NAME: ANIEBUE ESTHER NDIDIAMAKA

MATRIC. NO: 0219057004800 / 400LEVEL

DEPARTMENT: MICROBIOLOGY

COURSE: GENERAL TOXICOLOGY (MCB418)

LECTURER: PROF. M. D. MAKUT

ASSIGNMENT QUESTION

1 Outline and discuss the various factors that affect or may influence toxicity in living

organism

2 Describe the mechanism of drug toxicity in a biological system such as human cell or

tissue

3 Write detailed notes on toxins produced by

a. Bacteria

b. Fungi

c. Animals

d. Plants

4 Define the following terminologies used in toxicology

a. Toxin

b. Poison (toxicant)

c. Toxicity

d. Hazards

e. Risk

f. Acute exposure
 g. Chronic exposure

h. Synergistic effect

i. Additive effect

j. Antagonistic effect

ANSWERS TO QUESTION 1

1. Factors pertaining to chemical

(i) Chemical composition: The physico-chemical characteristics of a compound, such as

solubility, vapour pressure, ionization, functional groups, etc. are largely governed by the

chemical constitution of the compound. All these characteristics greatly affect the toxicological

properties of the substance. The nature of chemical reaction is determined by the functional

groups of the chemical and toxicity is a response of organisms consequent upon reaction between

chemical and certain part of the organism. The polar (hydrophilic) chemicals are not easily

soluble in lipids. Therefore, they cannot easily cross the membranous barriers and thus cannot

easily reach the target sites for appropriate action. However, non-polar or lipophilic substances

are highly soluble in lipids and other organic solvents.

(ii) Dose or concentration of chemical: A chemical induces toxic effects on account of its

interaction with appropriate receptors. The effect is directly related to concentration of the

chemical at the target site and concentration at the target site is often directly proportional to

dose/concentration of the chemical exposed. The lower doses of chemical cause less effects

whereas higher doses may cause pronounced effects. Thus, toxicity of chemical is directly

proportional to its dose/concentration.

(iii) Translocation of toxicant: There are two types of toxicants which are those producing local

effects and those producing systemic effects. In case of the latter group, effective translocation is

a key factor. Unless the toxicants are readily translocated to the specific sites, they cannot

produce adverse effects. During the course of translocation, some of the toxicants interact with

certain macromolecules present in the body of organisms and are then stored in certain relatively

inactive tissues, i.e. storage depots. The inefficient translocation and storage of toxicants in

inactive forms reduces their toxicity, while their effective translocation to the target in active

forms enhances their toxicity.

(iv) Bio-transformation of toxicants: Certain chemicals are normally inactive. During

translocation such chemicals are biocatalytically converted into active form in the body of

organisms with the help of certain enzymes. Consequently, the toxicity of that particular

chemical is increased. For instance, organophosphorodithionates and organophosphorodithioates

(having P=S group) are relatively poor inhibitors of AChE. Therefore, in the body of organisms,

they are first converted to their P=O forms, which are potent AChE inhibitors, hence highly

toxic. Thus, toxicant is not available at the target sites to produce adverse effects. This latter type

of biotransformation decreases the toxicity of xenobiotics.

(v) Chemical interactions: In nature often various chemicals may be present and the organisms

are not exposed to only one chemical, but to a variety of chemicals. These chemicals interact

with each other and such chemical interactions may have great toxicological significance.

However, in the laboratories, the organisms may be simultaneously or consecutively exposed to

two chemicals. These chemicals interact and affect the toxicity of each other. It may thus be

inferred that interaction of one chemical with another may:

(i) have no effect on their toxicity

(ii) increase their toxicity

(i) decrease their toxicity.

2. Factors pertaining to exposure: Toxicity of chemicals is greatly affected by various factors

pertaining to their exposures to the organisms. Some of the important factors related to exposures

are: exposure routes, exposure duration, and exposure systems.

(i) Exposure routes: The toxicant gain access to the body of organisms by dermal, oral and

inhalation exposures or by intraperitoneal, intramuscular, subcutaneous and intravenous

injections. The routes of exposure largely affect the toxicity of chemicals. A chemical produces

more rapid and greatest effect when given by intravenous route, because through this route the

chemical directly reaches in active form to the specific site and thus produces greatest effects.

