issue represents an important practical and
cost-effective challenge because underground environmental pollution is tough to
locate and eliminate by conventional extraction and excavation methods. Thus, there
is a necessity for a broader investigation of efficient, in situ remediation approaches
that uses the benefit of natural phenomena, such as bioremediation and natural atten-
uation. This chapter gives a prologue to the essential hypothesis and utilization of
contaminant transport demonstrating by numerical methods. In the wake of perus-
ing this section, the reader ought to have the capacity to choose a proper numerical
model for the circumstance under thought, run the code, and adjust it. The utiliza-
tion of numerical methods to tackle groundwater contaminant transport issues has
turned into a broadly utilized strategy in view of the intensive enthusiasm for ground-
water quality and the quick improvement of processing innovation, which has made
numerical simulations accessible to hydrogeologists and civil engineers. If suitably
applied, numerical models can give answers to the following questions.
∙ What is the relationship between solute concentration and location?
∙ What will be the time taken to reach a target level of solution concentration due
to remediation process?
∙ Will a remedial measure reach a targeted concentration (reduced) in a certain time?
∙ How might one recreate the historical data of contamination to discover the rela-
tionship between population, time of exposure, and concentration?
The chapter starts with fundamentals of groundwater flow. The concepts of ground-
water such as hydraulic head, Darcy’s Law, hydraulic conductivity are discussed.
Basics of transport processes, such as diffusion, advection that are involved in
groundwater contamination, are also discussed. Later, theory of the model equations,
assumptions considered, initial and boundary conditions are presented. In the latter

P. Vijay (✉) ⋅ M. Gopinath

Department of Chemical Engineering, University of Petroleum and Energy Studies,
Dehradun 248007, India
e-mail: pvijay@ddn.upes.ac.in

© Springer Nature Singapore Pte Ltd. 2018 7

S. J. Varjani et al. (eds.), Bioremediation: Applications for Environmental
Protection and Management, Energy, Environment, and Sustainability,
https://doi.org/10.1007/978-981-10-7485-1_2
8 P. Vijay and M. Gopinath

sections, analytical and numerical models are discussed in detail with few recent
advances in bioremediation modeling. The discussion presented here is considered
to be basics yet should provide ample background for the reader concerned with the
detailed workings of a numerical model.

Keywords Bioremediation ⋅ Fate and transport equations ⋅ FEM ⋅ Modeling

Solute transport

2.1 Basics of Flow of Groundwater and Transport

of Contaminants

2.1.1 Introduction

Groundwater includes the part of the underground water that fills the pore space com-
pletely. Formations that carry groundwater are divided into aquifers, high-
permeability hydraulic conductors, aquicludes, and low-permeability hydraulic iso-
lators. There is no sharp boundary between aquifer and aquiclude. Sometimes even
an transitional formation termed as the aquitard is formed, whose permeability is in
between the aquifer and aquiclude.
Groundwater arises in porous media, like rock, sediments, and soil, underneath
the ground surface. Once the entire pore space in a rock or soil is occupied with water,
the material is termed to be saturated. However, a medium is termed as unsaturated
when the pore space is partially filled with air and water. Water from snowmelt,
rainfall, lakes, and rivers penetrates through the pores and cracks of rocks and soil,
and it passes through vadose zone, as shown in Fig. 2.1. Water in the saturated and
vadose zones are termed as groundwater. Shallow root zones of the plants mainly
utilize soil water.
Groundwater is fundamentally held by capillary forces in between grains in the
vadose zone. Many pores are completely saturated even within the vadose zone.
The root zone reaches full saturation during substantial rainfall when the penetration
capacity is reached. Pores present within the capillary fringe are also fully saturated;
however, this zone has negative water pressure.
Ratio of volume of water and the volume of voids of the soil is termed as water
saturation.
At the water table or phreatic surface, the pressure of water (p) is atmospheric
pressure (1 atm). However, the water pressure becomes negative, above the water
table. Due to hydrostatic forces, it increases with depth in the saturated zone.
An aquifer (from Latin, aqua—water, fer—to bear) is defined as an underground
layer of water-bearing porous rock, rock fractures, or loosely packed constituents
(gravel, sand, or silt) from which groundwater can be extracted using a water well.
Discharge is termed as water moving out from an aquifer, whereas water moving in
to an aquifer is termed recharge. Aquifers which are entirely saturated, and under
2 Mathematical Modeling in Bioremediation 9

Fig. 2.1 Schematic representation of the vadose and saturated zones (http://www.westfield.ma.
edu/personalpages/draker/edcom/final/webprojects/sp07/watercycle/hydro_cycle.gif)

pressure from the layers above are called as confined. Unconfined aquifers, instead,
have the water table as an upper edge. Since unconfined aquifers are shallow, they
are exposed more to contamination from events on the ground (Fig. 2.2). Although
confined aquifers are believed to be protected by the aquitards. Yet, contamination of
a confined aquifer may happen through a constructed path such as a well or pollution
resulting in a recharge area.
Modeling of dissolved contaminant movement in groundwater includes two dif-
ferent processes: (1) flow of groundwater and (2) transport of contaminant. In the
dominion of flow of groundwater, parameters describing water flow in the aquifer
are focussed. In the domain of transport of contaminant, parameters depicting the
pore network of aquifer and interactions between different contaminants, the aquifer
solid, and contaminant concentrations in air or water are of interest. In this chapter,
fundamentals in both these domains are discussed.
This data forms the origin for the governing fate and transport equations described
later in this chapter. The dimensions of all physical and chemical terms will be
expressed as mass [M], length [L], and time [T].
10 P. Vijay and M. Gopinath

Fig. 2.2 Aquifer types and groundwater movement (http://www.in.gov/dnr/water/7258.htm)

2.1.2 Concepts of Groundwater

When following the flow paths of groundwater from a hill to an adjacent stream,
water discharges into the stream from all probable directions. This curving path can
be elucidated as a compromise amongst the force of gravity and the tendency of
water to flow laterally in the direction of the slope of the water table. The propensity
toward lateral flow is the result of the movement of water toward an area of lower
pressure, the stream channel. The resulting movement is neither directly downward
nor directly toward the channel, but relatively along the curving paths toward the
stream. Groundwater can move upwards or downwards.

