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HANDBOOK OF MICROBIAL
NANOTECHNOLOGY
This page intentionally left blank
 HANDBOOK OF
MICROBIAL
NANOTECHNOLOGY

Edited by

CHAUDHERY MUSTANSAR HUSSAIN

Department of Chemistry and Environmental Science, New Jersey Institute of Technology,
Newark, NJ, United States
Academic Press is an imprint of Elsevier
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be noted herein).
Notices
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understanding, changes in research methods, professional practices, or medical treatment may become necessary.

Practitioners and researchers must always rely on their own experience and knowledge in evaluating and using any
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Typeset by MPS Limited, Chennai, India

 Dedication

I would like to dedicate this handbook to

my beloved GOD

Meray Pyarey Allah (SWT)

This page intentionally left blank
 Contents

List of contributors xv 2. Cancer therapeutics with microbial

About the editor xix nanotechnology-based approaches 17
Preface xxi Linh B. Truong, David Medina Cruz, Hamed Barabadi, Hossein
Vahidi and Ebrahim Mostafavi
Acknowledgments xxiii
2.1 Introduction of cancer, current state,
1. Microbial nanotechnology
based treatments, and limitations 17
approaches for wound healing and 2.2 Introduction of nanoparticles, advantages,
infection control 1 properties, synthesis pathways, and the
Hamed Barabadi, Ebrahim Mostafavi, Linh B. Truong, emerging use of microbial synthesis 19
David Medina Cruz, Hossein Vahidi, Mohammad Ali Mahjoub, 2.3 Synthesis pathways and general
Omid Hosseini and Muthupandian Saravanan characteristics 20
2.4 Direct therapeutic mechanisms
1.1 Introduction 1 (nanoparticles as therapy) 23
1.2 Wound healing and infection control: 2.5 Indirect therapeutic mechanism 27
an insight 2 2.6 Current challenges and prospects in clinical
1.3 Use of nanotechnology in wound healing and translation 30
infection control 3 2.7 Conclusion 32
1.3.1 Current therapies and their References 32
drawbacks 3
1.3.2 Current nanoplatforms for wound
healing and infection control 3
3. Nanotechnological interventions
1.4 Microbial synthesis of nanomaterials 4 for the detection of pathogens through
1.5 Methods of microbial-based green synthesis of surface marker recognition 45
nanomaterials 5 Chandni Sharma, Mohini Verma, Shiwani Randhawa and
Amitabha Acharya
1.5.1 Bacterial-mediated synthesis of
nanoparticles 5
3.1 Introduction 45
1.5.2 Fungal-mediated synthesis of 3.2 Biomarkers exposed on the surface of
nanoparticles 5 microorganisms 46
1.5.3 Microalgal-mediated synthesis of 3.2.1 Surface proteins 47
nanoparticles 6 3.2.2 Carbohydrates 48
1.6 Microbially synthesized nanomaterials for 3.2.3 Glycoproteins 49
wound healing and infection control 6 3.2.4 Extracellular DNA 50
1.6.1 Gold nanoparticles 6 3.3 Conventional methods 51
1.6.2 Silver nanoparticles 7 3.3.1 Cell culture and colony counting based
1.6.3 Metal oxide nanoparticles 10 methods 52
1.7 Antibacterial mechanisms of metal-based 3.3.2 Microscopy 53
nanoparticles 11 3.3.3 Polymerase chain reaction 53
1.8 Conclusions and future outlook 12 3.3.4 Immunology-based detection
References 12 methods 54

vii
viii Contents

3.3.5 Flow cytometry 55 5.2.2 Classification based on the transducer

3.3.6 Biosensors-based detection 56 component 97
3.4 Switching from conventional to 5.3 Optical biosensors and methods 100
nanotechnological approach 57 5.3.1 Biofunctionalization strategies 101
3.4.1 Immunosensor 59 5.3.2 Polymer optical fibers 102
3.4.2 Surface-enhanced Raman 5.3.3 Immobilization of metal
spectroscopy-based biosensor 60 nanoparticles 102
3.4.3 Colorimetric Sensor 62 5.3.4 Aptamers as biorecognition
3.4.4 Fluorometric sensor 62 elements 102
3.4.5 Electrochemical sensor 63 5.4 Nanomaterial enhanced biosensors 103
3.4.6 Miscellaneous sensing platforms 65 5.4.1 Surface plasmon resonance-based
3.5 Conclusion and future prospects 68 biosensors 105
Acknowledgement 68
5.4.2 Fluorescence-based fiber optic
Abbreviations 68
biosensors 108
References 69
5.4.3 Raman spectroscopy and
surface enhanced Raman
4. An overview of microbial calcite spectroscopy 110
nanoparticle generation in self-healing 5.5 Conclusion 123
References 124
concrete: its potential, advantages, and
Further reading 132
limitations as a green building
material 79
Himanshi Saini and Lalita Ledwani 6. Utilization of flow cytometry in
nanomaterial/bionanomaterial
4.1 Introduction 79
4.2 Constituents of microbial concrete 80
detection 133
4.3 Implantation of healing agents and Ramakrishnan Geethalakshmi, SR Nivaz, GS Lekshmi,
Duraiarasan Surendhiran, Chaudhery Mustansar Hussain and
precipitation process inside the matrix 81 Abdul Razack Sirajunnisa
4.4 Performance and enhancement of bioconcrete
properties 84 6.1 Introduction 133
4.5 Potential of bioconcrete in the construction 6.2 Flow cytometer: principles and
industry 85 instrumentation 134
4.6 Advantages and disadvantages 87 6.2.1 Principle of flow cytometry 134
4.7 Conclusion 88 6.2.2 Instrumentation 135
References 88 6.3 Flow cytometry and its applications in
research 138
6.3.1 Immunophenotyping 138
5. Nanobiosensors for detection of 6.3.2 Cell sorting 138
bacteria: an overview of fiber-optics 6.3.3 Cell cycle analysis 139
and Raman spectroscopy based 6.3.4 Apoptosis 139
biosensors 91 6.3.5 Intracellular calcium flux 139
6.3.6 Analysis of microbiota 139
J. Nirgund, K.N. Purana, D. Selvakumar, N.S. Kumar and
S. Sil 6.4 Nanotechnology and flow cytometry 139
6.4.1 Imaging of nanoparticles in
5.1 Introduction 91 suspension 140
5.2 Biosensors for pathogen detection 94 6.4.2 Detection of nanoparticles 141
5.2.1 Classification based on biorecognition 6.5 Conclusion 142
element 95 References 142
 Contents ix
7. Utilization of Raman spectroscopy in 9.2 Microalgae production of metallic
nanomaterial/bionanomaterial nanoparticles 170
9.2.1 Microalgal synthesis of metallic
detection 145
nanoparticles 171
SR Nivaz, Ramakrishnan Geethalakshmi, GS Lekshmi,
9.2.2 Silver nanoparticles for use in antibiotic
Duraiarasan Surendhiran, Chaudhery Mustansar Hussain and
Abdul Razack Sirajunnisa applications 172
9.2.3 Microalgae production of gold and other
7.1 Introduction 145 metallic nanoparticles 173
7.2 Raman: principle and instrumentation 146 9.3 Microalgae production of biomolecules for
7.2.1 Principle: Raman scattering and pharmaceutical applications 174
shift 146 9.3.1 Bioassay-guided fractionation and other
7.2.2 Instrumentation 147 methods for determining
7.2.3 Variants in Raman spectroscopy 148 bioactivity 175
7.3 Detecting nanoparticles in cells using Raman 9.3.2 Anticancer bioactive compounds from
spectroscopy 151 microalgae 177
7.3.1 Surface-enhanced Raman 9.3.3 Antimicrobial bioactive compounds from
spectroscopy 151 microalgae 178
7.4 Detecting nanoparticles in cells using Raman 9.3.4 Microalgal drug discovery for other
spectroscopy 153 health applications 178
7.5 Conclusion 154 9.3.5 Phycotoxins as potential drugs 179
References 154 9.4 Microalgae as facilitating technologies 180
9.4.1 Genetic transformation of microalgae as
8. Nanotechnology based Pathogen drug factories 180
identification through surface marker 9.4.2 Microalgae as scavengers 182
identification 157 9.5 Summary and conclusions 182
References 184
Anamika Nayak and Debjani Dutta

