Practical Workbook Infectious Diseases and Preventive Medicine 2022

PRACTICAL WORK GUIDE IN
INFECTIOUS DISEASES AND

PREVENTIVE MEDICINE
VIRAL DISEASES - 4-th YEAR
DVM PhD DRAGOS COBZARIU
2022
Dr. Dragos Cobzariu
Practical Work Guide in

Infectious Diseases and
Preventive Medicine
VIRAL DISEASES - 4-th YEAR
Bucharest
2022
Published online: https://ro.scribd.com/
Tel. +40722258710
e-mail: dragoscobzariu@gmail.com
Coordinator: Head of works Dr. Dragoș Cobzariu

Scientific reference: Prof. univ. Dr. Daneș Doina
ISBN 978-973-0-37443-8.
© 2022 All rights to this edition are reserved.

Partial or complete reproduction of the text, on any medium,
without the written consent of the authors is prohibited and
will be sanctioned according to the laws in force.
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The practical work in the Discipline of Infectious Diseases and

INFECTIOUS DISEASES AND Preventive Medicine is based on all the knowledge you have
already acquired in the preclinical disciplines, and makes a
PREVENTIVE MEDICINE correlation of this necessary information, in order to understand
and not memorize these diseases.
Anatomy
Histology
Physiology
Semiology
Microbiology
Immunology
Pathophysiology
Epidemiology
Pathological anatomy
Practical work: Course holder :
Biochemistry and Biophysics
Dr. Dragos Cobzariu Prof.univ.Dr. Doina DANEȘ Nutrition
DVM PhD Dragoș Cobzariu 2022 1 DVM PhD Dragoș Cobzariu 2022 2
Notifications of labor protection, fire prevention and extinguishing

Entering the laboratories and classrooms is done in an organized manner, with the appropriate
Warning to Users-Students: protective equipment (gown, cap, protective gloves, apron).
The laboratory equipment is handled only with the approval and in the presence of the
teaching staff.
All documents placed in open access on the site the faculty site, and
presented, are protected by copyright. Students will not walk to the electricity and methane gas installation.
Food and drinks of any kind are not consumed in the work spaces (laboratories, necropsy
In accordance with the principles set out by the "Budapest Open Access rooms).
Initiative" (BOAI, 2002), the user of the site may read, download, copy,
Smoking is prohibited in public spaces (halls, hallways, restrooms).
transmit, print, search or link to the full text of these documents, dissect them
The handling of any material and biological preparation is done only under the supervision of
for them index, use data for software, or use it for any other legal (or copyright)
the teaching staff. It is forbidden to remove the pathological material from the work spaces.
purpose.
The activities during the practical work and the class hours as well as during the breaks must
Any use of the document for commercial purposes is strictly prohibited. comply with the norms of university ethics and deontology.
In addition, the user agrees to respect the moral rights of the author, mainly the The students will examine the animals in compliance with the restraint rules specified in
right to the integrity of the work and the right of paternity and this in any use Semiology only in the presence of the teaching staff.
that the user undertakes. During the trip to the farms and the teaching station, the consumption of alcoholic beverages
Thus, by way of example, when reproducing a document by extract or in its and smoking are prohibited.
entirety, the user will fully cite the sources as mentioned above. During the activities carried out on the farms and the didactic station as well as in the university
hospital, the rules of labor protection and P.S.I. will be respected. provided for each individual
Any use not explicitly authorized above (such as, for example, the modification objective.
of the document or its summary) requires the prior and express authorization of Any deviation from the rules of labor protection and P.S.I. as well as from the obligations and
the authors or their successors in title. duties of the students stipulated in the University Charter will be sanctioned according to the
DVM PhD Dragoș Cobzariu 2022 3
laws in force by the legal bodies. DVM PhD Dragoș Cobzariu 2022 4
PURPOSE OF PRACTICAL WORKS:

Establishing the diagnosis of suspicion and
confirmation in infectious diseases, and based on the
DIAGNOSIS IN confirmation, the establishment of prophylaxis,
INFECTIOUS DISEASES monitoring and control measures.
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Infectious disease-definition DIAGNOSIS OF INFECTIOUS DISEASES