2.1.2.1 Hydraulic Head

Hydraulic head is defined as the elevation to which water can rise naturally in a
well. Observation well or a monitoring well or a piezometer is an artificially created
well for measuring hydraulic head. For unconfined aquifers, the hydraulic head is
the same as the water table. However, for confined aquifers, it is not.
Fluid flow through porous and fractured media could be a mechanical method
within which the driving forces liable for the fluid flow basically ought to overcome
the resistance caused by viscous and resistance forces.
The general form of Phenomenological law is given by

𝜕(𝜌g)
+ ∇.(𝜌vg) = −∇.j + 𝜙g (2.1)
𝜕t g
2 Mathematical Modeling in Bioremediation 11

G—Property
G/mass ≡ g
g—Scalar or vector
j —flux
g
∇.j —you get a “minus” sign because area normal points outwards.
g

2.1.2.2 Continuity Equation

In particular, we have G = mass; therefore g = G/mass ≡ 1

By definition, our system is of constant mass, and therefore j is zero.
g
No generation of mass ⟶ 𝜙g = 0, we get

𝜕𝜌
+ ∇.(𝜌v) = 0 (2.2)
𝜕t
𝜕𝜌
+ v.∇𝜌 +𝜌∇.v = 0 (2.3)
𝜕t
⏟⏞⏞⏞⏟⏞⏞⏞⏟
D𝜌
Dt

Substituting Eq. 2.2 in Eq. 2.1, we get

𝜕g 𝜕𝜌
𝜌 + g + g∇.(𝜌v) + 𝜌v.∇g = −∇.j + 𝜙g (2.4)
𝜕t 𝜕t g

( )
:0
𝜕g 𝜕𝜌
  
𝜌 + g  + ∇.(𝜌v) + 𝜌v.∇g = −∇.j + 𝜙g (2.5)
𝜕t  𝜕t g

𝜕g
𝜌 + 𝜌v.∇g = −∇.j + 𝜙g (2.6)
𝜕t g

The above equation is used widely where density is constant.

𝜕(𝜌g)
+ ∇.(𝜌vg) = −∇.j + 𝜙g (2.7)
𝜕t g

G = Total momentum = mv; g = v

𝜕(𝜌v)
+ ∇.(𝜌vv) = −∇.𝜎 + 𝜌b
𝜕t =
𝜎= pI + 𝜏 (2.8)
=
⏟⏟⏟ ⏟⏟⏟
normal stress tangential shear stress
12 P. Vijay and M. Gopinath

𝜕v 𝜕𝜌
𝜌 +v + 𝜌v.∇v + v.∇(𝜌v) = −∇p − ∇.𝜏 + 𝜌b (2.9)
𝜕t 𝜕t

For constant density system, we get

( )
𝜕v
𝜌 + v.∇v = −∇p − ∇.𝜏 + 𝜌b (2.10)
𝜕t
⏟⏞⏞⏞⏞⏞⏞⏟⏞⏞⏞⏞⏞⏞⏟
Dv
Dt

The above equation is the “Equation of motion” for constant density system (Bird
et al. 1960).
Mechanical energy equation = v.(Equation of motion)
( ) ( )
𝜕 𝜌v2 𝜌v2
+ v.∇ = −v.∇p − v.∇.𝜏 + 𝜌b.v (2.11)
𝜕t 2 2

( ) ( )
𝜕 𝜌v2 𝜌v2 v
+∇ = −∇.(pv) + p∇.v − ∇.(𝜏.v) + 𝜏 ∶ ∇v + 𝜌b.v (2.12)
𝜕t 2 2

Omiting, time derivative term, replacing the last term (on the right-hand side)
by −g∇h, where h is the elevation, in the above equation, and then dividing by 𝜌
and then form the dot product with the unit vector s = v∕|v| in the flow direction.
When this is completed, the term involving the curl of the velocity field can be shown
to vanish (a good exercise in vector analysis), and (s ⋅ ∇) can be replaced by d∕ds,
where s is the distance along a streamline. Thus, we get
( )
d 1 2 1 dp dh
v =− −g (2.13)
ds 2 𝜌 ds ds

Integrating the above equation between two points leads to Bernoulli equation,
given as below.

1( 2 ) ( )
p
2
1
v2 − v21 + dp + g h2 − h1 = 0 (2.14)
2 ∫p1 𝜌

Hydraulic head is the total of elevation and also the pressure head. The hydraulic
head reflects the elevation of the top of a water column inside the aquifer relative to
some point of reference.
2 Mathematical Modeling in Bioremediation 13

Fig. 2.3 Schematic representation of hydraulic gradient in an aquifer (http://slideplayer.com/slide/

6104388/)

Hydraulic gradient could be a measure of the energy potential inflicting ground-

water to flow between two points in a formation. This is schematically represented
in Fig. 2.3.