The other exposure routes in descending order of toxicity are: inhalation>

intraperitoneal>subcutaneous>intramuscular>oral>topical.

(ii) Exposure duration: LC50 or LD50 values of toxicants decrease with increase in duration of

exposure. The values evaluated for long-term exposures are much less than those determined for

short-term exposures. The exposure to the same dose or concentration of a toxicant for different

periods has been reported to induce different grades of effects. The exposure for short duration

has less effect in comparison to long-term exposures. This simply indicates that the toxicity of

substances increases with increase in exposure period.

(iii) Exposure systems A variety of exposure systems may be used for the exposure of toxicants.

For instance, in aquatic medium, various exposure systems are used for the exposure of toxicants

to organisms, such as:

(a) Static system: Where toxicant is mixed in the water and the organisms are exposed to it in

still water.

(b) Recirculatory system: Where the toxicant solution is recirculated through certain pumps.

(c) Renewal system: Where the toxicant solution is renewed after certain interval.

(d) Flow through system: Where the toxicant solution flows into and out of test chamber

intermittently or continuously. These various types of exposure systems do affect the toxicity of

chemicals to the organisms to a great deal.

3. Factors pertaining to the surrounding medium: The factors of surrounding medium affecting

the toxicity of chemicals have been extensively worked out for the aquatic medium. The factors

related to the medium are physico- chemical characteristics of the water affecting the toxicity of

chemicals. Hence, those parameters are considered for this section.

(i) Water temperature: The water temperature is expected to greatly affect the toxicity of

xenobiotics. The increased water temperature increases the solubility of many substances, affects

the chemical form of some and governs the amounts of dissolved oxygen in water. Studies by

various investigators suggest that there is no single pattern of the effects of temperature on the

toxicity of chemicals to aquatic organisms. Temperature change in a particular direction may

increase or decrease or cause no effect on the toxicity of chemicals, depending on the chemical,

the species, the response and the particular procedure. Changes in water temperature may not

have much effects on chronic thresholds of chemicals, which are ultimately of great importance

to aquatic organisms. Several pesticides have been reported to be more toxic at higher

temperatures while some others show stronger lethal action at low temperatures. Similar findings
were also reported in case of insect larvae, fish species, mollusks, etc. in relation to pesticides

and several other chemicals.

(ii) Dissolved Oxygen: Freshwater can dissolve 14.6 mg/1 level of oxygen at 0ºC, which

gradually decreases to 9.1 mg/1 with increase in temperature upto 20ºC and reaches to the level

7.5 mg/1 at 30ºC. It is, therefore, obvious that increase in temperature decreases the dissolved

oxygen content of the water. Oxygen is essentially required for respiration by the organisms.

Any physiological change affecting the rate of respiratory flow of aquatic organisms may affect

the concentration of the toxicant at the gill surface and thus the toxicity of the chemical. Thus, it

might be expected that reduction in dissolved oxygen content of water imposes stress on the

aquatic organisms, which may greatly increase the toxicity of a chemical in water.

(iii) pH: pH may have greater effects on the toxicity of those chemicals that ionize under the

influence of pH. Usually undissociated forms of chemicals are more toxic to organisms, because

they easily penetrate the cell membranes. For instance, most of the pesticides do not easily

dissociate and owing to their lipophilic nature, they easily penetrate through the lipoprotein

bimolecular layer of cell membranes to induce the toxicity. pH has pronounced effect on the

toxicity of metals, because they ionize under the influence of pH. The ionic forms of metals are

more toxic to organisms than undissociated or molecular form. Therefore, changes in pH in

either direction may increase or decrease the ionic proportion and this in turn may increase or

decrease the toxicity of metallic compounds.

(iv) Salinity: The greatest differences in chemical characteristics of fresh water and seawater

may be expected to enormously affect the toxicity of chemicals. But, these differences do not

significantly alter the toxicity of chemicals. The tolerance between the close relatives of fresh

water and marine organisms are almost equal, if both are tested in their own environments. A
similar relationship has been established after the study of naphthalene toxicity to euryhaline

mummichog. The less mortalities of mummichogs in low salinities than in high salinities have

been ascribed to osmoregulatory dysfunction and increased uptake of naphthalene.

