ignou MEV-004

THE PEOPLE'S
UNIVERSITY Environmental
Indira Gandhi National Open University
School of Interdisciplinary and Toxicology
Trans-disciplinary Studies

INTRODUCTION TO TOXICOLOGY 1
 MEV-004
Environmental Toxicology
Indira Gandhi National Open University
School of Interdisciplinary and
Trans-disciplinary Studies

Block

1
INTRODUCTION TO TOXICOLOGY
UNIT 1
Introduction to Toxicants 5
UNIT 2
Chemical Toxicants 20
UNIT 3
Biological Toxicants 38
UNIT 4
Toxicity Assessment 58

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 PROGRAMME DESIGN AND EXPERT COMMITTEE
Dr. (Ms.) Shyamala Mani Dr. Rachna Agarwal Dr. Sushmitha Baskar Dr. Deeksha Dave
Professor, National Institute of Urban School of Vocational Education and Environmental Studies Environmental Studies, School
Affairs (NIUA) India Habitat Centre Training, Indira Gandhi National School of Interdisciplinary and of Interdisciplinary and Trans-
New Delhi Open University, Trans-disciplinary Studies disciplinary Studies,
New Delhi Indira Gandhi National Open Indira Gandhi National Open
Prof. R. Baskar University, New Delhi University, New Delhi
Department of Environmental Science Prof. Daizy R Batish
& Engineering, Guru Jambheshwar Department of Botany, Panjab Prof. Ruchika Kuba Dr. Shubhangi Vaidya
University of University, Chandigarh School of Health Sciences, Indira School of Interdisciplinary and Trans-
Science & Technology, Hisar Haryana Gandhi National Open University, disciplinary Studies, Indira Gandhi
Prof. M. Krishnan New Delhi National Open University
Prof. H.J. Shiva Prasad Vice Chancellor, Madurai Kamraj New Delhi
Professor of Civil Engineering University, Madurai, Tamil Nadu Prof. Nandini Sinha Kapur
College of Technology, G.B. Pant School of Interdisciplinary and Dr. Y.S.C. Khuman
University of Agriculture & Technology Dr. Chirashree Ghosh Trans-disciplinary Studies, School of Interdisciplinary and
Pant Nagar, Uttarakhand Department of Environmental Indira Gandhi National Open Trans-disciplinary Studies, Indira
Studies, University of Delhi, University, New Delhi Gandhi National Open University
Dr. T.K. Joshi New Delhi New Delhi
Director, Occupational & Dr. Shachi Shah
Environmental Programme, Centre Mr. Ravi Agarwal Environmental Studies,
Director, Toxic Link, Jangpura Dr. Sadananda Sahoo
for Occupational & Environmental School of Interdisciplinary and School of Interdisciplinary and
Health, Maulana Azad Medical Extension, New Delhi Trans-disciplinary Studies Trans-disciplinary Studies, Indira
College, New Delhi Prof. Jaswant Sokhi Indira Gandhi National Open Gandhi National Open University
School of Sciences, Indira Gandhi University, New Delhi New Delhi
Prof. Nilima Srivastava
School of Gender and Development National Open University, Dr. V. Venkat Ramanan
Studies, Indira Gandhi National Open New Delhi Environmental Studies
University, New Delhi Dr. B. Rupini School of Interdisciplinary and
Environmental Studies, School Trans-disciplinary Studies
Prof. S.K. Yadav Indira Gandhi National Open
School of Agriculture of Interdisciplinary and Trans-
disciplinary Studies, Indira Gandhi University, New Delhi
Indira Gandhi National Open
University, New Delhi National Open University,
New Delhi

BLOCK PREPARATION TEAM

Unit 1 Unit 2 Unit 3 Unit 4
Dr. Sushmitha Baskar, Dr. B. Rupini Prof. Rajesh Dhankhar Dr. Shachi Shah
Environmental Studies, SOITS, Environmental Studies, Department of Environmental Environmental Studies, SOITS,
IGNOU, New Delhi SOITS, IGNOU, Science, Maharshi Dayanand IGNOU, New Delhi
New Delhi University, Rohtak, Haryana

PROGRAMME COORDINATORS
Dr. B. Rupini Dr. Sushmitha Baskar Prof. Ruchika Kuba
Environmental Studies, School of Interdisciplinary Environmental Studies, School of Interdisciplinary School of Health Sciences, Indira
and Trans-disciplinary Studies, Indira Gandhi and Trans-disciplinary Studies, Indira Gandhi Gandhi National Open University,
National Open University, New Delhi National Open University, New Delhi New Delhi

COURSE COORDINATOR CONTENT EDITORS

Dr. Sushmitha Baskar, Prof. M.V. Usha Rani, Retd Professor, Department of Environmental Sciences, Bharathiar
Environmental Studies, University, Coimbatore, Tamil Nadu.
SOITS, IGNOU, New Delhi Dr. B. Rupini, Environmental Studies, SOITS, IGNOU, New Delhi
Dr. Sushmitha Baskar, Environmental Studies, SOITS, IGNOU, New Delhi
Prof. Ruchika Kuba, School of Health Science, IGNOU, New delhi

FORMAT EDITORS
Dr. B. Rupini Dr. Sushmitha Baskar
Environmental Studies, School of Interdisciplinary and Environmental Studies, School of Interdisciplinary and Trans-
Trans-disciplinary Studies, Indira Gandhi National Open disciplinary Studies, Indira Gandhi National Open University,
University, New Delhi New Delhi

Secretarial/Technical Assistance: Ms. Sonali, SOITS, IGNOU, New Delhi; Mr. Vikram, SOITS, IGNOU, New Delhi

PRINT PRODUCTION
Mr. S. Burman Mr. Sudhir
Mr. Y. N. Sharma
Deputy Registrar (P), IGNOU, New Delhi Asst. Registrar (P), IGNOU, New Delhi Section Officer (P) IGNOU, New Delhi

February, 2019
 Indira Gandhi National Open University, 2019
ISBN: 987-88-88498-87-6
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at www.ignou.ac.in
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COURSE INTRODUCTION
Welcome to the Post Graduate Diploma in Environmental and Occupational
Health (PGDEOH) of IGNOU. The curriculum prepared for this programme is
relevant and significant in the present day scenario. This programme is in
consensus with the mission of Environmental and Occupational Health which is
to prevent adverse health effects related to chemical and biological contaminants
in the Environmental and Occupational exposures through education, research
and service.

This core course is entitled “Environmental Toxicology”. This course consists

of 4 blocks comprising of 15 Units.

Block 1 deals with the introduction to the field of toxicology. The block deals
with the basic concepts of toxicology, types of toxicants and their effects on
human health. The importance of toxicity assessment and types of tests for
assessment is also described in detail. The block also describes how toxic
substances can enter the human body and cause deleterious effects. The different
types of toxicants namely chemical, biological toxicants and their effects have
also been described. Finally the block explains the toxicity assessment and the
types of tests that can be used for assessment of toxicity.

Block 2 deals with the basics of eco-toxicology and concepts related to

immunotoxicology, neurotoxicity and reproductive toxicity. The block also
describes toxic biotransformations and the various effects of toxicants on the
environment and human health. The concepts of detoxification and mechanisms
have been described with suitable examples.

Block 3 deals with environmental toxicity risk assessment. The block vividly
describes the acceptable limits of toxicants and how estimating health risks are
most essential. Finally the block focuses on toxic remediation of the air, water
and soil environments with suitable case studies.

Block 4 deals with environmental cytotoxicity and genotoxicity. The terms such
as mutagenecity, carcinogenecity and teratogenecity has been explained with the
mechanisms of action. The block summarizes with the prevention of cancers
and mutagens and explains the screening methods for the same.

All these Blocks will provide you with sufficient knowledge about the toxicants,
their mode of action and how toxicity risk assessments can be done.
Introduction to Toxicology
INTRODUCTION TO BLOCK 1
This block focuses on the concepts of toxicology, various toxicants found in the
environment and their effects on the environment and also on human health. The
quality of our environment is degrading and we are exposed to a variety of
pollutants in everyday life. These pollutants can have acute, chronic, sub-acute
and toxic effects on the living organisms as well as human beings. Even the
substances such as paints, detergents, cleaning solutions, cosmetics that we use
can have hazardous chemicals giving rise to toxic effects on our health. The
different types of toxicants namely chemical, biological toxicants and their effects
have also been described. The block finally describes the different tests used in
toxicity assessment.

Unit 1 deals with the definition of toxicants, the various sources of toxicants and
the mode of entry of toxic substances along with their action. The routes of
absorption of toxic substances such as skin, lung, gastrointestinal tracts; and the
distribution and storage of toxins in human tissues such as the plasma, kidney,
fat, bone, blood and the placenta is also described in detail.

Unit 2 deals with the different types of chemical toxicants and their effects on
the environment and human health. Chemical toxicants are released into the
environment in different ways, and they can be transported through several
pathways. The unit discusses that the chemical toxicants released into the
atmosphere exert adverse effects on humans and other terrestrial and aquatic
organisms through ingestion, occupational exposure, environmental exposure,
as well as accidental and intentional poisoning.

Unit 3 deals with biological toxicants and their effects of human health and
environment. Biological toxicants are chemical substances which are toxins
produced by living organisms. The unit details on food intoxication from members
in the animal and plant kingdom and how we can protect ourselves from these
pathogens.

Unit 4 deals with the toxicity assessment and the types and classification of
toxicity tests including exposure assessments. In addition to dose, other factors
may also influence the toxicity of the compound such as the route of entry, duration
and frequency of exposure, variations between different species and variations
among members of the same species. The unit finally discusses the potential
hazards to humans as acute, subchronic and chronic toxicity.

4
 Introduction to Toxicants
UNIT 1 INTRODUCTION TO TOXICANTS
Structure
1.0 Introduction
1.1 Objectives
1.2 Definition and Concepts
1.3 Sources of Toxicants
1.4 Mode of Action of Toxic Substances
1.5 Exposure Routes
1.6 Distribution and Storage of Toxins in Human Tissues
1.7 Let Us Sum Up
1.8 Key Words
1.9 References and Suggested Further Readings
1.10 Answers to Check Your Progress

1.0 INTRODUCTION
We are exposed to different substances in our everyday life and some of these
may be toxic to our health. The quality of our environment is also decreasing
and people are exposed to a variety of pollutants. These pollutants can have
acute, chronic, sub-acute and toxic effects on the living organisms as well as
human beings. Substances that we use in daily life starting from paints, detergents,
cleaning solutions, cosmetics etc. can have hazardous chemicals which can have
toxic effects on our health. Rachel Carson is considered the mother of
environmental toxicology. She published the book Silent Spring in 1962 which
discussed the toxic effects of the pesticide DDT. Living organisms can be exposed
to toxic substances at any stage of their life cycle. They can be accumulated in
the fatty tissues and lead to bioaccumulation. This can lead to biomagnifications
of specific toxicants. In this unit let us learn about toxins, toxicants, their types,
sources and the mechanisms of their action.

1.1 OBJECTIVE
After reading this unit, you should be able to:
 define toxins and toxicants;
 understand the different concepts and terminologies used in environmental
toxicology;
 explain the different sources of toxicants;
 describe the routes of exposure to toxicants;
 understand the mechanisms and site of toxic action by toxicants; and
 explain how toxins are stored in the various tissues of the human body.

5
Introduction to Toxicology
1.2 DEFINITIONS AND CONCEPTS
1.2.1 Definitions
Let us now learn about some definitions and terms commonly used in
environmental toxicology.
a) Environmental toxicology: It is a branch of science that deals with the
harmful effects of different physical, chemical and biological agents on
living organisms. It is multidisciplinary in nature.
b) Ecotoxicology: This is a sub-discipline of environmental toxicology. This
deals with the harmful effects of toxicants at ecosystem and population
levels.
c) Toxicant: Any toxic material or substance is termed as a toxicant. They are
hazardous and poisonous. Toxicants are generally man-made and artificial
products introduced into the environment due to human activity. They
include bisphenol, insecticides and a number of industrial chemicals.
d) Toxins: These are produced naturally by living organisms. For example,
toxins from the mushroom plant and toxin from the venom of snake are
natural toxins.
e) Poisons: They are toxicants that cause death or illness in very small doses.
f) Toxicologists are scientists who deal with the study of toxicants and toxins.
g) Xenobiotic is referred to a foreign substance entering the body. It is derived
from the Greek word ‘xeno’ meaning ‘foreigner’.
h) Toxicosis/ Poisoning/ Intoxication: Any disease produced by a toxicant.
i) Tolerance: The ability of an organism to show less response to a specific
dose of a chemical than it demonstrated on a previous exposure; refers to
acquired and not innate resistance.
j) LD 50: The dose that is lethal to 50% of a test sample or population.
Expression of toxicant concentrations is in ppb or ppm in feedstuff, water,
air, tissue etc. Other expressions of dose are maximum nontoxic dose,
maximum tolerated dose, approximate lethal dose.

1.2.2 Basic Concepts of Toxicology

Both toxins and toxicants are referred to as toxic substances. Toxic substances
can be classified as systemic toxins and organ toxins. Systemic toxins have effects
on the entire body. An example of systemic toxin include: potassium cyanide. It
affects each cell and organ of the body causing complete damage. Some toxicants
affect specific tissues or organs. They are known as organ toxins. They do not
cause damage to the entire body. An example of organ toxins include: (1) benzene
which targets blood-forming tissues and (2) Lead which targets the central nervous
system, kidney, and the hematopoietic system.

1.2.3 Historical Development of Toxicology

The historical development of toxicology started very early in human civilizations.
Our ancestors who were cave dwellers found plants and animals that had toxins
6 and these toxic extracts were used in hunting and warfare. The earliest medical
works of Hippocrates, Aristotle, and Theophrastus published during 400 to 250 Introduction to Toxicants
BC include the mention of poisons. Scriptures and records indicate that by 1500
BC hemlock, opium, arrow poisons, and certain metals were also used to poison
enemies or for state executions. Some of the notable poisoning victims include:
Socrates, Cleopatra, and Claudius. After Renaissance and Age of Enlightenment
some basic concepts fundamental to toxicology started to evolve. In toxicology
the study of Paracelsus (~1500AD) and Orfila (~1800 AD) are of importance.
Advances in toxicology took place during the time of Galen (AD 131–200) and
Paracelsus (1493–1541). Paracelsus in his study showed that specific chemicals
were responsible for the toxicity of a plant or animal poison. He also reported
that the dose of a chemical is an important factor for the human body’s response.
This is now studies as the dose-response relationship. It is an important concept
of toxicology. Hence, Paracelsus was one of the founders of modern toxicology.
His well known quote says that “All substances are poisons; it is the dose that
makes the poison”. Aspirin which is acetylsalicylic acid is a drug consumed by
people for medical ailments all over the world. It is relatively safe at recommended
doses but chronic use causes deleterious effects on the gastric mucosa, and it is
fatal at a dose of about 0.2 to 0.5 g/kg. In the 18th century ‘Ramazini’s Diseases
of Workers’ was published in 1700. He is known as the father of occupational
medicine. The incidence of scrotal cancer among those working in chimney
sweeps was observed by Percival Pott in 1775. Also Hill in 1761 said that nasal
cancer and snuff use is correlated. The founder of modern toxicology was Orfila,
a Spanish physician who worked at the University of Paris in the early 19th century.
He brought out the systematic correlation between the chemical and biological
properties of poisons of the time. He analyzed materials for poisons and showed
the effects of poisons on specific organs. He was responsible for identifying
toxicology as a separate science and he published the first book on toxicology in
1815. In the 20th Century descriptions on the DNA were given and this lead to an
advancement of knowledge on toxic effects of substances at the cellular and
DNA level. The Silent Spring published by Rachel Carson in 1962 was a landmark
publication influencing modern environmental toxicology. Her book served as a
catalyst for the establishment of the US Environmental Protection Agency and
she is regarded as the mother of the environmental movement. Recently with the
advent of molecular techniques in biology detailed studies have been done in
toxicology and xenobiotic metabolism.

1.3 SOURCES OF TOXICANTS

Toxic agents can be chemical, physical, or biological in nature and produce toxic
effects on the body. The different toxic agents include: chemical (cyanide),
physical (radiation) and biological (snake venom). There exist a number of
toxicants and they can be classified by various means. Classification may be by
exposure classes and by user classes. The different toxicants include the following:

1) Natural Pollutants: Toxic pollutants can also be released through natural

processes. For example, volcanoes emit particulate matter, sulfur dioxide,
hydrogen sulfide, and methane. Forest fires release smoke, unburned
hydrocarbons, carbon monoxide, nitrogen oxides, and ash. These can be
harmful to human health when inhaled. Dust storms release particulate
matter and oceans release aerosols in the form of salt particles. Plants
produce pollen and spores, which cause respiratory problems and allergic
7
Introduction to Toxicology reactions.

2) Anthropogenic Pollutants. These are pollutants introduced due to human

activity/ man-made activities. These substances come primarily from three
sources: (1) combustion; (2) industrial; (3) mining and drilling processes.

Let us now learn about some important man-made sources of toxicants in detail.
They include the following.

1) Air pollutants: Humans have been polluting the air and there are also
significant natural pollutants such as terpenes from plants, smoke from
forest fires, and fumes and smoke from volcanoes. Among air pollutants
there are gaseous pollutants like carbon dioxide, carbon monoxide,
hydrocarbons, hydrogen sulfide, nitrogen oxides, ozone and other oxidants,
sulfur oxides. There are also fine particulates in the air. The particulates
include dust (coal, ash, sawdust, cement), fumes <1 ìm in diameter that
come from chemical processes, mist droplets, smoke (0.05–1.0 ìm) resulting
from incomplete combustion of fossil fuels and aerosols.

2) Indoor Pollutants: These are produced from heating, cooking, pesticides,

tobacco smoking, radon, gases, microbes from people and animals. Materials
used for construction of buildings can give out gaseous indoor chemicals
that have serious health concerns. Carbon monoxide and polycyclic aromatic
hydrocarbons released from wood, crop residues, animal dung used for
cooking cause acute respiratory infections in poorly ventilated areas.

3) Water pollutants: Surface waters may be polluted from point and nonpoint
sources. Industrial effluents discharged into waters are an example of point
source. Fertilizer and pesticide application in agricultural fields that enter
surface waters through rainfall are an example on nonpoint source of
pollution. Industrial wastes discharged into waters contain organic and
inorganic wastes including hazardous chemicals. Toxic effects are seen in
humans when they consume this contaminated water.

4) Soil pollutants: When wastes are not properly disposed off then soil also
gets polluted. Soil contaminants include: domestic waste, solid wastes,
electronic wastes, municipal wastes, agricultural wastes that contain a
number of chemicals harmful to life. Further agricultural toxicants like
persistent pesticides that do not biodegrade remain in the soil of many years
and move into the food chain causing greater health impacts. The most
toxic hazardous pesticides are the organochlorine compounds such as DDT,
aldrin, dieldrin, and chlordane.

5) Heavy metals: Metals released from industrial activities cause toxicity. For
example, the heavy metals lead and arsenic are highly toxic and is found in
potable water in certain areas. Lead can enter water from lead pipes, lead
solder, lead toys, leaded gasoline, utensils and also paints containing lead.
Lead is used in cosmetics like lipsticks can regular usage can enter the
system and cause health effects. Lead induces neurological damage and
can penetrate the placental barrier and induce birth defects among children.
Arsenic toxicity is also a serious cause of concern especially in West Bengal
in India. It can leach into water from pesticide sprays, arsenic-containing
fossil fuels, and leaching of mine tailings and smelter runoff. Chronic high-
8
 level exposures can cause abnormal skin pigmentation, hyperkeratosis, nasal Introduction to Toxicants
congestion, abdominal pain and cancers. Cadmium enters the food chain
through industrial activities. It can accumulate in the tissues of aquatic
organisms. Cadmium contaminated rice in Japan caused the disease Itai-
Itai. The disease was characterized by severe kidney damage, painful bone
and joint problems. Mercury from industries manufacturing plastics, vinyl
chloride is also highly toxic to living beings. The Minamata disease in
Japan occurred due to consumption of mercury contaminated fishes resulted
in neurological disorders, paralysis, and mental disorientation.