2.1.2.3 Flow Through Porous Media

Wide range of engineers and scientists found interest in flow through porous media,
who identified the importance of groundwater flows. In 1856, Darcy had performed
numerous experiments based on the practical applications of flow through porous
media. Darcy’s experiment can be performed in laboratory using a similar experi-
mental setup shown in Fig. 2.4.
Started as one-dimensional flow, extended to three-dimensional flow which cov-
ers most of the issues encountered in groundwater flow and oil recovery process. He
found that the volumetric flow rate of discharged water flow is directly proportional
to the hydraulic gradient Δh∕Δx, cross-sectional area A. The proportionality con-
stant is termed as the hydraulic conductivity (which we will discuss in detail in the
following sections).
The empirical relationship of Darcy’s law can be mathematically (Whitaker 1986;
Greenkorn 1981; Permeability 2017) represented for 1-D case as,
Δh
Q = −KA (2.15)
Δx
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your-name, to despatch your pharisaical letter sealed with black
wax.”
“Young wrathful,” meekly rejoined Mr Samuel, “it was dark green
wax, most emblematic, as I said to your aunt, my dear spouse, of the
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“Black and green fiends dog you to Satan,” roared I. “What an ass
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Oh! Broadcroft, Lilliesacre, Kittleford, Cozieholm, and Oxgang,
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With some more stuff of this kind, I parted with my venerable aunt
and her smooth-tongued spouse. These petrifactions of humanity
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driven him mad. I wished the rogue at the bottom of the Red Sea, or
in the farthest bog of Connaught, paring turf and cultivating potatoes
—anywhere but where I now saw him. I could have eaten him up raw
and unsodden, without salt or pepper, where he stood—ground his
bones to dust, or spit upon him till he was drowned in the flood of
my spite. I did neither; but throwing myself again on the back of
Kate, off I scampered home, more like a fury than a man.
In my way there was not a rascal I met but seemed to my heated
imagination to know my misfortunes, and enjoy, with sly
satisfaction, their fearful consummation. Two fellows I cut smartly
across the cheek; they were standing coolly by the wayside, with their
hands in their pockets, interchanging winks, and thrusting their
tongues provokingly out like hounds on a hot day. They did not relish
the taste of my thong, and one of them made an awkward squelsh
into a ditch on receipt, head over heels, immensely to my heart’s
content.
It was evening when I reached the little village where my head-
quarters for some weeks had been established. To add to my
miseries, I found that Tom had, in my absence, with his usual
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Cross Keys, on the faith of my accession of property. When I rode
past the tavern, my ears were assailed with most extraordinary
sounds of festivity, and my head endangered by a shower of bottles
and glasses that his reckless boon companions were discharging
from the windows. Some of these windows, too, were illuminated
with multitudes of dips—the extravagant dog!—three to the pound.
And some coarse transparencies were flaunting in my face pithy
sentences, such as—“A Glorious Revolution,” “Splendid Victory,”
“Jubilee to Hopeless Creditors,” “Intelligence Extraordinary!!” &c.
Then, at every pause of the maddening din, the explosion of another
bottle of champagne smote my ear like a death-knell. Cork after cork
popped against the ceiling—crack, crack, they went like a running
fire along a line of infantry, while loud above the storm rose the
vociferations of my jolly friend, as he cheered them on to another
bumper, with all the honours, or volunteered his own song. Poor
Tom, he had only one song, which he wrote himself, and never failed
to sing to the deafening of every one when he was drunk. It was
never printed, and here you have as much of it as I remember, to
vary the melancholy texture of my story:—
SONG.
Fill a can, let us drink,
For ’tis nonsense to think
Of the cares that may come with to-morrow;
And ’tis folly as big
As the Chancellor’s wig,
To dash present joy with dull sorrow.
Hip! hip! hip! fill away;
Our life’s but a day,
And ’twere pity that it proved a sad one;
’Twas in a merry pin
Our life did begin,
And we’ll close it, brave boys, in a mad one!
Hip! hip! hip! &c.

Never shrink, boys, but stand,

With a can in each hand,
Like a king with his globe and his sceptre;
And though slack in your joints,
Yet thus armed at all points,
The devil himself can’t you capture.
Hip! hip! hip! fill aright,
Should he seek us to-night,
We’ll toss off the old rogue as a whetter;
When the hot cinder’s down,
Take my oath on’t, you’ll own,
That good luck could not furnish a better.
Hip! hip! hip! &c.

Dull sophists may say,

Who have ne’er wet their clay,
That merry old wine gives no bliss,
But the flask’s sparkling high,
Gives the dotards the lie,
Crying, kiss me, my roaring lads, kiss!
Hip! hip! hip! jolly boys!
He who quarrels with those joys,
Which the longer they’re sipped of grow sweeter,
May he live to be wise,
And then when he sighs
For a smack, let him choke with this metre.
Hip! hip! hip! &c.

This was followed with what Tom emphatically styled a grand crash
of melody; that is, overturning the table, and burying in one
indiscriminate ruin, bowls, bottles, glasses, and all things brittle.
My heart sickened at the riot, and, broken in spirit and penniless, I
retreated to my lodgings.
Here I had at least peace to ruminate over my prostrate fortunes;
but as meditation would not mend them, and next morning would
assuredly bring the dire intelligence of my aunt’s marriage, I, that
same night, made a forced march, anxious to secure a convenient
spot for rustication and retirement, till fortune should again smile, or
the ferocity of my creditors be somewhat tamed. Poor Tom! I had the
savage satisfaction of breaking up his carousal by a few cabalistic
words written in a strong half-text hand: “Stole away! Done up.—
Fooled and finished.—Run, if you love freedom, and hate stone walls.
You will find me earthed in the old hole.”
Next evening I was joined by my luckless shadow. He had a hard
run for it; the scent lay strong, and the pack were sure-nosed and
keen as razors. But he threw them out from his superior knowledge
of localities. After this we both became exceedingly recluse and
philosophical in our habits. We had the world to begin anew, and we
had each our own very particular reasons for not making a noise
about it.—Paisley Magazine.
 THE COURT CAVE:
A LEGENDARY TALE OF FIFESHIRE.

By Drummond Bruce.

Chapter I.

