Antibody probes

Antibody probes
Chapter can either be
1: Introduction
Polyclonal
or
Monoclonal in nature.

• .
 Concept of health and disease

Health: (helth)
Health is the state or condition of the body
and mind in which all the organs are in normal
structure and the body fluid are of such
composition that the body as whole functions
in a state of harmony.

It is a state of optimal physical, mental, and

social wellbeing, and not merely the absence
of disease and infirmity.
Disease: (Fr. dès – from, aise - ease)
Disease is the abnormal condition of body and mind of an
organism which is expressed with certain symptoms.
Any deviation from or interruption of the normal structure
or function of any part, organ, or system (or combination
thereof) of the body that is manifested by a characteristic
set of symptoms and signs and whose etiology, pathology
and prognosis may be known or unknown.
Etiology: (Gr. aitia - cause)
Etiology is the study or theory of the factors that cause
disease and the method of their introduction to the host;
the cause or origin of a disease or disorder.
Prognosis: (Gr. prognosis - foreknowledge)
Prognosis is the forecast as to the probable outcome of
an attack of disease, the prospect as to recovery from a
disease as indicated by the nature and symptoms of the
case.
 Causes of disease outbreak

When the disease spread from one fish to another or

one population to another, can be expressed clinically
which are caused by various infectious and non-
infectious agents is called disease outbreak. Simply it is
sudden eruption of disease.
Following factors are responsible to cause disease
outbreak in aquatic animals

1. Environmental Factors
A. Physical factors
i) Temperature: Protected low temperature decreases
the metabolic activity of fish. So, fish take small
amount of food and they become weak and then the
secondary infection happens.

ii) Excess of heat: Excess of heat creates haemolysis

i.e. breakdown of blood vessels and damage the blood
cells. As a result, hemorrhages occur which is visible
on the body surface.
iii) Excess of illumination: Some fishes are very
sensitive to illumination during the development of
their eggs and larvae. Excess of illumination increase
water temperature, causes haemolysis, affect
protoplasm and body colour does not develop.

iv) Pollution: mining wastes, mill wastes, sewage

pollution. Affect nervous system, physiological
difficulties and ultimate death.

v) Turbidity: Turbidity decreases the productivity of

water. Excess turbidity causes the problems of
respiration and affects the vision of fish. Fish may be
mechanically injured by suspended particles which
cause the secondary infection of fish.
B. Chemical factors
i) Lack of oxygen: Lack of O2 causes hypoxia (reduced
of DO) or anoxia (total loss of DO) and suffocation.

ii) pH: Highly acidic pH (below 5.5) of water damage the

gills, reduces the dissolved oxygen level, nutrient level
and cause disease is called acidosis. On the contrary,
highly alkaline pH (above 9.0) of water causes erosion
of gills and disease is called alkalosis.

iii) Sudden decrease in gas pressure: It causes

imbalanced osmotic pressure between fish body and
environment and so, gas bubbles are appeared on the
body surface which is called gas embolism. The gases
involved this process are mainly N2 and O2 freed from
blood through skin.
iv) Intoxication: intoxications of fish by insecticides,
pesticides, heavy metals, NH3, DDT, Fe, Mg, Pb, Zn, Ni, Al,
Cd etc. in water.

2. Hereditary factors
There are some hereditary diseases which are transmitted
from parents to the offsprings. eg. tumors, albinism,
tuberculosis, dropsy and various deformities such as double
head, defective fins, swim bladder and spinal column, colour
problem etc.

3. Glandular dysfunction
i) Pituitary dysfunction: It causes gill impermeability and
suffocation.
ii) Thyroid dysfunction: Decrease iodine content and causes
goiter in branchial region.
iii) Liver dysfunction: Causes metabolic disturbances.
iv) Pancreas dysfunction: Causes metabolic disturbances.
v) Sex gland dysfunction: Causes reproductive inhibition.
vi) Adrenal cortical tissue dysfunction: Causes
osmoregulatory inhibition.
4.Mechanical injury
Mechanical injury by angling, netting and rough
handling which creates a stressful condition for fish and
causes secondary infection.

5. Pathogenic organisms
Pathogenic organisms such as virus, bacteria, fungus
and many protozoan and metazoan parasites which are
very important for producing disease in fish. The
pathogens attack the fish directly or indirectly and
cause characteristics disorders.

