Microbiological Methods for Environment, Food and Pharmaceutical Analysis

515 Pages·2020·13.125 MB·English

by  Abhishek Chauhan, Tanu Jindal

Abhishek Chauhan
Tanu Jindal
Microbiological
Methods
for Environment,
Food and
Pharmaceutical
Analysis
Microbiological Methods for Environment, Food
and Pharmaceutical Analysis
Abhishek Chauhan • Tanu Jindal
Microbiological Methods for
Environment, Food and
Pharmaceutical Analysis
Abhishek Chauhan Tanu Jindal
Amity Institute of Environmental Amity Institute of Environmental
Toxicology, Safety and Management Toxicology, Safety and Management
Amity University Amity University
Noida, Uttar Pradesh, India Noida, Uttar Pradesh, India
ISBN 978-3-030-52023-6 ISBN 978-3-030-52024-3 (eBook)
https://doi.org/10.1007/978-3-030-52024-3
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Preface
Discovery of microorganisms and the field microbiology began with the invention
of the first microscope in the year 1665, and since then, exponential growth of this
science can be seen. In fact, the current world largely depends upon the various
aspects of microbes, organisms that are invisible to the naked eye. These tiny organ-
isms demonstrate their presence in both positive and negative facets of life.
Microorganisms including bacteria, yeast, molds, etc., are ubiquitous in nature and
their applications are as well omnipresent throughout. Right from the air, soil, and
water, that is, three main constituents of the Earth, the applicability of microbes can
be found in any means. This science is gaining importance globally as far as the
safety and quality of various commodities such as food, water and wastewater, air,
cosmetics, drugs and pharmaceuticals, and antimicrobial preparation such as paints,
emulsions, herbal preparations, and medical devices are concerned. Safety and
quality evaluation of these commodities is a foremost requirement in order to pro-
vide the best services to mankind and for the betterment of society.
Microbes are significant to all our lives in enormous ways. Sometimes, the influ-
ence of microorganisms on human life is beneficial, whereas at other times, it is
detrimental. For example, microorganisms are required to produce cheese, yogurt,
bread, alcohol, antibiotics, vaccines, vitamins, enzymes wine, beer, etc. Several
microbial products contribute to public health such as aiding in nutrition, interrupt-
ing the spread of disease, and improving the quality of life in the years ahead.
Undesirable presence of microorganisms in various commodities such as food,
water, drugs, and pharmaceutical products makes it essential to ensure the safety
and quality of these products, which is usually achieved by quality evaluation or
analysis by carrying out microbiological experiments.
The textbook Microbiological Methods for Environment, Food and
Pharmaceutical Analysis specifically aims at the ever-demanding thoughtful need
of a well-documented compilation of factual details. The book emphasizes specifi-
cally on applied aspects of industrial microbiology including water, soil, air, food,
and pharmaceuticals. Good Microbiological Laboratory Practices (GMLP) cover-
ing all aspects of Good Laboratory Management System is well written in a techni-
cal manner so that if applied, it would aid in creating a state-of-the-art laboratory.
v
vi Preface
Special elaborations have been given to management with respect to safety, house-
keeping, spillage, disposal, and avoidance of laboratory-acquired infections.
Importance of microbiological culture media and their composition has also been
given so as to inform the user about ingredients of the media having characteristic
behavior during experiment. Knowledge of sterilization and disinfection procedure
from the point of view safety is an utmost requirement in laboratory while working
with highly contaminated sample and handling pathogenic culture. A concise reve-
lation about major and minor instruments/equipment used in industrial microbiol-
ogy has been given. Microbiological methods have been written in simple, lucid,
and crisp language keeping in view the requirement of industries and research orga-
nizations working on quality evaluation of variety of products. A fairly up-to-date
“Index” in the textbook will surely enlarge the vision of its readers by giving an easy
access of subject-enriched, well-documented text.
The book is highly useful for industrialists, researchers, academicians, and stu-
dents from both undergraduate and postgraduate levels of microbiology, biotechnol-
ogy, botany, and pharmaceutical sciences. It would have significant contribution in
the effective knowledge of scientists/analysts/lab personnel/technical managers/
quality managers working with the industrial aspect of microbiology. The authors
are grateful that Springer Nature has enthusiastically agreed to publish this work
and appreciates their help and collaboration on this project.