(v) Water hardness; The total hardness of water has little effect on the toxicity of most of the

chemicals except for metals. The toxicity of ammonia, phenols, surfactants and pesticides has

been reported to be affected the least by the hardness of water. LAS, a surfactant, has been

reported to be 1.5 times as toxic to bluegills in hard water as in soft water. Metal toxicity is

greatly affected by the hardness of water. Many heavy-metals are more toxic to fish and other

aquatic organisms in very soft water in contrast to hard water. The increase in toxicity of metals

with respect to decrease in hardness of water has been attributed to changes in gill permeability

of fish caused by their calcium content.

(vi) Suspended and dissolved matter: Natural water often contains suspended and dissolved

matter including organic ligands and chelators. They may partly detoxify some of the xenobiotic

chemicals as a result of sorption or binding. The metals are chief examples that may be

detoxified by this mechanism. There is very scant literature on the effect of sorption and binding

of other non-metallic xenobiotics with suspended and dissolved matters. The addition of clay

suspension affects the toxicity of an insecticide, endrin, to fathead minnows. In contract, the

addition of soil suspension to endrin solution moderates its toxicity. The addition of clay

suspension at levels normally present in nature did not affect the toxicity of the surfactant, LAS.

Thus, it may be concluded that the toxicity of metals is often greatly decreased by the suspended

and dissolved matters present in water because of sorption and binding while other xenobiotics

are much less affected.

4. Factors pertaining to organisms: Toxicity of chemicals is evaluated against a particular

organism or a group of organisms. Therefore, the factors related to the organisms are very

important in the study of toxicity of xenobiotics. The important modifying factors pertaining to

organisms are outlined below:

(i) Test species: The toxicity of xenobiotics greatly varies with variation in test organisms, as the

tolerance to chemicals differs in different groups of organisms. Among the organisms of same

group, the toxicity of chemical varies with variation in species of the organisms. Even in

different individuals of same species, the toxicity of chemicals varies because of variation in

susceptibility owing to certain genetic factors. Thus, certain individuals of a species may be

susceptible to a chemical whereas the other may be resistant to the same chemical. Owing to

these variations, it is often suggested that to have a clear picture of the toxicity of a chemical, it

must be tested against one plant (may be an alga), one or two invertebrates and at least one

vertebrate.

(ii) Sex: The toxicity of chemicals differs with respect to sexes, because the males and females

differ in their responses due to hormonal and metabolic differences. Males in some species

biotransform compounds more rapidly than females, although this is not true for all species. For

instance, aldrin (an organochlorine pesticide) is much more toxic to male rats than to female rats,

because aldrin is biocatalytically converted to epoxide form, dieldrin, by the males and this

biotransformation product is more toxic than the parent compound. Contrary to this, parathion is

bioconverted to paraoxon at a faster rate by the female rat than the male, which is more toxic

than the parent compound. Therefore, female rat is more susceptible to parathion than the male.

Besides sex-wise metabolic differences, differences in routes of excretion also occur, which may

cause differences in susceptibility.

(iii) Age: Generally, the young animals are more susceptible to xenobiotics. For a majority of

chemicals, the youngs are 1.5 to 10 times more susceptible than the adults. The main reasons for

the susceptibility of young ones may be less resistance and lack of biotransformation enzyme

systems. It has already been reported that the newly born individuals do not posses the enzyme

systems catalyzing the biotransformation reactions. These enzyme systems develop gradually,

reach at peak at a certain stage, thereafter tend to decline. In accordance with biotransformation

reactions involving detoxication of xenobiotics, the susceptibility of organisms may also change.

But, it is not necessary that all chemicals are more toxic to the young ones. Certain CNS

stimulants including DDT are less toxic to young ones.

(iv) Life-stage: The toxicity of chemicals varies with different stages of the life-cycle.