8.1 Introduction 157 10. Regulations and risk assessment of

8.2 Nanotechnological advancement in pathogen microbial green nanotechnology 191
identification 158 Katya M. Aguilar-Pérez, Gustavo Ruiz-Pulido, Dora I. Medina,
8.2.1 Gold nanoparticle
based Roberto Parra-Saldivar and Hafiz M.N. Iqbal
detection 158
8.2.2 Silver nanoparticle
based 10.1 Introduction 191
detection 159 10.2 Microbial green synthesis of
8.2.3 Quantum dot
based detection 160 nanomaterials 194
8.2.4 Carbon nanotube
based detection 161 10.2.1 Intracellular synthesis 194
8.2.5 Magnetic nanoparticle
based 10.2.2 Extracellular synthesis 194
detection 162 10.3 Life cycle assessment of nanomaterials:
8.3 Trends and challenges 163 environmental and health risk assessment 195
8.4 Conclusion 164 10.4 Influencing factors in toxicity of green
Abbreviation 164 nanomaterials 196
References 165 10.5 Challenges on safety assessment 199
10.6 Global regulatory aspects on microbial green
9. Microalgae nanotechnology and nanotechnology 201
drug development 169 10.7 Conclusion and future trends/Green Nano
Jennifer R. McCall, Ariel P. Brown, Kathryn T. Sausman and
Policy recommendations 202
Samuel H. McCall IV Acknowledgments 204
Conflicts of interest 204
9.1 Introduction 169 References 204
x Contents
11. Nanoparticles as antibacterial agent for
dental restorative materials and their
antibacterial activity evaluation 209
Dasmawati Mohamad and Habsah Hasan
11.1 Introduction 209
11.1.1 Dental restorative materials 209
11.2 Nanoparticles as fillers in restorative
materials 210
11.3 Surface morphology and roughness of
restorative materials with different filler
sizes 211
11.4 A study of surface roughness relation with
bacterial accumulation 211
11.5 Nanoparticles as antibacterial agents and their
mechanisms 214
11.5.1 Nano zinc oxide particles 214
11.5.2 Nanosilver particles 214
11.5.3 Nanographene 215
11.6 Antibacterial evaluation technique 216
11.6.1 Disk diffusion 217
11.6.2 Broth dilution method 218
11.6.3 Time-kill method 221
11.7 Microscopy cellular structure of the
microbes 221
References 222
12. Green synthesis of nanomaterials 225
N.B. Singh
12.1 Introduction 225
12.2 Synthesis of nanomaterials 226
12.3 Green chemistry 228
12.4 Methods of synthesizing nanomaterials
employing green routes 229
12.5 Synthesis of nanomaterials with plants 230
12.5.1 Synthesis of silver
nanoparticles 230
12.5.2 Synthesis of gold NPs 231
12.5.3 Synthesis of ZnO NPs 232
12.5.4 Synthesis of CNT using leaf extracts
as a catalyst 236
12.5.5 Synthesis of reduced graphene
oxide
nickel oxide nanocomposites
using Psidium guajava L. (guava)
leaf extract 242
12.6 Synthesis of nanomaterials through
microbes 243
12.7 Microwave 248
12.8 Sonochemical method 249
12.9 Synthesis from waste 249
12.10 Synthesis of nanomaterials by using
solvents 250
12.11 Conclusions 253
References 253
13. Antimicrobial nanocoating for food
industry 255
Raciye Meral, Zafer Ceylan, Nazan Kutlu, Ali Kılıçer,
Abdullah Çağlar and Oktay Tomar
13.1 Introduction 255
13.2 Coating 256
13.2.1 Coating applications for food
safety 256
13.2.2 Nanocoating 257
13.3 Widely used nanocoating antimicrobial
materials 261
13.3.1 Nano-silver-based nanocoating 261
13.3.2 Encapsulated bioactive materials for
nanocoating in food safety 262
13.3.3 Probiotic bacteria and
nanocoating 265
13.3.4 Widely used wall materials for
nanoencapsulation of antimicrobial
materials 267
13.4 Some potential convertible materials for
obtaining nanomaterials 271
13.4.1 Pumice and perlite 271
13.5 Conclusion 273
References 274
14. Antiviral potential of green-
synthesized silver nanoparticles 285
Hamed Barabadi, Kamyar Jounaki, Elahe Pishgahzadeh,
Hamed Morad, Salar Sadeghian-Abadi, Hossein Vahidi and
Chaudhery Mustansar Hussain
14.1 Introduction 285
14.2 Use of nanotechnology for antiviral
therapeutics 287
14.2.1 Current therapies and their
drawbacks 287
14.2.2 Current nanoplatforms for antiviral
therapeutics 288
14.3 Bioengineering of silver nanomaterials using
biological resources 290
 Contents xi
14.3.1 Plant-mediated synthesis of silver 16.3 Application of polymer nanocomposites in
nanomaterials 292 food industry 332
14.3.2 Fungal-mediated synthesis of silver 16.4 Bio-based polymer applications in food
nanomaterials 294 industry (nanocomposite) 333
14.3.3 Bacterial-mediated synthesis of silver 16.5 Polyhydroxybutyrate 335
nanomaterials 295 16.5.1 Production of
14.3.4 Algal-mediated synthesis of silver polyhydroxybutyrate 335
nanomaterials 297 16.5.2 Polyhydroxybutyrate nanocomposite
14.4 Green-synthesized silver nanoparticles for films for packaging applications 337
antiviral therapeutics: A mechanistic 16.6 Bacterial cellulose 341
approach 297 16.6.1 Production of BC 342
14.5 Conclusions and future outlook 302 16.6.2 Bacterial nanocellulose for food
References 303 packaging applications 342
16.7 Polylactic acid 343
15. Microbial nanotechnology in 16.8 Curdlan, gellan gum, k-carrageenan, and
xanthan gum nanocomposite 344
food industry: antimicrobial
16.9 Microbial biopolymeric nanocomposites’ role
packaging 311 in the food packaging 345
G. Sivaprakash, R. Karthik Raja, K. Mohanrasu, G.H. Dinesh and 16.10 Safety and regulation issues of NPs 346
A. Arun
16.11 Conclusion 346
Acknowledgments 347
15.1 Introduction 311
References 347
15.2 Drawbacks of existing packaging
materials 312
15.3 Role of nano research in the food 17. Pathogen identification through
industry 313
15.4 Food packaging 314
surface marker recognition methods 355
15.4.1 Active packaging 315 V. Ananthi and A. Arun
15.4.2 Intelligent or smart nanosystem
17.1 Introduction 355
packing 316
17.2 Components of nanoparticles involved in
15.4.3 Antimicrobial food packing 316
pathogen detection 357
15.4.4 Nanocomposites in food packing 317
17.3 Biosensors 358
15.5 Shelf life or preservation of food
17.3.1 Electrochemical biosensors 360
material 319
17.3.2 Colorimetric biosensors 360
15.6 Toxicological and safety aspects of
17.3.3 Fluorescent biosensors 361
nanotechnology in food packaging 320
17.3.4 Surface-enhanced Raman scattering
15.7 Conclusion 323
biosensors 361
Acknowledgments 323
17.4 Pathogen identification through surface
References 324
marker recognition 361
17.4.1 Pathogen detection by metallic
16. Microbial bio-based polymer nanoparticles 362
nanocomposite for food industry 17.4.2 Pathogen detection by quantum
applications 331 dots 365
17.4.3 Pathogen detection by magnetic
K. Mohanrasu, R. Guru Raj Rao, V. Ananthi, G. Sivaprakash,
G.H. Dinesh, Angelin Swetha, J. Jeyakanthan and A. Arun nanoparticles 365
17.4.4 Pathogen detection by fluorescent
16.1 Introduction 331 polymeric nanoparticles 366
16.2 Nanotechnology applications in food 17.4.5 Recent advancements in pathogen
industry 332 detection by other nanoparticles 367
xii Contents

17.5 Future perspectives 367 19.6 Mechanism of nano-remediation 402

References 368 19.6.1 Chemical catalysis 402
19.6.2 Photo-catalysis 402
19.6.3 Adsorption 403
18. Microbial nanotechnology in 19.6.4 Sensor: surface enhanced Raman
cancer therapy 375 scattering substrate 404
Shama Parveen and Monisha Banerjee 19.7 Conclusion 405
19.8 Future outlook 405
18.1 Introduction 375 References 406
18.2 Microbial nanotechnology 375
18.2.1 Enzymes responsible for
synthesis 376 20. Enzymes incorporated nanotechnology
18.2.2 Mechanism of synthesis 376 for wastewater treatment 415
18.2.3 Microbial nanotechnology in T. Angelin Swetha, K. Mohanrasu, Abhispa Bora,
cancer 377 V. Ananthi and A. Arun
18.3 Microbial nanotechnology in
immunotherapy 378 20.1 Introduction 415
18.4 Microbial nanoformulations 379 20.2 Enzymes 416
18.4.1 Fungus-based nanoparticles 379 20.2.1 Properties of enzymes 417
18.4.2 Bacteria-based nanoparticles 380 20.3 Types of enzymes used for wastewater
18.4.3 Algae-based nanoparticles 380 treatment 417
18.4.4 Future prospects 380 20.3.1 Phenolic contaminants and related
Acknowledgments 381 compounds 417
References 381 20.3.2 Pulp and paper waste treatment 420
20.3.3 Treatment of pesticides 421
20.3.4 Treatment of cyanide wastes 422
19. Green synthesized nanomaterials for 20.3.5 Treatment of food-processing
greener environment 385 wastes 422
Sudip Nag, Arnab Pramanik and Maitree Bhattacharyya 20.3.6 Solid waste and sludge
treatment 423
19.1 Introduction 385 20.3.7 Removal of heavy metals 424
19.2 Environmental pollutants: chemical and 20.4 Enzyme integrated nanoparticle for wastewater
molecular classification 386 treatment 424
19.2.1 Aromatic and poly-aromatic 20.4.1 Magnetic nanoparticles 426
hydrocarbon 387 20.4.2 Gold and silver nanoparticles 426
19.2.2 Synthetic dyes 387 20.4.3 Chitosan nanoparticles 427
19.2.3 Heavy metals 389 20.4.4 Carbon nanotubes 427
19.2.4 Halogen containing compounds 391 20.4.5 Silica nanoparticles 428
19.3 Environmental pollutants: effect on human 20.5 Delivery of enzyme by using nanoparticle for
health 391 wastewater treatment 428
19.4 Limitation of conventional remediation 20.6 Applications of enzyme-based
techniques 393 nanomaterials 430
19.5 Nano-bioremediation: a modern approach of 20.6.1 Use of enzyme-based nanomaterials
using green synthesized nanomaterials in for the elimination of emerging
environmental remediation 393 pollutants 430
19.5.1 Metal-based nanoparticles 394 20.6.2 Disinfection 431
19.5.2 Carbon-based nanoparticles 399 20.7 Conclusion 433
19.5.3 Biopolymer-based Acknowledgements 433
nanomaterials 400 References 433
 Contents
21. Microbes incorporated nanomaterials
for water purification 439
Abhispa Bora, K. Mohanrasu, T. Angelin Swetha,
V. Ananthi, P. Kumar, Muthusamy Govarthanan and A. Arun
21.1 Introduction 439
21.2 Microbial synthesis of silver
nanoparticles 440
21.3 Mechanism of microbially synthesized
nanoparticles 441
21.3.1 Mechanism of metal nanoparticle
production by bacteria 441
21.3.2 Mechanism of metal nanoparticle
generation by fungi 443
21.3.3 Utilization of whole-cell versus cell
extracts for nanoparticle
production 444
21.4 Endophytic microbes—the biofactories of
nanoparticles synthesis 444
21.5 Applications of nanomaterials on wastewater
treatment/water purification 446
21.5.1 Zero-valent nanoparticles 446
21.5.2 Metal oxide nanoparticles 447
21.5.3 Carbon nanomaterials 448
21.5.4 An amalgamation of microorganisms
with electrospun nanofibrous webs for
water decontamination 450
21.6 Use of microbially manufactured silver
nanoparticles for water purification 451
21.7 Conclusion 451
Acknowledgment 452
References 452
22. Green nanotechnology for the
environment 461
Ramalingam Karthik Raja, Selcuk Hazir, Govindan Balasubramani,
Gurusamy Sivaprakash, Ebenezer Samuel James Obeth,
Thulasinathan Boobalan, Arivalagan Pugazhendhi,
R. Hari Krishna Raj and Alagarsamy Arun
22.1 Introduction 461
22.2 Goals of green technology 462
22.3 Current scientific and technological
advancements 463
22.3.1 Recent advancements in green
nanotechnology 463
22.3.2 The potential impact of
nanotechnology on green
technologies 466
xiii
22.3.3 Water clean-up nanomaterial
technology 466
22.3.4 Nanomaterials for construction
industry 467
22.4 Energy and environmental technology 468
22.4.1 Nanomaterials for energy
conversion 468
22.4.2 Nanomaterials for energy
storage 468
22.5 Nano-enhanced green technologies 469
22.6 Nanomaterials in chemical industry 469
22.7 Impact on environmental filtration and
remediation 470
22.7.1 Safe treatment, filtration and
desalination of water by cheap and
compact nanotechnology filters 470
22.8 Green nanotechnology for eco-friendly
agriculture 471
22.8.1 Nanofertilizers 471
22.8.2 Nanopesticides 472
22.9 Nanotechnology and air pollution 472
22.9.1 Nanostructured membranes and
catalysts 472
22.10 Nanotechnology for pollution
prevention 472
22.11 Toxicity, risk assessment, and
management 473
22.12 Conclusion 474
Acknowledgment 474
References 474
23. Atomic force microscopy as
multifunctional microbial imaging and
characterization platform 479
Marta Woźniak-Budych, Barbara M. Maciejewska,
Stefan Jurga and Karolina Wieszczycka
23.1 Antibiotic resistance 479
23.1.1 Classification on the basis of Gram
stain and bacterial cell wall 480
23.1.2 Bacterial pathogens and their
antibiotic resistance 481
23.1.3 The bacterial antibiotic
resistance 485
23.2 Multifunctional microbial identification and
imaging 487
23.3 Atomic force microscopy, multifunctional tool
for biological sample characterization 495
xiv Contents