6. Atypical cell growth

Sudden abnormal uncontrolled growth of cell or tissue
is called neoplasm.
7. Dietary deficiency factors

i) Carbohydrate deficiency
• Less metabolic activity.
• Less ovarian or egg development.
• Disturbance in swimming.
• Frequent rest, diabetes.
• Ultimately death.

ii) Fat deficiency

● Migratory fish stop migration.
● Egg laying capacity inhibited.
● Stop energy metabolism.
● Skin diseases.
● Imperfect scale formation.
● Less ovarian development.
iii) Protein malnutrition
• Stunted growth.
• Head enlargement.
• Eye protrusion (exophthalmia).
• Secondary infection.
• Paralysis.
• Malformation of scale, bone and muscle etc.

iv) Avitaminoses
• Vitamin A deficiency – Exophthalmia.
• Vitamin B1 deficiency – Loss of balance.
• Vitamin B2 deficiency – Photophobia.
• Vitamin D deficiency – Lower haemoglobin.
• Vitamin E deficiency – Anaemia.
• Vitamin K deficiency – Blood do not coagulate.
Host, pathogen and environmental
interaction towards disease production
HOST PATHOGEN
SPECIES , AGE INFECTIVITY

STRAIN VIRULENCE
HOST
NUTRITIONAL PATHOGENECITY
STATUS
VIABILITY
Disease

PHYSIOLOGICAL
CONDITION STRAIN

Pathogen
DISEASE

TEMPERATURE ENV.
OXYGEN CONC.

ALKALINITY

HARDNESS

OTHER
PARAMETER
Host + Pathogen + Environmental Factors

Weak and stress ----- Stage I

(Invasion)
Infection ----- Stage II

(Grow & multiplication)

Disease ------ Stage III
(Expressed with signs & symptoms)
Overview of diseases in Bangladesh
Usually the following diseases occurred in Bangladesh
Infectious diseases
1. Viral diseases
i. Disease name: Spring viremia of carp (SVC)
Causative agent : Spring viremia of carp virus (SVCV)
Host : Cyprinus carpio
2. Bacterial diseases
i) Disease name: Columnaris
Causative agent : Flavobacterium columnare
Host : All freshwater species, world wide.

ii) Disease name: Motile aeromonus septicemia/

Haemorrhagic septicemia/Red sore disease/Infectious
dropsy
Causative agent : Aeromonas hydrophila, A. sobria, A.
caviae.
Host : All freshwater species, world wide.
iii) Disease name: Edwardsiellosis
Causative agent : Edwardsiella tarda
Host : All freshwater catfish, world wide.

iv) Disease name: Vibriosis

Causative agent : Vibrio anguillarum
Host : Freshwater, brackish water & marine species,
world wide.

v) Disease name: Bacterial gill disease (BGD)

Causative agent : Flavobacterium branchiophilum
Host : Freshwater & brackish water species, world
wide.

vi) Disease name: Bacterial hemorrhagic septicemia

Causative agent : Pseudomonas flurescens
Host : Freshwater species, world wide
3. Fungal diseases
i) Disease name: Epizootic Ulcerative Syndrome (EUS)
Causative agent : Aphanomyces invadans, others are
Saprolegnia sp., Achlya sp., virus, bacteria and
parasites etc.
Host : All freshwater species, world wide.

ii) Disease name: Saprolegniasis

Causative agent : Saprolegnia parasitica, S. declina,
Saprolegnia sp.
Host : All freshwater species, world wide.

iii) Disease name: Branchiomycosis

Causative agent : Branchiomyces sanguinis and
B. demigrans.
Host : All freshwater species, world wide.
4. Parasitic diseases
i) Disease name: Ichthyopthiriasis / Ich / white spot
disease
Causative agent : Ichthyopthirius multifiliis
Host : All freshwater species, world wide.

ii) Disease name: Ichthyobodosis

Causative agent : Ichthyobodo necatrix
Host : All freshwater species, world wide.

iii) Disease name: Chilodonellosis

Causative agent : Chilodonella cyprini, C. piscicola
and C. hexascha.
Host : All freshwater species, world wide.

iii) Disease name: Trichodiniasis

Causative agent : Trichodina nigra, T. pediculatus
Host : All freshwater species, world wide.
iv) Disease name: Gyroductylosis (skin fluke disease)
Causative agent : Gyroductylus elegans & G. gracilis
Host : Carp species.

v) Disease name: Dactylogyrosis (Gill fluke disease)

Causative agent : Dectylogyrus vastator & D. solidus
Host : Carp species.

vi) Disease name: Ligulosis

Causative agent : Ligula intestinalis
Host : Carp species.

vii) Disease name: Argulosis

Causative agent : Argulus foliaceus, A. siamensis &
A. bengalensis
Host : Carp species.
viii) Disease name: Lerneasis
Causative agent : Lernaea bengalensis, L.cyprinaecea
& L. chackoensis.
Host : Carp species.

ix) Disease name: Ergasilosis

Causative agent : Ergasilus lizae, Ergasilus sp. and
E. sieboldi.
Host : Carp species.

x) Disease name: Piscicolosis

Causative agent : Piscicola geometra
Host : Carp species.
Non infectious diseases
A number of non infectious diseases occurred in
Bangladesh