Noida, India Abhishek Chauhan
Tanu Jindal
About the Book
The current book has been intended with various microbiological methods pertain-
ing to water, soil, air, food, and pharmaceutical analysis with respect to quality and
safety evaluation of products. As per the requirements of different sectors, this text-
book has been divided into several chapters of current microbiological research
interest. Chapters 1 and 2 introduce a detailed knowledge of good microbiological
and management practices, Chap. 3 is designed to provide the essential information
pertaining to various types of microbiological culture media and their types and role
of ingredients. Chapter 4 describes various methods of sterilization and disinfec-
tion. Microbiological experiments required several major and minor equipment;
therefore, Chap. 5 has been designed in order to provide the necessary information
with respect to all required instrumentations and equipment. Chapter 6 describes
methods of sampling for water, soil, air, food, and pharmaceutical products. The
Chaps. 7, 8, and 9 demonstrate the standard laboratory procedures for carrying out
the analysis of water, soil, air, food, and pharmaceuticals. Methods written in this
book are significantly sub-headed to make it more user-friendly with better accu-
racy for evaluation of the products for betterment of mankind and society. Chapter 10
describes various methods relating to biochemical and molecular characteristics of
microorganisms in order to identify the microbe effectively. The script is also
equipped with relevant tables, figures, and photographs. The textbook specifically
aims at the ever-demanding thoughtful need of a well-documented compilation of
factual details. The book is written in simple, lucid, and crisp language keeping in
view the requirement of industries and research organizations working on quality
evaluation of a variety of products.
Abhishek Chauhan
Tanu Jindal
vii
Contents
1 Introductory Analytical Microbiology . . . . . . . . . . . . . . . . . . . . . . . . . 1
1.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
1.2 Analytical Microbiology: Various Disciplines . . . . . . . . . . . . . . . . . 2
1.3 Food Microbiology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
1.3.1 Microbiological Analysis of Food and Food Products. . . . . 3
1.3.2 Microbial Contamination of Food and Food Products:
Major Factors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
1.4 Environmental Microbiology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
1.4.1 Water Microbiology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
1.4.2 Air Microbiology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
1.4.3 Soil Microbiology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
1.5 Pharmaceutical Microbiology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
1.6 Accreditation for Microbiological Laboratories . . . . . . . . . . . . . . . 11
1.7 Quality Assessment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
1.7.1 Internal Quality Assessment . . . . . . . . . . . . . . . . . . . . . . . . 11
1.7.2 External Quality Assessment . . . . . . . . . . . . . . . . . . . . . . . . 12
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
2 Good Microbiological Laboratory Practices . . . . . . . . . . . . . . . . . . . . 15
2.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
2.2 Safety Management . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
2.2.1 Biosafety Level (BSL) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
2.2.2 Avoidance of Laboratory-Acquired Infections . . . . . . . . . . 16
2.3 Entry to Microbiology Lab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
2.4 Housekeeping Management . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
2.5 Spillage Management . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
2.6 Disposal Management . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
2.6.1 Containers for Waste Material in Microbiology Lab . . . . . . 20
2.6.2 Discard Bins, Bags and Jars . . . . . . . . . . . . . . . . . . . . . . . . . 20
2.6.3 Good Disposal Practices (GDP). . . . . . . . . . . . . . . . . . . . . . 21
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
ix
x Contents
3 Microbiological Culture Media: Types, Role and Composition . . . . . 23
3.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
3.2 Types of Media. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
3.3 Consistency-Based Classification of Media . . . . . . . . . . . . . . . . . . . 24
3.3.1 Liquid (Broth) Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
3.3.2 Semisolid Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
3.3.3 Solid Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
3.4 Composition-Based Classification of Media . . . . . . . . . . . . . . . . . . 25
3.4.1 Chemically Defined Media (Synthetic Media) . . . . . . . . . . 25
3.4.2 Complex Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
3.5 Function-Based Classification of Media . . . . . . . . . . . . . . . . . . . . . 26
3.5.1 All Purpose Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
3.5.2 Selective/Differential Media . . . . . . . . . . . . . . . . . . . . . . . . 26
3.5.3 Enrichment Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
3.5.4 Reducing Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
3.6 Culture Media: Check Points . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
3.7 Culture Media: Role of Ingredients (Table 33.1.1) . . . . . . . . . . . . . 28
3.8 Culture Media: Compositions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