23.4 Challenges 501 24.5 Microbial nanotechnology in energy usage or

Acknowledgment 502 consumption 527
Credit: authorship contribution statement 502 24.5.1 Current status of energy usage 527
References 502 24.5.2 Nanotechnology for energy
consumption 527
24. Role of microbial nanotechnology in 24.5.3 Role of microbial nanotechnology in
energy devices 517 energy consumption 528
24.6 Microbial nanotechnology in energy
Shareefraza J. Ukkund, Bhavna Alke, Syed Noeman Taqui and
Usman Taqui Syed storage 529
24.6.1 Bioelectrochemical systems 529
24.1 Introduction 517 24.6.2 Supercapacitive microbial fuel
24.2 Microbial nanotechnology in energy cells 529
sources 519 24.6.3 Nanotechnology and supercapacitive
24.2.1 Nanotechnology in energy microbial fuel cells 529
sources 519 24.7 Biofuels 531
24.2.2 Microbes as energy sources 520 24.7.1 Biohydrogen 531
24.3 Microbial nanotechnology in energy 24.7.2 Biodiesel 534
conversion 520 24.7.3 Bioethanol 536
24.3.1 Microbial photoelectrochemical 24.7.4 Biomethane 538
system 523 24.8 Conclusion 541
24.4 Microbial nanotechnology in energy References 541
distribution 523
24.4.1 Nanotechnology in energy
distribution 525 Index 549
 List of contributors

Amitabha Acharya Biotechnology Division, Science Talent Search, Kolkata, West Bengal,
CSIR-Institute of Himalayan Bioresource India
Technology, Palampur, Himachal Pradesh, Thulasinathan Boobalan Department of
India; Academy of Scientific and Innovative Microbiology, Alagappa University,
Research (AcSIR), Ghaziabad, Uttar Pradesh, Karaikudi, Tamil Nadu, India
India
Abhispa Bora Bioenergy and Bioremediation
Katya M. Aguilar-Pérez Tecnologico de Laboratory, Department of Microbiology,
Monterrey, School of Engineering and Alagappa University, Karaikudi, Tamil
Sciences, Monterrey, Mexico City, Mexico Nadu, India
Bhavna Alke LAQV/REQUIMTE, Department
Ariel P. Brown Clinical Research Program,
of Chemistry, Faculty of Science and
School of Nursing, University of North
Technology, NOVA University of Lisbon,
Carolina Wilmington, Wilmington NC, USA
Caparica, Portugal
Abdullah Çağlar Faculty of Agriculture and
V. Ananthi Department of Molecular Biology,
Natural Science, Kocaeli University, Kocaeli,
Madurai Kamaraj University, Madurai, Tamil
Turkey
Nadu, India
Zafer Ceylan Van Yüzüncü Yıl University,
T. Angelin Swetha Bioenergy and
Faculty of Tourism, Department of
Bioremediation Laboratory, Department of
Gastronomy and Culinary Arts, Van, Turkey
Microbiology, Alagappa University,
Karaikudi, Tamil Nadu, India David Medina Cruz Department of Chemical
Engineering, Northeastern University,
A. Arun Bioenergy and Bioremediation
Boston, MA, United States
Laboratory, Department of Microbiology,
Alagappa University, Karaikudi, Tamil Nadu, G.H. Dinesh Department of Microbiology,
India Alagappa University, Karaikudi, Tamil
Govindan Balasubramani Aquatic Animal Nadu, India
Health and Environmental Division, ICAR- Debjani Dutta Department of Biotechnology,
Central Institute of Brackishwater National Institute of Technology, Durgapur,
Aquaculture, Chennai, Tamil Nadu, India West Bengal, India
Monisha Banerjee Molecular and Human Ramakrishnan Geethalakshmi Bionanomaterials
Genetics Laboratory, Department of Laboratory, Centre for Biotechnology, Anna
Zoology, University of Lucknow, Lucknow, University, Chennai, Tamil Nadu, India
India Muthusamy Govarthanan Department of
Hamed Barabadi Department of Pharmaceutical Environmental Engineering, Kyungpook
Biotechnology, School of Pharmacy, Shahid National University, Daegu Campus,
Beheshti University of Medical Sciences, Gyeongbuk Province, South Korea
Tehran, Iran R. Guru Raj Rao Structural Biology and
Maitree Bhattacharyya Department of Bio-Computing Lab, Department of
Biochemistry, University of Calcutta, Kolkata, Bioinformatics, Alagappa University,
West Bengal, India; Jagadis Bose National Karaikudi, Tamil Nadu, India