6) Nitrates and phosphate: These arise from contamination due to fertilizers,

discharge from sewage treatment plants, leachate from septic tanks, manure
and detergents which are hazardous. They leach into the soil and drinking
water. Nitrates in drinking water cause adverse health effects. It occurs due
to the formation of: (1) nitrosamine and (2) methemoglobinemia. The
nitrates are converted to nitrites by bacteria in the intestine. Thereafter nitrite
ions combine with hemoglobin to form methemoglobin. This reduces the
oxygen-carrying capacity of the blood and leads to the blue-baby syndrome.
This is seen in young or new born children who have ingested nitrate
containing water or milk foods.

7) Petroleum and oil pollutants: Shore animals, such as crabs, shrimp, mussels,
and barnacles, are also affected by the toxic hydrocarbons (oil and petroleum
compounds) ingested by them.

8) Volatile organic compounds (VOCs): They include halogenated solvents

and petroleum products. They are used in industries involving degreasing,
dry cleaning, paint, and in the military. The most important VOCs include:
trichloroethylene, toluene, benzene, chloroform, tetrachloroethylene, 1,1,1-
trichloroethane, ethylbenzene, trans-1,2-dichloroethane, xylene,
dichloromethane, and vinyl chloride. They can move quickly in
groundwaters. Exposure to high levels can result in headache, impaired
cognition, kidney toxicity, cancer and reproductive disorders.

9) PCB organic compounds: These include polychlorinated biphenyls (PCBs),

phenols, cyanides, plasticizers, solvents, and numerous industrial chemicals.
They are used as coolants in electrical transformers. PCB’s are stable,
lipophilic, and are broken down slowly only in tissues. Due to these
properties they accumulate to high concentrations in fish and waterfowl.

10) Solvents: Many types of solvents are used which have systemic toxic effects
on nervous system and the blood. Benzene is used in the rubber, canning,
printing, shoe manufacturing industries. Benzene affects the hematopoietic
tissue in the bone marrow and is an immunosuppressant. Benzene exposure
results in decrease of white blood cells, red blood cells and platelets.
Continued exposure causes severe bone marrow damage, aplastic anemia
and leukemia. Other toxic solvents include: aliphatic hydrocarbons,
halogenated aliphatic hydrocarbons, aliphatic alcohols, glycols, glycol ethers
and aromatic hydrocarbons.

11) Asbestos: This material is used in insulation, roofing for houses, asbestos
cements, brake linings, electrical appliances and coating materials. The
inhalation of asbestos causes ‘Asbestosis’, a respiratory disease characterized 9
Introduction to Toxicology by scarring of lungs, fibrosis, calcification, and also leads to lung cancer.

12) Therapeutic drugs: Generally all therapeutic drugs are toxic and produce
hazardous effects at some dose. This depends on many factors like: dose,
nature of the drug, individual (genetic) variation, diet, age, etc. The side
effect of chloroquinol, an antidiarrhea drug used in Japan in 1960 caused
stiffness of the joints accompanied by damage to the optic nerve. Birth
defects or teratogenesis can be caused by drugs particularly, thalidomide.
Dermatitis is a common side effect of topically applied corticosteroids.
Toxic effects on the blood are caused by chlorpromazine. Hemolytic anemia
is caused by methyldopa and megaloblastic anemia is induced by
methotrexate. Eye toxicity and glaucoma is caused by thioridazine and
systemic corticosteroids. There are many more drugs that cause toxicity to
organs.

13) Drugs of abuse: They include depressants of central nervous system

(ethanol, methaqualone, secobarbital), stimulants of central nervous system
(cocaine, methamphetamine, caffeine, nicotine; opioids - heroin, mependine,
hallucinogens - lysergic acid diethylamide (LSD), phencyclidine (PCP),
and tetrahydrocannabinol, the most active ingredient of marijuana. More
important is that these drugs are synthesized illegally and are contaminated
with compounds of unknown origin and highly toxic and hazardous to
health.

14) Biological toxicants: Some naturally occurring substances that cause toxicity
include plant, animal, algal, fungal and microbial toxins. They include many
phytotoxins and mycotoxins. It is pertinent to understand that a toxin is a
toxicant produced by a living organism and is not used as a synonym for
toxicant. All toxins are toxicants, but all toxicants are not considered as
toxins. Toxins are the metabolic products used for defense against pathogens.
These natural products are used in beneficial pharmaceutical purposes and
also in biological warfare. For example, Aflatoxins are products of
Aspergillus flavus. It is fungus that contaminates grain, maize, peanuts,
and so on. Aflatoxin B1 is the most toxic and is reported to have carcinogenic
effects.

15) Cosmetics: Cosmetics induce allergies and contact dermatitis. Lipsticks

contain lead at varying concentrations. Hair dyes contain resorcinol which
is toxic. In ladies, some of the fibroids are related to regular use of hair
dyes. Thioglycolates and thioglycerol used in cold-wave lotion and
depilatories have toxic effects. Sodium hydroxide and formaldehyde used
in hair straighteners also are reported to exert toxicity. Further paraffin wax
is used in some lotions which hinders skin breathing. Parabens and sodium
laureth sulfates used in shampoo and body cleansers show toxicity. All
these cosmetics are toxic in nature affecting human health.

These were some of the sources of toxicants that have harmful effects on human
health. Now let us see the mode of action of some toxicants.
Check Your Progress 1
Note: a) Write your answer in about 50 words.
b) Check your progress with possible answers given at the end of the
10 unit.
 Introduction to Toxicants
1) Describe the various sources of toxicants.
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2) Write short notes on (a) therapeutic drugs and (b) drugs of abuse as toxicants.
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3) Define environmental toxicology. Write a brief account of the historical

development in toxicology.
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1.4 MODE OF ACTION OF TOXIC SUBSTANCES

1.4.1 Levels of Toxic Action
The mode of action of toxicants can be at the cellular and molecular level and
also at the organ level. It depends on the following: (1) uptake, (2) distribution,
(3) metabolism, (4) mode of action, and (5) excretion of the substance involved.
The different levels and mode of action of toxicants are: (a) Molecular toxicology:
This includes effects at biochemical and molecular levels. It involves the enzymes
that metabolize xenobiotics and the generation of reactive intermediates. It also
involves the interaction of xenobiotics with macromolecules and gene expression
in metabolic activities inducing toxic pathways. (b) Behavioral toxicology: This
involves toxic effects on the peripheral and central nervous systems and the
endocrine glands. (c) Nutritional toxicology: This involves the toxic effects of
diet. (d) Carcinogenesis: It includes the effects that lead to proliferation of cells
in an uncontrolled manner. (e) Teratogenesis: It involves chemical, biochemical,
and molecular events that lead to toxic effects during development of the foetus.
(f) Mutagenesis: It involves toxic effects that occur on the genetic material and
11
Introduction to Toxicology the inheritance of these effects causing mutagenecity. (g) Organ toxicity: This is
the toxic effects at the organ level. They are neurotoxicity - brain, hepatotoxicity
- liver, nephrotoxicity - kidney, reproductive toxicity depending on the organ
involved.

1.4.2 Sites of Toxic Action

Toxicants can have different mechanisms and sites for initiating their toxic effects.
Some of the mechanisms are explained below.

a) Enzyme Inhibition/Activation: One important site of toxic action for metals

is the interaction with enzymes. It results in enzyme inhibition or activation.
Two mechanisms are significant. Enzyme inhibition can occur due to
interaction between metal and sulfhydryl groups on the enzyme, or the metal
can also displace an essential metal cofactor of the enzyme. For example, in
the zinc-dependent enzyme ä-aminolevulinic acid dehydratase, lead can
displace zinc thereby inhibiting the synthesis of heme.

b) Subcellular Organelles: Toxic metals have the ability to disrupt the structure
and function of a number of subcellular organelles. The enzymes associated
with the endoplasmic reticulum may be inhibited and metals can be
accumulated in the lysosomes. Also respiratory enzymes in the mitochondria
can be inhibited and metal inclusion bodies are produced in the nucleus.

c) Carcinogenic effects: Some chromium compounds, arsenic, nickel, cadmium

cause carcinogenic effects. It may be due to the interaction of the metallic
ions with DNA.

d) Kidney: It is the main excretory organ of the body and a common target
organ for metal toxicity. Cadmium and mercury are potent nephrotoxicants.

e) Nervous System: The nervous system is also a common target of toxic organic
metal compounds. For example, methylmercury is lipid soluble. It can readily
go across the blood-brain barrier and enter the nervous system. Also organic
lead compounds are mainly neurotoxicants.

f) Endocrine and Reproductive System: The reproductive organs are under

neuroendocrine and hormonal control. Hence any toxicant that alters any of
these processes can affect the reproductive system. Certain metals can act
directly on these organs. Acute exposure to cadmium can cause testicular
injury. Toxic exposures to lead can cause testicular degeneration, inhibition
of spermatogenesis, and the disease Leydig-cell atrophy.

g) Respiratory System: The system can be affected by inhalation of metal dust

that can cause irritations and inflammation of the respiratory tract. Chronic
exposure can give rise to fibrosis (aluminum dust exposure) or carcinogenesis
(arsenic, chromium, nickel metal exposures).

1.5 EXPOSURE ROUTES

The common absorption routes include the: skin, lung and the gastrointestinal
tracts. The skin is the largest organ in the human body and is a physical barrier to
the absorption of toxic substances. The other major routes are the respiratory
12
and gastrointestinal tract. These give less resistance to toxicant absorption than Introduction to Toxicants
the skin. The respiratory tract is the most rapid route for entry and the skin is the
least rapid route for entry. This difference is due to thickness of the membranes.
Let us now see them in detail.

a) Skin: The skin is a complex multilayered tissue. It has a relatively large

surface area exposed to the environment. The factors like skin anatomy,
physiology, and biochemistry vary among species and within species. So
these factors can influence absorption. Our skin is permeable to many
toxicants. Dermal exposure to pesticides and industrial chemicals can result
in severe systemic toxicity. The human skin is 3 mm thick and the epidermis
provides the greatest resistance to toxicant penetration. Most of the systemic
absorption occurs at the capillary loops located at the epidermis-dermis
junction. Certain other factors like air flow, temperature and humidity etc.
can also influence dermal absorption.

b) Lung: The respiratory system includes nose, mouth, pharynx, trachea, and
bronchus which can reduce the toxicity of airborne particulate substances.
There is little or no absorption in these structures. But the cells lining the
respiratory tract absorb agents that can cause toxicological effects. The
absorption site is the alveoli-capillary membrane that is very thin (0.4–1.5
ìm). It allows for rapid exchange of gases/vapors. The ‘residual volume’ is
the amount of air retained in the lung despite maximum expiratory effort.
Hence toxicants may not be cleared out immediately due to the slow release
from the residual volume. The rate of entry of some vapor-phase toxicants
is controlled by the alveolar ventilation rate. The site of deposition of particles
in the respiratory tract is dependent on several factors. They include:
aerodynamic behavior of the particles, particle size, density, shape,
hygroscopicity, breathing pattern, and lung airway structure. The particle
sizes less than 10 - 20 ìm which get through the nasopharyngeal regions and
reach the alveoli are medically significant.

c) Gastrointestinal tract: This is lined by a layer of columnar cells and protected

by mucous. It gives minimal resistance to toxicant penetration. Most of the
absorption occurs in the intestine and then in the stomach. Also buccal and
rectal absorption can further occur in some cases. Absorption is through
passive diffusion mostly, except for nutrients; glucose, amino acids, and
drugs. The smaller the particle size of the toxicant, the greater is the
absorption. For absorption in this tract the chemical should be in an aqueous
form. Bacterial endotoxins, large particles of azo dyes and carcinogens are
absorbed by endocytotic mechanisms. Gastrointestinal tract motility is
important in the absorption of any toxicant. Excessive rapid movements of
the gut can reduce absorption by reducing residence time. Increased blood
flow after a meal can result in absorption of several drugs.

1.6 DISTRIBUTION AND STORAGE OF TOXINS IN

HUMAN TISSUES
The quantity of toxicant that reaches the target tissue like bone, fat and so on is
dependent upon the amount of toxicant absorbed, the distribution in the body,
the metabolism and the rate of excretion of the toxic substance. The cell membrane
13
Introduction to Toxicology is selectively permeable. Hence only some substances can pass through. This
depends on the molecular weight of the substance, lipid solubility and so on.
Most toxic substances pass through cell membranes by diffusion and 75% of the
cell membranes are composed of lipids. The electric charge on the toxicant is
also an important factor for diffusion. So, non-ionized toxic substances diffuse
easily than the ionized ones. Phagocytosis plays an important role in the
disposition of particulates that enter the respiratory tract. Asbestos dust and silica
dust are absorbed by the process phagocytosis and are engulfed by WBC’s in the
respiratory tract. Water soluble substances are absorbed in the small intestine by
pinocytosis.

a) Plasma: In humans the plasma protein binding can vary between and within
chemical classes. It is also species specific. The systems of human beings
bind acidic drugs more extensively than any other species. Further there
are also other variables that can alter plasma protein concentrations.
Pregnancy, malnutrition, carcinogenesis, liver abscess, kidney disease, and
age can reduce serum albumin. The á1-glycoprotein concentrations can
increase with age, inflammation, infections, obesity, kidney failure and
stress. These characteristics bring about changes in the body temperature,
in the acid-base balance and alter chemical protein-binding characteristics.

b) Liver and Kidney: Both these organs have a high affinity for toxic substances
and store more toxicants than any other tissue in the whole body. Lipophilic
substances like organochlorine pesticides and organic solvents like
trichloroethane, methyl chloroform, are readily absorbed in the liver. They
can remain in the liver for long periods if they are not biotransformed into
water-soluble substances. The liver is a major storage site for water-insoluble
toxic heavy metals. Some toxicants can be stored in the liver also. For
example, the antimalaria drug quinacrine accumulates in the liver as result
of reversible intracellular binding. The concentration in the liver can be
several thousand times than that of plasma. In the kidney certain large
molecules such as proteins do not easily pass through the walls of Bowman’s
capsule. Further, unbound metals like cadmium and mercury can be
reabsorbed in the cells of the proximal convoluted tubule. Inside the cell
they bind to metallothionein resulting in concentration of the toxicants in
the kidneys. The kidneys store ten times the amount of cadmium found in
the liver, and it can be stored for 10 years or more. Finally bioaccumulation
in the kidneys cause complete renal failure.

c) Lung: Some particulate material like asbestos, fiberglass toxicants <1µm

remain in the lungs. They give rise to respiratory diseases.

d) Fat tissue: Toxicants such as lipophilic pesticides, polychlorinated biphenyls

and lipid soluble gases accumulate in high concentration in the fat tissue.
The adipose tissue (fat) acts as a reservoir for lipophilic persistent organic
pesticides, a number of drugs and pollutants. They can sequester these
pollutants and then slowly release them into the bloodstream later.

e) Bone: Toxicants can be stored in the bone and it can become a reservoir
which allows the slow release of chemicals such as lead. Calcium is an
important component of bone. Lead can easily replace calcium and is stored
in the bones. The effects can be acute or chronic depending on how the
14
 toxicant is suddenly released or mobilized from these depots. Also perfusion Introduction to Toxicants
of tissues is an important factor for toxicant storage and distribution. The
organs like heart, kidney, liver and brain are well perfused than fat and
bone. In the fat and bone slow elimination of toxicants occurs. Bone can
store toxicants for 10 to 20 years. The toxicants stored in the bone may not
be toxic to the bone. It becomes toxic when it is released slowly resulting
in nerve damage and so on.

f) Blood: The circulatory system and components in the blood stream are
primarily responsible for the transport of toxicants to the target tissues or
reservoirs. Erythrocytes and lymph also transport the toxicants. Absorption
also depends on physiological factors and physicochemical properties of
the drug. Hence a reversible movement of toxicants occurs between blood
and tissues. Most toxicants pass by simple diffusion down a concentration
gradient from the blood to tissues. Tissue mass, blood flow, molecular
weight, lipid solubility are also important factors for toxicant distribution.
Water soluble toxins are stored here. Also heavy metals like arsenic, thallium,
cadmium and chlorinated pesticides, bisphenyl A targets the blood. The
toxic substances upon entering the bloodstream bind with plasma proteins
such as albumin, transferrin, globulin, and lipoproteins. Most toxic
substances are known to bind with the plasma protein albumin.

g) Passage of toxicants across placenta: Toxicants can cross the placental

barrier and affect the growing foetus by simple diffusion. It occurs readily
and easily when the toxicants are lipid-soluble. So the foetus is exposed to
almost all drugs even if low lipid solubility drugs are consumed by the
mother.

h) Blood-brain barrier (BBB): This limits the distribution of toxicants into

the central nervous system and cerebrospinal fluid. There are several
membranes the toxicant has to cross in order to get into the cerebrospinal
fluid. Certain diseases like meningitis disrupt this barrier and allow
antibiotics and toxicants to penetrate.

Toxic substances are absorbed and distributed in our body. They are also
eliminated, biotransformed and detoxified by the liver and kidneys. Further details
will be discussed in the following units.
Check Your Progress 2
Note: a) Write your answer in about 50 words.
b) Check your progress with possible answers given at the end of the
unit.
1) Describe the various routes of exposure to toxicants.
.....................................................................................................................
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15
Introduction to Toxicology 2) Where are toxic substances stored in the human body?
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1.7 LET US SUM UP

In this unit we have studied about the various toxicants, their mode of action and
sites of action. We have also learnt about the routes of exposure through which
toxic substances can enter our healthy body. The toxicants can be absorbed,
distributed and also stored in the various tissues and organs of the human body.
If the toxic substances are not detoxified or biotransformed, they can remain in
the organs causing organ failure and disease. Hence environmental toxicology is
a very important aspect that deals with effects of toxicants on human health.

1.8 KEY WORDS

Environmental Toxicology : It is a branch of science that deals with the
harmful effects of different physical, chemical
and biological agents on living organisms. It is
multidisciplinary in nature.
Toxicant : Any toxic material or substance is termed as a
toxicant. They are hazardous and poisonous.
Toxicants are generally man-made and artificial
products introduced into the environment due
to human activity. They include bisphenol,
insecticides and a number of industrial
chemicals.
Toxins : These are produced naturally by living
organisms. For example, toxins from the
mushroom plant and toxin from the venom of
snake are natural toxins.
Xenobiotic : is referred to a foreign substance entering the
body. It is derived from the Greek word ‘xeno’
meaning ‘foreigner’.

Toxicosis/ Poisoning/ : Any disease produced by a toxicant.

Intoxication
Organ toxicity : Toxic effects at the organ level. They are
neurotoxicity - brain, hepatotoxicity - liver,
nephrotoxicity - kidney, reproductive toxicity
depending on the organ involved.