A few years before the pride of Scotland had been prostrated by

English bows and bills, on the disastrous day of Flodden, the holding
of Balmeny, in the county of Fife, was possessed by Walter Colville,
then considerably advanced in years. Walter Colville had acquired
this small estate by the usual title to possession in the days in which
he lived. When a mere stripling, he had followed the latest Earl of
Douglas, when the banner of the bloody heart floated defiance to the
Royal Stuart. But the wavering conduct of Earl James lost him at
Abercorn the bravest of his adherents, and Walter Colville did not
disdain to follow the example of the Knight of Cadzow. He was
rewarded with the hand of the heiress of Balmeny, then a ward of
Colville of East Wemyss. That baron could not of course hesitate to
bestow her on one who brought the king’s command to that effect;
and in the brief wooing space of a summer day, Walter saw and loved
the lands which were to reward his loyal valour, and wooed and
wedded the maiden by law appended to the enjoyment of them. The
marriage proved fruitful; for six bold sons sprung up in rapid
succession around his table, and one “fair May” being added at a
considerable interval after, Walter felt, so far as his iron nature could
feel, the pure and holy joys of parental love, as his eye lighted on the
stalwart frames and glowing aspects of his boys, and on the mild blue
eyes and blooming features of the young Edith, who, like a fair pearl
set in a carcanet of jaspers, received an added lustre from her
singleness. But alas for the stability of human happiness! The truth
of the deep-seated belief that the instrument of our prosperity shall
also be that of our decay, was mournfully displayed in the house of
Walter Colville. By the sword had he cut his way to the station and
wealth he now enjoyed; by the sword was his habitation rendered
desolate, and his gray hairs whitened even before their time. On the
field of Bannockburn—once the scene of a more glorious combat—
three of his sons paid with their lives for their adherence to the royal
cause. Two more perished with Sir Andrew Wood, when Steven Bull
was forced to strike to the “Floure and Yellow Carvell.” The last,
regardless of entreaties and commands, followed the fortunes of the
“White Rose of York,” when Perkin Warbeck, as history malignantly
continues to style the last Plantagenet, carried his fair wife and
luckless cause to Ireland; and there young Colville found an untimely
fate and bloody grave near Dublin.
Thus bereft of so many goodly objects of his secret pride, the heart
of Walter Colville naturally sought to compensate the losses which it
had sustained in an increased exercise of affection towards his
daughter. The beauties of infancy had now been succeeded by those
of ripening maidenhood. The exuberant laugh, which had so often
cheered his hours of care or toil, while she was yet a child, had given
place to a smile still more endearing to his time-stricken feelings;
face and form had been matured into their most captivating
proportions, and nothing remained of the blue-eyed, fair-haired
child, that had once clung round his knee, save the artless openness
of her disposition, and the unsullied purity of her heart. Yet, strange
to tell, the very intensity of his affection was the source of bitter
sorrow to her who was its object, and his misdirected desire to secure
her happiness, threatened to blench, with the paleness of secret
sorrow, the cheek it was his dearest wish to deck with an ever-during
smile of happiness.
Edith Colville was but an infant when her three brothers fell at
Sauchie, and had scarcely completed her eighteenth year, when the
death of her youngest brother made her at once the object of her
father’s undivided regard, and of pursuit to many who saw and were
smitten with charms in the heiress of Balmeny, which had failed to
attract their attention while her brother yet stood between the
maiden and that heritage. But the heart they now deemed worth the
winning was no longer hers to give. The death of her mother while
she was yet a child, had left her her own mistress long before the
period when maternal care is most essential; and Edith’s love was
sought and won by one who had little but youth and a warm heart to
recommend him.
Arthur Winton was the orphan son of a small proprietor in the
neighbourhood, who, having been deprived of the best part of his
property by what he conceived the injustice of King James III., and
the rapacity of his favourite Cochrane, was easily induced to join the
insurgent nobles who wrought the destruction of that monarch. He
was, however, disappointed in his expectations of personal reward,
having fallen in the conflict; and his son was too young to vindicate
his claim in an age so rude as that of which we write.
Walter Colville, whose family had been so sadly thinned in the
battle we have mentioned, though they had fought on the other side,
naturally bore no goodwill to the boy; but his younger son, who was
nearly of the same age, viewed him with different feelings. He was
much about the house of Balmeny; and, to be brief, he won the
affections of the young Edith long before she knew either their
nature or their value. Until the departure of young Walter Colville,
Arthur’s visits were attributed by the old man to his friendship for
his son, but when Edith had unhappily become his heiress, he at
once attributed them to their proper cause. A stern prohibition of
their repetition was the consequence, and the lovers were henceforth
reduced to hurried and sorrowful meetings in secret.
On the morning wherein we have chosen to begin the following
veritable narrative, the youthful pair had met unobserved, as they
imagined, in a shady corner of Balmeny wood, and had begun, the
one to lament, and the other to listen, when the sudden apparition of
the angry father checked the pleasing current of their imaginings.
He drew his sword as he approached, but the recollection of his
seventy years, and his now enfeebled arm, crossing his mind, he
replaced the useless weapon, and contented himself with demanding
how the youth had dared thus clandestinely to meet his daughter.
Arthur attempted to allay his anger, and to plead his passion as he
best could; but the grim and angry frown that sat on Walter Colville’s
brow, as he listened to him, soon showed how vainly he was
speaking, and he ceased in confusion.
“Have you finished, young master?” said Colville, with a sneer.
“Then listen: you are not the wooer I look for to Edith. I should
prefer him something richer, something wiser, and something truer
to the king, than any son of your father is likely ever to prove; so set
your heart at rest on that matter. And you giglot, sooth! to your rock
and your chisart. But stay; before you go, tell this gallant gay to prowl
no longer about my dwelling. By St Bride, an he does, he may chance
to meet a fox’s fate!”
“Dear father,” said the weeping girl, “upbraid us not. Never will I
disobey you, never be his, without your own consent.”
“Hold there,” replied Colville, smiling grimly, “I ask no more.” And
he led away the maiden, who dared not so much as steal a parting
look.
Arthur Winton bore this fiat of the old man, and the dutiful
acquiescence of his daughter (though he doubtless thought the latter
pushed to the very extreme of filial obedience), if not with
equanimity, at least with so much of it as enabled him to leave the
presence of his mistress and her father with something like
composure. He wandered slowly to the beach, which lay at no great