xv
xvi List of contributors
Habsah Hasan Microbiology and Parasitology
Department, School of Medical Sciences,
Health Campus, Universiti Sains Malaysia,
Kubang Kerian, Kelantan, Malaysia
Selcuk Hazir Department of Biology, Faculty
of Arts and Science, Adnan Menderes
University, Aydin, Turkey
Omid Hosseini Shahid Beheshti University of
Medical Sciences, Tehran, Iran
Chaudhery Mustansar Hussain Department of
Chemistry and Environmental Science, New
Jersey Institute of Technology, Newark, NJ,
United States
Hafiz M.N. Iqbal Tecnologico de Monterrey,
School of Engineering and Sciences,
Monterrey, Mexico city, Mexico
J. Jeyakanthan Structural Biology and Bio-
Computing Lab, Department of
Bioinformatics, Alagappa University,
Karaikudi, Tamil Nadu, India
Kamyar Jounaki Department of Pharmaceutical
Biotechnology, School of Pharmacy, Shahid
Beheshti University of Medical Sciences,
Tehran, Iran
Stefan Jurga NanoBioMedical Centre, Adam
Mickiewicz University, Poznan, Poland
N.S. Kumar Department of Environmental
Protection, Defence Bioengineering and
Electromedical Laboratory (DEBEL),
Bengaluru, Karnataka, India
P. Kumar Department of Animal Health and
Management, Alagappa University,
Karaikudi, Tamil Nadu, India
Nazan Kutlu Faculty of Engineering, Food
Engineering Department, Van Yüzüncü Yıl
University, Van, Turkey; Institute of
Science, Van Yüzüncü Yıl University, Van,
Turkey
Ali Kılıçer Faculty of Engineering, Geological
Engineering Department, Van Yüzüncü Yıl
University, Van, Turkey
Lalita Ledwani Manipal University Jaipur,
Jaipur, Rajasthan, India
GS Lekshmi Advanced Nanomaterials
Laboratory, Centre for Nanoscience and
Technology, Anna University, Chennai,
Tamil Nadu, India
Barbara M. Maciejewska NanoBioMedical
Centre, Adam Mickiewicz University,
Poznan, Poland
Mohammad Ali Mahjoub Department of
Pharmaceutics, School of Pharmacy, Shahid
Beheshti University of Medical Sciences,
Tehran, Iran
Jennifer R. McCall Clinical Research
Program, School of Nursing, University of
North Carolina Wilmington, Wilmington
NC, USA; SeaTox Research Inc, Wilmington
NC, USA
Samuel H. McCall, IV SeaTox Research Inc,
Wilmington NC, USA
Dora I. Medina Tecnologico de Monterrey,
School of Engineering and Sciences,
Monterrey, Mexico City, Mexico
Raciye Meral Faculty of Engineering, Food
Engineering Department, Van Yüzüncü Yıl
University, Van, Turkey
Dasmawati Mohamad Biomaterials Unit,
School of Dental Sciences, Health Campus,
Universiti Sains Malaysia, Kubang Kerian,
Kelantan, Malaysia
K. Mohanrasu Department of Microbiology,
Alagappa University, Karaikudi, Tamil
Nadu, India; Bioenergy and Bioremediation
Laboratory, Department of Microbiology,
Alagappa University, Karaikudi, Tamil
Nadu, India
Hamed Morad Department of Pharmaceutics,
Faculty of Pharmacy, Mazandaran University
of Medical Sciences, Sari, Iran; Ramsar
Campus, Mazandaran University of Medical
Sciences, Ramsar, Iran
Ebrahim Mostafavi Stanford Cardiovascular
Institute, Stanford University School of
Medicine, Stanford, CA, United States;
Department of Medicine, Stanford
University, School of Medicine, Stanford, CA,
United States
Sudip Nag Department of Biochemistry,
University of Calcutta, Kolkata, West Bengal,
India
 List of contributors xvii
Anamika Nayak Department of India; Academy of Scientific and Innovative
Biotechnology, National Institute of Research (AcSIR), Ghaziabad, Uttar Pradesh,
Technology, Durgapur, West Bengal, India India
J. Nirgund Department of Environmental Gustavo Ruiz-Pulido Tecnologico de
Protection, Defence Bioengineering and Monterrey, School of Engineering and
Electromedical Laboratory (DEBEL), Sciences, Monterrey, Mexico City, Mexico
Bengaluru, Karnataka, India Salar Sadeghian-Abadi Department of
SR Nivaz Advanced Nanomaterials Pharmaceutical Biotechnology, School of
Laboratory, Centre for Nanoscience and Pharmacy, Shahid Beheshti University
Technology, Anna University, Chennai, of Medical Sciences, Tehran, Iran
Tamil Nadu, India Himanshi Saini Manipal University Jaipur,
Ebenezer Samuel James Obeth Department of Jaipur, Rajasthan, India
Microbiology, Alagappa University, Muthupandian Saravanan Department of
Karaikudi, Tamil Nadu, India Medical Microbiology and Immunology,
Roberto Parra-Saldivar Tecnologico de Institute of Biomedical Sciences, College of
Monterrey, School of Engineering and Health Science, Mekelle University, Mekelle,
Sciences, Monterrey, Mexico city, Mexico Federal Democratic Republic of Ethiopia; AMR
and Nanomedicine Laboratory, Department
Shama Parveen Molecular and Human
of Pharmacology, Saveetha Dental College,
Genetics Laboratory, Department of Zoology,
Saveetha Institute of Medical and Technical
University of Lucknow, Lucknow, India
Sciences (SIMATS), Chennai, India
Elahe Pishgahzadeh Department of
Kathryn T. Sausman Clinical Research Program,
Pharmaceutical Biotechnology, School
School of Nursing, University of North Carolina
of Pharmacy, Shahid Beheshti University of
Wilmington, Wilmington NC, USA
Medical Sciences, Tehran, Iran
D. Selvakumar Department of Environmental
Arnab Pramanik Jagadis Bose National Science
Protection, Defence Bioengineering and
Talent Search, Kolkata, West Bengal, India
Electromedical Laboratory (DEBEL),
Arivalagan Pugazhendhi Innovative Green Bengaluru, Karnataka, India
Product Synthesis and Renewable
Chandni Sharma Biotechnology Division,
Environment Development Research Group,
CSIR-Institute of Himalayan Bioresource
Faculty of Environment and Labour Safety,
Technology, Palampur, Himachal Pradesh,
Ton Duc Thang University, Ho Chi Minh
India; Academy of Scientific and Innovative
City, Viet Nam
Research (AcSIR), Ghaziabad, Uttar Pradesh,
K.N. Purana Department of Environmental India
Protection, Defence Bioengineering and S. Sil Department of Environmental
Electromedical Laboratory (DEBEL), Protection, Defence Bioengineering and
Bengaluru, Karnataka, India Electromedical Laboratory (DEBEL),
R Hari Krishna Raj Department of Bengaluru, Karnataka, India
Biotechnology, Padmavani Arts & Science N.B. Singh Department of Chemistry and
College for Women, Salem, Tamil Nadu, India Biochemistry, School of Basic Sciences and
R. Karthik Raja Department of Microbiology, Research and Research Development Cell,
Alagappa University, Karaikudi, Tamil Sharda University, Greater Noida, Uttar
Nadu, India Pradesh, India
Shiwani Randhawa Biotechnology Division, Abdul Razack Sirajunnisa Bionanomaterials
CSIR-Institute of Himalayan Bioresource Laboratory, Centre for Biotechnology, Anna
Technology, Palampur, Himachal Pradesh, University, Chennai, Tamil Nadu, India
xviii List of contributors
G. Sivaprakash Department of Microbiology,
Alagappa University, Karaikudi, Tamil
Nadu, India
Duraiarasan Surendhiran Department of Food
Science and Technology, Pukyong National
University, Busan, South Korea
Usman Taqui Syed LAQV/REQUIMTE,
Department of Chemistry, Faculty of Science
and Technology, NOVA University of
Lisbon, Caparica, Portugal; Department of
Chemical and Environmental Engineering,
Institute of Nanoscience of Aragon (INA),
University of Zaragoza, Zaragoza, Spain
Syed Noeman Taqui Department of
Chemistry, University of Malaya, Kuala
Lumpur, Malaysia
Oktay Tomar Faculty of Agriculture and
Natural Science, Kocaeli University, Kocaeli,
Turkey
Linh B. Truong Department of Chemical
Engineering, Northeastern University,
Boston, MA, United States
Shareefraza J. Ukkund Department of Nano-
Technology, Srinivas Institute of Technology,
Mangalore, Karnataka, India; Centre for
Nanoscience & Technology, College of
Engineering and Technology, Srinivas
University, Mangalore, Karnataka, India
Hossein Vahidi Department of Pharmaceutical
Biotechnology, School of Pharmacy, Shahid
Beheshti University of Medical Sciences,
Tehran, Iran
Mohini Verma Biotechnology Division, CSIR-
Institute of Himalayan Bioresource
Technology, Palampur, Himachal Pradesh,
India; Academy of Scientific and Innovative
Research (AcSIR), Ghaziabad, Uttar Pradesh,
India
Karolina Wieszczycka Institute of Chemical
Technology and Engineering, Poznan
University of Technology, Poznan, Poland
Marta Woźniak-Budych NanoBioMedical
Centre, Adam Mickiewicz University,
Poznan, Poland
 About the editor
Chaudhery Mustansar Hussain, PhD, is an
adjunct professor and director of laborato-
ries in the Department of Chemistry &
Environmental Science at the New Jersey
xix
Institute of Technology (NJIT), Newark,
New Jersey, United States. His research is
focused on the applications of nanotechnol-
ogy and advanced materials, environmental
management, analytical chemistry, smart
materials and technologies, and other vari-
ous industries. Dr. Hussain is the author of
numerous papers in peer-reviewed journals
as well as a prolific author and editor of
around hundred books, including scientific
monographs and handbooks in his research
areas. He has published with Elsevier,
American Chemical Society, Royal Society
of Chemistry, Springer, John Wiley & Sons,
and CRC Press.
This page intentionally left blank
 Preface
Nanotechnology has shown great prom-
ise in various technical disciplines includ-
ing nanomedicine, nano-drug delivery
systems, fuel cell catalysts, self-assembled
polymer films, nanofabrication, miniaturi-
zation, imprint lithography, and microelec-
tronics. Similarly microbiology is related to
nanotechnology at a number of levels.
Several bacterial entities are nano-machines
in nature. These bacteria also create bio-
films by the process of self-assembly where
controlled as well as ordered building
blocks are formed. Moreover, the formation
of virus capsids is a typical procedure of
molecular recognition and self-assembly at
the nanometer level. As a result microbial
nanotechnology is predicted to be a main
driver of industry and business in this cen-
tury and will significantly impact all
aspects of society. Likewise microbial nano-
technology is taking part in creating devel-
opment and innovation in various sectors.
Despite the participation of microbial nano-
technology in modern development, there
are some hindrances. The lack of informa-
tion, the possibility of adverse impacts on
the environment, human health, safety, and
sustainability are still a challenge which are
addressed in this handbook. Special atten-
tion is paid to those approaches that are
green and sustainable for industrial devel-
opments. This proposed handbook dis-
cusses about recent advancements in
microbial nanotechnology arena.
To apprehend inclusive impression of
microbial nanotechnology and to provide
the reader a logical and expressive repre-
sentation, the book is divided into different
xxi
chapters. Chapter 1 discusses about micro-
bial nanotechnology-based approaches for
wound healing and infection control.
Cancer therapeutics with microbial
nanotechnology-based approaches are
explored in Chapter 2. Chapters 3
5 are on
nanotechnological interventions for the
detection of pathogens, microbial calcite
nanoparticle generation in self-healing con-
crete, and nano-biosensors for the detection
of bacteria. Utilization of flow cytometry and
Raman spectroscopy in bio-nanomaterial
detection is carried out in Chapters 6 and 7.
Pathogen identification through surface
marker recognition, microalgae nanotechnol-
ogy, and drug development are topics for
Chapters 8 and 9. Regulations and risk
assessment of microbial green nanotechnol-
ogy is discussed in Chapter 10. Nanoparticles
as antibacterial agent for dental application,
green synthesis of nanomaterials, antimicro-
bial nanocoating, and antiviral potential of
green synthesized silver nanoparticles are
described in Chapters 11
14. Then antimicro-
bial packaging, microbial bio-based polymer
nanocomposite for food industry applications,
pathogen identification through surface
marker recognition, and microbial nanotech-
nology in cancer therapy are discussed in the
next three chapters. Chapter 19 onwards are
based on green synthesized nanomaterials for
greener environment, enzymes-incorporated
nanotechnology for wastewater treatment,
microbes-incorporated nanomaterials for
water purification green nanotechnology for
environment, and AFM as multifunctional
microbial imaging and characterization
platform. The last chapter describes the
xxii Preface
applications of microbial nanotechnology in
energy devices.
Overall, this book is intended to be a ref-
erence guidebook for experts, researchers,
and scientists who are searching for a new
and modern development in microbial nano-
technology. The editor and authors are well-
known researchers, scientists, and specialists
from various universities and industry.
On behalf of Elsevier, I am very delighted
with all authors for their outstanding and
enthusiastic hard work in making of this
book. Very special acknowledgments to
Patricia Osborn (acquisition editor), Chezca
Cabural (editorial project manager), and
Kumar Anbazhagan (production manager)
at Elsevier, for their devoted support and
help during this project. In the end, I offer
my sincere thanks to Elsevier for publishing
the book.
Chaudhery Mustansar Hussain
 Acknowledgments