INVOLVES TWO STAGES:
Uncontrolled disruption of one or more functions of an
organism (with or without clinical - lesion expression) as a
result of body penetration of a foreign entity capable of
multiplying and giving birth to identical entities.  SUSPICION DIAGNOSIS
BACTERIA
Gram positive and negative Rickettsia and Chlamidia
Spirochete  CONFIRMATORY DIAGNOSIS
PRIONS
VIRUSES
MYCOPLASMAS
Within the SUSPICION-ASSUMPTION-mandatory diagnosis, three large groups of EPIDEMIOLOGICAL DIAGNOSIS

investigations are described:
EPIDEMIOLOGYCAL,
CLINICAL,
ANATOMOPATHOLOGICAL.
As part of the CONFIRMATION-CERTAINTY-diagnosis, a sufficient number of It studies the origin, evolution and extinction of mass
CONFIRMATION examinations necessary for the identification of the ANTIGEN or
ANTIBODIES must be performed (sometimes a single examination is sufficient) in morbid processes
order to finally pronounce a positive or negative result.
At this stage, depending on the suspicion, additional examinations are performed: Work instrument: EPIDEMIOLOGICAL INQUIRY - It is
bacterioscopical,
bacteriological, mandatory when you suspect an infectious disease.
virological,
serological,
histopathological,
biological-bioprobe,
PCR
CONCLUSION-The diagnosis of an infectious disease is made by isolating and
identifying the pathogen involved-the antigen, as well as the (immunological) traces
of its passage through/or in contact with the body-antibodies.
EPIDEMIOLOGICAL INQUIRY
It is defined as a complex of investigations aimed to: EPIDEMIOLOGICAL INQUIRY
• Determining the conditions in which the disease
occurred
WORKING METHODS:
• The source of the pathogen
Collecting anamnetic data;
• Transmission ways
Investigation of the veterinary sanitary records;
• Factors that favored the occurrence of the disease
Examination of the outbreak.
• The extension of the disease
• Other data necessary for the DEVELOPMENT OF

THE DISEASE CONTROL PROGRAM.
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EPIDEMIOLOGICAL INQUIRY EPIDEMIOLOGICAL INQUIRY

At the end of the epidemiological investigation, it must be established :
 Origin and pathways for the introduction of the disease- endogenous
 Date of the disease occurrence and if it was announced from the first
or exogenous;
suspicion;
 Clinical and anatomo-pathological aspects that created the suspicion of  Establishment of suspected contamination areas by the disease
disease as well as subsequent investigations required to confirm the outbreak studied (through vehicles, humans, objects or animals);
disease;
 Identifying the favoring factors of the onset (occurrence) and
 Degree of the disease expansion into a territory, evolution of the disease;
 Species and animals categories affected,

 Other specific local issues related to the occurrence of the disease.
 Number of sick or suspected to be contaminated cases.
SOURCES OF INFECTION
PRIMARY SOURCES:
SECONDARY SOURCES:
Allow multiplication of the pathogen:
Are only support for the pathogens
- Living receptive organisms to the disease:
- without sensitivity Outdoor Environment:
- with sensitivity but reduced by vaccination Fixed supports: soil, meadows, roads, buildings,
- sensitive inventory items (stalls, gullies, tools)
- Carcasses of dead animals Mobile supports: water, air (aerosols), vehicles,

humans, domestic and wild animals
- Animal products
LIVING RECEPTIVE ORGANISMS

LIVING RECEPTIVE ORGANISMS
Organisms WITH SENSITIVITY to the disease but
Organisms WITHOUT SENSITIVITY allows the development REDUCED BY VACCINATION
of the pathogen up to a certain level, but without expressing
the symptomatology. In general, vaccination protects animals and does not allow
the development of targeted pathogen in the animal.
Epidemiological importance:
- no. pathogens/individual vs. no. individuals/population Vaccination protects the animals from the clinical expression
of the disease but not from an infection (vaccination
- maintain the disease into the flock. suppresses sensitivity but not receptivity).
A vaccinated flock can be infected in a silent manner, the

infection being detectable only at the individuals who have
not been vaccinated.
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LIVING RECEPTIVE ORGANISMS CARCASSES OF DEAD ANIMALS
SENSITIVE ORGANISMS WITH CLINICAL SIGNS of the Frequent are sources of infection.
disease.
Destruction the carcasses of the dead animals is