16
 Introduction to Toxicants
1.9 REFERENCES AND SUGGESTED FURTHER
READINGS
Abel, P. D., ed. Water Pollution Biology. London: Taylor and Francis, 1996.
Costa, D. L. Air pollution. In Casarett and Doull’s Toxicology: The Basic Science
of Poisons, 6th ed., C. D. Klaassen, ed. New York: McGraw-Hill, 2001,
pp. 979–1012.
Doull, J. Recommended limits for occupational exposure to chemicals. In Casarett
and Doull’s Toxicology: The Basic Science of Poisons, 6th ed., C. D.
Klaassen, ed. New York: McGraw-Hill, 2001, pp. 1155–1176.
Hodgson, E., and R. C. Smart, eds. Introduction to Biochemical Toxicology, 3rd
ed. New York: Wiley, 2001. Hodgson, E., R. B. Mailman, and J. E.
Chambers, eds. Dictionary of Toxicology, 2nd ed. London: Macmillan,
1998. Klaassen, C. D. ed. Casarett and Doull’s Toxicology: The Basic
Science of Poisons, 6th ed. New York: McGraw-Hill, 2001. Timbrell, J.
A. Principles of Biochemical Toxicology, 3rd ed. London: Taylor and
Francis, 2000. Wexler, P. Information Resources in Toxicology, 3rd ed.
San Diego: Academic Press, 2000.
Hoffman, D. J., B. A. Rattner, G. A. Burton, and J. Cairns, eds. Handbook of
Ecotoxicology, 2nd ed. Boca Raton: Lewis, 2002.
Holgate, S. T., J. M. Samet, H. Koren, and R. Maynard, eds. Air Pollution and
Health. San Diego: Academic Press, 1999.
Larson, S. J., P. D. Capel, and M. S. Majewski, eds. Pesticides in Surface Waters.
Chelsea, MI: Ann Arbor Press, 1998.
Thorne, P. S. Occupational toxicology. In Casarett and Doull’s Toxicology: The
Basic Science of Poisons, 6th ed., C. D. Klaassen, ed. New York:
McGraw-Hill, 2001, pp. 1123–1140.

1.10 ANSWERS TO CHECK YOUR PROGRESS

Answers to Check Your Progress 1
1) Your answer should include the following points:
 Air pollutants
 Indoor Pollutants
 Water pollutants
 Soil pollutants
 Heavy metals
 Nitrates and phosphate
 Petroleum and oil pollutants
 Volatile organic compounds (VOCs)
 PCB organic compounds
17
Introduction to Toxicology  Solvents
 Asbestos
 Drugs of abuse
 Biological toxicants
 Cosmetics
2) Your answer should include the following points:
 therapeutic drugs
 drugs of abuse as toxicants.
3) Your answer should include the following points:
 Environmental toxicology: It is a branch of science that deals with the
harmful effects of different physical, chemical and biological agents on
living organisms. It is multidisciplinary in nature.
 The historical development of toxicology started very early in human
civilizations. Our ancestors who were cave dwellers found plants and
animals that had toxins and these toxic extracts were used in hunting
and warfare.
 Paracelsus in his study showed that specific chemicals were responsible
for the toxicity of a plant or animal poison. He also reported that the
dose of a chemical is an important factor for the human body’s response.
Hence, Paracelsus was one of the founders of modern toxicology. His
well known quote says that “All substances are poisons; it is the dose
that makes the poison”.
 In the 18th century ‘Ramazini’s Diseases of Workers’ was published in
1700. He is known as the father of occupational medicine.
 The founder of modern toxicology was Orfila, a Spanish physician who
worked at the University of Paris in the early 19th century.
 Rachel Carson, 1962, Silent Spring.

Answers to Check Your Progress 2

1) Your answer should include the following points:
 The common absorption routes include the: skin, lung and the
gastrointestinal tracts. The skin is the largest organ in the human body
and is a physical barrier to the absorption of toxic substances. The other
major routes are the respiratory and gastrointestinal tract. These give
less resistance to toxicant absorption than the skin. The respiratory tract
is the most rapid route for entry and the skin is the least rapid route for
entry. This difference is due to thickness of the membranes.
 Skin
 Lung
 Gastrointestinal tract

18
2) Your answer should include the following points: Introduction to Toxicants

 The quantity of toxicant that reaches the target tissue like bone, fat and
so on is dependent upon the amount of toxicant absorbed, the distribution
in the body, the metabolism and the rate of excretion of the toxic
substance. The cell membrane is selectively permeable allowing the
passage of some substances depending on the molecular weight of the
substance, lipid solubility and so on.
 Plasma
 Liver and Kidney
 Lung
 Fat tissue
 Bone
 Blood
 Passage of toxicants across placenta
 Blood-brain barrier (BBB)

19
Introduction to Toxicology
UNIT 2 CHEMICAL TOXICANTS

Structure
2.0 Introduction
2.1 Objectives
2.2 Classes of Chemical Toxicants
2.3 Exposure Classes
2.3.1 Types of Air Pollutants
2.3.2 Sources of Air Pollutants
2.3.3 Examples of Air Pollutants
2.4 Water and Soil Pollutants
2.4.1 Examples of Pollutants
2.5 Types of Classes
2.5.1 Food Additives
2.5.2 Detergents
2.5.3 Cosmetics
2.6 Key Words
2.7 Let Us Sum Up
2.8 References and Suggested Further Readings
2.9 Answers to Check Your Progress

2.0 INTRODUCTION
Toxicology is a branch of science which discuss about the source, physical and
chemical properties, absorption and pharmacological activity. Chemical
toxicology deals with the nature and reactions of toxic substances which involves
the origin, exposure and degradation. Any substance whose physiological action
gives adverse effects on health is a toxicant. Toxicity of a chemical is determined
by many factors like dose, exposure route, and the individual susceptibility
(response). Number of synthetic chemicals is produced on the global market and
many other chemicals released as by-products. The vital elements like Fe, Cu,
Zn, Mo, and Co,V, Mn etc., for biological systems may also show adverse effects
above certain concentrations. Elements like As, Sb, Cd, Hg, Be, Al and Pb etc.,
exist in the atmosphere are not essential for biological system exhibit toxicity
even at low concentrations.

2.1 OBJECTIVES
After studying this unit you will be able to:
 classify chemical toxicants;
 define toxicity, toxicant;
 describe various factors of route of exposure;
20
 describe toxic chemicals at home, food; Chemical Toxicants

 differentiate between domestic and occupational toxicants and

 describe effects of drug and their management.

2.2 CLASSES OF CHEMICAL TOXICANTS

The chemical toxicants categorized under several classes based on their chemical
composition and their mode of action. Any condition or disease that results from
the exposure to a toxicant is known as toxicosis. The word toxicosis used as an
alternative for poisoning or intoxication.

The broad classification of chemical toxicants is 1.Exposure classes 2. User

classes.

Exposure Classes: The toxicants under this class are present in domestic and
occupational environment. (Ex: food, water, air, soil).

User Classes: This type of toxicants include drugs of abuse, theraupeutic drugs,
agricultural chemicals, food additives, metals, solvents and combustion products.

Dear learner we have studied the chemical toxicants in air, water and soil pollution
units in course 1.In this unit we will discuss about other toxic exposures in our
daily lives.

2.3 EXPOSURE CLASSES

The pollutants under exposure class are air, water and soil pollution has studied
extensively in block 2, course 1. However, in this unit we will discuss some
important points about them.

2.3.1 Types of Air Pollutants

Air pollutants are classified into following ways. They are:

Gaseous Pollutants: These pollutants are gases and vapors at normal temperature
and pressure as well as vapors. The toxic air pollutants of greatest concern are
carbon monoxide (CO), hydrocarbons, hydrogen sulfide (H2S) nitrogen oxides,
ozone (O3), sulfur oxides, and CO2.The concentrations of pollutants generally
expressed as micrograms per cubic meter (ìg/m3) or for gaseous pollutants as
parts per million (ppm) by volume in which 1 ppm = 1 part pollutant per million
parts (106) of air.

Particulate Pollutants: These are fine solids or liquid droplets that are suspended
in air. They exist in various forms. In the form of dust where the particle size is
about 100 µm in diameter and released into the atmosphere directly from
substances like coal dust, ash, sawdust, cement dust, grain dust. In the form of
fumes it exists as suspended solids with size less than 1 µm in diameter, released
from metallurgical processes. In the form of mist, it exists as liquid droplets
suspended in air with a size of less than 2.0 µm diameter. In the form of smoke
it exists as solid particles from incomplete combustion of fossil fuels with a size
of 0.05–1.0 µm diameter. In the form of aerosol it exist as a liquid or solid
particles suspended in air or in any another gas with a size of <1.0 µm diameter.
21
Introduction to Toxicology 2.3.2 Sources of Air Pollutants
Natural Pollutants: Examples of some of the natural pollutants are volcanic
eruptions emits particulate matter as well as various gases like sulfur dioxide,
hydrogen sulfide, and methane. Huge quantities of pollutants from forest and
prairie fires release unburned hydrocarbons, CO, nitrogen oxides, and ash in the
form of smoke. Dust storms produce particulate pollutants and aerosols in the
form of salt particles produced by oceans. Plants and trees also produce particulate
pollutants in the form of produce pollen and spores which cause respiratory and
allergic reactions. By the atmospheric reactions with volatile organic compounds
released by the trees produce blue haze over forested mountain regions.

Anthropogenic Pollutants: Anthropogenic pollutants released in to the

atmosphere from three sources:
i) by burning fossil fuel for heating and power, or exhaust emissions from
transportation vehicles that use gasoline or diesel fuels,
ii) industrial processes,
iii) mining and drilling.
The pollutants released from combustion are fly ash, smoke, sulfur, CO, CO2
and nitrogen oxides. Combustion of coal and oil releases large amounts of sulfur
and its oxides that contribute to the formation of acidic deposition. In addition to
the fossil fuel combustion automobile exhaust include smoke, lead particles,
CO, nitrogen oxides, and hydrocarbons. Industries emit pollutants like sulfuric,
acetic, nitric, and phosphoric acids in effluents, solvents, resins, gases like chlorine
and ammonia, and metals.

2.3.3 Examples of Air Pollutants

Carbon Monoxide: Once carbon monoxide enters into human body it readily
combines with hemoglobin in the blood to form carboxyhaemoglobin, which
prevents the transfer of oxygen to tissue thereby affects the cardiovascular
function. The CO concentrations e”100 ppm it can cause headaches, dizziness,
nausea, and breathing difficulties. Above 700ppm level is always fatal. Sulfur
Oxides is released in to the atmosphere by industrial combustion of coal because
coal containing the highest levels of sulfur. The oxides of sulfur enter the
respiratory tract there by create irritation. Nitrogen Oxides found in
photochemical smog which is also a respiratory irritant that leads to pulmonary
edema and hemorrhage. Ozone is an oxidizing gas that is formed by
photochemical reaction in the atmosphere. The ozone present in the troposphere
is harmful and known as bad ozone. Whereas the ozone present in the stratosphere
filters the incoming UV radiation known as good ozone. Good ozone is destroyed
by chemical compounds like chlorofluorocarbons (CFCs). Lead is one of the
harmful particulates in air pollutants which impair renal function, nervous system,
reduce the development of red blood cells, and impair the nervous system that
leads to mental retardation and blindness. Particles like dust and fibers from
coal, clay, glass, asbestos, and minerals develop lung fibrosis. The most common
disease observed in the coal miners is pneumoconiosis, silicosis is observed in
inhaling silica-containing dusts and asbestosis from asbestos fibers are common
industrial pollution diseases.

22
 Chemical Toxicants
2.4 WATER AND SOIL POLLUTANTS
Surface water is polluted by various point or nonpoint sources. Effluents from
an industrial plant or a sewage-treatment plant are a point source and pesticides
and fertilizer runoff carried by rainwater into various water bodies is an example
of a nonpoint source. Industrial contaminants such as organic waste, solvents,
and inorganic wastes like toxic metals pollute the soil and water. In addition
industrial accidents may lead to severe local contamination. In addition to the
above mentioned pollutants pesticides, fertilizers, detergents, and metals
are important pollutants released from urban areas. Perpetual fertilizers and
pesticides which are applied directly to the soil in the course of action they move
from the soil to the water and make the way to enter the food chain. In another
way they leach out of the soil or runoff through rain and flow into the water
systems.

2.4.1 Examples of Pollutants

There are various types of pollutants which show toxic effects on environment
and human beings. Some of them are discussed in this unit.

Metal Toxicants: Metals toxicants mainly classified into three classes depend
on their nature.
1) Metals that are suspected to be a carcinogen,
2) Metals that transport readily in soil, and
3) Metals that proceed through the food chain.

Lead: The sources of lead in water are from lead pipes and lead solder. Lead soil
pollution is from seepage of lead from fallout from leaded gasoline and hazardous-
waste sites. Lead poisoning has been common in children, particularly in older
housing units and inner city dwellings, in which children may consume chips of
lead contaminated paint. The toxicity of lead mainly damages hematopoietic
system and the nervous system. At low levels of exposure, hyperactivity, decreased
attention span, mental deficiencies, and impaired vision is observed in children.
At high levels, encephalopathy occurs in both adults and children. Lead damages
the arterioles and capillaries which lead to cerebral edema and neuronal
degeneration.

Arsenic: Arsenic contamination is due to the leaching of pesticide sprays, released

by combustion of arsenic containing fossil fuels, smelter and mine runoff. Toxic
level exposures can produce abnormal skin pigmentation, hyperkeratosis, nasal
congestion, and abdominal pain. Epidemiologic studies have connected chronic
arsenic exposure to various cancers, including skin, lungs, and lymph glands in
humans.

Cadmium is released from industrial effluents and enters into the water bodies
untreated. This cadmium contaminated water is used for irrigation. The toxicity
of cadmium is recognized in Japan after outbreak of the disease Itai-Itai. The
people who suffer with this disease have combination of severe kidney damage
and painful bone and joint and recognized that occurs in areas where rice is
23
Introduction to Toxicology contaminated with high levels of cadmium. The aquatic organisms can accumulate
cadmium in their tissues, leading to increased concentrations in the food chain.

Mercury: Mercury poison is recognized after Minamata tragedy in Japan. In

Japan, effluents from a chemical and plastics plant containing mercury were
released into Minamata Bay. Bacteria in the aquatic sediments converted the
mercury into methyl mercury and that was absorbed by aquatic animals.
Consumption of mercury contaminated fish and shellfish by the people suffered
with mercury poisoning, or Minamata disease that resulted in death.

Fertilizers: Not only the metal contamination observed in soil and water pesticides
are also of major concern. The most toxic pesticides are organochlorine derivatives
like DDT, aldrin, dieldrin, and chlordane due to their stability and persistence
they can accumulate in food chains.

Fertilizers containing nitrates discharge from sewage treatment plants, and

leachate from septic systems and manure leach from soils and enter water bodies.
Phosphate fertilizers have a tendency to be absorbed and get accumulated. The
increase in the nutrients like phosphates, leads to “algal blooms” or eutrophication,
in water bodies. The algal bloom chokes off light penetration and lessens the
atmospheric reoxygenation of the water that creates anaerobic conditions and
finally death of many aquatic organisms occurs. The adverse health effects from
nitrates in drinking water mainly are nitrosamine formation and
methemoglobinemea. In human beings by the action of intestinal bacteria the
nitrates will be converted to nitrites. These nitrite ions combine with hemoglobin
to form methemoglobin, which decreases the oxygen-carrying capacity of the
blood and resulting in anemia or blue-baby disease. It is particularly affected to
young babies who consume water and milk-formula prepared with nitrate contains
water. Some of the nitrosamines are known carcinogens. Oil and petroleum is
everlasting pollutants where it is produced by vehicular oil emission or spillage
from oil tankers. Oil slicks are very common and responsible for the deaths of
many birds in the marine environment. Sea animals like crabs, shrimp and mussels
are also affected by the toxic hydrocarbons they consume.

VOC: Volatile organic compounds (VOCs) are a group of halogenated solvents

and petroleum products, which are used in large quantities by a many industries,
like degreasing, dry cleaning and paint. The properties of VOCs allow them to
move quickly into groundwater, and contaminate. The exposure of VOCs can
cause headache, impaired cognition, and kidney damage and acute levels of
exposure causes cancer, particularly childhood leukemia.

PCBs: Polychlorinated biphenyls (PCBs), phenols, cyanides, plasticizers, organic

solvents, and other toxic chemicals that are used as coolants in transformers and
are also by-products of the plastic, lubricant, rubber, and paper industries released
into the water and contaminates the aquatic animals there by enters the food
chain. They are stable, lipophilic in nature and metabolite very slowly in tissues
cause tissue damage and responsible for death.

Dioxins: Dioxins are another group of toxic chemical released from industrial
accidents and through use of the herbicide 2, 4, 5-T, that contaminates water and
soil.
24
Check Your Progress 1 Chemical Toxicants

Note: a) Write your answer in about 50 words.

b) Check your progress with possible answers given at the end of the
unit.
1) Define the following
i) Toxicity
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ii) Toxicant
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iii) Toxicosis
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2) What are user and exposure class of chemical toxicants?
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2.5 TYPES OF CLASSES

Dear Learners, let us know about types of classes in the following sentences:

2.5.1 Food Additives

Food additives are compounds which are deliberately added to the food products
to improve the colour, texture, flavor, appearance and preservation. According
to the WHO/FAO joint expert committee, “food additives are non-nutritive
substances added intentionally to food, generally in small quantities, to improve
its appearance, flavor, texture or storage properties. In view of the safety 25
Introduction to Toxicology requirement Joint FAO/WHO codex Alimentarious commission 1973 has
proposed six general principles to be food additive.
1) All proposed food additives must be tested, evaluated toxicologically in all
aspects with cumulative, synergetic and potentiating effects.
2) Food additives that are under safe level of intended use will be endorsed.
3) They should be re-evaluated periodically in view of use and safety.
4) Confirmation from CA Commission.
5) Justification should be based on :
 Preservation and reduction at nutritional quality.
 Special dietary food products.
 Monitoring of quality, stability and organoleptic properties.
 Quality of raw material for food products.
6) Temporary or Permanent approval.

2.5.2 Detergents
1) Detergents are the substances containing molecules which are amphiphilic
in nature with a hydrophilic head group and a hydrophobic hydrocarbon
tail. The detergents are classified into anionic, cationic, nonionic or
amphoteric on the basis of hydrophilic head group. On the basis of chemical
characteristics detergents are of two kinds.

2) Phosphate containing detergents

3) Surfactant containing detergents.

 Phosphate containing detergents are highly caustic in nature where as

surfactant detergents are very toxic in nature. The phosphates detergents
soften the hard water there by suspend dirt in water. The surfactant detergents
increase the detergent properties like wetting, foaming, dispersing and
emulsifying and helps in removing dirt.

 A large number of surfactant molecules are associated with other components

to augment the detergency by which removal of dirt is difficult because of
the strong attraction between dirt particles and the fabric but the penetration
and adsorption of surfactant molecules onto the and fabric interface becomes
very poor.

 Detergents comprise various molecules of surfactants. Surfactants are surface

active promoter that is heterogeneous long chain molecule with hydrophilic
and hydrophobic components. Surfactants properties like wetting,
emulsifying, dispersive, foaming and foaming control ability can be modified
by altering the hydrophobic and hydrophilic part. On the basis of this
character and ionic (electrical charge) properties in the water surfactants are
classified as i. Cationic surfactants ii. Anionic surfactants iii. Nonionic
surfactants iv. Amphoteric surfactants.fig.1, table 1.

26
 Hydrophilic Hydrophobic Chemical Toxicants

Non-iconic

anionic

cationic

Amphoteric

Figure 2.1: Properties in the water surfactants

Cationic Surfactants: Cationic surfactants comprises of a positively charged

nitrogen atom and one long chain hydrophobic substituent. Examples are
quaternary ammonium compounds with the general formula R4N+X– , where X–
is a chloride ion and R is an alkyl group, alkyl trimethyl ammonium chloride,
where R contains C8-18 atoms like dodecyl trimethyl ammonium chloride
(DDTMAC), dialkyl dimethyl ammonium chloride (DADMAC), and alkyl
dimethyl benzyl ammonium chloride. These are commonly used in detergents as
softeners.