I would like to acknowledge Chaudhery Ghazanfar Hussain for his dedicated support
during the compilation of this handbook.

xxiii
This page intentionally left blank
 C H A P T E R

1
Microbial nanotechnology
based
approaches for wound healing and
infection control
Hamed Barabadi1,*, Ebrahim Mostafavi2,3,*, Linh B. Truong4,
David Medina Cruz4, Hossein Vahidi1, Mohammad Ali Mahjoub5,
Omid Hosseini6 and Muthupandian Saravanan7,8
1
Department of Pharmaceutical Biotechnology, School of Pharmacy, Shahid Beheshti
University of Medical Sciences, Tehran, Iran 2Stanford Cardiovascular Institute, Stanford
University School of Medicine, Stanford, CA, United States 3Department of Medicine, Stanford
University School of Medicine, Stanford, CA, United States 4Department of Chemical
Engineering, Northeastern University, Boston, MA, United States 5Department of
Pharmaceutics, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran,
Iran 6Shahid Beheshti University of Medical Sciences, Tehran, Iran 7Department of Medical
Microbiology and Immunology, Institute of Biomedical Sciences, College of Health Science,
Mekelle University, Mekelle, Federal Democratic Republic of Ethiopia 8AMR and
Nanomedicine Laboratory, Department of Pharmacology, Saveetha Dental College, Saveetha
Institute of Medical and Technical Sciences (SIMATS), Chennai, India

1.1 Introduction

The National Nanotechnology Initiative described the term nanotechnology as “the

understanding and control of matter at dimensions between approximately 1 and 100 nm,
where unique phenomena enable novel applications.” Besides, the U.S. Environmental
Protection Agency defined the term nanotechnology as “the creation and use of structures,
devices, and systems that have novel properties and functions because of their small size”

* These authors contributed equally to this study.

Handbook of Microbial Nanotechnology

DOI: https://doi.org/10.1016/B978-0-12-823426-6.00009-7 1 © 2022 Elsevier Inc. All rights reserved.
2 1. Microbial nanotechnology
based approaches for wound healing and infection control

(Khare et al., 2014). Nanotechnology is an emerging area of science that has the potential
to revolutionize a diverse range of fields involving biology, medicine, chemistry, physics,
agriculture, electronics, etc (Mostafavi, 2019). Nanomaterials have large surface area to vol-
ume ratios with specific optical, chemical, mechanical, and thermal properties and are
being developed for use in various applications (Ahmed et al., 2013; Khare et al., 2014).
Nanotechnology is emerging as an innovative tool in the area of wound healing and infec-
tion control. Not surprisingly, nanomaterials have been extensively studied for their ulti-
mate performance in wound healing and infection control. Commonly, the nanomaterials
that have wound healing properties could be categorized into two groups. The first group
refers to the nanomaterials that have direct wound healing activities such as metallic nano-
particles (MNPs), and the second group refers to the nanomaterials that have indirect
wound healing activities and play the role of nanocarriers such as liposomes and micelles
(Salimi & Mohammadipanah, 2021). Nanocarriers can be engineered for targeted drug
delivery systems to release their payload drug at the specific location to not only protect
the drug from the environment and inactivation but also reduce the drug adverse effects
along with controlling the drug release rate (Baranyai et al., 2021; Edis et al., 2021; Salimi
& Mohammadipanah, 2021; Shah et al., 2021). Among different nanomaterials, MNPs have
been attractive as effective alternatives to treat antibiotic-resistant bacterial infections
(Singh et al., 2020). Due to the sustained release of metal ions from the MNPs, they have
successfully been applied for the treatment of wounds. For instance, nanocrystalline
silver-coated antimicrobial barrier dressings represented significant efficacy for wound
healing and infection control through surgical wounds and burns (Khare et al., 2014).
Importantly, among different physicochemical and biological techniques for the synthesis
of MNPs, biological methods have attracted significant attention due to their advantages
over traditional physicochemical methods (Singh et al., 2020). The biological approach for
the fabrication of MNPs involves the use of biological resources such as microorganisms,
plants, algae, enzymes, etc., as reducing and capping agents in the process of MNP pro-
duction. In this chapter, we have addressed the microbial-mediated synthesis of various
MNPs and highlighted their recent advances for wound healing and infection control.

1.2 Wound healing and infection control: an insight

Wound healing is a complex and highly regulated process of restoring damaged tissue
that includes a series of continuous, and occasionally overlapping events involving hemo-
stasis, inflammation, proliferation, epithelization, maturation, and remodeling of the scar
tissue (Tocco et al., 2012). Wounds impose a great cost on society with an average annual
cost of $2.8 billion in 2014 and are expected to rise to $3.5 billion in 2021 worldwide.
Based on the 2018 market research report, the global market for wound care products
reaches over $15 billion by 2022 (Barroso et al., 2020). Wounds are categorized into either
acute or chronic. Accelerated wound healing is predictable for acute wounds. However,
any perturbation and disturbance through the wound healing phases may prolong the
wound healing stages and the wounds may be susceptible to infections, which are named
chronic wound healing (Naskar & Kim, 2020; Parani et al., 2016). The colonization of
biofilm-producing pathogenic bacteria at the site of the wound avoids the healing process

Handbook of Microbial Nanotechnology

 1.3 Use of nanotechnology in wound healing and infection control 3
(Naskar & Kim, 2020). The incidence of chronic wounds is on the rise involving pressure
ulcers, diabetic ulcers, and vascular ulcers (Hamdan et al., 2017; Parani et al., 2016). It is
estimated that there are around 300 million chronic and 100 million traumatic wound
patients worldwide who contribute major costs to healthcare systems (Barroso et al., 2020).
The risk of a wound being infected with opportunistic or pathogenic bacteria depends on
the individual susceptibility of a patient to acquire an infection such as an underlying dis-
ease as well as the ability of the wound to heal (Parker, 2000). The major risk factors of
chronic wounds include age, venous insufficiency, immunocompromised status, periph-
eral arterial diseases, and peripheral neuropathy (Parani et al., 2016). In addition, chronic
wounds cause patients to suffer, decrease the quality of life of patients, and may lead to
death in extreme situations (Barroso et al., 2020). The most common inhabitants of human
skin are bacteria that live on the superficial layers of the epidermal stratum corneum.
Although these resident bacteria seem harmless on healthy skin, they may have the ability
to become pathogenic if they have the chance to penetrate the skin due to a wound
(Parker, 2000). The parameters of an ideal healing system include: (1) moisture the wound;
(2) stimulate healing mechanisms; (3) reduce infection; (4) mimic the extracellular matrix
feature; and (5) reduce the wound scar (Barroso et al., 2020).

1.3 Use of nanotechnology in wound healing and infection control

1.3.1 Current therapies and their drawbacks

The standard chronic wound care involves swabbing for infection, cleaning, dressing, and
in some cases debridement of the wound. The gold standard in chronic wound management
is split-thickness autograft. Wound dressings based on biocompatible and biodegradable
polymers (collagen, silicon, chitosan, and hyaluronic acid) are another Food and Drug
Administration (FDA)-approved approach for chronic wound healing treatment. The quantity
of donor skin available, severe donor site morbidity, infection, and pain are the limitations of
split-thickness autograft treatment. Besides, comorbidities were found in high-risk patients fol-
lowing the wound repair with hyaluronic acid grafts. The other limitations of wound repair
with hyaluronic acid grafts involve arterial occlusion and wound site infection (Dreifke et al.,
2015). There are few FDA-approved treatment modalities for treating chronic skin wounds
because some of them are not user-friendly to use. For instance, some of the conventional
dressings applied for wound healing are usually dry and are uncomfortable. It is estimated
that 44%
70% of patients with chronic wounds with the abovementioned treatments remain
unhealed (Naskar & Kim, 2020). In addition, the other wound healing techniques such as
growth factor delivery and skin dressing are required to be improved to heal wounds prop-
erly without delaying and address the possible barriers to wound healing ranging from infec-
tion to hypoxia (Dreifke et al., 2015; Han & Ceilley, 2017).