Generally, the early life-stages or immature stages are more susceptible to toxicant exposures

than the late stages or mature individuals. The fries and fingerlings of fishes are most sensitive

stages. A particular stage of the lifecycle may be particularly susceptible to toxicants and

exposure of toxicants at this stage appreciably affects the results. For instance, the time of

moulting is particularly susceptible in case of aquatic arthropods.

(v) Size: The toxicity of chemicals is also affected by the size of the organisms. Often larger

sized individuals are more resistant to toxicants and this has been found true in case of certain

fishes.

(v) Acclimation The animals acclimated to sub lethal levels of a toxicant may become more

tolerant or more weakened, depending upon the mode of action of toxicant and the types of

detoxifying mechanism of the animals. For instance, acclimation of trout to sub lethal (0.22

mg/1) level of arsenic for three weeks, increased the threshold of LC50 by a factor of 1.5. While

acclimation to one-third of the lethal level of cyanide to fish rendered them to become more
sensitive by a factor of one-third in the first week. The tolerance improved to the original level

by the end of three weeks.

ANSWER TWO

Drug toxicity in biological systems, such as human cells or tissues, can occur through various

mechanisms, often resulting from the interaction between the drug and cellular components

1. Direct Interaction with Cellular Components:

Protein Binding: Drugs may bind to specific proteins within cells, altering their structure or

function. This can interfere with essential cellular processes, such as enzyme activity, receptor

signaling, or ion channel function. For example, some drugs may bind to ion channels in cardiac

muscle cells, leading to arrhythmias or cardiac toxicity.

DNA Damage: Certain drugs or their metabolites can directly interact with DNA, leading to

DNA adduct formation, DNA strand breaks, or interference with DNA replication and

transcription. This can result in genetic mutations, cell cycle arrest, or apoptosis, contributing to

tissue damage or carcinogenesis.

2. Disruption of Cellular Processes:

Mitochondrial Dysfunction: Drugs may disrupt mitochondrial function, leading to impaired

ATP production, increased reactive oxygen species (ROS) generation, and mitochondrial

membrane damage. This can result in cellular energy depletion, oxidative stress, and activation

of cell death pathways.

Protein Synthesis Inhibition: Some drugs may inhibit protein synthesis by targeting ribosomes

or other components of the protein synthesis machinery. This can disrupt normal cellular

functions and lead to cytotoxicity.

Membrane Damage: Drugs may disrupt the integrity of cellular membranes, leading to leakage

of ions, metabolites, or cellular contents. This disruption can interfere with cellular homeostasis,

signaling pathways, and cell viability.

3. Indirect Effects and Cellular Responses:

Oxidative Stress: Drugs may induce oxidative stress by promoting ROS generation or inhibiting

antioxidant defenses. Oxidative stress can damage cellular macromolecules such as proteins,

lipids, and DNA, leading to cellular dysfunction and death.

Inflammation: Some drugs may induce an inflammatory response by activating immune cells or

releasing pro-inflammatory mediators. Chronic inflammation can contribute to tissue damage

and organ dysfunction.

Apoptosis or Necrosis: Drugs may induce programmed cell death (apoptosis) or uncontrolled

cell death (necrosis) in affected cells. This can occur through various signaling pathways,

including those involving caspases, Bcl-2 family proteins, and death receptors.

4. Metabolic Activation and Detoxification:

Metabolic Activation: Some drugs are metabolized by cellular enzymes into reactive

intermediates that can covalently bind to cellular macromolecules, leading to cellular damage.

This process, known as metabolic activation, is often associated with drug-induced

hepatotoxicity or idiosyncratic reactions.

Detoxification Pathways: Cells possess detoxification pathways, such as phase I and phase II

metabolism, which can metabolize and eliminate drugs or their metabolites. Dysfunction of these

pathways can lead to impaired drug metabolism and increased susceptibility to toxicity.

5. Genetic and Epigenetic Factors:

Genetic: variations in drug-metabolizing enzymes, transporters, or drug targets can influence

individual susceptibility to drug toxicity. Polymorphisms in genes encoding these proteins can

affect drug metabolism, distribution, and response, potentially altering toxicity outcomes.

Epigenetic modifications: such as DNA methylation or histone acetylation, can also influence

drug toxicity by regulating gene expression patterns and cellular responses to toxic insults.