Anionic Surfactants: Soaps are examples of anionic surfactants comprise of

sodium, potassium, or ammonium group. The most commonly used hydrophilic
groups are carboxylates, sulphates, sulphonates and phosphates. The anionic
surfactants are effectively used to clean oily sand clay l suspension with different
degrees on the basis of their chemical composition.

Nonionic Surfactants: This type of surfactants is nonionisable in solution and

is effectively used in removing oily soils by the process of emulsification and
solubilization. These can be mixed with anionic surfactants for cleaning oil soils.
Examples are ethylene oxide, alcohol ethoxylates, alkyl phenol ethoxylates, fatty
acid ethoxylates, monoalkanolamide ethoxylates, sorbitan ester ethoxylates, fatty
amine ethoxylates and ethylene oxide-propylene oxide copolymers, glycol esters,
glycerol esters, glycosides, sucrose esters, amine oxides and sulfonyl surfactants.

Amphoteric Surfactants: Amphoteric surfactants contain both cationic and

anionic groups. Examples are the N-alkyl betaines, like laurylamidopropyl-
dimethylbetaine.The. Amphoteric surfactants are pH dependent. In acidic
solutions, they act as a cationic surfactant, whereas in alkaline solutions they
behave like an anionic surfactant. Amphoteric surfactants are also termed as
zwitterionic compounds. They are chemically stable and soluble in water and
have affinity with other surfactants. The surface activity is depends on the
separation of charge between the groups and maximum at the isoelectric point
where it exist as zwitterion. Examples are N-alkyl amino propionates, amino
dipropionate, alkyl imidazolines.These are also pH dependant. The pH affects
their wetting, detergency, foaming abilities.

Along with these the surfactants contain;

Builders- to enhance the cleaning ability. Eg: Citrate,phosphate,sodium carbonate

etc. 27
Introduction to Toxicology Antiredeposition Agents: to prevent removed soil from redepositing on cleaned
fabrics.Ex: Carboxymethyl cellulose, sodium polyacrylate, polyethylene glycol
polymers.

Zeolite: to sequester multivalent metal ions and prevents the anionic surfactants
from precipitating out of the solutions.

Alkaline Agents: to remove oily soil containing fatty acids. Ex: Sodium carbonate
and sodium silicates.

Processing aids: to provide required physical properties.Ex: sodium sulfate,

water, alcohol, xylene sulphonate. Sodium sulfate provides crisp and free-flowing
texture; alcohols regulate the viscosity and prevent product separation.

Colorants & Fragrances: to fetch uniqueness to the product especially blue

colorants provide a desirable blue/white color to white fabrics. Fragrance conceals
the chemical odour of the detergents.

Oxygen bleaches: to remove stain and soil by bleaching action. They contain
inorganic peroxygen compounds like sodium perborate tetrahydrate and sodium
percarbonate and hydrogen peroxide.

Enzymes: to break down complex dirt molecules, especially proteins like blood
and grass. Examples: Protease, lipase and cellulose.

Table 2.1: Properties in the water surfactants

Type Commercial and domestic Major industrial examples

examples
Anionic Sodium linear alkylbenzene Petroleum sulphonates;
sulphonate (LABS); sodium linosulphonates; naphthalene
lauryl sulphate; sodium sulphonates, branched
lauryl ether sulphates alkylbenzene sulphonates; linear
alkylbenzene sulphonates; alcohol
sulphates
Cationic Stearalkonium chloride; quaternary ammonium
benzalkonium chloride compounds; amine compounds

Non-ionic Dodecyl dimethylamine alkylphenol ethoxylates; alcohol

oxide; coco diethanol-amide ethoxylates; EO/PO polyol block
alcohol ethoxylates; linear polymers; polyethylene glycol
primary alcohol esters; fatty acid alkanolamides
polyethoxylate
Amphoteric Cocoamphocarboxyglycinate; Betaines; imidazolines
cocamidopropylbetaine

Toxicity
The effluents containing surfactants from different sources released into water
bodies and shows adverse effects on humans and ecosystems. The degree of
toxic effect of surfactants to aquatic plants depends on its concentration. If the
concentration of surfactants is high the growth of algae and other microorganisms
28
in water will diminish slowly, resulting in less productivity of water bodies, Chemical Toxicants
thereby impair the food chain of aquatic organisms. This is kind of toxicity
increase the rate of membrane permeability thereby disintegrate the structure by
material exosmose. In aquatic animals the surfactants enter through skin piercing,
gills and animal feeding and thereby enter the food chain. The phosphorous in
waste water inhibit the degradation of other toxic constituents. Surfactants
stimulate emulsification and dispersion there by reduces the efficiency of sewage
treatment. Long term use of surfactant causes skin irritation and allergy.
Surfactants like sodium dodecyl benzene sulfonate (SDBS) absorbed by the skin
and enter into the blood stream that can cause damage to the internal organs.

Types of Food Additives

According to Federal food, drug and cosmetic Act, five categories of compounds
are associated with human food that includes GRAS.
Cat-I : Whose GRAS stature was reaffirmed.
Cat-II : No evidence of toxic hazard.
Cat-III : Additional studies are required for safety.
Cat-IV : Incomplete reaffirm safety. (Evidence of toxicity was reported)
Cat-V : No biological studies available.

Food Colors
Food colors are the natural pigments that are present in the fresh foods which
disappear and show toxic effects due to the adverse physical and chemical
processing methods. It decreases the visual perception of food product. At about
75% of food products are processed in developed countries. All Food
manufacturers replace lost colour and appearance by additives. Food colors are
used to restore the original appearance, uniformity and intensity of colour, preserve
the flavor and to protect light sensitive vitamins, attractive appearance to colorless
gelatin desserts and to identification.

Colorants Subject to Certification

They are very pure chemical with standardized color strengths. The raw materials
used are ‘Coal tar’ dyes or by products of the petroleum industry. Where as
certified colors are accessible as water soluble dyes or insoluble pigments. All
soluble dyes are available as primary colors or addititive mixtures /with other
certified colors.

Colorants Exclude from Certification

They will be classified as natural colors. According to colour additives amendment
act of 1960 they or subject to surveillance by FDA.

They are (i) Non-Synthetic (ii) Nature identical (iii) Inorganic colour, which are
extracted from animal, plant or mineral source.

Colourants Subject to Certification: The delisted by FDA are:

1) FD & C Red No.1: (Ponceau 3R, color index No. 16155). This is a
disodium salt of 1 – Pseudocumylazo – 2- naphthol -3,6- disulfonic acid,
29
Introduction to Toxicology which is a dark red color dissolved in H2O . It is proved to be a liver
carcinogen.

2) FD & C Red No. 2: Amaranth color index no. 16185. Amaranth is reddish
brown powder which is soluble in H2O gives megenta red or bluish red
colour. It is also proved to be a carcinogenetic.

3) FD & C Red No. 3: Erythrocine, color index no. 45430 xanthine group of
dyes. It is a brown coloured powder and soluble in H2O that yields red
colour and fluorescence with 95% alcohol. Proved to be thyroid tumor,
blood and gene mutations.

4) FD & C Red No. 4: Ponceau SX, color index no. 14700 originally
approved food colour in butter and margarine. It is proved to be chronic
follicular cystitis with hematomatoury Projections in to the Urinary bladder,
hemosiderotic.

5) FD & C Red No. 32: (oil Red Xo, color index No. 12140) It is brownish
red powder soluble in oil. Used to colour Oils, Fats, Waxes, Greases, acrylic
emulsions, colour the oranges.It is proved to be cathartic,growth
retardant,damage to liver & heart tissue.

6) FD & D Red No. 40: Allura Red AC. Color index No. 16035 used in
cosmetics, drugs and food (soft drinks & cotton candy). Reported to be
hyper active agent and growth retardant.

7) Citrus Red No. 2: (Solvent Red 80, color index No. 12156) It belongs to
monoazodye group. Used to colour the skin of oranges to prepare orange
marmalade. Proved to be adenocarcinoma,limphosarcoma and bladder
cancer.

8) FD & C Green No. 3: (Fast green FCF color index No. 42053). Belongs
to triphenyl methane group of dys. Reddish or brownish violet color, soluble
in H2O. It induce sister chromatid exchanges in bone marrow cells and
produce sarcoma.

9) FD & C Blue No. 2: (Indigotin, Indigo carmine. Colour index No. 73015).
It belongs to indigoid family of synthetic dyes soluble in H2O yielding
blue solutions proved to be innocuous and produce tumors at the site of
application.

10) FD & C Yellow No. 3 & 4: Belong to monoazo group with yellow AB &
yellow OB with colour index No. 11380, 11390. They used to colour
oleomargarines. Proved to be liver and bladder carcinogens.

11) FD & C Yellow No. 5: (Tartrazine, CI no. 19140) orange yellow powder,
soluble in H2O. Proved to be allergic.

12) FD & C Yellow No. 6: (Sunset yellow FCF, color index no. 15985).
Orange red powder soluble in H2O gives orange yellow solution. It is
proved to be allergic.

Toxicological characteristics of colorants except from certification are Annato

extract, Anthocyamius, Dehychated beets, Chlorophyls, Caramel, Tumeric &
Cakolene etc.
30
Acidulate and Sequestrates. Chemical Toxicants

Acidulant are the food additives which are added as preservatives, chelating
agent and anti oxidant synergist, flavouring agent’s viscosity & melting modifiers
and to control pH.
Examples of Acidulant :-
Inorganic Acids: Phosphoric acid and its derivatives, HCl & H2SO4.
Organic Acids: Citric acid, benzoic acid, sorbic acid, butyric acid and caprylic
acid.
According to the FAO, 0.5% of phosphates are the tolerable level in the diet
without any adverse effect. Higher levels may be tolerated if the other ions like
Ca, Mg, and K maintained at required levels otherwise it produce adverse effects
on physical and chemical characteristics and off flavors as well in food items.
The approved dietary intake of phosphorous is <30mg/kg body wt/day in the
nutrition of human beings. HCl and H2SO4 are not directly used as an acidulant
but used in hydrolysis of proteins, starch and corn syrups. HCl & H2SO4 are
corrosive to all body tissues. Inhalation causes lung damage and skin contact
results in necrosis.Vinegar is a aqueous solution of acetic acid. It is used as
acidifier, flavor enhancer, pickling agent and pH controlling agent. It is absorbed
in the gastro intestinal tract and used up in oxidative metabolism, formation of
glycogen intermediates of carbohydrates and synthesis of fatty acids and
cholesterol. Acetic acid in H2O or organic solvents is strongly corrosive to the
skin causes tissue damage and produce canker sores. Lactic acid is present in
pickles, beer, buttermilk and cheese. It is used as acidifier, antimicrobial agent,
curing agent, flavoring and carrier agent. According to FAO/WHO the permissible
limit of D (-) isomer is 100mg/kg/body/day.

Adipic Acid: It is one of the most important of aliphatic dicarboxylic acid used
as leavening agent, neutralizing and flavoring agent. The permissible limits are.
Baked items – 0.05%
Non-alcoholic beverages – 0.005%
Condiment and relishes – 0.5%
Dairy products – 0.45%
Fats & Oils – 0.3%
Frozen dairy desserts – 0.0004%
Gelatin & puddings – 0.55%
Meat products – 0.3%
Above these limits it causes intestinal hemorrhage.
Allyl isothiocyanate: It is used as spice seasorings and condiments. It is formed
from sinigrin by crushing the moistened mustard seeds.It is proved to be skin
irritant and at high concentrations results in epithelial hyperlasia and uleers.

Cinnamyl Anthranilate: Is a synthetic flavoring agent used to provide grape or

cherry flavor to beaverages, candy, puddings, and chewing gum at 1000 pm
concentrations. Above this limit it is thought to be carcinogenic.

31
Introduction to Toxicology Menthol: It is a synthetic and natural constituent of peppermint oil used as
flavoring agent in candies, chewing gum. It can cause sensitization reactions
like urticaria. At high concentrations it causes heart fibrillation.

Monosodium Glutamate (MSG): MSG is liberally used in flavor enhancer in

meat products. It provides Umami sensation generally called as a 5th basic taste.
At high concentrations, it causes Chinese restaurant syndrome (CRS) in sensitive
population with symptoms like upper body tightness, warmth and feeling pressure.

Myristicin: Nutmeg oil, mace oil contains < 4% Myristicin. It has psychomimetic
and narcotic properties. It is thought to cause headache, abdominal pain and
nausea at high doses. At elevated levels cause liver damage & death.

Yeast: Yeast is used to ferment the baked food items. At high levels in food
products causes high uric acid levels in blood. The urate oxidase enzyme is
absent in human beings. During the fermentation of yeast, pharmacologically
active amines and tyramine formed which are responsible for higher BP.

Parabenes: Parabenes are the esters of p-hydroxy benzoic acid (PHB) used an
antimicrobial agent in food products, cosmetics and also known as paracepts or
PHB esters. These are used in malt beverages and non carbonated soft drinks. At
high concentrations, it can causes dermatitis.

Polycyclic aromatic hydrocarbon (PAH): PAH enters into the human food
chain by (i) Polluted air on food crops. (ii) Heat processing of foods like roasting,
smoking and grilling. iii.Preparing the food products above 400 0C resulting in
significant formation of PAH like benzo [] pyrene, dibenzo (,) anthraene,
dibenzo [,] pyrene and benzo[] fluro anthrene. All these compounds are
potent carcinogens.

2.5.3 Cosmetics
Cosmetic products are the substances to be applied to the external parts of the
human body including teeth and oral cavity for cleaning purpose, appearance,
protecting and maintaining in good condition. Cosmetics are classified into two
types.

1) Leave-on 2) Rinse-off

Cosmetics which are leave-on category can be intended to lasting for certain
period on the skin like perfumes, cosmetics used for decoration, body and face
creams. A rinse-off cosmetic is a product one which should be rinsed off after a
period of time like shampoos, soaps, shower gels, ointments and toothpastes.

Recent years, cosmetics, and many other beauty products for personal care that
do not fall within cosmetic regulation are noticed as emerging pollutants because
they are constantly released into the terrestrial and aquatic environment. They
are found to be persistent, bioactive, and bio-accumulate with potential adverse
ecological and environmental impact. Substances like perfluroalkyl compounds,
parabenes, organic UV filters and microplastics.

1) UV Filters
Cosmetic products like sunscreens and skin lotions contain benzophenones
32 (BPs) have ultraviolet (UV) filter properties which absorb UV-A (315–400
 nm) and UV-B (280–315 nm) radiation. They also used as additive in plastics, Chemical Toxicants
printing inks, shampoos, perfumes and photographic films to prevent UV
light damage. The Benzophenones are highly lipophilic in nature and
bioaccumulate in the human body by crossing dermal tissue. Studies proved
that BP UV filters could be detected in the plasma, bile and urine after
application. These filters also found in the surface waters there by enter in
to the food chain. The concentration of BPs in sewage sludge exceeds 10
mg/kg of dry matter proved to be mammary cancer cell proliferation. The
photo stability of UV filters plays an important role to absorb UV light.

2) Inorganic Filters
The inorganic filters present in sunscreen are TiO2 and ZnO. The nano
TiO2 and ZnO particles are almost customarily present in sun blocker
formulations because they nanoscale enhance skin retention, acceptance for
consumers, and the UV depletion properties. By the process of immersion
or abrasion after application these compounds released into the environment.
The sewage treatment plants (WWTPs) expel most of the TiO2 present in
the sewage, but a small amount is released into natural water bodies where
it aggregate and remain in suspension. The residues of TiO2 released from
sunscreen form aggregates submicron level which remains in suspension in
fresh water, and in water bodies with high salt concentrations like sea they
aggregate and progressively settle down and detained in the sediment.

By the photocatalytic reactions of UV radiation the nano TiO2 and ZnO

generates reactive oxygen species likes OŔ2", ŔOH, and H2O2. The
photoactivity of these species causes cellular damage and environmental
toxicity. To prevent the production of ROS, TiO2nanoparticles present in
sunscreen are coated with silica and alumina and/or doped with manganese
or vanadium, even though sometimes these coatings do not withstand contact
with water.

These results suggest that the normal recreational activities in coastal resorts
can result in the production of significant amount of H2O2 and consequent
damage to or death of marine coastal phytoplankton; this could have
reverberations on the marine food web, which relies on these
microorganisms. The ZnO is extremely toxic to the aquatic animals like
zebra fish, marine algae and sea urchins.

3) Parabenes
Parabenes are alkyl esters of the para-hydroxybenzoic acid which are used
as preservatives due to their antimicrobial activity against yeasts, molds,
and bacteria and their chemical stability, low toxicity, and low cost. They
are extensively used in cosmetics, including powders foundations, , lipsticks,
eye shadows, mascara, lip glosses and nail polishes, and in pharmaceuticals
and personal care products such as lotions, sunscreens, cleansers, shampoos,
deodorants, hair care products, and toothpaste. They are proved to have the
potential to produce contact dermatitis, irritation, or photo contact dermatitis.
4) Triclosan

Triclosan (TCS), 5-chloro-2-(2, 4-dichlorophenoxy) phenol is a preservative

used personal care products like hand soaps, shampoos, detergents,
33
Introduction to Toxicology toothpastes, sunscreen, and deodorants. The domestic sewage contains most
of the TCS and enters into the aquatic environment in spite of the treatment.
During sewage treatment TCS is converted into chlorinated derivatives that
are more persistent and toxic to the aquatic life. It also enters into the
terrestrial environments because of the sewage sludge application on farm
land as a fertilizer. TCS is stable and lipophilic in nature due to which it
bioaccumulate in algae, plants, earthworms, marine mussels, snails,
amphibian larvae, fish, and marine mammals causes adverse ecological
effects and alters benthic bacterial community composition, exhibits
teratogenic effect. TCS has potential to impair thyroid function, endocrine
disruption, oxidative stress, and liver carcinogenesis.

5) Plastic Microbeads
Plastic microbeads are pieces of plastic, spherical in shape, size varying
from <mm-1mm. They are widely used in soaps, face wash, toothpaste,
exfoliating scrubs and anti-ageing creams because exfoliating debris from
the skin by replacing natural exfoliating materials like pumice, oatmeal,
apricot husks. The microbeads are made by different types of plastic material.
They are

Polyethylene (PE), polymethyl methacrylate (PMMA), Nylon, Polyethylene

terephthalate (PET) and Polypropylene (PP).

When these products are washed down after use the microbeads in the
products pass through sewage systems. Because of nonbiodegradable nature
and very small size they enter into rivers and canals and finally into the sea
and ocean that contribute to the plastic pollution. Because of the small size
and large surface area plastic microbeads absorb POPs and other pollutants
in aquatic environment. Aquatic species consume these microbeads by
mistake since they are not able to distinguish between food and microbeads
there by enter into the food chain and regularly consumed by people. The
harmful toxic chemicals which are added during the manufacture of plastic
like plasticizers and flame retardants which are drain out into water bodies
and produce adverse effects by polluting them. These plastic microbeads
block the intestine in marine animals, impair reproductive ability and reduce
the growth rate.
Check Your Progress 2
Note: a) Write your answer in about 50 words.
b) Check your progress with possible answers given at the end of the
unit.
1) Define food toxicology
.....................................................................................................................
.....................................................................................................................
.....................................................................................................................
.....................................................................................................................
.....................................................................................................................

34 .....................................................................................................................
2) What are the types of food additives? Chemical Toxicants

.....................................................................................................................
.....................................................................................................................
.....................................................................................................................
.....................................................................................................................
.....................................................................................................................