1. Environmental diseases
i. Acidosis
ii. Alkalosis
iii. Gas bubble disease

2. Nutritional diseases
i. Nutritional gill disease
ii. Thyroid tumor
iii. Protein malnutrition/deficiency disease
iv. Lipid/fatty acid deficiency or excess
v. Carbohydrate deficiency or excess
vi. Avitaminosis
vii.Hypervitaminosis
3. Hereditary diseases
i. Albinistic opthalmia
ii. Albinism
iii. Cystic condition
iv. Auto immune disease
v. Diabetes
vi. Tumor
Impact of aquatic animal diseases
Aquatic animal health therefore has major social
and economic impacts on the people, businesses,
communities and economies that rely on aquatic
animal production. Given the diversity of species,
systems and business models within aquaculture,
these socio-economic impacts clearly vary widely.

Socio-economic impact of disease

At a macroeconomic level, seafood (mainly
shrimp) is the most traded food commodity
globally, with transboundary disease a barrier to
international trade that can have severe
consequences for exporting countries like
Bangladesh.
Disease impact in open water fishery
i. Open water fishes affected by diseases
ii. Other aquatic organisms also affected by diseases
iii. Pen and cage cultured fishes will be disease affected
iv. Different types of pathogens will be aggregated in
open water bodies
v. Water quality will be deteriorated
vi. Healthful environment will be disrupted
vii. Open water fish, shrimp and other aquatic animals
production will be reduced
viii.Supply of aquatic animals in the market will be
reduced
ix. Natural breeding ground will be hampered
x. Livelihood of open water depended fishermen will be
affected
xi. Coastal shrimp fishery will be hampered severely by
disease outbreak
xii. The affected aquatic animals may carry/transfer
Disease impact in hatchery
i. Brood fish become unhealthy
ii. Reduce the use of fishes which have disease history
iii. Lower quantity of eggs is production
iv. Fertilization rate is lowered
v. Consequently fry production is decreased
vi. Survival rate of fry is declined
vii. Fry quality also become deteriorated
viii.Mortality of fry is increased during transportation
ix. Goodwill of hatchery become questionable and faith
of fry purchaser is reduced
x. Ultimately hatchery owner become loser.
Disease impact in pond culture fishery
i. It increases the mortality of fry in nursery and grow-
out ponds
ii. Pond ecosystem hampered
iii. Survival rate of juvenile is declined
iv. Lower quality brood produced
v. Early harvest may be necessary in some cases, either
due to compulsory controls or to avoid further losses
in the presence of disease
vi. Fish culture technique become risky
vii. Extra costs for disease control such as medicines
viii.Pond productivity reduced
ix. Lowered biomass produced
x. Direct losses can result from increased mortality
xi. Consequently people loss their interest in aquaculture
Overall disease impact
i. National fish production reduced
ii. Product condemnation and lower product prices due to
disease lesions
iii. National nutritional requirement not fulfilled
iv. Disease outbreaks may also damage public
perceptions of aquaculture, with negative
consequences for consumer demand for aquaculture
products
v. Export quantity and earnings reduced
vi. Employment opportunity declined.
Concepts of immunology and serology
Immunology is defined as the study of the molecules,
cells, organs, and systems responsible for the recognition
and disposal of foreign material. Immunology began as a
branch of microbiology. The study of infectious disease
and the body’s response to them has a major role for the
development of immunology. More over, the concept of
germ theory of disease has contributed to the field of
immunology.

Edward Jenner first studied the response of the body to

foreign substances in 1796. He observed that dairy maids
who had naturally contracted a mild infection called
cowpox seemed to be protected against smallpox, a
horribly disfiguring disease and a major killer.
 Preliminary Concepts
● Disease problems have grown proportionally
with the intensive or expansive culture of
aquaculture species
● Why?
1) Increased stocking densities (lower profit
margins)
2) Infected carriers (largely broodstock)
3) Infected facilities (GMPs being followed?)
4) Poor nutrition (we are way behind)
5) Substandard water quality (traditional)
● Biggest problem: greater susceptibility via
weakening of resistance under intensive culture
conditions
 The Immune Response
● For fish, response to a foreign agent is rather
similar to that of mammals; shrimp, very
rudimentary
● Response can be highly specific (a specific
antibody for a specific antigen) is known as the
immune response.
● The immune system “scans” the body to identify
any substance (natural/synthetic or living/inert)
that it considers foreign
● Differentiates between “self” and “non-self”
● Works with several types of white blood cells,
located throughout the body, that work together in
a highly integrated way
 Definitions
● resistance: any type of barrier within
the host that allows it to resist the
pathogen
● innate or natural immunity: attributed
to inherited ability to produce antibodies
without stimulation by antigens
● acquired immunity: host is stimulated
by contact with antigens
● passive immunity: acquired through the
use of antibodies from other animals
(vaccination)
● we will add another term today,
tolerance
Immune
Response
System