ANSWER THREE

A Bacterial Toxins

Bacterial toxins are typically classified under two major categories: exotoxins or endotoxins.

Exotoxins are immediately released into the surrounding environment whereas endotoxins are

not released until the bacteria is killed by the immune system. The release of toxins into the

surrounding environment, regardless of when released, results in the disruption of metabolic

pathways in the host eukaryote. These metabolic pathways include damaging cell membranes,

disrupting protein synthesis, inhibiting neurotransmitter release, or activating the host immune

system. The mechanisms of action by which toxins disrupt eukaryotic cell processes are

dependent on the target. For example, the bacteria Listeria monocytogenes, associated with food-

borne illnesses, specifically targets cholesterol by producing a pore-forming toxin protein,

listeriolysin O. This exotoxin affects intracellular processes and creates unregulated pores within

the cell membranes of the host. Another example of an exotoxin includes an enterotoxin

produced by the bacteria Staphlycoccal aureus. S. aureus can producestaphylococcal enterotoxin

B (SEB), associated with intestinal illness, which promotes activation of the immune system.

Upon activation of the immune system, the release of large amounts of cytokines, inflammatory

related molecules, causes significant inflammation. Lastly, an example of an endotoxin, includes

the protein lipopolysaccharide (LPS) produced by gram-negative bacteria. The LPS is a

component of the bacteria’s outer membrane and promotes structural integrity. Upon destruction

of the membrane by an immune response, the LPS is released and functions as a toxin.

B. Fungi Toxins

Fungi toxins which is also called Mycotoxins are the classes of toxins produced by fungi.

Mycotoxins are numerous and production of a specific mycotoxin is not restricted to one specific

species. Mycotoxins are secondary metabolites that are toxic to humans and produced by fungi.

There are various types of mycotoxins including, but not limited to, aflatoxins, ochratoxins,

citrinin, and ergot alkaloids.

i. Aflatoxins: are a type of mycotoxin that are produced by certain strains

of Aspergillus fungi. The aflatoxins are further broken down into types: AFB1, AFB2,

AFG1, and AFG2. These strains are present in a wide range of agricultural commodities

associated with tropic and subtropic zones. These commodities include species of peanuts

and corn. The most potent toxin is AFB1 and it is associated with carcinogenic effects.
 ii. Ochratoxin: is a type of toxin produced by both Penicillium and Aspergillus species.

Ochratoxins are further classified in types A, B and C and differ in structure. Ochratoxins

have demonstrated carcinogenic properties and are often found in beverages such as beer

and wine, as the fungal species which produce ochratoxins are often found on the plants

used to produce these products.

iii. Citrinin: Citrinin is a mycotoxin that has been isolated in numerous species of

both Penicillium and Aspergillus. Many of these fungal species are utilized in food

processing and are often found in foods including cheese, wheat, rice, corn, and soy

sauce. Citrinin is known to function as a nephrotoxin, indicating it has toxic effects on

kidney function.

iv. Ergot Alkaloids: are specific compounds that are produced as toxic alkaloids

in Claviceps, a group of fungi associated with grasses, rye, and related plants. The disease

caused by ingestion of this fungi is called ergotism. Ergotism is characterized by

detrimental effects on the vascular system in particular, including vasoconstriction of

blood vessels resulting in gangrene, and eventually, limb loss if left untreated.

Additionally, ergotism can present as hallucinations and convulsions as ergot alkaloids

target the central nervous system. Due to the vascular system effects of ergot alkaloids,

they have been used for medicinal purposes.

C. ANIMAL TOXINS

Animal toxins are a complex mixture of polypeptides, enzymes and chemicals which can cause

cellular injury. Several mechanisms are involved in pathophysiology of venom poisoning. Direct
injury can be induced by chemicals, enzymes and polypeptides. Polypeptides exert their effect

through action on ion channels and receptors on the cell membrane. The action of polypeptides

upon ion channels and receptor sites of excitable membranes through signal transduction causes

the release of neurotransmitters which results in neuromuscular effects. Enzymes can cause

membrane lysis, pore formation, cytoskeletal destruction, and release inflammatory and

vasoactive mediators with action on the coagulation pathway at various levels. Proteolytic

enzymes and phospholipase A2 are important causes of cellular injury and coagulopathy.