2.6 KEY WORDS

Acute Effect : Effect of finite duration occurring rapidly (usually in
the first 24 h or up to 14 d) following a single dose or
short exposure to a substance or radiation

Additive Effect : Consequence that follows exposure to two or more

physicochemical agents which act jointly but do not
interact: The total effect is the simple sum of the effects
of separate exposures to the agents under the same
conditions

Adverse Effect : Change in biochemistry, physiology, growth,

development morphology, behavior, or lifespan of an
organism which results in impairment of functional
capacity or impairment of capacity to compensate for
additional stress or increase in susceptibility to other
environmental influences.

Air Pollution : Presence of substances in the atmosphere resulting ither

from human activity or natural processes, in sufficient
concentration, for a sufficient time and under
circumstances such as to interfere with comfort, health,
or welfare of persons or to harm the environment.

Bioaccumulation : Progressive increase in the amount of a substance in an

organism or part of an organism that occurs because the
rate of intake exceeds the organism’s ability to remove
the substance from the body.

Contaminant : 1. Minor impurity present in a substance. 2. Extraneous

material inadvertently added to a sample prior to or
during chemical or biological analysis 3. In some
contexts, as in relation to gas cleaning equipment, used
as a synonym for “pollutant”, especially on a small scale.
4. Unintended component in food that may pose a hazard
to the consumer

Detergency : The term ‘detergency’ is used to describe the process of

cleaning by surface active agent. Detergency can be
defined as removal of unwanted substance (soil) from a
solid surface brought into contact with a liquid. The word
‘soil’ in connection with textile surfaces most frequently 35
Introduction to Toxicology denotes the unwanted accumulation of oily and/or
particulate materials on the surfaces or interior of fibrous
structure

Poison : Substance that taken into or formed within the organism

impairs the health of the organism and may kill it.

Photostability : The term photostability means resistance to permanent

structural and functional changes under the infl uence
of solar energy.

2.7 LET US SUM UP

Chemical toxicants are released into the environment in different ways, and they
can transport through many pathways and show adverse effects on the
environment. A toxicant present in the environment at a given point in time, it
can be stationary, it can be transported, or it can be transformed into another
chemical species. The life cycle of a chemical depends on the physicochemical
properties, characteristics of the environment to which it is released. The chemical
toxicants released into the atmosphere exerts adverse effects on humans and
other terrestrial and aquatic organisms like intentional ingestion, occupational
exposure, environmental exposure, as well as accidental and intentional poisoning.

2.8 REFERENCES AND SUGGESTED FURTHER

READINGS
Hemond, H.F., and E.J.Fechner. Chemical fate and transport in the environment.
New York Academic Press,1994

Mackay, D.,W.Y.Shiu, and K.C. Ma.Physical-Chemical Properties and

environmental fate and degradation hand book, CRC Press 2000.

Rand, G.M., ed. Fundamentals of aquatic Toxicology:Part II Environmental

Fate,Washington DC;Taylor and Francis,1995.

2.9 ANSWERS TO CHECK YOUR PROGRESS

Answers to Check Your Progress 1
Your answer should include the following points.

1) a) Toxicity is the amount of poison that, under a specific set of conditions,

will cause a detrimental effect (agents are usually compared on a mg/
kg basis, toxicity is not the condition produced by the toxicant).

b) Toxicant: Any substance whose physiological action gives adverse

effects on health is a toxicant.

c) Toxicosis: It is the condition or disease state that results from exposure

to a toxicant. The term toxicosis is often used interchangeable with
the term of poisoning or intoxication.

36
2) Exposure classes are the toxicants under this class are present in domestic Chemical Toxicants
and occupational environment. (Ex: food, water, air, soil).Where as User
classes are the type of toxicants include drugs of abuse, therapeutic drugs,
agricultural chemicals, food additives, metals, solvents and combustion
products.
Answers to Check Your Progress 2
Your answer should include the following points.

1) It deals with natural contaminants, food and feed additives, and toxic and
chemo-protective effects of compounds in food. Food Toxicology is involved
in delivering a safe and edible supply of food to the consumer. During
processing, a number of substances may be added to food to make it look,
taste, or smell better. Fats, oils, sugars, starches and other substances may
be added to change the texture and taste of food. All of these additives are
studied to determine if and at what amount, they may produce adverse effects.
A second area of interest includes food allergies. Almost 30% of the American
people have some food allergy. For example, many people have trouble
digesting milk, and are lactose intolerant. In addition, toxic substances such
as pesticides may be applied to a food crop in the field, while lead, arsenic,
and cadmium are naturally present in soil and water, and may be absorbed
by plants. Toxicologists must determine the acceptable daily intake level
for those substances

2) According to Federal food, drug and cosmetic Act, five categories of

compounds are associated with human food that includes GRAS.
Cat - I - Whose GRAS stature was reaffirmed.
Cat – II - No evidence of toxic hazard.
Cat – III - Additional studies are required for safety.
Cat – IV - Incomplete reaffirm safety. (Evidence of toxicity was reported)
Cat – V - No biological studies available.

37
Introduction to Toxicology
UNIT 3 BIOLOGICAL TOXICANTS
Structure
3.0 Introduction
3.1 Objectives
3.2 Types of Biological Toxicants and Food Intoxication
3.2.1 Types of Biological Toxicants
3.2.2 Food Intoxication
3.3 Classification of Toxicants Present in Food
3.3.1 Naturally Occurring Toxicants
3.3.2 Toxicants from Microorganisms
3.4 Microbial Agents: Symptoms, Effects on Health and Management
3.4.1 Bacterial Agents
3.4.2 Fungal Agents
3.4.3 Viral Agents
3.5 Endotoxins and Enterotoxins
3.5.1 Endotoxins
3.5.2 Enterotoxins
3.6 Let Us Sum Up
3.7 Key Words
3.8 References and Suggested Further Readings
3.9 Answers to Check Your Progress

3.0 INTRODUCTION
The term toxin was first used by organic chemist Ludwig Brieger. Biological
toxins are unique biological molecules that are mainly used for protection and
predation which affect cells or organs or an entire system of a target species.
Most commonly all biological toxins are proteins and especially are capable of
modulating biological process by a number of ways, interacting with the biological
receptors and blocking their activities. Some of the toxins are nonspecific, but
most of them have the target specificity. Biological toxins are mainly biological
agents which involve bacterium, virus, protozoan, parasites or fungus. There are
about 1,200 different types of potential bio agents. Biological toxins are toxic
substances which are produced by plants, animals and microorganisms that can
cause harmful effect when ingested, injected, inhaled or absorbed. Biological
toxins may be stated as “any toxic substances occurring in nature produced by
animals, plants or microbes (pathogenic bacteria), such as bacteria, fungi,
flowering plants, insects, fish, reptiles or mammals.”

3.1 OBJECTIVES
After examining this unit, you must be able to;
 give a list of different type of biological toxicants;
38
 discuss suitable examples of bacterial, viral and fungal toxins; Biological Toxicants

 list the diseases caused by the biological toxicants;

 describe the measures of bacterial, viral and fungal toxicants;
 discuss the various types of virulent and prevalent biological agents;
 describe the classification of Toxicants present in food and
 give an outline of preventive measure by biological agents and relevant
measure that can be used to control exposure.

3.2 TYPES OF BIOLOGICAL TOXICANTS AND

FOOD INTOXICATION
Biological toxins are poisonous substances produced by living organisms from
emergent microscopic species to well-developed species. Normally, toxins are
neither dermal nor considered a volatile exposure hazard. Toxins are usually
much more noxious than any of the other chemical agents. Characteristics of
selected biological toxins are shown in Table 3.1.

Table 3.1: Characteristics of Selected Biological Toxins (Thomas et al., 2000)

Source Toxin LD50 (µG/kg) Required Notes

Detection
Capabilitya
Fungus
Fusarium Trichothecenemy- 25 to 500 40 mg/m3 Nonlethal and
species cotoxins (inhalation) (air) delayed effects.
(‘’yellow rain”)

Inhalation,
ingestion, dermal.

1600 oral 40 ppm Very stable

repeated small
doses are
cumulative
Bacteria
Clostridium Botulinium A, B, ~ 0.02 0.1 mg/m3 Comes under
botulinium C, D, E (inhalation) most potent
Known toxins

1 (oral) 0.02 ppm Delayed

(water or Lethaleffect .
food)

Persists in food
and water.

It takes around
12 hoursin air for
its break down in.
39
Introduction to Toxicology
Clostridium Gangrene-causing 0.1 to 5 0.3 mg/m3 Delayed action.
perfringens enzyme

Low mortality,
but very
debilitating.

Clostridium Tetanus toxin ~3 N/A Delayed action.

tetani

Heat sensitive and

unstable.

Cornyebacteri- Diptheria toxin 0.03 N/A lethal.

umdiptheria

Rapid acting.

Staphylo- Staphylococcus 0.4 (aerosol 0.058 mg/ Rapid acting.

coccus enterotoxin A, B, ED50) m3
aureus C, D, E (Toxicity
is for type B)

0.3 (oral Severely

ED50) incapacitating.

20 (aerosol Symptoms persist

LD50) for almost 24 to
48 hours

3 mg/m3 Can be lethal.

Large-scale
production
feasible.

0.007 ppm Very stable.

Plants
Ricinusco- Ricin 1,000 150 mg/m3 Lethal, delayed
mmunis (air) action.

20 ppm Easily produced.

(water)

Persistent
Animals
Palythoa Palytoxin 0.08 to 0.4 0.006 Stable
(soft corals) ppm(water)

0.035 mg/ Lethal and rapid

m3 (air) acting.
Conusmagnus ~0.1 ppm Highly stable.
(fish-hunting (water)
40 cone snails)
 Biological Toxicants
Easily
synthesized.

Conusgeo- Conotoxins 3 to 6 ~0.6 mg/m3 mostly used as

graphus (air) aerosols.

Phyllobatesa- Batrachotoxin 0.1 to 0.2 0.015 mg/ Water soluble.

urotaenia m3 (air) Rapid acting and
and lethal.
Phyllobates-
terribilis

Very stable.

Can be
synthesized.

a
Assumption : (For 70 Kg adult person ) Air= 0.016 m3/min for half hour breathing,
water= 3 Lingestion or Food: 3 kg ingestion.
b
IP = intraperitoneal injection doses to mice.

Dear Learners, let us now learn about types of biological toxicants in the following
sentences:

3.2.1 Types of Biological Toxicants

 Aitoxin: It is honey bee venom and it is injected via the sting.
 Cyanotoxins: These toxins are produced by cyanobacteria.
 Cytotoxins: These are the substances which are toxic at the cell level (kills
individual cells).
 Haemotoxins: These substances are carried in bloodstream and target red
blood cells.
 Mycotoxins: These toxins are produced by various fungi.
 Necrotoxins: These substances cause tissue destruction via cell death and
are carried in the bloodstream.
 Neurotoxins: These substances affect the nervous system.

3.2.2 Food Intoxication

Food intoxication is a type of food-borne illness which is caused by ingesting
exotoxins formed by organisms such as fungi, bacteria or other microbes or by
having the food which is toxic in nature for animals or humans. Food intoxication
can lead to illness very rapidly and person can become very sick. About 300,000
people are reported to be hospitalizations every year and the mortality rate is
around 2000-4000 person.

Microbial food borne illnesses or food poisonings mainly falls into one of two
categories that is Food Poisoning Infection vs. Food Poisoning Intoxication. The
first is food infection; in this case the microorganism itself grows inside the
body. The second is food intoxication, where a chemical or natural toxin is present
in the food. Most of the bacterial food poisonings are mostly food infections.
Symptoms and Signs may start after few hours of intake of contaminated food,
41
Introduction to Toxicology or symptoms may begin after few days or even weeks later. Sickness due to food
poisoning generally persists from a few hours to several days.

3.3 CLASSIFICATION OF TOXICANTS PRESENT

IN FOOD
1) Naturally Occurring Toxicants
2) Toxicants from Microorganisms

3.3.1 Naturally Occurring Toxicants

Carbohydrates, sugars, proteins and vitamins are natural chemicals obtained from
food that are essentials for growth and health. But some food items contain natural
toxins which are potentially harmful. The cause for the presence of natural toxins
is always not known. In some food items, a toxin is available as a naturally
occurring pesticide to protect them from insect attack.

i) Lathyrism
 This is a crippling disease accompanied by paralysis of the leg muscles
occurring mostly in adults who consume large quan-tities of the seeds of
L. sativus or other lathyrus species for a long period
ii) Ackee Fruit Poisoning
 The poisonous properties of the fruit are due to unusual amino acid,
hypoglycin A, B. Both have strong hypoglycemic action resulting in coma
and death.
iii) Goitrogens
 Many food stuffs contain organic com-pounds which have goitrogenic
proper-ties.
 The red skin of groundnut contains phe-nolic glycosides which possess
goitrogenic properties.

iv) Pressure Amines

 The pressure amine foods are mainly plan-tains (green and ripe), juices
of pineapple and tomato, banana, lemon etc.
v) Argemone Seed Oil Poisoning
 The toxic substance in argemone seed is sanguinarine.
vi) Fruit Seeds and Pits
 Inner stony pit (kernel) of peaches and apricots and seeds of apple and
pear contain a naturally occurring substance known as amygdalin.
Amygdalin causes discomfort or illness by releasing hydrogen cyanide
in the gut. It can be lethal if consumed too much in a short duration of
time.
Check Your Progress 1
Note: a) Write your answer in about 50 words.
b) Check your progress with possible answers given at the end of the unit.

42
1) What do you understand by biological Toxicants? Biological Toxicants

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2) What are the types of Biological Toxicants?
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3) What is food intoxication? Give the classification of toxicants in food.
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3.3.2 Toxicants from Microorganisms

Anthrax : It is an infectious disease showing acute nature of onset
which is caused by a spore-forming bacterium known
as Bacillus anthracis. It is generally acquired after being
in contact with any anthrax-infected animals or by anthrax-
contaminated animal products.

Avian Flu : Avian influenza is a disease of birds which is highly

contagious and is currently epidemic amongst poultry in
Asia. Despite the uncertainties, poultry experts agree that
immediate culling of infected and exposed birds is the
first line of defense for both the protection of human health
as well as for the reduction of further losses in the
agricultural sector.

Hantavirus : Hantaviruses are mostly transmitted to humans from the

dried droppings, urine or saliva of mice and rats. Workers
who work in animal laboratory and in infested buildings
are at higher risk to this disease.

Molds : Molds produce and release millions of spores which are

small enough to be air, water, or insect-borne which may
have negative impacts on the human health such as
asthmas, allergic reactions and causes other respiratory
problems.

43
Introduction to Toxicology Mycotoxins : Mycotoxins are metabolites of fungus. Some of the
mycotoxins are highly toxic to the animals and are
potentially toxic even to human beings also. Recently
concern is related to their presence in many food and
theiritems carcinogenic properties.

Patulin : Patulinis mostly produced by the P. expansum. P.

expansum is related especially with a range of moldy
vegetables and fruits, in particular figs and rotting apples.
It is not available in apple beverages, like cider because it
is destroyed by the fermentation process. It is reported
that the damage of immune system of animals has been
reported but it has not shown to be carcinogenic.

Penicillium : The yellow discol-ouration in rice has been reported in

Islandieum Japan by the contamination of rice by Penicillium
islandieum during storage and develops toxic symptoms
in man.

Plague : World Health Organization reported that1,000 to 3,000

cases of plague are reported every year. A bioterrorist
release of plague can cause rapid spread of the pneumonic
form of the disease and this will have devastating
consequences.

Smallpox : It is a highly contagious disease which is unique to

humans. It is estimated that not >20 percent of the
population has any immunity from the previous
vaccination.

Tularemia : Tularemia is well known as “deer fly fever” or “rabbit

fever” and it is extremely infectious. It is an attractive
weapon for use in bioterrorism because relatively only
few bacteria are required to cause the disease.

3.4 MICROBIALAGENTS: SYMPTOMS, EFFECTS

ON HEALTH AND MANAGEMENT
Dear Learners let us now learn about the various microbial agents which can act
as a biological toxicants, it can be a bacterium, virus, protozoan, parasite, or
fungus that can transmit infection ranging from mild to severe, and in some
cases may even be life-threatening. They affect blood, skin, gastrointestinal tract
or any organ of the body. Therefore, it is important to know their symptoms and
how to prevent infections caused by them.

3.4.1 Bacterial Agents

They are defined by its virulence or ability to induce disease, mode of transmission
incubation period, and case mortality rates. Some bacteria form spores when
they find themselves in a hostile environment. The bacterium replicates its genetic
material, through a process known as sporulation, and then it surrounds it by a
thick coating. The bacterium released its water and its metabolism ceases in the
44 spore form. In spore form the bacterium can survive in extreme temperature,
radiation, and lack of nutrients, air, and water for extended periods of time and Biological Toxicants
they are revived again when nutrients are abundant. A spore is ideal as biological
weapons agent due to its virtual indestructibility.

a) Salmonellosis
Salmonella food poisoning results from ingestion of any food containing
appropriate strains of this genus in significant numbers. The salmonella are
gram negative small, non-spore forming rods that are indistinguishable from
Escherichia coli under the microscope or ordinary nutrient media.
Symptom/Effects
The salmonella has incubation period of about 12-36 hours. Salmonella does
not release any direct toxin product rather the bacterium induces the responses
in the infected animal, this may result in appearance of symptoms. The clinical
signs includes diarrhea, which may be watery, greenish and foul smelling.
Other findings include muscle weakness, moderate fever or prostration. The
main symptoms are gastrointestinal, which includes nausea, vomiting, bloody
diarrhea with mucous or abdominal cramps with possibility of rose spots. In
young children and in older age .These symptoms can be more severe. In
most of the cases symptoms resolves within 2-3 days without any
complications.
Management
There is an urgent requirement to develop new methods to control the food
poisoning and spoilage by salmonella, ordinary farms by instituting bio-
security and bio-containment practices in addition to enhanced food storage
practices, food preparation and processing method. Effective heat processing
of animal origin food products includes pasteurization of milk and eggs,
poultry thermal processing and irradiation of meat; vaccination of food
producing animals and egg-producing flocks, following good hygiene
practices during food production. Food service establishment, safe food
preparation practices, which includes through cooking, boiling of milk and
reheating of food, adequate refrigeration, prevention of cross contamination
of food; cleaning and disinfection of food preparation surfaces; exclusion of
pets and other animals from food handling areas are good practices. The
vulnerable group person should avoid raw milk, eggs or under cooked meat
and poultry products, and foods containing raw egg or unclean vegetables.

b) Staphylococcus aureus
Saureus is gram positive cocci. It occurs in the form of singles, tetrads, short
chains and irregular grape like clusters. Only those strains of Staphylococcus
which produces enterotoxins can cause food poisoning. The one who handles
the food with an active lesion or carriage later they initiate the infection.
Symptom/Effects
The characters of food poisoning caused by Staphylococcus aureus appear
in around 2-4 hours, because of shorter incubation period of bacteria. The
onset of symptoms is sudden and is characterized by vomiting and diarrhea
without any fever. This illness can remains for less than 12 hours. In severe
cases masked pallor, dehydration and collapse may need treatment including
intravenous infusion. The intoxication is characterized by short incubation
periods where illness is the results of ingestion of the toxin in the food. 45
Introduction to Toxicology Management
Food borne bacterial illness by bacteria can be commonly controlled and
prevented by proper cooking as well as storing. For example adequate
refrigeration of food, personal hygiene, adequate cooking and heating are
important. The control measures also include; a) educating people who
prepare or handle food, so that they can have proper measures; b) prohibiting
persons from handling food who have skin lesions; c) keeping food at 4
degree centigrade or lower to prevent bacterial multiplication and the
formation of toxin. Food items should not be left at room temperature for
longer times.

c) Clostridium botulinum
It is a gram positive, anaerobic and spore bearing bacilli, which has wide
distribution range in soil, decaying vegetations, sediments of lakes and ponds.
Seven different strains of the organisms (A-G) are classified based on
serologic specificity and other neurotoxin. Most reported human outbreaks
are associated with fish and sea food products.