distance, as if he had hoped to inhale with the cool breeze that
floated from off the waters, some portion of the calmness in which
they then lay bound, his mind occupied in turning over ill-assorted
plans for the future, ever broken in upon by some intruding
recollection of the past. The place where he now walked was one well
calculated, according to the creed of those who believe in the power
exercised over the mind by the face of external nature, to instil
soothing and tranquillizing feelings. It was a smooth grassy lawn,
forming the bottom of a gentle eminence, undulating and stretching
downwards to the pebbly beach, among whose round white stones
the quiet waters of the Firth fell kissingly. The view was bounded to
the north by the rising eminences we have mentioned, and shut in on
the west by the woody promontory which is still crowned by Wemyss
Castle. To the eastward several rocky eminences stretch into the
Firth, the more distant still increasing their seaward march until the
bay is closed by the distant point of Kincraig. Before him lay the
silver Firth, and, half-veiled in distance, the green fields and hills of
Lothian, terminated by the picturesque Law of North Berwick, and
the great Bass, frowning like some vast leviathan awakening from his
sleep. One or two white-sailed barks lay motionless upon the water.
The effect of the whole was so stilling and sedative, that Arthur, half
forgetting his recent disappointment, stretched himself upon the
sward, and abandoned himself to contemplation.
While he lay thus chewing the cud of sweet and bitter fancy, the
sounds of distant song and merriment occasionally broke upon his
ear. He at first regarded them as the mere offspring of imagination,
but at length the choral swell of a seemingly joyous ballad, followed
by a hearty, far-reverberated shout, convinced him that the merry-
making was real, and at no great distance. He started to his feet in
some alarm, for his first impression was that the Good Neighbours
were holding their revels near him, and he well knew the danger of
being detected as a prying overlooker of their mystic merriment. A
moment served to dissipate this fear. The voices which he had
listened to were too rough and boisterous ever to be mistaken for the
singing of those tiny minstrels, whose loudest notes never exceeded
in sound the trumpet of the bee. There was no fairy ring round the
spot on which he had lain, nor was the hour either the “eye of day” or
that of midnight, at which, as is well known, the elfin power was
most formidable. After looking and listening for some time, he
ascertained that the sounds proceeded from a cave, which we have
not yet mentioned, but which forms a striking ornament to the
beach, and an object of considerable interest to the geologist, having
been doubtless formed long before the Forth had found its present
modest limits. Being anxious to dispel the feelings that now preyed
on his peace, by a diversion of whatever kind, he walked towards the
place. As he approached, the mirth was renewed with increased
vehemence, and he perceived, at the western entrance of the cave, a
female, from whose swarthy hue and singular habiliments he at once
divined the nature of its present inmates. The woman, whose
features were stern and somewhat repulsive, wore a long gown, of
some coarse dark-coloured material, which fell almost to her feet,
having short wide sleeves, which left the arms at perfect liberty, and
coming up to the neck, was there fastened with a golden brooch. Her
head-dress consisted of a red and yellow coloured shawl, twisted
fantastically into a conical shape. Pendants of gold hung from her
ears, and rings of the same metal, in many of which were set rubies
and other sparkling gems, garnished her tawny fingers. Arthur at
once recognised an Egyptian or gipsy in the dark-featured damsel
who stood before him, and hesitated a moment whether he should
pursue the determination of mixing with the revellers within, to
which his eager desire of escaping from his present unhappy feelings
had prompted him. The Egyptians were in those days of a much
darker character than the remnants of their descendants, which, in
spite of press-gangs and justice-warrants, still linger amongst us.
Murder among themselves was a thing of everyday occurrence, and
desperate robberies, committed upon the king’s lieges, by no means
rare. The present gang, from their vociferation, seemed in a state of
excitement likely to remove any little restraints which the fear of the
law’s vengeance might at another time have imposed on them, and
the features of the woman, contrary to their custom, wore no look of
invitation, but rather seemed to deepen into a warning frown the
nearer he approached the door at which she was posted. On the other
hand, the honour of the race, to such as trusted them, was
proverbial. His curiosity to know more intimately the manners of a
people so remarkable as the Egyptians then were, and still are—
perhaps a latent wish of being able to extract from their prophetic
powers some favourable auspice to his almost expiring hopes—or
that nameless something which at times impels us to court the
danger we at other times shun with care—all conspired to induce him
to enter the cave, and he accordingly attempted to do so. In this,
however, he was opposed by the gipsy, who, stepping exactly in his
way, waved her arm in a repelling attitude; and, seeing him
disinclined to obey this silent injunction, coming still closer to him,
whispered, “Get you gone; your life will be endangered if you enter
here.”
Before Arthur could reply to this injunction, she who gave it was
suddenly attacked by a man, who, issuing from the entrance, struck
her a smart blow across the shoulders with a staff which he carried,
and then, with a scowling look and angry accent, spoke a few words
to her in a language which Arthur understood not. She muttered
something in reply, and proceeded towards the beach. “The woman
is mad at times, young sir,” said the man, now addressing Arthur.
“Heed her not, I beseech you. We are only a few wandering puir
folks, making merry, and if you wish to share our revelry, enter, and
welcome. Some of our women may be able to read your weird, should
you so incline; you have nothing to fear.”
Arthur was by no means satisfied either that the woman was mad,
or that the man meant him fairly; but as he could not now retreat
without betraying his fear to the dark searching eye which the gipsy
bent on him, and was besides conscious that he possessed a well-
proved sword, and considerable skill and strength in the handling of
it, he signified his wish to join the merrymaking, and followed the
gipsy into the cave.
On entering he found himself in the interior of a high-roofed
cavern, of considerable extent, partly exposed to the seaward side by
two arched openings between the lofty recesses of rock which
support the roof, that towards the east being the smaller and lower of
the two; and the other rising in height nearly to the roof, affording a
view of the Firth, and admitting light to the place.
The inhabitants of the cave had ranged themselves along the north
and inner side. Nearest the western entrance, stretched on sacks,
sheepskins, cloaks, and other nondescript articles of clothing, sat, or
rather lay, ten or twelve men, with rather more than double that
number of women, all busily engaged in drinking; farther off, some
ragged crones were busily superintending the operation of a wood
fire on a suspended pot; while, farther off still, a few barebacked
asses, and a plentiful variety of worse clad children, were enjoying