1.3.2 Current nanoplatforms for wound healing and infection control

The current nanoplatforms for wound therapy are classified as two types, including (1)
nanomaterials that possess intrinsic characteristics that promote wound treatment and (2)

Handbook of Microbial Nanotechnology

4 1. Microbial nanotechnology
based approaches for wound healing and infection control

nanomaterials that play the role of delivery vehicles for therapeutic agents. The first type
involves metallic/metal oxide nanomaterials (e.g., silver, gold, copper, etc.) and nonmetal-
lic nanomaterials [e.g., carbon-based nanomaterials like graphene and metalloid-based
nanoparticles (NPs) like silica]. The second type of nanomaterials involves nanospheres,
nanocapsules, nanoemulsions, colloids, liposomes, micelles, vesicles, solid lipid nanoma-
terials, and nanofibers (Bhattacharya et al., 2019; Hamdan et al., 2017; Salimi &
Mohammadipanah, 2021). The first type of nanomaterials due to their antibacterial proper-
ties are ideal options for integration in wound dressings (Salimi & Mohammadipanah,
2021). In a study, it was reported that ultrafine Ag/AgCl NPs coated on graphene exhib-
ited major antibacterial activity with enhanced in vivo regeneration of the epidermis that
make it a candidate to repair burn wounds (Zhou et al., 2016). In another study, it was
reported that chitosan
Ag/ZnO composite dressing exhibited significant in vitro blood
clotting and antibacterial activity as well as enhanced wound healing and promoted ree-
pithelialization and collagen deposition in mice model wounds that made it a
suitable dressing for wound care application (Lu et al., 2017). The advantages of the sec-
ond type of nanomaterials include the sustained and controlled release of drugs, increas-
ing the solubility, and bioavailability of drugs, increasing the half-life of the drug, and
protecting the protein-based drugs from inactivation by proteolytic enzymes at the site of
the wound (Salimi & Mohammadipanah, 2021). The insufficient delivery of antibiotics in
chronic wounds through conventional antibiotic therapy was a major drawback that the
nanocarriers can overcome this problem by the delivery of antibiotics to a specific site
with optimum concentration. The unique physicochemical properties of nanomaterials
originated from their high surface area to volume ratios as well as their ultrafine size
caused the increased use of nanomaterials in wound healing owing to their significant role
in penetrability, active drug delivery, and cellular responses (Naskar & Kim, 2020).

1.4 Microbial synthesis of nanomaterials

Microbial-mediated synthesis of MNPs provides a simple, low-cost, eco-friendly

approach that, most importantly, does not use any hazardous chemical in the MNP pro-
duction process (Mohd Yusof, Rahman, et al., 2020). Microbial synthesis of MNPs defines
as the use of microorganisms including fungi, bacteria, yeasts, and algae for biofabrication
of MNPs. The advantages of this approach are related to the microbes including rapid
growth rate, simpler cultivation, and their capacity to grow at atmospheric temperature
and pressure conditions (Ghosh et al., 2021). Broadly, there are two pathways for the bio-
fabrication of MNPs including extracellular and intracellular routes. In the extracellular
pathway, the secreted or membrane-bound enzymes/proteins are involved in the metal
ion reduction and capping, whereas in the intracellular method, the metal ions are trans-
ported through the microbial cells and the NPs are synthesized in the presence of enzymes
(Dhanker et al., 2021; Ghosh et al., 2021). In the intracellular route, the chemical reaction
and ultrasound are required to draw out the MNPs from inside of the cells, whereas in the
extracellular pathway, the MNPs are formed out of the cells and through a supernatant
that is free of microorganisms. Hence, the extracellular method seems more favorable for
the green synthesis of MNPs owing to the ease of harvesting in the case of large-scale

Handbook of Microbial Nanotechnology

 1.5 Methods of microbial-based green synthesis of nanomaterials 5
production scenarios (Dahoumane et al., 2017; Sachin & Karn, 2021; Zhang et al., 2011).
Several reducing NAD(P)H-dependent enzymes have been stated for reducing the metal
ions and converting them to their nanoforms. These enzymes involve nitrate reductase,
cysteine desulfhydrase, sulfite reductase, and glutathione (Sachin & Karn, 2021). Control
of the distribution and monodispersity of MNPs is challenging in biological synthesis. The
major parameters that affect the particle size of MNPs involve the type of microorganism,
growth medium, and synthesis conditions (Zhang et al., 2011).

1.5 Methods of microbial-based green synthesis of nanomaterials

1.5.1 Bacterial-mediated synthesis of nanoparticles

Bacteria are prokaryotic microorganisms with a significant ability to reduce metal ions to
form MNPs. Although all bacteria do not have this potential, a wide range of bacteria have
been reported to have the ability to synthesize MNPs through intra- and/or extracellular path-
ways. Bacterial-mediated synthesis of MNPs is a reliable, eco-friendly, and nontoxic approach
(Dhanker et al., 2021; Iravani, 2014; Majeed et al., 2021; Mostafavi, 2021). Nitrate reductase
enzymes have been reported to be principle reducing agents in the bacterial route for the syn-
thesis and stabilization of MNPs (Ali et al., 2019). A study reported the extracellular synthesis
of zinc oxide NPs using the Bacillus licheniformis with an average size of 100 nm with hexago-
nal morphology (Abinaya et al., 2018). Likewise, spherical-shaped cadmium sulfide NPs were
fabricated extracellularly using Pseudomonas aeruginosa in the range of 20
40 nm (Raj et al.,
2016). Alternatively, extracellular biosynthesis of gold NPs (AuNPs) was reported using
Bacillus subtilis in the range of 20
25 nm with spherical morphology (Srinath et al., 2018).

1.5.2 Fungal-mediated synthesis of nanoparticles

Fungi are eukaryotic organisms that exist as unicellular or multicellular. The yeast and
fungal spores are microscopic, whereas mushrooms are large and conspicuous (Bochdansky
et al., 2017; Weber, 2016). Fungi can produce a large number of proteins and enzymes that
some of which have excellent potential for the bioreduction of metal ions and form MNPs
(Guilger-Casagrande & Lima, 2019). Fungal-mediated synthesis of MNPs is more favorable
due to the advantages involving simple culturing procedures, remarkable growth rate, sig-
nificant stability with high nanoparticle concentrations, and large quantities of extracellular
enzyme secretions (Feroze et al., 2020). The extracellular NAD(P)H-dependent nitrate reduc-
tase enzymes are considered to play a major role in the fungal-mediated synthesis of MNPs
by electron transfer during the conversion of NAD(P)H to NAD(P)1 (Guilger-Casagrande &
Lima, 2019). A study reported the biogenic synthesis of spherical-shaped selenium NPs
using the supernatant of Penicillium chrysogenum with an average hydrodynamic size of
24.65 nm (Vahidi et al., 2020). Likewise, titanium dioxide NPs were fabricated using the
Aspergillus flavus in the range of 62
74 nm with spherical and oval shapes (Rajakumar et al.,
2012). Alternatively, biogenic zirconium NPs were synthesized using the supernatant of
Penicillium aculeatum, Penicillium notatum, and Penicillium purpurogenum with spherical mor-
phology below 100 nm (Golnaraghi Ghomi et al., 2019).

Handbook of Microbial Nanotechnology

Another random document with
no related content on Scribd:
of any adhesions that may Fig. 305.—Inguinal hernia in a
exist. young pig.
Fourth stage. Resection
of the sac and obliteration of
the peritoneal orifice by suture and ligature.
Fifth stage. Suturing of the muscles and skin, and application of
a surgical dressing.
In practice, the deep sutures should be of bichromatised catgut or
silk, and the skin sutures of catgut ligature or aseptic silk.

INGUINAL HERNIA IN YOUNG PIGS.

One of the most frequent forms of hernia which the practitioner is

called on to treat is inguinal hernia in young pigs. Although this
allows little tendency towards strangulation it is always desirable to
operate, as otherwise the patients develop badly. There is no
difficulty in this, though the animals must be cast and placed on their
backs, the hind quarters being raised (Fig. 305).
First stage. A longer or shorter cutaneous incision over the neck
of the hernia and along its greater curvature.
Second stage. Isolation of the hernial sac, consisting of the
dilated internal sheath.
Third stage. Direct reduction of the hernia without opening the
sac, provided no adhesions occur, or, in the event of adhesions, after
incision of the sac.
Fourth stage. Torsion of the hernial sac and of the testicular cord
up to the inguinal ring. Application of a catgut or silk ligature around
the sac and cord at the level of the inguinal ring.
Fifth stage. Fixation of the ligature to the lips of the ring. Suture
of the skin wound, and drainage of the wound with a strip of
iodoform gauze.

IMPERFORATE ANUS.

This anomaly of development, which is not uncommon, presents

two different degrees of development.
 In the first degree the rectum is well formed, and extends as far as
the skin below the base of the tail.
In the second the rectum is incomplete or non-existent, the
floating colon terminating in a blind end at the entrance to the pelvis.
In calves, lambs, and young pigs very often imperforate anus is not
diagnosed until the second or third day after birth. Defæcation
cannot occur, and death is inevitable unless an artificial anus be
established.
First Degree.—The patient loses appetite, the abdomen remains
distended, and on examination of the anal region a doughy swelling
is felt, which projects backwards when the animal strains. The
operation is quite elementary, and always proves successful.
First stage. The skin beneath the tail is incised vertically; the
rectal cul-de-sac projects towards the incision.
Second stage. The rectal cul-de-sac is punctured, the contents
are removed, and the rectum and skin united by a few sutures. An
anus is thus established, though there is no sphincter.
Second Degree.—The general symptoms are similar, though very
often the little patient shows symptoms of atrophy or arrest in
development. The operation is somewhat complicated.
First stage. Vertical incision through the skin at the base of the
tail.
Second stage. Digital exploration of the cavity of the pelvis after
breaking down of the layers of connective tissue, and search for the
blind end of the floating colon. When discovered, the colon is
grasped between the jaws of a clamp or large forceps with smooth
jaws, and gently drawn towards the opening.
Third stage. Puncture of the blind end of the colon, and suture of
the latter to the cutaneous wound, as in the former case.
A third condition may exist, where the extremity of the colon
remains within the abdomen. Operation by way of the pelvis then
proves unsuccessful. If considered advisable, an opening may be
made through the right flank, so that the floating colon may be
brought to the surface and an artificial anus produced in this region.
An incision 1 or 2 inches in length is made below the haunch, to
allow of the introduction of the index finger, with which the loop is
 sought. The colon is
withdrawn, and the
operation thenceforth is
as above described.

PROLAPSUS AND
INVERSION OF THE
RECTUM.