Symptom/Effects
C. botulinium have 12 - 36 hours of incubation period. The most common
features include vomiting, thirst, dryness of mouth, constipation, ocular
paresis (blurred-vision), difficulty in speaking, breathing and swallowing.
Death occurs due to respiratory paralysis within 7 days. Clinically, botulism
recognized as a lower motor neuron disease resulting progressive flaccid
paralysis.

Management
Preformed toxin in food can completely destroyed by exposure to a
temperature of 80oC for 30 minutes or boiling for 10 minutes. Therefore all
homo canned low acid foods should be boiled before tasting for consumption.
Never taste food if it has an odor shows gas formation. Prevention of food
born botulism also depends on ensuring effective control of commercially
and home canned foods are destroying all C. botulism spores. This requires
cooking at 121°C or higher. Home canned vegetables should be boiled and
stirred for at least 3 minutes prior to serving to destroy botulism toxins.
Foods with apparent off odors or suspected odor should not be opened.

d) Clostridium perfringens
Clostridium perfringens is anaerobic spore bearing,gram-positive bacilli that
is present abundantly in the environment, vegetation, sewage and animal
feces. Organisms which produce type A enterotoxins commonly causes
perfringens food poisoning.

Symptom/Effects
The incubation period of Clostridium perfringens is about 8-24 hours. The
illness is characterized by acute abdominal pain, vomiting and diarrhea. The
classic symptoms of C. perfringens type A food poisoning are diarrhea with
lower abdominal cramps. Vomiting is not common, and fever is rare.
Symptoms typically occur within 8-24hours after ingestion of temperature
abused foods containing large number of vegetative cells of the bacteria.
46 The illness is self-limiting and the patient recovers within 8-24 hours.
 Management Biological Toxicants

The C. perfringens gastroenteritis syndrome often occurs in institutional

cafeterias thus giving proper attention towards the leading causes of food
poisoning can prevent bacterial intoxication. Cooking at temperatures below
100oC will allow the survival of the spores. The cooking process blocks
oxygen, creating real anaerobic conditions in foods like pies, gravies, strew
and rolls of cooked meat and in poultry carcass. Therefore, prevention of
vegetative cells can be done by cooking food properly and this is the most
practical way of preventing C. perfringens food borne illness.

e) Escherichia coli
Certain strain of E. colishows characteristics of hemolytic activity on blood
agar. It is motile with peritrichous flagella and often fimbriae.

Symptom/Effects
The incubation period of Escherichia coli about is 72-120 hours. The clinical
symptoms initially may be diarrhea including abdominal cramps, which may
turn into grossly bloody diarrhea in a few days. There is however, no fever.
The symptoms of E. coli septicemia are mainly referable to bacteremia, and
toxemia where the effect of bacteria localization is seen in variety of tissue
spaces throughout the body.

Management
The prevention of food borne illness caused by E.coli can be the same as
thatof other food borne illness caused by bacteria. However the consequences
in young children may be more and special precaution is needed. The
sensitivity of this organism is such that cases should not occur when food
properly cooked. In the cases of ground beef, the recommendation is that it
should be cooked to 160oF or the core temperature should be brought to a
minimum of at least 155oC for at least 15 second and that the juices are
clear. Once cooked, the food meals should not be held for more than 3-4
hours at temperatures between 40oF to 140oF.

3.4.2 Fungal Agents

Fungi are found almost everywhere. Approximately 1.5 million different types
of fungi species are found on the Earth, out of which only about 300 have the
capability of making people sick. Often the common fungi found in our
environment cause the fungal disease. Most of the fungi are not so very dangerous;
however some can be very harmful for our health.

a) Aspergillus
The most common pathogenic species are Aspergillus fumigatus and A.flavus
they causes allergic disease. Some species of Aspergillus causes disease of
grain crops, e.g. in crops of maize, where they synthesize toxins (mycotoxins
including aflatoxin). The group of diseases which are caused by infection of
Aspergillus are known as Aspergillosis.

Symptoms
The symptoms include fever, cough, fatigue pain in chest and bones or
breathlessness, and cough. Usually, only patients with weakened immune 47
Introduction to Toxicology systems or with other lung infections are susceptible. Removing patients from
high risk areas and the use of well fitted masks are two approaches that are
commonly used as anti-fungal prophylaxis. Prevention of Aspergillus
inhalation can be by nasal amphotericin B spray.

b) Cryptococcus
Cryptococcus neoformans can cause intense form of meningitis and meningo-
encephalitis in patients with HIV infection and AIDS. Those who are immune
suppressed should avoid contact with birds, digging and dusty activities in
areas heavily contaminated with bird droppings.

Symptoms
The symptoms of the infection depend on the parts of the body that are
affected. The majority of symptoms of cryptococcosis occur in the lungs or
the brain, or both. Cryptococcal infection may cause a pneumonia-like illness,
with fever, coughing and shortness of breath. There may appear skin lesions
as well. Another common form of cryptococcosis is central nervous system
infection, such as meningoencephalitis. Symptoms may include fever,
headache, or change in mental status.

c) Histoplasma
Histoplasmacapsulatum can cause histoplasmosis in humans, dogs and cats.
The fungus is most prevalent in the Americas, India and southeastern Asia.
It is endemic in certain areas of the United States. Infection is usually due to
inhaling contaminated air. Avoiding areas with bird and bat droppings may
provide some protection. Wearing a respirator face mask can provide
protection for workers in contaminated areas. Spraying soil with water before
working the soil may help prevent release of spores into the air

Symptoms
Not everyone who inhales the fungal spores becomes sick. When illness
occurs, the signs and symptoms appear in 3 to 17 days after exposure. The
symptoms are similar to pneumonia and include fever, chills, sweats, a
dry cough, malaise and chest pains. Joint pains can also be experience by
some affected peoples. In absence of any treatment, the advancement of
disease may develop weight loss, fatigue and shortness of breath in the
affected people.

d) Pneumocystis
Pneumocystis jirovecii (or Pneumocystis carinii) can cause a form
of pneumonia in people with weakened immune systems, such as the elderly,
premature children, and AIDS patients.

Symptoms
The symptoms of Pneumocystis include difficulty breathing, chest pain,
chills,cough, fatigue (tiredness) and fever. In people with HIV/AIDS,
Pneumocystis symptoms usually develop over several weeks and include a
mild fever. In people with weakened immune systems (not due to HIV/AIDS),
PCP symptoms usually develop over a few days, usually with a high fever.

48
e) Stachybotrys Biological Toxicants

Stachybotryschartarum or “black mold” can cause respiratory damage and

severe headaches. It frequently occurs in houses in regions that are chronically
damp.

Symptoms
Toxic black mold causes serious symptoms and health problems such as
mental impairment, breathing problems, damage to internal organs and
sometimes even death. The most common black mold symptoms and health
effects are associated with a respiratory response. Chronic coughing and
sneezing, irritation to the eyes, mucus membranes of the nose and throat,
rashes, chronic fatigue and persistent headaches can all be symptomatic of
black mold exposure or black mold poisoning. Often compounded by allergic
reaction to the black mold spores, these symptoms can include nausea,
vomiting, and bleeding in the lungs and nose

f) Coccidioides
Coccidioides causes Coccidioidomycosis also known as San Joaquin Fever
or Valley Fever. Inhalation of dust contaminated with Coccidioides spores
introduces the fungus to the lungs.

Symptoms
Symptoms of Valley fever may appear between 1 and 3 weeks after a person
breathes in the fungal spores. About 60% of all infected people (without
immunosuppression) have no symptoms and do not seek medical care. About
30%-35% of people who develop symptoms have flu-like symptoms (fever,
cough, malaise, and chills) that resolve over about two to six weeks without
treatment. Some may develop additional symptoms such as shortness of
breath, night sweats, headaches, sputum production, and joint and muscle
pains (symptoms resembling pneumonia).

g) Candida
Candidiasis is a common type of fungal infection caused by different species
yeast in the Candida family. Candida yeast thrives on moist surfaces of the
body and is a common cause of vaginal infections. It can also cause an
infection of the mouth or throat, known as thrush.

Symptoms
A candidiasis infection of the skin appears as a clearly defined patch of red,
itchy skin, often leaking fluid. Scabs and pustules may be seen around the
edge of the rash. It will usually be found in areas such as the groin, the folds
of the buttocks, between the breasts, toes, or fingers, and in the navel. It may
be hard to see on people with darker skin. In oral thrush white patches can
form on the tongue and gums. If the white patches are wiped away the tissue
beneath may bleed. It may become difficult to eat and the corners of the
mouth may crack.

Prevention and Management of Fungal Agents

To reduce or remove the risks, various steps are needed and it will depend upon
the distinct biohazards but there are a many common actions that can be applied:
49
Introduction to Toxicology  Many biological agents are communicated via air, such as toxins of mouldy
grain or exhaled bacteria. For those it is important to avoid the formation
of aerosols and dusts, including when cleaning or during maintenance.

 Good housekeeping, use of relevant warning signs and hygienic working

strategy are main elements of safe and healthy working conditions

 For survive or resist heat, many microorganisms have developed

mechanisms of dehydration or radiation, for example by producing spores.
Decontamination measures used for clothing, waste and equipment are
most appropriate hygienic measures for the workers that should be included.

3.4.3 Viral Agents

Viruses are very small cellular microorganisms, each of them being 15 to 400
nm in diameter and mostly containing only a single kind of nucleic acid. They
may induce various types of diseases in plants, humans, and animals. These
strictly intracellular parasites also have cellular specificity (cell tropism). The
Host organism is strongly responsible for their replication; viruses cannot be
multiplied outside the host. Different kinds of food products that get contaminated
as they are processed are commonly responsible for infecting people who eat
them. Contamination may happen through:
 Contact with animal and human faeces, or water that has been contaminated
with the faeces and is being used for irrigation and washing, etc.
 Contact with \hands or objects that have been soiled by faeces.
 Contact with contaminated water having vomit in it.
 Contact with the objects present in an environment where an infected person
had previously been present.
 Aerosol that contains virus since it is produced by an infected person. The
extent and incidence of contamination may vary between products.
a) Norovirus and Sapovirus
In year 2002, four genera of Caliciviridae were renamed means they are
getting new names. One of them is e Norovirus, which was earlier known
as Norwalklike viruses, and is small and round in structure; and another one
is Sapovirus, which was earlier known as Sapporolike viruses. The viral
genome of these two viruses contains single-stranded plus sense RNA. The
clinical indication of a Norovirus infection, is comparatively very mild. The
symptoms shown in this viruses are diarrhoea and vomiting, and in extreme
cases, convulsion. If Norovirus is introduced in a population or community,
it may then be followed by a further spread of the disease. This is caused by
the highly-infectious nature of the virus, which may result in extensive
secondary infections, and leads to 50% of the contacts. In today’s time, this
virus is at present known to be the root cause of almost all outbreaks of
nonbacterial gastroenteritis. Saproviruses primarily affect children below
the age of five. In Argentina, mixed infections with Noroviruses, Sapoviruses
and astroviruses were recorded in the faeces of children affected by acute
gastroenteritis.

50
b) Enterovirus Biological Toxicants

The enteroviruses present in human are pervasive, enterically transmitted

viruses that give rise to a severe range of illnesses in childrens as well as
infants. Choice of enteroviruses is employed because of their ability to rapidly
multiply in the intestine.
c) Hepatovirus
Hepatitis A virus (HAV) is the sole member of the genus Hepatovirus belongs
to the family Picornaviridae. HAV act as a single antigenic type; it has four
human genotypes, out of which three genotypes naturally affect the other
primates that can be discriminated like non-human primates and chimpanzee.

d) Hepatitis E virus
The virus is transmitted by the faecal-oral route with faecal contaminated
drinking water being the usual vehicle. Direct contamination is rare. Its
primary targets are young adults, between the age of 15 and 30, and the
overall death rate is between 0.5 and 3.0%. The disease is usually mild,
however, pregnant women suffer a high fatality rate from fulminant hepatic
failure.

e) Astrovirus
Astroviruses are small, 28 nm diameter non-enveloped, single-stranded RNA
viruses which constitute the only members of the familyAstroviridae. Human
astrovirus is a remarkable cause of acute diarrhea among children, result in
the outbreaks of diarrhea and occasionally in hospitalization. Disease caused
by Astrovirusis generally sensitive than that of the disease caused by
rotaviruses. However, recurrent co-infection of astrovirus with caliciviruses
and rotavirus in childhood, diarrhea complicates the epidemiology. Infections
are more common in winter.

f) Rotavirus
Rotaviruses are segmented double- stranded RNA viruses which belong to
the family Reoviridae; which may explain the presence of mixed infections
and their genetic variability. Rotaviruses can live for weeks in recreational
and potable water and for at least four hours in the hand of humans. Mostly
rotaviruses are found in waste water and can be concentrated by shellfish;
may be linked with infectious disease that is followed by seafood
consumption. The viruses are not destroyed by or relatively resistant to
commonly used hard surface disinfectants and best hygienic hand- wash
agents. It may be transmitted by contaminated hands and surfaces, faecal-
oral contact and possibly by and respiratory spread. Oral-faecal transmission
is easily transmitted by deficient sanitary conditions. In group A, Human
rotaviruses, are considered the main root cause of viral gastroenteritis in all
infants and most of the young children.

g) Flavivirus
Flavivirus is an enveloped virus with so many minuscule protuberances
having a size of 40-60 nm in diameter and it belongs to genus flavivirus.
Their genome consists of positive-sense RNA, single-stranded and non-
segmented. In flavivirus genus, many viruses are designated as B group
51
Introduction to Toxicology arboviruses. According to antigenic relationships, diverse antigenic groups
have been recognized midst the flavoviruses: Tick-borne encephalitis virus
group (TBE) or Yellow fever virus group or Dengue group etc. These
arboviruses may be present all over the world. Their vectors are particularly
known as gnats of the tick-borne encephalitis virus group.

h) Hantavirus
Hantavirus is a RNA-virus which belongs to the family Bunyaviridae and
mainly found in droppings, urine and saliva of infected deer mice and many
wild rodents. It is identified as etiologic agents of two humans diseases.
They may also cause a serious but rare lung disease called Hantavirus
pulmonary syndrome (HPS) and “hemorrhagic fever with renal syndrome”
of varying severity which is a human disease known as European
Hantaviruses.

i) Foot-and- Mouth Disease virus

The main agent which may be responsible for Foot-and- Mouth Disease
(FMD) virus is a RNA virus that belongs to the genus Aphthovirus of family
Picornaviridae. Foot and Mouth Disease is a zoonosis, which is a
transmissible disease to humans, but it may cross the species barrier with
more difficulty and with less effect.

j) Aichi Virus
The type species of a new genus, Kobuvirus, of the family Picornaviridae,
was first recognized in 1989 as the cause of oyster-associated nonbacterial
gastroenteritis in man Aichi strain is a new type of small round virus which
may mostly produces diarrhea in patients having the age group of 15 to 34
years, almost 50 to 76% of them possess neutralizing antibody.

Management of Viral Agents

Most of the food borne viral outbreaks can be identified or traced to food which
can be manually handled by an infected food handler, instead of industrially
processed foods. The viral contamination of food can be identified anywhere in
the process which is start from the farm to fork, but most of the viral infections
caused by food can be traced back to infected persons who handle food that is
not heated or otherwise that can be treated afterwards. Therefore, then emphasis
should be on personal hygiene during their preparation. During food pre-
processing, if viruses are present, then residual viral infectivity may be present
after some industrial processes. There are many other methods of inactivating
viruses except sufficient heating that exist within a food are relatively inconstant,
but viruses present in water and on the exposed surfaces can also be inactivated
with ultraviolet light or also with strong oxidizing agents. Therefore, it is important
that adequate surveillance should be given to good manufacturing practice (GMP)
and good agriculture practice (GAP) to avoid viruses introduction onto the raw
material and also into the food-manufacturing environment, and move to HACCP
to make sure adequate control over viruses that may be present during the
manufacturing process.
1) Heighten awareness about the presence and spread of these viruses by food
handlers.
52 2) Optimize and standardize methods for the detection of food borne viruses.
3) To emphasize on the consideration of viruses in all stages of production, Biological Toxicants
processing, and distribution, firstly set up the food safety quality control
and strictly apply different management systems (GHP, GMP, and HACCP)
and inspection of hygiene is more frequent of food manufacturers.

3.5 ENDOTOXINS AND ENTEROTOXINS

Toxins are having diverse nature; it includes small molecules, peptides,
lipopeptides, cyclic peptides, alkaloids, Carbamate alkaloids, organophosphates,
proteins, etc. There are two kinds of toxins called as Endotoxins and Enterotoxins.
The important difference between Enterotoxin and Endotoxin is that Enterotoxin
is a poisonous substance which may be released into or produced in intestines
of bacterial cells while Endotoxin is a poisonous substance which may be
produced within the bacterial cell while.

3.5.1 Endotoxins
Endotoxin is a poisonous substance which may present inside a bacterial cell
that may released when the disintegration of bacterial cell occurs. They are
lipopolysaccharides which are detected in the outer membrane of gram-negative
bacteria. The outer membrane is distinctive to gram-negative bacteria. Hence,
endotoxins are always related with gram-negative bacteria. Some of the species
of gram-negative bacteria such as Endotoxin is a poisonous substance which
may present inside a bacterial cell that comes out when the disintegration of
bacterial cell occurs Escherichia coli, Salmonella, Shigella, Pseudomonas,
Haemophilus influenza, Neisseria, Bordetella pertussis and Vibrio cholera are
known as endotoxin producers.

3.5.2 Enterotoxins
An enterotoxin is a protein exotoxin liberated or released by a microorganism
that may target intestine. Enterotoxins may be produced in or released into the
intestines. Enterotoxins belong to the Exotoxin category. They are proteins as
well as act as enzymes. Enterotoxins are those which may help in pore-forming
toxins. Hence, they can creating pores in the epithelial cells of the intestine wall.
The enterotoxins can increase the permeability by chloride ions in the cells of
intestinal mucosa, it leads to secretory diarrhea. E. coli and Staphylococcus aureus
are the two bacterial species which can generate such conditions by the
enterotoxins.

Activities
 Identify the poisonous substances produced by plants, animals and
microorganism which can induce harmful effect when exposed to
tissues.
 Identify the food borne diseases by the organism which are toxic to
human and animals.
 Find out the toxicity rate of the nearby areas where you are residing.
 Find out the toxicity level of the plant, animal and microorganism of
your surroundings.