their common straw.
Arthur was immediately introduced to the company of carousers,
some of whom received him with a shout of welcome, but others with
evident dissatisfaction; and he overheard, as he seated himself, what
seemed an angry expostulation and reply pass between his conductor
and one of the party. This individual, who was evidently the chief of
the gang, was an aged man, with a beard of silver gray, which, as he
sat, descended to his lap, entirely covering his breast. His head was
quite bald, with the exception of a few hairs that still struggled for
existence behind his ears, and this, added to the snowy whiteness of
his eyebrows, and the deep wrinkles in his brow and cheeks, would
have conferred an air of reverence on his countenance, had not the
sinister expression of his small and fiery-looking eyes destroyed the
charm. On each side of him sat a young girl—the prettiest of the
company; and the familiar manner in which they occasionally lolled
on the old man’s bosom, and fondled with his neck and beard,
showed the intimate terms on which they lived with him. The rest of
the men were of various ages, and though all of them were marked
with that mixed expression of daring recklessness and extreme
cunning which has long been “the badge of all their tribe,” they
attracted (with one exception) little of Arthur’s attention. Of the
women, the very young ones were extremely pretty, the middle-aged
and old ones, more than equally ugly. Young and old, pretty and ill-
favoured, all were alike deficient in that retiring modesty of
expression without which no face can be accounted truly lovely, and
the want of which darkens into hideousness the plainness of homely
features. They joined freely in the draughts, which their male
companions were making from the horns, which, filled with wine and
ale, circulated among the company, and laughed as loud and joked as
boldly as they did.
Arthur seated himself in silence, and, somewhat neglectful of the
kindness of the female who sat next him, occupied himself in
surveying the motley group before him. His eye soon rested on a man
seated next the damsel who occupied the place immediately to the
left of the chief, and the moment he did he became anxious and
interested. The individual was a man of rather more than middle
height, of a muscular, though by no means brawny frame. His
countenance was ruddy, and of a pleasant mirthful expression; his
eyes were full, of a dark hazel colour; his nose, though prominent,
gracefully formed, and his mouth small and piquant. His beard was
of a dark auburn hue, and he wore moustaches of the same colour.
He was dressed in a hodden-gray doublet and hose, which were
fastened round his body by a strong leathern girdle, from which hung
a broad sword of the two-edged shape. The manner of this individual
was evidently different from those of his present companions, and
that from the very pains which he took to assimilate it. There was all
their mirth without their grossness, and his kind, affable demeanour
to the female part of the company differed widely from the blunt and
sometimes brutal behaviour of his comrades.
“Who is that on the left of the old man?” whispered Arthur to the
man who had introduced him.
“That—that’s his favourite dell,” replied the man.
“Nay, I mean not the woman—the man upon her left.”
 “Why, I know not—he’s none of us—strayed in like you to share the
revelry, I fancy,—though, if he takes not better care of his eyes and
hands, an inch or two of cold iron will pay his reckoning. I think he
dallies too much with the mort.”
The cool, even tone in which this annunciation of probable murder
was uttered, rendered the communication more startling to Arthur
than if it had been made with a vindictive exclamation or suppressed
groan; and he looked anxiously and steadily on the stranger, whose
gallant bearing more and more attracted him. The latter had
observed him more than once bending his eyes on him, and was not
apparently pleased with the strictness of his scrutiny. Twice, when
their eyes met, the stranger had checked a rising frown by emptying
the horn which he held in his hand; the third time he set it down
untasted, and, fixing on Arthur a look of calm commanding dignity,
which seemed more native to him than aught around, exclaimed, in a
deep and powerful accent,—
“Friend, wherefore peer you so steadily this way? If you have aught
to say, out with it—if not, reserve your ogling for some of the fair eyes
near you.”
Arthur felt abashed beneath the rebuke which his solicitude for
this individual had exposed him to, and he could only mutter in reply
something about the young damsel beside him.
“Ah! ah!” replied the stranger, resuming his good humour, “it is to
her your looks were sent? Soul of Bruce! but she is well worthy of
your wonder. Never—and I have seen many bright eyes—have I
lighted on a pair so witching.” Then, turning to the object of these
praises, he took her hand, and whispered in her ear something,
which, though inaudible to those present, was evidently of no
unpleasing nature, as her dimpling cheek unquestionably testified.
The patriarch had viewed, for some time, with ill-dissembled
anger, the approaches of the stranger to the temporary sovereign of
his affections. But whether he thought them becoming too close, or
was enraged at the placidity with which they were received, his
indignation now burst out, and as is usual in matters of violence, the
weight of his vengeance fell heaviest on the weaker individual. He
smote the girl violently on the cheek, and, addressing the stranger in
a voice hoarse with passion, poured forth a torrent of words which
were to Arthur utterly unintelligible.
 The stranger, who did not seem to understand the expressions of
this address, could not, however, mistake its meaning. The language
of passion is universal—and the flashing eye and shrivelled brow of
the Egyptian chief were too unequivocal to be misunderstood. He
remained silent but a moment, and then, drawing from his bosom a
purse, apparently well-filled, he took out a golden Jacobus, and
proffered it to the patriarch, as a peace-offering to his awakened
anger. The fire of indignation fled from the old man’s eyes as they
lighted on the gold, but they were instantaneously lighted up by a
fiercer and more deadly meaning. Arthur could observe significant
looks circulating among the men, who also began to speak to one
another in a jargon unintelligible to him. He felt convinced that the
purse which the incautious stranger had produced had determined
them to destroy him; and, prepossessed with this idea, he saw at
once the necessity of the keenest observation, and of the danger
which attended his scrutiny being detected. He pretended to begin to
feel the influence of the potations in which he had indulged, and
apparently occupied himself in toying with the willing dell who sat
beside him. He now perceived one or two of the men rise, and
proceed to the several openings of the cave, evidently to see that no
one approached from without, or perhaps to cut off retreat. He saw,
too, that they plied the stranger and himself with wine and ale; and,
more convincing than all, he perceived on the darkening brow and
gleaming eye of the hoary Egyptian, the awakening excitement of a
murderous design. The stranger, in the meantime, apparently
unconscious of the peril he was in, began again to bandy kind words
and looks with the favourite of the chief. The old man looked grimly
on, but did not now seem to wish to interrupt the dalliance. Suddenly
he drew his hand from his bosom. It was filled with a dagger, which