3.8.1 Actinomycetes Isolation Agar . . . . . . . . . . . . . . . . . . . . . . . 30
3.8.2 Alkaline Peptone Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
3.8.3 Ammonifying Bacteria Isolation Medium . . . . . . . . . . . . . . 31
3.8.4 Anaerobic Agar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
3.8.5 Antibiotic Assay Medium No-1 . . . . . . . . . . . . . . . . . . . . . . 31
3.8.6 Antibiotic Assay Medium No-12 . . . . . . . . . . . . . . . . . . . . . 32
3.8.7 Antibiotic Assay Medium No-19 . . . . . . . . . . . . . . . . . . . . . 32
3.8.8 Antibiotic Assay Medium No-11 . . . . . . . . . . . . . . . . . . . . . 33
3.8.9 Baird–Parker Agar Medium . . . . . . . . . . . . . . . . . . . . . . . . . 33
3.8.10 BG-11 Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
3.8.11 Biotin Assay Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
3.8.12 Bismuth Sulphite Agar (BSA) . . . . . . . . . . . . . . . . . . . . . . . 35
3.8.13 Brain–Heart Infusion Broth . . . . . . . . . . . . . . . . . . . . . . . . . 35
3.8.14 Blood Agar Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
3.8.15 Baird Parker Agar Medium . . . . . . . . . . . . . . . . . . . . . . . . . 36
3.8.16 Brilliant Green Agar (BGA). . . . . . . . . . . . . . . . . . . . . . . . . 36
3.8.17 Brilliant Green Bile Lactose (BGBL) Broth . . . . . . . . . . . . 37
3.8.18 Buffered Peptone Water (BPW) . . . . . . . . . . . . . . . . . . . . . . 37
3.8.19 Buffered Sodium Chloride Peptone Solution . . . . . . . . . . . 38
3.8.20 Cetrimide Agar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
3.8.21 Chloramphenicol Yeast Extract Glucose Agar (CYGA) . . . 39
3.8.22 Columbia Agar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
3.8.23 Cook Meat Medium (CMM) . . . . . . . . . . . . . . . . . . . . . . . . 40
3.8.24 Deoxycholate Citrate Agar (DCA). . . . . . . . . . . . . . . . . . . . 40
3.8.25 Differential Reinforced Clostridia Broth . . . . . . . . . . . . . . . 40
3.8.26 Enterobacteria Enrichment Broth . . . . . . . . . . . . . . . . . . . . 41
3.8.27 Eosin Methylene Blue (EMB) Agar . . . . . . . . . . . . . . . . . . . 41
Contents xi
3.8.28 Ethyl Violet Azide Dextrose Agar . . . . . . . . . . . . . . . . . . . . 42
3.8.29 Fluid Thioglycollate Medium (FTM) . . . . . . . . . . . . . . . . . 42
3.8.30 Glucose Agar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
3.8.31 Glucose-Salt-Teepol Broth . . . . . . . . . . . . . . . . . . . . . . . . . . 43
3.8.32 GN Broth . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
3.8.33 Glucose Yeast Extract Agar . . . . . . . . . . . . . . . . . . . . . . . . . 44
3.8.34 Hugh-Leifson Medium. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
3.8.35 K-Agar Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
3.8.36 KG (Kim Goepfert) Agar Base . . . . . . . . . . . . . . . . . . . . . . 45
3.8.37 Lactose Broth . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
3.8.38 MacConkey Broth . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
3.8.39 Mac Conkey Agar (MCA) . . . . . . . . . . . . . . . . . . . . . . . . . . 47
3.8.40 Malt Extract Broth . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
3.8.41 Mannitol Salt Agar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
3.8.42 Mannitol Yeast Polymyxin Agar (MYP Agar) . . . . . . . . . . . 48
3.8.43 Minimal Agar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
3.8.44 Motility Test Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
3.8.45 MR-VP Medium (Glucose Phosphate Broth) . . . . . . . . . . . 49
3.8.46 Mueller Hinton Broth (MHB) . . . . . . . . . . . . . . . . . . . . . . . 50
3.8.47 Muller Hinton Agar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
3.8.48 Nitrate Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
3.8.49 Nutrient Agar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
3.8.50 Nutrient Broth . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
3.8.51 Osmophilic Agar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
3.8.52 Osmophilic Dilution Blanks . . . . . . . . . . . . . . . . . . . . . . . . 52
3.8.53 Pantothenate Assay Medium . . . . . . . . . . . . . . . . . . . . . . . . 52
3.8.54 Pantothenate Inoculum Broth . . . . . . . . . . . . . . . . . . . . . . . 53
3.8.55 Peptone Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
3.8.56 Plate Count Agar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
3.8.57 Rappaport Vassiliadis Medium (RV) . . . . . . . . . . . . . . . . . . 54
3.8.58 Sabouraud Dextrose Agar . . . . . . . . . . . . . . . . . . . . . . . . . . 55
3.8.59 Sabouraud Dextrose Broth . . . . . . . . . . . . . . . . . . . . . . . . . . 55
3.8.60 Selenite Cystine Broth (SCB) . . . . . . . . . . . . . . . . . . . . . . . 55
3.8.61 Simmon’s Citrate Agar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
3.8.62 Skim Milk Agar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
3.8.63 Soyabean Casein Digest Broth . . . . . . . . . . . . . . . . . . . . . . . 57
3.8.64 Soybean Casein Digest Agar . . . . . . . . . . . . . . . . . . . . . . . . 57
3.8.65 Starch Agar. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
3.8.66 TC-SMAC Agar Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
3.8.67 Tergitol – 7 Agar (T-7) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
3.8.68 Tetrathioanate Broth (TTB) . . . . . . . . . . . . . . . . . . . . . . . . . 59
3.8.69 Thiosulphate-Citrate-Bile Salts-Sucrose Agar (TCBS) . . . . 59
3.8.70 Top Agar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
3.8.71 Triple Sugar-Iron Agar Medium . . . . . . . . . . . . . . . . . . . . . 60
3.8.72 Urease Broth Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60