53
Introduction to Toxicology Check Your Progress 2
Note: a) Write your answer in about 50 words.
b) Check your progress with possible answers given at the end of the unit.
1) What are bacterial agents and viral agents?
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2) Write the methods for prevention of fungal agents.
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3) What are endotoxins and enterotoxins?
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3.6 LET US SUM UP

Biological Toxicants are chemicals that are toxin produced by living things in
nature. Different type of living things such as bacteria in the animal gut, various
plants which may produce potent toxins such as spore-forming bacteria producing
anthrax toxin, ricin, red algae producing saxitoxin, certain insects and animals
that may produce toxic venom, etc.; the list is very long. A terrorist or a nation
state can potentially harvest and purify a toxin to be used against an enemy,
perhaps by poisoning a water supply or by releasing a dust/aerosol in an airplane
passenger compartment or inserting a fine powder in a letter. However, most
poisonings occur accidentally when people eat contaminated food. Food
intoxication is a type of food-borne illness which may be caused by ingesting the
exotoxins made by different types of organisms such as bacteria, fungi etc or by
consuming/taking the food items that are naturally toxic to humans and animals.
Food intoxication leads to onset of illness which is usually very rapid and people
feel sick. Every year around 300,000 peoples are reported to be hospitalizations
and the fatality is almost 2000-4000.
Microbial food poisonings / food borne illnesses mainly fall under the one of the
two categories that is known as Food Poisoning Infection vs. Food Poisoning
Intoxication. The first is food infection, where the microorganism itself grows
inside body. The second is food intoxication, where a chemical or natural toxin.
54 Most bacterial (microbial) food poisonings are mainly food infections. Signs
and Symptoms may start within hours after eating the contaminated food, or Biological Toxicants
they may begin days or even weeks later. Sickness caused by food poisoning
generally lasts from a few hours to several days. The steps required to remove or
reduce the major risks will be contigent upon the specific biohazard, but there
are many common actions that may be tried or applied like various biological
agents may be communicated through air, such as exhaled toxins of mouldy
grain or bacteria. For those it is important to avoid the formation of aerosols and
dusts, including when cleaning or during maintenance. Secondly, use of relevant
warning signs, good housekeeping and hygienic working strategy are major key
components of safe and the healthy working conditions. In some instances
preventive measures can include vaccination that can be given to workers on a
voluntary basis.

3.7 KEY WORDS

Endotoxin: It is a poisonous substance which may present inside a bacterial
cell that may released when the disintegration of bacterial cell
occurs. They are lipopolysaccharides which are detected in the
outer membrane of gram-negative bacteria.

Rotaviruses: They are segmented double- stranded RNA viruses which belong
to the family Reoviridae.

Mycotoxins: Mycotoxins are metabolites of fungus. Some of the mycotoxins

are highly toxic to the animals and are potentially toxic even to
human beings also.

3.8 REFERENCES AND SUGGESTED FURTHER

READINGS
Ananthanarayan, R. and C. K. J. Paniker (2009): Ananthanarayan and Paniker’s
Textbook of Microbiology.8th Edition. Universities Press.
Borriello, S. P.; Murray, P. R. and G. Funke (2007): Topley& Wilson’s
Microbiology and Microbial Infections. 10th Edition, Vol. 2.
Dey, N.C. (1965): Medical Bacteriology. 4th Edition. Allied Agency, Calcutta.
Kalia, A. N. (2009): Textbook of Industrial Pharmacognosy. CBS Publishers &
Distributors: 188-203.
http://www.differencebetween.com/ difference-between- endotoxin and-vs-
enterotoxin (Retrieved on 07 July, 2017).
Madigan, M. T.; J. M. Martinko and J.Parker (2003): Brock Biology of
Microorganisms. 10th Edition. Pearson Education.
Nester, E.; Anderson, D. and Jr. C. E. Roberts (2011): Microbiology: A Human
Perspective 77thEdition. McGraw Hill Company, New York, America.
Prescott, L. M. (1999): Microbiology.4th Edition. McGraw-Hill Company.
Suresh, M. X. and R. B. Kumar (2014): A Review on Biological Toxins: Their
Pharmacological Significance and Structural Importance. Int. J. Pharm.
Sci. Rev. Res., 28(2): 128-136. 55
Introduction to Toxicology Thomas, E. M.; Huey, B. M.; Downing, E. and L. M. Duffy (2000): Strategies to
Protect the Health of Deployed U.S. Forces (Detecting, characterizing,
and Documenting Exposures). National Academy Press, Washington,
D. C.: 59.

Tortora, G. J. (2016): Microbiology: An Introduction. 8th Edition. Pearson

Education.

3.9 ANSWERS TO CHECK YOUR PROGRESS

Answers to Check Your Progress 1
Your answer should include the following points:
1) Biological toxicants are the poisonous substances which are released by the
living organisms either for their defense like in the bees, ants, wasp or for
predation like in the spiders, snakes, scorpion etc. Biological toxins are
non–replicative, non-infectious material that can be hazardous in a very
small quantity.
2) Biological toxicants are of various types depending upon the specific target
they attack like Necrotoxins (cause tissue destruction via cell death),
Neurotoxins (affect the nervous system), Haemotoxins (carried in the
bloodstream and target RBCs), Cytotoxins, (kills individual cells). Biological
Toxins can be classified according to their production source like,
Cyanotoxins (produced by cyanobacteria), Mycotoxins (produced by fungi),
Aitoxin (injected via the sting).
3) Food intoxication results when a person eats food containing toxins that
cause illness. Toxins are produced by harmful microorganisms, the result of
a chemical contamination, or are naturally part of a plant or seafood. Some
bacteria, all viruses, and all parasites cause foodborne illness via infection.
Food intoxicants can be classified as: (a) Naturally occurring toxicants (b)
Toxicants from microorganisms.
Answers to Check Your Progress 2
Your answer should include the following points:
1) Bacteria are free-living organisms that are one celled, microscopic having
size range from 1µm-10µm. Bacterial agents are defined by their ability to
induce disease, incubation period, mode of transmission and case fatality
rates. Salmonellosis, Clostridium botulinum, Clostridium perfringers,
Staphylococcus aureus etc. are some of the examples of disease causing
harmful bacterial agents.

Viruses are small (15-400nm diameter), unicellular stern intracellular parasite

which are not replicateoutside a particular living cell, viruses are not having
its own metabolism as it is a inert particles. The more the time they survive
in the environment which is infectious, the more the chance of transmission
and to proliferate infection.Viral agents are the most known pathogens
transmitted through food. Some of the viral agent causing various diseases:
Enterovirus, Hepatovirus, Astrovirus, Flavivirus, Rotavirus, Aichi virus etc.

2) The steps required needed to remove or reduce the risks will depend upon
the specific fungus and their mode of transmission and infection, however
56
 there are a number of common act that can be tried or applied like hygienic Biological Toxicants
working procedures, avoid the formation of aerosols, use of relevant warning
signs and dusts, good housekeeping, and decontamination methods used
for waste, appliances and clothing and use suitable hygienic measures.

3) Endotoxin is a substance which is poisonous in nature and may be present

inside the cells of bacteria that comes out when the disintegration of bacterial
cell occurs. Endotoxins are composed of lipopolysaccharides having high
molecular weight which are detected in the outer membrane of the gram-
negative bacteria. Endotoxins are substances which are heat stable and non-
soluble in nature. Hence, it cannot be tear down by boiling.

An enterotoxin is a protein exotoxin released by a microorganism that targets

intestines. Enterotoxins are released or produced in or into intestines.
Different types of bacterial species are capacity of producing enterotoxins.
Enterotoxins belong to the Exotoxin category. They are proteins and can act
as enzymes. Enterotoxins help in formation of pore-forming toxins.

57
Introduction to Toxicology
UNIT 4 TOXICITY ASSESSMENT
Structure
4.0 Introduction
4.1 Objectives
4.2 Overview of Toxicity Assessment
4.3 Routes of Exposure
4.4 Toxic Effect
4.4.1 Acute Effects
4.4.2 Chronic Effects
4.5 Dose Response Assessment
4.6 Dose - Response Curve
4.7 LD50 and LC50
4.8 Assessing Toxicity
4.8.1 Assessing Acute Toxicity
4.8.2 Assessing Chronic Toxicity
4.9 Let Us Sum Up
4.10 Key Words
4.11 References and Suggested Further Readings
4.12 Answers to Check Your Progress

4.0 INTRODUCTION
Toxicity may be expressed in a variety of forms depending on the chemical which
is involved, the nature of the population exposed, and the conditions of exposure.
We can define toxicity as the ability of a substance to cause adverse effects in
living organisms. This ability to cause adverse effect is dependent upon several
conditions. For example, it will depend on the quantity or the dose of the substance
and whether the effects of the chemical are toxic, nontoxic or beneficial. In
addition to dose, other factors may also influence the toxicity of the compound
such as the route of entry, duration and frequency of exposure, variations between
different species (interspecies) and variations among members of the same species
(intraspecies). The potential hazards to humans may be acute, subchronic and
chronic. The more specific types of toxicity that are determined include
carcinogenicity; teratogenicity and reproductive toxicity, mutagenicity and
neurotoxicity.

Toxicity testing is necessary to provide some basis for the regulation of substances
those humans and other living things come into contact. It is used to determine
the safety of range of chemicals used for example cosmetics, pharmaceuticals,
food additives, pesticides, chemicals, additives and consumer products. A toxic
effect can result from a natural or a manufactured substance and manifest a variety
of symptoms which may be both immediate and long-term. Toxicity testing
introduces a variety of methods and rates of exposure to the test organism, which
58
help in formulating a more accurate assessment of the risk of harm that the test Toxicity Assessment
substance may pose to human health and the environment. Toxicity assessment
is characterization of the toxicological properties and effects of a chemical, with
special emphasis on establishment of dose-response characteristics. This unit
focuses on toxicity assessment and role of toxicity assessment in toxicological
studies.

4.1 OBJECTIVES
After completing this unit, you will be able to:
 understand toxicity assessment and the routes of exposure,
 describe the dose response relationship,
 explain the lethal dose and lethal concentration, and
 discuss the various methods for toxicity assessment.

4.2 OVERVIEW OF TOXICITY ASSESSMENT

The term toxicity refers to the inherent potential of a substance to cause systemic
damage to living organisms. All substances are toxic in quantity. Many therapeutic
medications are acutely toxic, but beneficial when used at the appropriate level.
Vitamin D, table salt, oxygen, and water are toxic in quantity. Thus, the mere
presence of a substance does not automatically imply harm. This is why toxicity
assessment is concerned with the type and degree of harm caused by differing
amounts of a substance. A toxicity assessment is a tool to investigate the potential
for a substance to cause harm and how much causes what kind of harm. Toxicity
assessment is a major component of risk assessment.

A toxicity assessment provides information on how much of a chemical causes

what kind of harm. If the toxicity assessment is based on an animal study, the
degree of harm to humans must be extrapolated using mathematical models based
on a variety of assumptions. Thus the toxicity assessment provides only an
estimate of the harm to humans. As more toxicity studies on a particular chemical
are conducted-dose-response studies on different species of animals for example,
or epidemiological and in vitro (test tube) studies-scientists become more
confident in their characterization of the toxicity of the substance. The risk
assessor’s job is to determine the real world risk to humans of a substance by
combining information on toxicity and exposure. This job is made more
complicated if data are collected from many different studies, but the results will
be more likely to reflect the best estimates scientists can make.

4.3 ROUTES OF EXPOSURE

There are many routes by which a substance can enter the body. The major routes
are discussed below:

1) Inhalation: The major route of entry for most chemicals in the form of
vapors, gases, mists, or particulates is inhalation. Once inhaled, chemicals
are either exhaled or deposited in the respiratory tract. The deposited chemical
can, damage through direct contact with tissue or the chemical may diffuse
into the blood through the lung-blood interface.
59
Introduction to Toxicology 2) Absorption: The absorption of the chemical may be either through skin
(dermal) or through eye. Dermal contact can cause redness or mild dermatitis
or more severe effects like destruction of skin tissue or other debilitating
conditions. Many chemicals can also cross the skin barrier and be absorbed
into the blood system. Once absorbed, they may produce systemic damage
to internal organs.

3) Transformation:Many chemicals are metabolized or transformed via

chemical reactions in the body. In some cases, chemicals are distributed and
stored in specific organs.

4) Ingestion: Chemicals that inadvertently get into the mouth and are swallowed
do not generally harm the gastrointestinal tract itself unless they are irritating
or corrosive. Chemicals that are insoluble in the fluids of the gastrointestinal
tract (stomach, small, and large intestines) are generally excreted. Others
that are soluble are absorbed through the lining of the gastrointestinal tract.
They are then transported by the blood to internal organs where they can
cause damage.

5) Injection: Substances may enter the body if the skin is penetrated or

punctured by contaminated objects. Effects can then occur as the substance
is circulated in the blood and deposited in the target organs.

6) Storage: Storage may reduce metabolism and therefore, increase the

persistence of the chemicals in the body. The various excretory mechanisms
(exhaled breath, perspiration, urine, feces, or detoxification) rid the body,
over a period of time, of the chemical. For some chemicals elimination may
be a matter of days or months; for others, the elimination rate is so low that
they may persist in the body for a lifetime and cause deleterious effects.

4.4 TOXIC EFFECT

Toxic effect is defined as an adverse change in the structure or function of an
experimental animal as a result of exposure to a chemical substance.” Such
changes may be effected via acute, sub chronic or chronic exposure studies.
Acute toxicity tests measure the immediate effects of exposure with an estimated
time for peak effect of approximately eight hours after the initial exposure.
Toxicity tests usually focus on:
1) cytotoxicity (damages cells),
2) mutagenicity (alters genetic materials),
3) carcinogenicity (causes cancer)
4) teratogenicity (causes birth defects).
Toxic effects are classified as either acute or chronic.

4.4.1 Acute Effects

Acute effects happen very rapidly after a single exposure has occurred (food
poisoning, breathing fumes from a chlorine spill). Sweating, nausea, paralysis,
and death are examples of acute effects. In acute effect, there is a rapid and
serious response to a high but short lived dose of toxic chemical. Acute poisons
60
interfere with essential physiological processes, leading to a variety of symptoms Toxicity Assessment
of distress and, if the interference is severe, to death.

Acute toxicity is relatively easy to measure. At high enough levels, the effects of
toxins on bodily function are obvious and fairly consistent across individuals
and species. These levels vary enormously for different chemicals. Almost
everything is toxic at some level, and the difference between toxic and nontoxic
chemicals is a matter of degree. The most widely used index of acute toxicity is
LD50, the lethal dose for 50% of a population. This number is obtained by
graphing the number of deaths among a group of experimental animals, usually
rats, at various levels of exposure to the chemical, and interpolating the resulting
dose response curve to the dose at which half the animals die. The dose is generally
expressed as the weight of the chemical per kilogram of body weight, on the
assumption that toxicity scales inversely with the size animal.

4.4.2 Chronic Effect

Chronic effects happen only after repeated long-term exposure (cigarette smoking,
eating foods with low levels of contaminants, breathing polluted air). Cancer,
organ damage, reproductive difficulties, and nervous system impairment are
examples of chronic effects. In a chronic effect, in which the dose is relatively
low but prolonged, and a time lag occurs between initial exposure and the full
manifestation of the effect. Chronic toxins have more subtle effects, often setting
in motion a chain of biochemical events that leads to disease states, including
cancer.

Chronic effects are much more difficult to evaluate, especially at the low exposure
levels that are likely to be encountered in the environment. In an experimental
setting, the lower the dose, the fewer the animals that show any particular effect.
To obtain statistically significant results, a study might have to include a
prohibitively large number of animals. The only available recourse is to evaluate
effects of a series of high doses, and then to extrapolate the dose response curve
to the expected incidence at low doses. But extrapolation may have to extend
over several orders of magnitude, and there is no assurance that the actual dose
response function is linear. The biochemical mechanisms that control effects
may be different at high and low doses. The controversy over this issue is
especially heated in the context of animal testing for cancer.
Chronic effects fall into two categories: carcinogenic effects and non-carcinogenic
effects.
Examples of non-carcinogenic chronic effects:
 Organ damage: Cirrhosis of the liver from long-term alcohol consumption;
emphysema from long-term tobacco smoking.
 Reproductive difficulty: Decreased fertility from the pesticide DBCP
(dibromochloropropane).
 Nervous system impairment: Mental retardation in people exposed to high
levels of lead during early childhood
Check Your Progress 1
Note: a) Write your answer in about 50 words.
b) Check your progress with possible answers given at the end of the unit.
61
Introduction to Toxicology 1) What do you understand by toxicity assessment?
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2) Enlist the different routes of exposure of toxic chemicals in the body.
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4.5 THE DOSE RESPONSE ASSESSMENT

The amount of a chemical that an organism (such as a person) is exposed to is
called the dose, and the severity of the effect of that exposure is called the response.
A dose response assessment is a scientific study to determine the relationship
between dose and response, and how much dose is correlated with how much
response. Response can be expressed as measured or observed incidence, percent
response in groups of subjects (or populations), or as the probability of occurrence
within a population. The process of characterizing the relation between the dose
of an agent administered or received and the incidence of an adverse health
effect in exposed populations and estimating the incidence of the effect as a
function of human exposure to the agent. It takes into account the intensity of
exposure, age pattern of exposure, and possibly other variables that might affect
response, such as sex, lifestyle, and other modifying factors. A dose response
assessment usually requires extrapolation from high to low dose and from animals
to humans.

The second step in the assessment of risks to humans from potentially toxic
agents, in which the relationship between the dose levels to which animals or
humans are exposed and the health effect responses at each dose level are
characterized quantitatively. Often the close response assessment is based on
high dose experimental animal studies and applied to humans who are exposed
at much lower doses. The process of characterizing the relation between the
dose of an agent administered or received and the incidence of an adverse health
effect in exposed populations and estimating the incidence of the effect as a
function of human exposure to the agent.

4.6 DOSE-RESPONSE CURVE

Dose response curve is a mathematical relationship between the dose administered
or received and the incidence of adverse health effects in the exposed population;
toxicity values are derived from this relationship. The characteristics of exposure
to a chemical and the spectrum of effects caused by the chemical come together
in a correlative relationship that toxi-cologists call the dose response relationship.
62
This relationship is the most fundamental and pervasive concept in toxicology. Toxicity Assessment
To understand the poten-tial hazard of a specific chemical, toxicologists must
know both the type of effect it produces and the amount, or dose, required to
produce that effect. The relationship of dose to response can be illustrated as a
graph called a dose response curve. There are two types of dose-response curves:
one that describes the graded responses of an individual to varying doses of the
chemi-cal and one that describes the distribution of responses to different doses
in a population of individuals. The dose is represented on the x-axis and response
on the y axis. An important aspect of dose-response relationships is the concept
of threshold.

For most types of toxic responses, there is a dose, called a threshold, below
which there are no adverse effects from exposure to the chemical. The human
body has defenses against many toxic agents. Cells in human organs, especially
in the liver and kidneys, break down chemicals into nontoxic substances that
can be eliminated from the body in urine and feces. In this way, the human body
can take some toxic insult (at a dose that is below the threshold) and still remain
healthy. The identification of the threshold beyond which the human body cannot
remain healthy depends on the type of response that is measured and can vary
depending on the individual being tested. Thresholds based on acute responses,
such as death, are more easily determined, while thresholds for chemicals that
cause cancer or other chronic responses are harder to deter-mine. Even so, it is
important for toxicologists to identify a level of exposure to a chemical at which
there is no effect and to determine thresholds when possible.

The horizontal axis indicates the dose in mg/kg of body weight, while the vertical
axis is the percent of maximum response. For a very low dose there is no or little
response. The response increases with the dose until the maximum response is
reached and increasing the dose has no additional effect.

4.7 LD50 AND LC50

Lethal dose (50) or LD (50) is standard measure of the toxicity of a material that
will kill half of the sample population of a specific test animal in a specified
period through exposure via ingestion, skin contact, or injection. LD50 is
measured in micrograms (or milligrams) of the material per kilogram of the test-
animal’s body weight; lower the amount, more toxic the material. Used in
comparison of toxicities, LD50 values cannot be directly extrapolated from one
species to the other or to humans. Also called median lethal dose, LD50 (originally
abbreviated as DL50 for dosis letalis, 50%) is a test used in animal experiments.
It was designed by the British pharmacologist J W Trevan in 1927. In other
words LD50 is the dose of any substance tested required to kill half the number
(50%) of test animals. The test shows how much of a substance must be taken
before it becomes deadly.

Example: A reported “rat oral LD50 of 50 mg/kg” means that half of the rats
that ingested a dose of 50 milligrams of the substance per kilogram of body
weight died within 14 days.