he raised high, evidently with the intention of slaying the unguarded
stranger, who was too much occupied with the eyes and hands of the
beauty to perceive his villanous intention.
Arthur, who at the moment was lifting to his mouth the ponderous
pewter “stoup,” or flagon, containing the ale on which the Egyptians
were regaling, saw the wretch’s intent, and on the impulse of the
moment flung the vessel at the lifted hand. His aim was fortunately
true; the villain’s arm fell powerless by his side, while the dagger flew
to a considerable distance. Arthur then rose, and crying hastily to the
stranger to defend himself, drew his blade and made towards him.
 The stranger had perceived the intended blow, though, entangled
as he at the moment was, he would unquestionably have fallen a
victim to it. He now leaped hastily up, and exclaiming loudly, “Morte
de ma vie!—Treason!” drew out his sword, and looked for the foe.
Arthur now joined him, and, setting their backs to the rocky wall of
the cave, they prepared to defend themselves against the enraged
gipsies, who, now shouting wildly, drew from under their cloaks long
sharp knives, which they brandished furiously in their faces.
The stranger swept his sword around him in a manner that proved
him a practised master, and Arthur manfully seconding him, the
Egyptians were kept completely at bay, for none seemed daring
enough to trust himself within the sweep of the stranger’s sword, or
that of his new companion. But it was only while they could keep
their backs to the rocky wall that they could hope to cope with their
savage enemies, who, though they did not come near enough to stab,
surrounded them as nearly as they could, and yelled and shouted like
so many disappointed fiends. There was apparently no means of
escape, though there might be of resistance, as the moment they
quitted the wall their backs would have been exposed to the daggers
of the infuriated assassins. Arthur perceived, too, to his dismay, that
sure means were taken to render their length of sword unavailing.
Several women were clambering up the rock behind them carrying
large blankets and other cloths, clearly for the purpose of throwing
over their swords and themselves, and thus yielding them up a
fettered prey to these ruffians. All hope of escape died in his bosom
as he discovered the well-laid design, and he was about to rush on
the savages, and at least sell his life dearly, when he observed the
women who carried the blankets pause and look upwards. He too
looked up, and saw, with a consternation that for a moment
unmanned him, an immense fragment of loose rock in the very act of
being removed from its immemorial resting-place, and precipitated
on their heads.
“Holy Virgin! help us, or we are lost!” exclaimed the youth; and the
prayer had hardly left his lips ere the threatened engine of their
destruction was converted into the means of their immediate escape.
The ponderous stone dropped so far directly on its fatal errand, that
Arthur instinctively crouched beneath the apparently inevitable
blow; but encountering a few feet only above his head a projecting
piece of rock, it rebounded from the side of the cave in a slanting
direction, and, falling clear of its intended victims, smote to the earth
the hoary head of the patriarch. He fell beneath the huge fragment,
which hid from their sight the face and neck of the Egyptian; but the
convulsive writhings of the unhappy man, which for a moment
contorted his frame, only to leave it in utter stillness, told plainly that
his long career had ceased, and that the man of blood had become
the victim of his own pitiless design.
The Egyptians, panic-struck by this sudden death-blow, set up a
loud and stunning wail, as they crowded round the body of their
chief; but the stranger and Arthur stayed not to observe their farther
demeanour, and, taking advantage of the opening among their
enemies, which was now afforded them, sprang out of the cave, and
ascended at the top of their speed to the brow of the eminence
behind it.
They continued their rapid walk for some time in silence, induced,
no doubt, by the tumultuous nature of their feelings, and the violence
of their present exertion. At length, having entered a few yards into a
wood, which then decorated the place, though soon after to be
converted into keel and timbers for the “Great Michael,” the stranger
halted, and, taking Arthur by the hand, said breathlessly,—
“By Saint Andrew, young sir, you have done us this day good
service. I never thought to have been so indebted to a pint-stoup,
trow me.”
“But what sorrow tempted you, man,” replied Arthur, rather
crossly, “to play the fool with the old villain’s dearie in yon wild sort
of fashion; and, above all, what induced you to flourish your well-
filled purse in the eyes of those who love gold better than anything
else save blood?”
“Whim—chance—fate—I thought at one time. It is long since
cunning men have told me that I shall die for a woman, and, by the
Bruce’s soul! I thought the hour had come. As for my Jacobuses, I
rejoice I saved them from the filching crew, as they will serve for an
earnest—a poor one, to be sure—of my thankfulness to my brave
deliverer;” and so saying, he drew from his bosom the purse which
had excited the fatal cupidity of the Egyptians, and gracefully
proffered it to the youth.
 Arthur had all along suspected—nay, felt assured—that his
companion was of a rank superior to his appearance; and, had it not
been so, his present conduct would have convinced him.
“Whoever you are, sir,” said he, “that in this lowly guise speak the
language and the sentiments of a noble-born, your own heart will, I
know, convince you that I dare not accept your gold. The service I
rendered you I would have rendered to the poorest carle in Fife, but
were it ten times greater than it was, it must not be repaid with coin.”
“All are not carles who wear hodden gray and blue bonnets with
you, I find,” replied the stranger, smiling approvingly. “But come, if
gold cannot repay the service you have done me, tell me what can.”
“Nothing in your power to perform,” replied Arthur, calmly.
“Try,” continued the stranger; “I bear with me a talisman which
can command all objects which men in general desire. Choose, then
—wealth, worship, or a fair wife!”
There was something so frank, open, yet condescending, in the
tone and appearance of this extraordinary stranger, that Arthur
could not resist their fascinating influence, and although he could
not imagine that any interference on the part of his new friend would
produce the slightest change in the stern sentence of Walter Colville,
he communicated to him a general outline of his present situation.
The stranger listened attentively to the detail—then demanded
how far distant the dwelling of Colville was; and, on being informed
of its near vicinity to the spot on which they then stood, declared his
intention of immediately proceeding thither and using his influence
in Arthur’s behalf.
The latter opposed this resolution but faintly; for, though he was,
as we have said, utterly at a loss to conceive how his cause was to be
benefited by the proffered kindness of the stranger, yet a vague and
almost latent hope of still obtaining Edith never entirely forsook him.
He conducted the stranger through the wood, therefore, by the
path which led most directly to the house of Balmeny. On reaching
the skirt of the forest, it was agreed that the former should proceed
alone to the dwelling of Colville, and that Arthur should remain
where he was, and await the result.
 Chapter II.