Indirectly, animal enzymes, especially phospholipase A2 and metalloproteases, can induce the

release of vasoactive mediators and proinflammatory cytokines which can lead to hemodynamic

alterations and cause organ injury. Cardiovascular symptoms are therefore common. They can

also trigger the inflammatory process with generation of adhesion molecules, complement

activation, acute phase proteins, free radicals, increased vascular permeability and increased

blood viscosity. Local reactions include pain, swelling and redness. Hemodynamic changes may

result if the process is severe. Immunologic reaction usually manifests in the form of allergic

response such as skin rashes and edema. In the severe form, anaphylaxis with respiratory and

cardiovascular symptoms may occur.

D PLANT TOXINS

Plant toxins are a diverse group of chemical compounds produced by plants as a defense

mechanism against herbivores and pathogens. These toxins can affect various vital organs and

the central nervous system, with the most dominant being neurotoxins, cytotoxins, and metabolic

toxins. Some examples of toxic plants found in the wild or cultivated include:

i. Monkshood (Aconitum napellus): Contains aconitine, a potent neurotoxin

ii. Castor bean (Ricinus communis): Contains ricin, one of the most toxic substances known
iii. Deadly nightshade (Atropa belladonna): Contains tropane alkaloids that can cause intense

digestive and nervous system disturbances

ANSWER FOUR

a) Toxin: A toxin is a poisonous substance produced within living cells or organisms,

including plants, animals, bacteria, and fungi. Toxins can cause harm to other organisms

when they are ingested, inhaled, or come into contact with the skin.

b) Poison (toxicant): A poison, also known as a toxicant, is any substance that, when

ingested, inhaled, or absorbed, is capable of causing injury, illness, or death to an organism.

Poisoning can occur through various routes of exposure, including ingestion, inhalation,

and dermal contact.

c) Toxicity: Toxicity refers to the degree or extent of harm caused by a toxic substance to an

organism or biological system. It is typically assessed by evaluating the adverse effects of

the substance on various endpoints, such as mortality, morbidity, organ damage, or

biochemical alterations.

d) Hazards: Hazards refer to the inherent properties or characteristics of a substance that

make it capable of causing harm under certain conditions. These may include physical

hazards (e.g., flammability, reactivity), health hazards (e.g., toxicity, carcinogenicity), and

environmental hazards (e.g., aquatic toxicity, bioaccumulation).

e) Risk: Risk in toxicology refers to the probability or likelihood that exposure to a hazardous

substance will result in harm to human health or the environment. It is determined by

considering factors such as the toxicity of the substance, the level and duration of exposure,

and the susceptibility of exposed individuals or ecosystems.

f) Acute exposure: Acute exposure refers to short-term or brief exposure to a toxic substance

over a relatively short period, typically lasting hours to days. Acute exposure can result in

immediate adverse effects, ranging from mild symptoms to severe toxicity or death.

g) Chronic exposure: Chronic exposure refers to long-term or repeated exposure to a toxic

substance over an extended period, often weeks, months, or years. Chronic exposure can

lead to cumulative effects, resulting in persistent or delayed health effects, such as chronic

diseases, organ damage, or cancer.

h) Synergistic effect: A synergistic effect occurs when the combined effect of two or more

substances is greater than the sum of their individual effects. In toxicology, synergistic

effects can increase the toxicity of a mixture of substances, leading to more severe or

unexpected adverse effects than would be predicted based on the effects of each substance

alone.

i) Additive effect: An additive effect occurs when the combined effect of two or more

substances is equal to the sum of their individual effects. In toxicology, additive effects are

observed when substances with similar toxic mechanisms or modes of action produce

effects that are simply additive when combined.

j) Antagonistic effect: An antagonistic effect occurs when the combined effect of two or

more substances is less than the sum of their individual effects. In toxicology, antagonistic

effects can occur when one substance mitigates or counteracts the effects of another

substance, resulting in reduced toxicity or efficacy of the mixture.