The LD/50 test is used to determine the acute toxicity of a substance. This is the
dose at which the test substance is lethal to 50% of the test animals. During the
test period the animal forcibly inhales, ingests or is otherwise exposed to the
63
Introduction to Toxicology substance. Often the animals involved experience acute distress including “pain,
convulsions, discharge, diarrhea and bleeding from the eyes and mouth.”

LC stands for “Lethal Concentration”. LC values usually refer to the

concentration of a chemical in air but in environmental studies it can also mean
the concentration of a chemical in water. According to the OECD (Organization
for Economic Cooperation and Development) Guidelines for the Testing of
Chemicals, a traditional experiment involves groups of animals exposed to a
concentration (or series of concentrations) for a set period of time (usually 4
hours). The animals are clinically observed for up to 14 days. The concentration
of the chemical in air that kills 50% of the test animals during the observation
period is the LC50 value. Other durations of exposure (versus the traditional 4
hours) may apply depending on specific laws.

The LD50 and LC50 are specific cases of the generalized values LDn and LCn.
The LDn is the dose of a toxicant lethal to n% of a test population. The LCn is
the exposure concentration of a toxicant lethal to n% of a test population. Thus,
the LD50 is the statistically derived single dose of a chemical that can be expected
to cause death in 50% of a given population of organisms under a defined set of
experimental conditions. Similarly, the LC50 is the statistically derived exposure
concentration of a chemical that can be expected to cause death in 50% of a
given population of organisms under a defined set of experimental conditions.

Another important value that may be derived from the relationship shown is the
threshold dose or concentration, the minimum dose or concentration required to
produce a detectable response in the test population. The threshold value can
never be derived with absolute certainty and therefore the lowest observed effect
level (LOEL) or the NOEL have normally been used instead of the threshold
value in deriving regulatory standards. There is a move to replace these values
by the benchmark dose (BMD). This is defined as the statistical lower confidence
limit on the dose that produces a defined response (called the benchmark response
or BMR, usually 5 or 10%) in a given population under defined conditions for
an adverse effect compared to background, defined as 0%.

In assessing the significance of LD50 or other toxicological values, it is necessary

to note the units used in expressing dosage. Normally dosage is expressed in mg
kg_1 body weight, but it may be expressed as mg cm_2 body surface area as this
has been shown in a number of cases to permit more accurate extrapolation
between animals of different sizes and from test mammalian species to humans.
For biocides, selective toxicity is the key property, since they are to be used to
kill pests with minimal harm to other organisms. Selective toxicity depends upon
differences in biological characteristics that may be either quantitative or
qualitative. Minimizing the amount of pesticide used and targeting its application
is crucial to avoid harm to non-target organisms. On a body weight basis, it is
assumed for toxicity data extrapolation that humans are usually about 10 times
more sensitive than rodents. On a body surface area basis, humans usually show
about the same sensitivity as test mammals, i.e. the same dose per unit of body
surface area will give the same given defined effect, in about the same percentage
of the population. Knowing the above relationships, it is possible to estimate the
exposure to a chemical that humans should be able to tolerate. The toxicity
rating can be different for different animals. Table 4.1 gives the toxicity classes
given by Hodge and Sterner Scale.
64
 Table 4.1: Toxicity Classes: Hodge and Sterner Scale Toxicity Assessment

Routes of Administration
Oral Inhalation Dermal Probable
LD50 LC50 LD50

Toxicity Commonly (single dose (exposure of (single Lethal

Rating Used Term to rats) rats for 4 application Dose for
mg/kg hours) ppm to skin of Man
rabbits)
mg/kg

1 Extremely 1 or less 10 or less 5 or less 1 grain

Toxic (a taste,
a drop)

2 Highly Toxic 1-50 10-100 5-43 4 ml (1 tsp)

3 Moderately 50-500 100-1000 44-340 30 ml

Toxic (1 fl. oz.)

4 Moderately 50-500 100-1000 44-340 30 ml

Toxic (1 fl. oz.)

5 Practically 5000 – 10,000 – 2820 – 1 litre

Non-toxic 15,000 100,000 22,590 (or 1 quart)

6 15,000 or 100,000 22,600 or 1 litre 15,000

more more (or 1 quart) or more

Source: Nancy M. Trautmann, 2001

Table 4.2: Toxicity Categories Used for Human Poisons

Toxicity LD 50 Probable Lethal Example

Category (mg/kg) Dose for 70 kg Compounds
Human Adult

Super toxic <5 <0.35 g Botulin Aflatoxin

Extremely toxic 5¨ 0.35³.5 g Cyanide Vitamin D

(calciferol)

Very toxic 50@ 3.5 g Nicotine Caffeine

Moderately toxic 500µ,000 35è g Aspirin

(acetylsalicyclic
acid) Salt (sodium
chloride)

Slightly toxic 5,000±5,000 350±,050 g Ethanol

Trichloroethylene

Practicallynontoxic >15,000 >1,050 g Sugar (sucrose)

Source: Nancy M. Trautmann, 2001

65
Introduction to Toxicology The toxicity of a chemical depends on many factors, including whether it gets
broken down, is stored in the body, or is excreted.

4.8 ASSESSING TOXICITY

All quantitative toxicity assessments are based on the dose-response concept.
With the increase in the dose (exposure), the response (toxicity) also increases.
The smaller the dose needed to cause an effect, the more potent (toxic) the
substance is. For all compounds other than cancer-causing agents (carcinogens),
it is assumed that there is a dose below which no effect occurs (a threshold). This
is similar to a drug where too small of a dose has no beneficial effect. For
carcinogens, it is often assumed that even the smallest dose can cause an effect.
Although the dose response concept is used in all types of toxicity assessments,
it is used somewhat differently for each of them. The toxicity assessment
commonly involves two steps: hazard identification and dose response
assessment. The outcome of a toxicity assessment is usually expressed as a toxicity
value, such as a reference dose (RfD) or cancer slope factor (CSF), which
incorporates the findings of the hazard and dose-response assessments and safety
factors that address uncertainties in the assessment. A toxicity assessment may
also conclude that a toxicity value cannot be developed because of inadequate
data.

 Acute toxic effects are estimated by LD50 studies or observation of

accidental exposures. Acute toxicity is assessed using observations of
accidental human exposures or by conducting LD50 tests on experimental
animals, usually rodents.

 Chronic toxic effects are estimated by dose-response studies on animals.

 Carcinogenic effects are estimated by a type of dose-response study called

a carcinogenesis bioassay.

Toxicity Testing
Toxicity can be evaluated in whole organisms (in vivo) or using molecules or
cells (in vitro). The main advantage of toxicity testing is that it detects toxic
compounds based on their biological activity, and as such does not require a
prior knowledge of the toxicant to identify its presence (unlike chemical analysis).
Identification of the toxic component is required. Once a suspected toxicant is
identified, modelling approaches (in silico) can be used to predict its toxicity
based on the physico-chemical properties of the compound and its likely fate
and transport in the environment.

Direct Toxicity Assessment

1) In Vivo Bioassays

Conventional toxicity testing relies on direct toxicity assessment in whole

organisms (algae, shrimp, sea urchins, fish, rats, etc.).The organisms are
exposed to the chemical(s) or mixture(s) of interest and monitored for any
sign of adverse health effect. This can be either a gross morphological effect
(such as weight loss, visible lesions, death) or more subtle biochemical
markers, these being either biomarkers of exposure (an indicator of the
66 internal dose, such as a metabolite in urine) or biomarkers of effect (an
 indicator of a health effect, such as enzyme activity). The duration of the Toxicity Assessment
exposure depends on the type of toxicity detected or being monitored, from
short-term acute effects (96 h or less), sub-acute (a couple of days), sub-
chronic (a couple of weeks) to chronic effects (a significant portion of the
organism’s life expectancy). Depending on the species used, in vivo toxicity
testing is generally seen as the most relevant predictor of human health
effects. This is because in vivo tests include a measure of absorption,
distribution, metabolism and excretion, all of which could modulate the
toxicity of the sample.

2) In Vitro Bioassays (Bioanalytical Methods)

In vitro bioassays have been in use for drug discovery by the pharmaceutical
industry for decades. In in vitro bioassays, molecules (e.g. enzymes) or
whole cells are exposed to the chemical(s) or mixture(s) of interest and
monitored for specific responses. However, for chemically-induced toxicity
the initial interaction of the chemical at the molecular or cellular level is a
necessary (but not sufficient) prerequisite for toxicity This is because toxicity
occurs at the site of interaction of the toxicant (which can be either the
parent compound or a metabolite) and the target biomolecule (“primary
effect”). Organisms, however, have defense and detoxification mechanisms
to cope with a certain degree of primary toxicity, and it is only when those
defense mechanisms overcome, the observable toxicity occurs (“secondary
effect”). This means that in vitro toxicity is likely to occur at significantly
lower doses than in vivo effects but also means that a substance can be
toxic in vitro but not in vivo. A variety of toxic effects can be monitored in
vitro, from basal toxicity (cytotoxicity) and reactive toxicity (interaction
with protein or DNA, which can then lead to carcinogenicity) that can
potentially affect all cells, to specific toxicity that may only affect certain
cells or organs (e.g. endocrine effects, neurotoxicity, immunotoxicity, liver
toxicity, etc.). Typically, in vitro tests are carried out on specific cell types
depending on the endpoint of interest. Some assays can be more variable
than others, and thorough quality assurance / quality control procedures
such as consistent use of positive and negative controls, monitoring of assay
performance with control charts, quantification of detection limits,
determination of reproducibility and robustness, use of inter-assay samples,
intra- and inter-assay duplication and adoption of Good Laboratory Practices
ensure the production of reliable high-quality data. Each type of bioassay
has its advantages and limitations, and no single assay can provide a
complete assessment of the biological activity of a sample. Therefore a
battery of bioassays is required to rigorously assess the potential of a sample
to cause biological effects in exposed organisms. In vitro assays are generally
high-throughput short-term (<1 week) assays that provide a quick
measurement of potential toxicity in a sample. These methods are presently
at different stages of development and not all are presently suitable for
inclusion in a monitoring program.

In Direct Toxicity Assessment

In silico approaches
Some of the shortcomings of in vitro bioassays, particularly the lack of integration
of toxicokinetics, can be partly overcome by combining them with computer (in
silico) modelling using structure-activity relationships (SAR). In SAR, the 67
Introduction to Toxicology chemical structure and other physico-chemical properties of the substance (once
it is known) can be used to predict its toxicokinetics. In silico methods are very
useful in the absence of other toxicological data, but are based on data from
other chemicals and as such should be viewed with appropriate caution

4.8.1 Assessing Acute Toxicity

Acute Toxicity Tests
Acute toxicity standard tests though appears to be simple, routine methods but
they should be performed strictly in accordance with standard requirements,
observing recommended conditions and using indicated organisms. The tests
are conducted either to determine the concentration of a test material (a chemical,
or effluent) or the level of an agent (e.g., temperature or pH) that produces a
deleterious effect on a group of test organisms during short-term exposure under
controlled conditions. These tests are performed to determine the concentration
of a particular chemical that would elicit a specific response or measurable
endpoint from a test species in a relatively short period of time, usually from two
to seven days. These tests are also applied in order to compare the sensitivity of
different species or organisms at different stages of their development, to find
the relationship with other variables that can affect overall results. They are also
widely used for the assessment of the degree of potential harm to the environment
caused by industrial discharges from the already existing or prospective factories
or/and the application of new industrial processes. These tests must meet the
following main requirements:
 They must be applicable to a wide range of chemicals
 They must be short, reproducible, sensitive, cheap and of low variability
 They must be standardized, accepted by the regulatory and scientific
community.

4.8.2 Assessing Chronic Toxicity

Chronic Toxicity Tests
Chronic toxicity tests are more complex and the effects are studied for longer
periods of time. The aim of these tests is to determine the concentration of a test
material/ substance (e.g., a chemical, or effluent) that produces an adverse effect
on a group of test organisms during long-term exposure under controlled
conditions. Usually, chronic tests are conducted with a set of species of different
phylogenetic level and different stage of development. Unlike acute toxicity tests,
chronic toxicity studies evaluate not only mortality, but also investigate endpoints
such as individual growth/growth rate, abnormal development, hatching time
and success, reproduction (the total number of young individuals) and vitality of
offspring, behaviour of individuals, physiological parameters, or histology. Using
these methods, NOEC and LOEC can be calculated. NOEC (no observed effect
concentration) is the highest concentration of the test substance that does not
cause any observed and statistically significant adverse effects on the exposed
organisms compared with controls. LOEC (the lowest observed effect
concentration) is the lowest concentration of the substance used in a test that has
statistically significant adverse effect on the exposed population of test organisms
compared with controls. As legislation aims at preventing effects by reducing
the amount of emitted chemicals, NOECs play a major role in applied
68 ecotoxicology. Obtained results can be calculated to estimate MATC (Maximum
allowable toxic concentration), which is the geometric mean of LOEC and NOEC. Toxicity Assessment
Three categories of tests are commonly used to predict the chronic effects of
toxic chemicals on aquatic organisms and data from these categories of tests can
be used to estimate PNEC (Predicted no effect concentration). Chronic toxicity
tests may be divided into: Full life cycle tests, i.e. those which evaluate the effects
of a series of chemical concentrations on the reproduction, growth, survival and
other parameters of one or more generations of a population of test organisms.
Full life cycle tests are often applied to algae or invertebrate species which may
be performed for more than one complete life cycle. However, they are not carried
out on fish, as their performance requires long duration (till 1.2 years). Partial
life cycle or sensitive life stage tests (embryolarvae of fish), i.e. those which
evaluate survival, morphological, physiological parameters, hatching success,
development abnormality etc. Duration of partial life cycle tests on fish can be
as short as 7 days or as long as 60 days. However, the early life stage of fish
(embryo/larval period) is the most sensitive period of fish life cycle. Therefore,
embryo/larval tests are used for the more exact evaluation of substance toxicity
to fish.
Chronic Toxicity Can Be Divided Into Two Categories:
 Cancer (carcinogenic toxicity).
 All other effects (non-carcinogenic toxicity).
Cancer is in a separate category because public concern about it is so great.
People want to know if even one person in a million persons who are exposed to
a substance will get cancer. To discover this, researchers conduct a specific type
of dose-response study- a carcinogenesis bioassay. Non-carcinogenic effects are
usually assessed with a different type of dose-response study.

Non-Carcinogenic Assessment
Non-carcinogenic chronic toxicity is assessed by studies to determine the smallest
dose that causes any detectable effect. Scientists assess non-carcinogenic chronic
toxicity by administering varying amounts of a substance (dose) to laboratory
animals and noting the effects (responses), if any, at each dose. Essentially, the
scientists look for the smallest dose that causes any detectable effect. This smallest
dose is called the Lowest Observable Effect Level (LOEL).

Carcinogenesis Bioassay
Scientists assess carcinogenic toxicity very differently than they assess non-
carcinogenic toxicity. This is in response to public fear about cancer. People
want to know if even one in a million individuals will get cancer from exposure
to a suspected carcinogen. To find this out with any degree of confidence by
traditional dose-response studies, scientists would have to use several million
test animals. The impracticality of such experiments has led to the development
of the carcino-genesis bioassay. With a carcinogenesis bioassay, scientists are
not looking for the safe level of exposure (NOEL). Rather, harm is assumed, and
they are looking for the incidence, or risk, or harm. The carcinogenesis bioassay
is a method of testing substances for carcinogenic effects that utilizes high-dose
studies on laboratory animals to look for even the rare case of cancer. It is not
necessarily the best scientific approach to assess the carcinogenic effects of
chemicals. Instead it is a way to respond to public concerns by generating
carcinogenic risk values with large margins of safety.
69
Introduction to Toxicology Example of A Carcinogenesis Bioassay: A carcinogenesis bioassay was
performed for benzene on both rats and mice. Both sexes of each species got
leukemia at the high doses administered. Extrapolating the cancer incidence at
high dose to low dose and from rodents to humans resulted in the risk estimate
that a benzene dose of 1 mg/kg/day will result in 3 cancers per 100 people exposed
daily for a lifetime to that dose. This dose is much higher than anyone would be
exposed to in the environment under normal conditions.
Check Your Progress 2
Note: a) Write your answer in about 50 words.
b) Check your progress with possible answers given at the end of the unit.
1) Discuss the dose response relationship with the help of dose response curve?
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2) What is LC?
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4.9 LET US SUM UP

A toxic effect can result from a natural or a manufactured substance and manifest
a variety of symptoms which may be both immediate and long-term. Toxicity
testing introduces a variety of methods and rates of exposure to the test organism,
which help in formulating a more accurate assessment of the risk of harm that
the test substance may pose to human health and the environment. Toxicity
assessment is characterization of the toxicological properties and effects of a
chemical, with special emphasis on establishment of dose-response
characteristics. The guiding principles of toxicity testing is to check the effect of
the test substances on laboratory animals and its direct toxic effect on human
and secondly, the exposure of laboratory animals to high doses in order to evaluate
its possible hazard on human that are exposed to much lower dose.

4.10 KEY WORDS

Cute : Occurring over a short time, usually a few minutes
or hours.

Acute Exposure : One or a series of short term exposures generally

lasting less than 24 hours

70
Administered Dose : The amount of a substance given to a test subject Toxicity Assessment
(human or animal) in determining dose-response
relationships, especially through ingestion or
inhalation.

Chronic : Occurring over a long period of time (more than 1

year).

Chronic Exposure : Long term exposure, usually lasting one year to a

lifetime.

Dose Response : The relationship between the dose of a pollutant

and the response (or effect) it produces on a
biological system.

Dose Response : The amount of a chemical that an organism (such

Assessment as a person) is exposed to is called the dose, and
the severity of the effect of that exposure is called
the response. A dose response assessment is a
scientific study to determine the relationship
between dose and response, and how much dose
is correlated with how much response

4.11 REFERENCES AND SUGGESTED FURTHER

READINGS
Duffus, J.H, Worth. H.G.J. (eds), 2006 Fundamental Toxicology, ISBN:978-
085404-614-0, Springer.

Michael J. Derelanko, Carol S. Auletta, 2014. Handbook of Toxicology, Third

Edition, March 7, by CRC Press ISBN 9781439890134.

Nancy M. Trautmann, 2001. Assessing toxic risk, Assessing Toxic Risk, Student’s
edition (Cornell Scientific Inquiry Series) .

Steven G. Gilbert, 2004. A Small Dose of Toxicology - The Health Effects of

Common Chemicals. CRC Press, p266.

4.12 ANSWERS TO CHECK YOUR PROGRESS

Answers to Check Your Progress 1
Your answer should include the following points:
1) Toxic effect is defined as an adverse change in the structure or function of
an experimental animal as a result of exposure to a chemical substance.”
Such changes may be effected via acute, sub chronic or chronic exposure
studies. Acute toxicity tests measure the immediate effects of exposure with
an estimated time for peak effect of approximately eight hours after the
initial exposure. Subchronic toxicity tests occur over a period of weeks,
while chronic affects tests measuring long term exposure last several months.
2) Inhalation, Absorption, Transformation, Ingestion , Injection, Storage

71
Introduction to Toxicology Answers to Check Your Progress 2
Your answer should include the following points:
1) Dose response curve is a mathematical relationship between the dose
administered or received and the incidence of adverse health effects in the
exposed population; toxicity values are derived from this relationship.
2) LC stands for “Lethal Concentration”. LC values usually refer to the
concentration of a chemical in air but in environmental studies it can also
mean the concentration of a chemical in water. According to the OECD
(Organization for Economic Cooperation and Development) Guidelines for
the Testing of Chemicals, a traditional experiment involves groups of animals
exposed to a concentration (or series of concentrations) for a set period of
time (usually 4 hours). The animals are clinically observed for up to 14 day.

72