● Made up of two cellular systems: 1) cell-mediated immunity (T

cells) and 2) humoral antibody system (B cells)
● Both work by identifying antigens (foreign proteins or
glycoproteins)
Immunology : That branch of biomedical science
concerned with the response of the organism to
antigenic challenge, the recognition of self and not self,
and all the biological (in vivo), serological (in vitro), and
physical-chemical aspects of immune phenomena. It
encompasses the study of the structure and function of
the immune system (basic immunology); immunization,
organ transplantation, blood banking, an immunology
(clinical immunology); laboratory testing of cellular and
humoral immune function (laboratory immunology); and
the use of antigen- antibody reactions in other laboratory
tests (serology and immunochemistry).

Immunity: The condition of being immune; the

protection against infectious disease conferred either by
the immune response generated by immunization or
previous infection or by other non-immunologic factors.
Antigen: [antibody + Gr. Genan – to produce]
Any substance which is capable, under appropriate
conditions, of inducing a specific immune response and of
reacting with the products of that response, that is with
specific antibody or specifically sensitized T-lymphocytes,
or both. Antigens may be soluble substances, such as
toxins and foreign proteins, or, particulate, such as
bacteria and tissue cells. Abbreviated Ag.

Immunogen: A substance capable of inducing an

immune response, in most contexts synonymous with
antigen; in some contexts immunogen is used to draw a
distinction with substances capable of reacting only with
antibody ( antigens or haptens) or to denote a form of an
antigen that induces an immune responses as opposed to
a tolerogen, a form that induces tolerance.
Antibody: An immunoglobulin molecule that has a
specific amino acid sequence by virtue of which it
interacts only with the antigen that induced its synthesis
in cells of the lymphoid series (especially plasma cells), or
with antigen closely related to it. Antibodies are classified
according to their mode of action as agglutinins,
bacteriolysins, hemolysins, opsonins, precipitins etc.
Abbreviated Ab.

Immune response: The reaction of the immune system

to foreign substance.

Immunogenecity: The property that endows a

substance with the capacity to provoke an immune
response, or the degree to which a substance possesses
this property.
Serology
Testing for the presence of a specific antigen
using specific antibody (antiserum). Examples:
ELISA blood test for HIV, home pregnancy test.

Beginning of Serology
►Serology as a science began in 1901.
Austrian American immunologist Karl
Landsteiner (1868-1943) identified groups
of red blood cells as A, B, and O. From that
discovery came the recognition that cells of all
types, including blood cells, cells of the body, and
microorganisms carry proteins and other
molecules on their surface that are recognized by
cells of the immune system.
Purpose of Serological Tests
►Serological tests may be performed for
diagnostic purposes when an infection is
suspected, in rheumatic illnesses, and in many
other situations, such as checking an individual's
blood type. Serology blood tests help to diagnose
patients with certain immune deficiencies
associated with the lack of antibodies.
Serology

►The branch of
laboratory medicine
that studies blood
serum for evidence
of infection and
other parameters by
Evaluating antigen-
antibody reactions
in vitro
Serology
►Serology is the
scientific study of
blood serum. In
practice, the term
usually refers to
the diagnostic
identification of
antibodies in the
serum
We can detect
antigens too
Major serological/ immunological
techniques

1. Agglutination
2. Indirect fluorescence antibody test (IFAT)
3. Direct fluorescence antibody test (DFAT)
4. Enzyme linked immunosorbent assay (ELISA)
5. Enzyme labeled antibody technique (ELAT)
6. Western blot
7. Immunohistochemistry (IHC)
At a microeconomic level,
i. Losses due to disease have financial implications for
aquaculture businesses,
ii. Direct losses can result from increased mortality,
iii. Reduced productivity,
iv. Product condemnation and lower product prices due to
disease lesions,
v. Early harvest may be necessary in some cases, either
due to compulsory controls or to avoid further losses
in the presence of disease, and
vi. Control costs such as medicines,
vii. Increased labour and depopulation of infected sites,
viii.Disease outbreaks may also damage public
perceptions of aquaculture, with negative
consequences for consumer demand for aquaculture
products.
For example, in Bangladesh, carp and other freshwater
species such as catfish are produced in large scale,
independently owned freshwater ponds. This type of
aquaculture production is mainly for domestic and local
consumption and delivers benefits for communities, such
as creating employment opportunities and diversifying
and increasing household income. A number of diseases
affect pond aquaculture in Bangladesh, with epizootic
ulcerative syndrome considered to be the greatest threat.
On average, economic losses to farmers from fish disease
have been estimated at US $344 (approximately 15% of
production value) per annum (Faruk et al., 2004).

Due to disease outbreak employment opportunity

reduced