This condition occurs

in young pigs in various
degrees. The necessity
for reduction depends
on the extent to which
tearing or gangrene of
the mucous membrane
has progressed. The
inverted portion is
carefully washed, freely
dressed with some non-
irritant fatty substance
Fig. 306.—1. Prolapse of the rectum and such as vaseline, and
vagina; 2, schema showing the relations progressively pushed
of the layers of the rectum in prolapse; 3, back with the thumbs
first phase, showing manner of fixing the and index fingers of
superposed layers of tissue by inserting both hands applied flat
four sutures—the left index finger is on either side of the
inserted into the rectum in order to anus. To facilitate
manipulate the parts; 4, interrupted reduction it is best to
sutures inserted around the bowel after check the animal’s
amputation. expulsive efforts by
placing a gag in the
mouth.
In more aggravated cases, when prolapsus of the rectum has
returned several times and the mucous membrane is gangrenous in
places so that such a complication as peritonitis of the pelvic cavity is
to be feared, it is better to amputate the prolapsed portion.
 The animal is secured either standing or lying down, and a large
enema is administered to remove the contents of the rectum. The
herniated portion of bowel is carefully examined, for it sometimes
happens that loops of intestine have become lodged in the dilated
peritoneal sac, produced by displacement of the rectum. In such
cases reduction should be effected before anything more is done, and
for this purpose the patient’s hind quarters should be lifted or even
suspended.
The operation for removal comprises two stages:
(1.) Fixation of the two layers of bowel by the passage of either two
or four sutures about ½ an inch behind the anus.
(2.) Circular amputation of the sutured tissues; insertion of
interrupted silk sutures through the lips of the wound; reduction.
The patient is restricted to milk diet for a week. Laxative gruels, etc.,
may then be given.
The complication to be feared is peritonitis of the pelvic cavity
owing to the sutures tearing out and allowing infective material to
pass from the bowel into the cavity.
Slight cases of prolapsus might possibly be treated by the injection
in lines of melted paraffin wax beneath the mucous membrane of the
last part of the bowel. The injection is made by means of a large
syringe provided with a long needle, the needle being gradually
withdrawn as the melted wax is expressed. Four “pillars” of wax are
usually injected at equidistant points. As they solidify they support
the bowel and prevent the recurrence of the prolapse. The operation,
however, is delicate, and scarcely to be recommended in pigs.
Moreover, in man, in whom it has chiefly been practised, the
deferred results have not always proved satisfactory.
 CHAPTER V.
RESPIRATORY APPARATUS.

TREPHINING THE FACIAL SINUSES.

This operation is necessary when pus, tumours, or parasites exist

within the sinuses, and in some cases where tumours form within the
nasal cavities, etc.

TREPHINING THE HORN CORE.

This cavity is opened in front, at the base of the horn, about ¾ of

an inch above the keratogenous band.

FRONTAL SINUS.

The frontal sinus may be trephined at one of two points, that is,
either towards its highest or lowest extremity.
In the former case the point selected is in the direction of the axis
of the horn core, about ¾ of an inch nearer the middle line than the
base of the horn itself.
The animal should be cast.
First stage. A V-shaped incision ¾ of an inch long on each side is
made through the skin and subjacent tissues, exposing the bone.
Second stage. The skin and periosteum are dissected away and
reflected upwards.
Third stage. Trepanation.
The lower portion of the cavity is trephined within the angle
formed by a transverse line uniting the upper margin of the orbits
and the inner margin of the super-orbital foramen.
The stages are precisely the same as those above described.

MAXILLARY SINUS.
 In adult animals the maxillary sinus is opened immediately above
the maxillary tuberosity. In the young the point selected is ¾ of an
inch higher.

TRACHEOTOMY.

In bovine animals tracheotomy is only performed in urgent cases,

in order to ward off asphyxia or to facilitate some other operation on
the upper air passages. It is performed exactly as in the horse, the
animal either standing or lying down. In the former case, the animal
may be placed in the trevis, but two strong assistants holding the
animal’s head and nose by means of “bulldogs” are often sufficient.
To prevent the animal from striking out with the front legs, a rope
is passed above and around the knees in the form of the figure 8; the
animal is backed into a corner, and operation is then quite safe.
Large animals must be cast or placed in the trevis.
The seat of operation should be washed, shaved, and disinfected.
The operation may be divided into four stages.
First stage. Vertical median incision about 2 inches long through
the skin at the height of the upper third of the trachea.
Second stage. Separation with a blunt director of the muscles
covering the trachea. Incision through the pretracheal connective
tissue.
Third stage. Circular or elliptical opening through the trachea of
a size corresponding to that of the tracheotomy tube.
Fourth stage. Insertion of the tracheotomy tube.
 CHAPTER VI.
GENITO-URINARY ORGANS.

In the domestic ruminants the penis exhibits a peculiar S-shaped

curve, situated in the subpubic region (Fig. 226), so that when
operation on the urethra, or even on the extremity of the penis,
becomes necessary the organ must first of all be withdrawn.
The manipulation is as follows:—
The animal having been fixed by the head and front legs in a
standing position, and if possible thrust against a wall, the operator
stands on its left side. With his right hand he seizes the penis and the
skin immediately in front of the scrotum and pushes them forward in
the direction of the opening of the sheath.
The extremity is nipped between the first fingers of the left hand,
and to prevent the glans slipping or escaping when the right hand is
removed (for the purpose of taking a fresh hold of the body of the
penis further back) the operator may reverse the free extremity of the
penis so that it forms a loop, and thus secure a firmer hold. With the
right hand the skin is thrust backward, a new portion of the sheath
fixed, and the organ again pushed forward. In this way the penis is
gradually extended. When the animal is cast, this manipulation is
much easier.

URETHROTOMY IN THE OX.

Urethrotomy consists in incising the urethra, usually for the

purpose of extracting a foreign body or calculus which impedes
micturition. In the ox, calculi may become fixed either in the intra-
pelvic portion of the urethra, though this is very rare; in the ischial
curvature, or more commonly at some point in the S-shaped curve of
the penis; or sometimes even within the sheath itself.
Urethrotomy is performed in the ischial or scrotal region,
according to the point where the obstruction exists.

ISCHIAL URETHROTOMY.
 Urethrotomy is performed in the ischial region either to displace
or indirectly to abstract a foreign body fixed in the membranous
portion of the urethra, or directly to remove one from the spongy
portion opposite the ischial curve.
Calculi fixed in the intra-pelvic region are detected by rectal
exploration.
The exact position of the foreign body is determined by inspection
and palpation, whilst distension of the urethra by urine may be noted
even before more striking symptoms appear.
The urethra can be incised by one of three methods.
The animal should be secured, if possible, in the standing position.
The first method, which dates back to very early times, consists
in puncturing the urethra at one stroke with the fleam or lancet, and
opening it more freely, after introducing a grooved director. This
method is very useful where rupture of the bladder is imminent.
The extraction of a calculus fixed in the ischial region, or the
manipulation of an obstruction at any other point, can afterwards be
undertaken.
Second method. A second method consists in incising the
subcutaneous tissues, layer by layer, until the urethra is reached at
the ischial arch.
The operation is terminated by puncturing the urethra and
enlarging the incision in an upward direction after passing a grooved
director. This method minimises hæmorrhage and urinary
infiltration. By previously injecting cocaine, the operation may be
made practically painless.
Third method. Puncture of the urethra by a single stroke with a
straight bistoury at the ischial arch.
The opening is enlarged in an upward direction with the same
instrument.

SCROTAL URETHROTOMY.

Scrotal urethrotomy is necessary when the calculus is situated in

one of the S-shaped curves of the penis or nearer the glans.
 The operation is facilitated by casting the animal and withdrawing
the penis from the sheath, but as there is considerable danger of
rupturing the bladder when casting an animal with marked
distension of that organ, the more serious operation should be
preceded by puncturing the urethra with a fleam at the ischial arch.
By repeated moderate traction on the extremity of the glans, the S-
shaped curve can be obliterated and the anterior portion of the penis
withdrawn beyond the sheath.
One of two conditions may exist.
First case. Where the calculus is in the anterior, extra-prepubic
portion, it is removed through an incision made directly over it. After
extraction and disinfection, one or two sutures are inserted.
Second case. Should the calculus be situated in that portion of
the penis which remains within the sheath after the fullest
withdrawal of the organ, it is necessary to proceed as follows:—
(1.) The skin covering the sheath, the subcutaneous tissue, and the
mucous membrane are first incised for a length of from 1¼ to 1¾
inches.
(2.) The penis is drawn through this opening; an incision is made
directly over the calculus, dividing the fibrous layer, erectile tissue
and mucous membrane of the urethra; the parts are disinfected and
the wounds closed with sutures.
With ordinary antiseptic precautions little danger is to be feared.
Even should infiltration of urine occur, the operator need not be
unduly anxious, for, provided the parts are punctured or scarified
early, recovery usually follows.

PASSAGE OF THE CATHETER AND URETHROTOMY IN THE

RAM.

Obstruction of the urethra in rams is more commonly caused by

deposits of gravel than by single large calculi. It is generally found in
highly-fed animals, in which gravel accumulates and becomes
massed together at some point in the canal, often near the free
extremity, where it forms a plug, causing complete retention of urine.
In other cases this retention is due to a mass of sediment formed by
vesical mucus and fine gravel which collects about the neck of the
bladder.
Three operations have been advised for the removal of this
condition:—
(1.) Section of the Appendix of the Penis.—When the disease
is just appearing the sedimentary material may be collected at the
anterior extremity of the penis behind the appendix. The shepherds
in such cases remove the extremity of the penis. The resistance
disappears, the plug formed of gravel yields to the pressure of urine,
and micturition occurs as usual. Excision of the appendix, however,
incapacitates the ram for service.
(2.) Passage of the Catheter.—Passage of the catheter has been
recommended for the removal of deposits of gravel in the urethra,
but it seems a very questionable method.
Should it be determined on, the animal must be placed on its back.
The penis is then withdrawn and the double S-shaped curve is
obliterated. An incision is made over the canal behind the appendix
and a soft gutta-percha sound is passed. The sabulous accumulation
is thus dispersed.
(3.) Urethrotomy.—Scrotal urethrotomy may be performed as in
the ox.
Fig. 307.—Passing the catheter in the cow.