The stranger set out on his voluntary mission at a rapid pace, and
soon arrived at the house. The door stood open, and he entered with
the careless sauntering air of one entirely indifferent as to the
welcome he might be greeted with. He found Colville seated
apparently in no very pleasant humour, and his daughter, bustling
about among the servant-maidens, wearing on her flushed cheek and
suffused eye undoubted symptoms of the sorrow with which the
morning’s adventure had afflicted her.
“Give you good-e’en, gudeman of Balmeny,” said the stranger,
seating himself, without waiting an invitation, on the bench opposite
Colville.
“The same to you, neebour,” said the landlord, in a tone that had
little of welcome in it.
A few moments’ silence now ensued, Colville evidently waiting
with some impatience for the tidings which the other seemed in no
haste to communicate to him. But this could not last.
“Have you anything to tell, ask, or deliver, friend?” at last said
Colville.
“This bright-e’ed maiden is the bonny lass of Balmeny, I’m
thinking,” was the unreplying answer.
“That is my daughter, truly,” said the landlord, becoming more
and more impatient; “does your coming concern her?”
“That it does,” replied the stranger. “There’s an auld bye word, that
‘foul fish and fair daughters are nae keeping ware.’ This fair May is
the object of my visit; in short, gudeman, I come awooing.”
At the sound of this magnetic word, a universal commotion arose
in the dwelling of Colville. The maiden, who was its object, surveyed
the stranger with indignation and surprise; the servants whispered
and tittered among each other; and Colville seemed for a moment
about to give vent to the feelings of his anger, when the current of his
feelings suddenly changed, and, directing a look of malicious joy to
his daughter, he addressed the stranger—
“Welcome, wooer—welcome. Come, lasses, set meat and drink
before this gentle here; as the auld Earl of Douglas said, ‘It’s ill
arguing between a fu’ man and a fasting.’”
The order was obeyed with great readiness by the serving maidens,
who set before the stranger the household bread and cheese, and a
bicker of no scanty dimensions, containing the reaming ale for which
Scotland has been so long famous. There was a malicious merriment
twinkling from every eye as the scene went on; for all knew well that
the over-strained kindness of the host was soon to be converted into
outrageous and overwhelming abuse of the guest. The stranger,
however, seemed either not to notice or to slight these indications.
He partook heartily of the good cheer set before him, and amused
himself by returning with good-humoured smiles the stolen looks of
the simpering maidens. He looked in vain, however, for Edith, who
had retired from the place.
“And now,” said Colville, who began to think the stranger
somewhat more at ease than he could have wished, “Your name,
wooer?”
“My name?” said the stranger, somewhat embarrassed.
“Ay, your name—all men have a name. Knaves [laying an
emphasis on the word] many.”
“True, gudeman, true. My name, then, is Stuart—James Stuart. I
hope it pleases you?”
“The name is the best in the land,” said the old man, touching his
bonnet. “As to the wearer—hem!—‘a Stuarts are no sib to the king’, ye
ken. What countryman are you?”
“I was born at Stirling,” said the stranger.
“Ay, ay, it may be, it may be,” replied Walter Colville; “but, to bring
the matter to a point, what lands and living hae ye, friend?”
“Sometimes less, sometimes more,” replied the stranger, “as I
happen to be in the giving or the taking humour. At the lowest ebb,
however, I think they are at least worth all that ever called a Colville
master.”
 “Faith, and that’s a bauld word, neebour,” cried Colville, bitterly
—“and one that, I’m jalousing, you’ll find it difficult to make gude.”
“At your own time it shall be proved, gudeman; but it is not for
myself I come to woo the bonny lass of Balmeny. I am, thanks to a
wise old man who sits in Windsor, wived already.”
“And who, in Beelzebub’s name, may you be blackfit for?”
demanded Colville, rising in wrath.
“Give your daughter to the youth I shall name, and I will, on her
wedding-day, fill you up one lippie with the red gold, and five
running o’er with silver.”
“Give her! To whom?”
“To one who loves her dearly; and, what is more, is dearly loved in
return, old man.”
“Who is he?” reiterated Colville.
“One who is worthy already of the hand of the best ae daughter of
any laird in Fife; and who, ere to-morrow’s sun sets, will be wealthier
than yourself.”
“Who—who—who is he?” cried the old man, stamping in a
paroxysm of rage.
“Arthur Winton!” said the stranger.
The anger of Colville, when this unpleasing name was uttered,
almost overwhelmed him.
“Out of my doors, you rascally impostor,” at length he was able to
exclaim; “out of my doors! Swith away to the minion who sent you
here, an you would wish not to taste the discipline of the whip, or to
escape being worried by the tykes.”
To the stranger, the anger of the old man, instead of fear, seemed
only to occasion merriment. He laughed so heartily at the violence
into which the rage of his host seduced him, that the tears actually
stood in his eyes—conduct that naturally increased the passion which
it fed on. The servants stood looking on in silent wonder; and Edith,
startled by the noise of the discordant sounds, returned to the place
in wonder and alarm.
An unexpected termination was suddenly put to the scene by the
entrance of Arthur Winton. His cheek was flushed with haste; and he
was so breathless that he could hardly exclaim,—
 “Save yourself, sir stranger, by instant flight; the Egyptians have
tracked our path hither, and are pursuing us here with numbers ten
times exceeding those we encountered in the cave.”
“Let them come,” said the stranger, with a smile; “Egyptians
though they be, they cannot eat through stone walls or oaken doors.
We will carouse within while they howl without, and drink the dirige
of their chief.”
Arthur said nothing, but looked doubtingly at Colville.
“And do you really imagine, worthy youth, and no less worthy
blackfit, that I am to have my house sieged, my cattle stolen, and my
corn carried off, to shield you from the consequences of your
drunken brawls? Not I, by the cat of the blessed Bride. Out of my
doors, ye caitiffs,—they can but slay you, and the whittle has crossed
the craig of mony a better fellow than any of ye twasome is likely to
prove. Begone, I say.”
“Nay, my dear father,” said Edith, imploringly, “do not drive them
forth now; the Egyptians are approaching the house—they cannot
escape.”
“And they shall not stay here,” replied the old man, harshly, the
tone of agony in which Edith’s entreaties were uttered recalling all
the bitterness of his feelings against Arthur.
“At least, Walter Colville,” said Arthur, “save this stranger. He
cannot have offended you. It was on my errand he came hither. I will
go forth alone. Perhaps one victim may suffice.”
“Nay, brave youth,” said the stranger, “we go together. Farewell,
old man. You are a Scot, and yet have betrayed your guest. You are a
Colville, and the first of the line that ever turned his back upon a
Stuart at his utmost need.”
The tone and sentiment of these words had a powerful effect on
Walter Colville. A momentary confusion rested on his countenance,
and then, with a smile ill put on, he said,—
“Come, come, sirs; I but joked wi’ ye. Did you really think that
Walter Colville would abandon to his enemy any who have bitten his
bannock, and kissed his cup as you have done? Na, na; here you are
safe while the auld wa’s stand. Sit down. I’ll go above and look out for
the landloupers.”