Ischial urethrotomy is impracticable in very fat animals, but when

the obstruction is about the neck of the bladder, and the animal’s
condition admits of it, this operation may be performed.
The patient is fixed on its back, and a metallic or gutta-percha
sound is passed into the urethra. The tissues are incised layer by
layer in the direction of the sound. Once the urethra has been opened
the soft magma may be washed out of the bladder by a free injection
of boiled water or similar aseptic liquid.

Fig. 308.—Catheter for cows.

PASSAGE OF THE CATHETER IN THE COW.

It sometimes becomes necessary to examine the bladder of the

cow.
 There is an obstacle, however, to the introduction of the sound into
the urethral canal. The meatus urinarius is covered by a little valve
which springs from the lower wall and forms behind the real opening
of the urethra a cul-de-sac, into which the point of the catheter is apt
to pass. The instrument usually employed is of gutta-percha, glass,
or, better still, of metal, as more easily sterilised (Fig. 308). It is held
like a pen, and is directed along the floor of the vagina as far as the
opening of the meatus, being guided by the index finger of the left
hand, which has previously been introduced. The point being very
slightly depressed, it enters the cul-de-sac. It is then only necessary
to reverse the movement, that is to say, raise the point, whilst gently
pressing forward; a slight resistance is felt and the sound enters the
bladder. If necessary the little valve may be held down by gently
pressing on it with the point of the left index finger.
It is sometimes an advantage to expose the seat of operation. In
such cases the lips of the vulva and the walls of the vagina may be
separated by retractors or by the use of a speculum.

CASTRATION.

Castration is performed for the purpose of removing the

reproductive power, either by obliterating the testicle or ovary or by
suppressing their functions.
In ruminants, the testicles are elongated and placed in a vertical
position, the upper portion of the scrotum presenting a constriction
and the whole scrotal mass resembling in shape a cone with its base
downwards.

CASTRATION OF THE BULL AND RAM.

These two animals, when destined for slaughter, are usually

castrated either at birth or at latest two or three months afterwards.
In Normandy, in Franche-Comté, and in England breeders castrate
young bulls by torsion of the cord.
Two incisions about 1¼ to 1¾ inches in length are made on the
lower extremity of the scrotum. The testicles are enucleated and the
testicular cords seized with two pressure clamps, with which torsion
is effected. In the South of France, in Auvergne, and in the Limousin,
bulls intended for working are not castrated until after the lapse of
some months, on account of the influence which the testicles have on
the development of the bones and muscles. Such animals are only
operated on at the age of from six months to a year, and as a rule the
method employed is that of bistournage.

BISTOURNAGE.

This method of castration has been practised from time

immemorial. It consists essentially of torsion of the testicular cord,
and aims at obliterating the vessels which it contains, and thus
bringing about atrophy of the organs served by them.
The Bull.—The animal is operated on in the standing position.
The head is fixed to a post or ring somewhat high up, in order to
check movement of the hind legs. The hind legs are also partially
secured by means of ropes or two pieces of webbing passed in a
running noose about the hocks and fixed above the knee. No
preliminary disinfection is practised, because no wound is made.
Manual Technique. The operation comprises four stages:
First stage. The operator, standing behind the animal’s hocks,
grasps the testicular cords with the hands, immediately above the
testicles, and by exercising strong pressure, thrusts the latter to the
extreme base of the scrotum. The movement is next reversed; seizing
the base of the scrotum with the right hand, he draws it smartly
downwards, whilst he places the left hand above the right, and
thrusts the testicles towards the abdomen. If the testicles do not rise
sufficiently high, the right hand is slipped between these and the left
hand, and the testicles are thus thrust upwards towards the lower
inguinal rings, slightly dilating the latter.
After this manipulation has been repeated two or three times, the
scrotum, etc., become more pliable and the testicles more easily
displaced. The second stage of the operation is thus facilitated.
Second stage. The second stage of operation may be effected by
one of two methods.
Fig. 309.—Bistournage. First phase. Manipulating the scrotum.
Fig. 310.—Bistournage. First
phase. Thrusting the testicles
upwards; manipulating the
scrotum.
 Fig. 311.—Bistournage. Second phase.

Old method: The oldest method consists in allowing one of the

testicles to rise towards the inguinal ring and to turn the other in a
vertical plane. If, for instance, it is desired to turn the right testicle,
the cord is grasped between the thumb and index finger of the left
hand (Fig. 311), the lower part of the scrotum is seized with the right
hand, and the object then is to slide the point of the testicle along the
dorsal surface of the fingers (Fig. 311). Simultaneously the operator
presses on the base of the testicle with the thumb of the left hand,
thus causing a rotary movement in a vertical plane; the tail of the
epididymis becomes uppermost. A certain empty space separates the
testicle from the base of the scrotum.
Third stage. Torsion of the cord. The testicle having been
rotated, the cord must be twisted so that the vessels may be
obliterated. The left hand continues grasping the cord, which is then
brought in front of the testicle, whilst with the right hand the testicle
is pushed backwards and is made to describe a semi-circle. The cord
was previously on the left side; it is now on the right, and
simultaneously the testicle passes from right to left.
In completing the turn
the hands must not be
changed, and, above all,
must not let go their hold;
and the cord is pushed
forwards and towards the
right with the right hand,
whilst the testicle is pushed
backwards and to the left
with the left hand. The
cord and the testicle
resume their original
position; one complete
turn has been effected.
These manipulations are
repeated several times, and
the cord soon assumes the
appearance of a large,
hard, tense string. To
ensure obliteration seven
or eight turns should be
made in the case of the bull
Fig. 312.—Bistournage. Third phase. and four or five in that of
the ram.
Torsion of the right testicle being complete, the gland is thrust
towards the upper part of the scrotum and the left testicle is
submitted to the same manipulation, the position of the hands,
however, being reversed.
Fourth stage. Fixation of the testicles in the inguinal region.
Both testicles having been thrust upwards as far as possible into the
inguinal region, the scrotum is ligatured below them. Tape or thick
cord should be used, to guard against gangrene of the lower portions
of the scrotum. A considerable œdematous swelling soon occurs, and
when at the end of twenty-four or forty-eight hours infiltration is
well developed, the ligature should be removed.
 Dubourdieu has described a different method, in which the testicle
is rotated in a horizontal plane. The position of the hands is then
different. The left testicle, for instance, being at the base of the
scrotum, the cord is grasped with the right hand opposite the base of
the testicle, and the tail of the epididymis and the testicle are held
with the whole hand whilst being rotated. If care is taken to fix the
cord with the right hand, rotation is more rapid and easier than in
the preceding method.
Difficulties
in Operation.
—Bistournage
is highly
commended in
France on
account of its
avoiding all the
complications
resulting from
sanguinary
operations.
Nevertheless it
presents great
difficulties,
particularly in
bulls of from
two to three
years of age, in
which the
testicles are
hard to
manipulate on Fig. 313.—Bistournage. Second phase.
account of their Dubourdieu’s method.
size, the
thickness of the
connective tissue, and sometimes because of abnormal adhesions. In
such cases the preliminary manipulation alone sometimes extends
over half an hour.
 Bistournage is of
doubtful efficacy
when the testicles
are small and round,
because after the
ligature has been
applied the testicular
cord tends to
untwist, and the
shape of the testicles
readily lends itself to
such movements. If
untwisting occurs,
the operation fails.
Consequences
of the Operation.
—The operation is
often followed by
more or less violent
attacks of colic; the
animal may suffer
Fig. 314.—Bistournage. Second phase. for five or six hours,
Dubourdieu’s method. after which it
recovers.
If torsion has been
clumsily performed, or if the ligature becomes displaced, the testicle
may descend and the cords untwist; the latter then appear to have
lost the firm, tense consistence which they presented after operation.
To prevent slipping of the ligature and untwisting of the cord,
Guittard suggests the use of an iron needle, with which the scrotum
is pierced through the median line, just beneath the testicles when at
their highest position; above this is placed the ligature, which then
cannot possibly slip.
The Basque operators, in order to avoid untwisting, exercise
vigorous traction from above downwards after rotating the testicle.
In this way ruptures occur which diminish the elasticity of the cord
and the epididymis, and tend to check the untwisting of the former.
 When the operation has succeeded the testicles gradually atrophy.
They do not disappear completely, and may sometimes be found
several years later of the size of a hazel-nut or a chestnut and of
fibrous consistence. It need scarcely be said that in the event of
bistournage failing, cutting operations can always be resorted to.

MARTELAGE.

The process of martelage consists in mutilating with a mallet the

testicular cord whilst still covered by all its envelopes. This
mutilation injures the walls of the arteries, causing the formation of a
clot, which cuts off the supply of blood to the testicle and causes the
gland to atrophy.
The practice is very ancient.
The animal is fixed by the horns as if for bistournage, and the
limbs are secured by two strips of webbing or two ropes, as in the
former case, though some practitioners neglect the latter precaution.
Two cylindrical rods the size of broomsticks and a wooden mallet
or farrier’s hammer are the instruments employed.
The method, however, is barbarous, cruel, and of doubtful value. It
would never be countenanced in England.

CASTRATION BY CLAMS.

Castration of bulls by means of clams has been practised in many

different forms.
Castration by the Exposed Method.—The operation is the
same as in the horse, the scrotum being incised on either side, and
the dartos, connective tissue, tunica vaginalis scroti, and tunica
vaginalis testis being divided. Short clams are applied to the cord,
and the lumen of the arteries is completely obliterated in five to six
days, when the clams can be removed.
Instead of an incision being made for the removal of each testicle,
the scrotum and dartos may be divided in the middle line, after
which incisions may be made to the right and left respectively,
exposing the fibrous tissue and enabling the testicles to be