Can eliminate infectious agents in different phases of the one of the most important ways of fight and
disease evolution:
eradication of infectious diseases.
- incubation,
-clinical expression, If additional examinations are required, all necessary

measures are taken (non-specific and specific) to
- convalescence. avoid the release of the pathogen.
PRODUCTS OF ANIMAL ORIGIN PRODUCTS OF ANIMAL ORIGIN

They generally come from clinically healthy animals.
Survival time of the pathogen
Products from diseased animals are banned from ↓
consumption based on the sanitary examination or are INFECTIVITY PERIOD OF THE PRODUCTS
directed to be used after conditioning.
Carrier → inappropriate processing in kitchens or public Environmental factors with inactivation properties action
catering establishments (UV, heat, dryness, dampness)
↓
↓
Pathogen replication
↓ progressive reduction of infectious units.
Collective food poisoning
RECEPTIVITY OF ORGANISMS TRANSMISSION WAYS

The receptivity of organisms is influenced by a series of DIRECT - through direct contact with the organism which
determining and favorizing factors. spread the pathogen:
DETERMINING FACTORS
- Horizontal - transcutaneous and mucosal route
INTRINSIC or inborn-are dependent on the body and cannot be (digestive, respiratory, conjunctivitis, genital);
modified: species, race, age, sex, individual.
EXTRINSIC are those external to the body and at this level you can act - Vertical - from mother to baby animal (hen embryo).
to interrupt the epidemiological chain:
PATHOGENS, SHELTER, FOOD, EXPLOITATION SYSTEM,
CAREGIVER. INDIRECT - by indirect contact
FAVORABLE FACTORS-are represented by those elements that -contaminated food, water, air, vehicles, humans and
"help" the major factors of receptivity in the expression of the disease, non-receptive animals.
but which alone cannot trigger the morbid process.
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EPIDEMIOLOGICAL DYNAMICS
Disease evolution in (population, time, space)
SPORADIC-disease that evolves with isolated cases in a herd, in an indefinite
period of time, usually in certain small areas and in certain environmental
conditions. These cases appear to be unrelated to each other.
ENZOOTIC-disease with slow evolution-months, years in the herd, involving a small
number of animals, without a tendency to spread outside the outbreak-farm SUSPICION DIAGNOSIS
and with a trailing character in the herd.
EPIZOOTIC-a disease involving a large number of animals in a relatively short time CLINICAL AND
with a tendency to spread outside the herd, including several neighboring ANATOMOPATHOLOGICAL
farms, in a short period of time, on geographical areas as large as counties or
provinces. EXAMINATION
PANZOOTIC-disease with very rapid evolution-days, including in a relatively short
time a large number of animals, or species, on large areas such as countries or
continents, COVID, FMD example.
ANAZOOTIC DYNAMICS-is a particular case of enzootic evolution in which, based
on the epidemiological investigation, the evolution of the disease is found in a
large number of animals, within a farm, in a relatively short time, but which
recognizes a single source of infection, for example poisoning with botulinum
toxin.
CLINICAL
DIAGNOSTIC GENERAL (MAIN) EXAMINATION METHODS:
Measures prior to clinical examination, -Inspection,

-Palpation,
Ensuring labor protection measures- -Percussion,
protective equipment (boots, gown, gloves); -Listening,
-Thermometry
Hand washing and disinfection;
SPECIAL METHODS (COMPLEMENTARY)
Training of support staff;
-Physical: measurements, some laboratory exams (VSH),
The examination is done in the presence of the owner -Endoscopic, ultrasound, radiological, scraping, surveys;
or the caregiver; -Biochemical and Hematological: qualitative and quantitative;
-Biological.
The examiner will determine the approach plan and
contention as well as the examination plan.
CLINICAL OBSERVATION SHEET ANATOMOPATHOLOGICAL EXAMINATION

Following the anatomopathological examination, common data are obtained can be:
Clinical observation. non-specific lesions (refers only to the main types of diseases),
characteristic lesions for an organ (points to a group of diseases),

THE OBJECTIVE EXAM
pathognomonic lesions (Babeş-Negri corpuscles).
APPARATUS AND SYSTEMS EXAMINATION
Conditions for performing a accurate anatomopathological examination:
to be executed immediately after death or slaughter

in specially places
in compliance with labor protection measures.
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EXTERNAL EXAMINATION OF THE CORPSE:

Assessing the state of maintenance,
The external appearance (cadaveric changes,

skin, mucous membranes, natural openings,
horns, hooves, hair).
The necropsy technique involves:
Positioning and skinning will be performed before the internal examination.
Informative stage Large animals:
-position suspended from the rear train,
-left or right dorso-lateral position with the opposite biped suspended.
Descriptive stage
Medium and small animals are usually positioned dorsally, tying the limbs to the necropsy
table or tray if possible.
External examination of the corpse
Skinning is not done in animals suspected of anthrax, rabies or snort and is more difficult in
pigs due to adipose tissue.
Internal examination of the corpse
Skinning involves making several lines of incision:
A line joining the pubis with the anterior extremity of the mandible passing along the white
line,
Two lines perpendicular to it that join the metacarpal regions
and respectively metatarsals of the limbs, on their internal face.
INTERNAL EXAMINATION OF THE CORPSE:

Evisceration of small animals-done in bloc (all organs are removed in one
piece).
THE EVISCERATION
The opening order of the cavities:
Evisceration of large animals-performed on anatomical parts:
1. Pelvico-abdominal
2. Thoracic
Oral-cervical-thoracic piece
3. Pericardial
(tongue+larynx+pharynx+esophagus+trachea+lungs+ heart)
4. Head cavities
Stomach and spleen (before they are removed, the permeability of the
A buttonhole is made at the level of the white line, it is examined through the
choledochal canal is tested)
buttonhole if there are collections at this level (it is collected, examined to
establish its nature), the incision is continued up to the xiphoid appendix and then
The greater mesentery, the duodenum, the jejunum, and the ileum.
on either side of the hypochondrium .
The small mesentery, and the large intestine as a whole.
Opening the chest cavity can be done in several ways:
Liver
Large animals-suspended from the rear train-by cutting the diaphragm;
-dorso-lateral decubitus-by cutting the opposite hemithorax, only the first and last ribs
The kidneys and ureters remain attached to the abdominal cavity (they
are left in the anatomical position;
are removed only if changes are necessary, in order to do additional
Small animals-supine by raising the sternal plastron (sectioning the sterno-costal
examinations, including the renal decapsulation test).
joints) or by raising the sternum and ribcage (sectioning the ribs as close as
possible to the spine).
ANATOMOPATHOLOGICAL EXAMINATION IN BIRDS
A buttonhole is performed behind the xiphoid appendix, continuing with an HARVESTING, PACKAGING AND SHIPPING
incision line, the ribs, coracoid processes and clavicular branch are sectioned, PATHOLOGICAL MATERIAL
including the muscular structure.
The sternal plastron is removed by bending it forward. Harvesting must comply with a number of general rules:
A first inspection of the organs in the pericardial cavity is done.
Collected samples must be representative for the respective pathological process
THE EVISCERATION (representativeness);
-heart and liver Sufficient amount of pathological material is collected (sufficiency) to allow complete
-esophagus, proventriculus, ventricle, duodenum and pancreas laboratory analyses;
-spleen Avoiding sample contamination (sterile instruments, compliance with asepsis rules);
-the intestinal mass, the cloaca, the anal orifice, and the oviduct. Harvesting is done only from fresh corpses;
Samples for the bacteriological examination must come from animals that have not
In the thoraco-abdominal cavity remain: the lungs, been treated with antibiotics, otherwise it is specified in the ACCOMPANYING NOTE
ovary or testicles and kidneys which antibiotics were used, which other treatments were carried out and on what
(which are extracted, only if they are modified). date.
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Samples are kept in a refrigerator (2 to 8 degrees Celsius) until shipping.

SAMPLES TRANSPORTATION
Samples collected for bacteriological and virological examinations:
-sick or alive animals. The corpses are packed in plastic bags that are placed in wooden boxes
-whole carcasses of birds and small animals, in bags soaked with 3% tannic with sawdust.
acid or 1% sublimate.
-whole organs or parts of organs, long bone with unopened marrow. Whole organs and parts of organs, if they are rich in liquids, are packed in
-pathological fluids: secretions, excretions, pus. plastic bags that are later placed in hermetically sealed plastic or glass
-blood or organ smears. containers, if they are poor in liquids, they will be wrapped in parchment
paper.
Biological sample in the bacteriological examination, the same measures
specified above, and for virology, blood, pathological humors are collected in a Smears are wrapped two by two with the side used for performing the
thermos with ice or 50% glycerin. smear on the outside, the wrapping is done in paper.
For the histopathological examination, 5-10 mm slices of the organ are DISPATCH
collected in a 10% formalin solution.
Shipping is done with:
1. OFFICIAL ADDRESS and
2. ACCOMPANYING NOTE
completed and signed by the veterinarian.
Newcastle Disease - birds:

Harvested:
-tracheal and cloacal tampons
-the lung
-the Provençal
-brain
-blood or serum
Required laboratory analyses:
(specific to a viral disease):
-virus isolation
-immunofluorescence
-seroneutralization
Birds pasteurelosis CHOLERA

Harvested:
-unopened long bone
-blood
-liver
-heart with intact pericardial sac
-fingerprint smears on the surface of the heart
Required laboratory analyses:
-bacteriological examination
-bacterioscopic examination
-mouse bioassay to determine pathogenicity
DVM PhD Dragoș Cobzariu 2022 39
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CONFIRMATORY METHODS:
 Isolation and identification of the CAUSATIVE AGENT,

CONFIRMATORY DIAGNOSIS  Highlighting IMMUNOLOGICAL RESTRUCTURING,
1. ISOLATION AND IDENTIFICATION OF THE  Highlighting the specific ANATOMOPATHOLOGICAL LESIONS,
PATHOGEN
 Inoculation-BIOPROBE: mice. rat
2. IMMUNOLOGICAL RESTRUCTURING
 BIOMOL: POLYMERASE CHAIN REACTION
RECORDING
DVM PhD Dragos Cobzariu 2022 1 DVM PhD Dragos Cobzariu 2022 2
ISOLATION AND IDENTIFICATION OF THE CAUSATIVE ISOLATION AND IDENTIFICATION OF THE

AGENT CAUSATIVE AGENT
1. BACTERIOLOGICAL EXAMINATION
1. BACTERIOLOGICAL
EXAMINATION
Requires an initial direct bacterioscopy examination, for visualizing
bacteria in smears from pathological material after coloring :
GOALS bacterioscopic examination:
 Usual methods: Gram, Methylene blue form,
size (the largest are rotting bacillus) grouping mode,
 Special methods: Ziehl-Nielsen, Fontana-Tribondeau,
Coster-Koslovski presence, number and layout of the cilia,
Methods used for highlight certain cellular organisms:
the presence of the capsule and the spore ability,
Capsule: Giemsa, Toluidine blue
Spore: Green of Malachit tinctorial affinity.
Cilia: Casares-Gill
INSULATION AND IDENTIFICATION OF THE CAUSATIVE

AGENT
2. BACTERIOLOGICAL EXAMINATION
It assumes seeding on culture media. 2. BACTERIOLOGICAL EXAMINATION
Cultivation conditions and Cultural characters.

CULTURAL CHARACTERS
CULTIVATION CONDITIONS
Usual culture environments: broth and agar,
Differentiates bacteria according to: Special culture environments:
• respiratory type: aerobic, anaerobic, aerobic optional, Isolation: serum, blood, glycerin, potato, egg
• temperature (36-39°C for those that parasite humans and Enrichment: Kauffman-Muller, Chapmann
animals) Selective: Lowenstein, Sabouraud
• requires the presence of growth factors Differential: Wilson-Blair for salmonella
• time of crop development.
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THE OBJECTIVES OF THE EXAMINATION OF CULTURAL

CHARACTERISTICS:
- liquid media:
- turbidity, EXAMINATION OF BIOCHEMICAL PROPERTIES
- deposits
- surface formations,  fermentation of sugars
- the pigment.  the hemolysis test
- solid media:  the indole test
- how to develop,  highlighting hydrogen sulfide
- size, shape,  oxidase test
- the surface,
- edges,
- the color,
- opacity of colonies.
For bacterial species with antigenically differentiated populations, the

antigenic structure is defined by group and type serums:
 bacterial typing
 pathogenicity testing
Identification of antigenic characters - identification of an antigenic
structure based on positive reactions with a known specific serum.
Determination of pathogenecity by performing in vivo or in vitro tests.
Sensitivity to environmental factors (physical, chemical, biological):
Halophilic (grows in or can tolerate saline conditions).
Sensitivity to certain antibiotics.

Sensitivity to specific bacteriophages.
Antibiogram
Fagotipizare Salmonella spp.
The mechanism of the antibiogram implies
inhibiting the development of a microbial culture
contacted with different types of antibiotics.
VIRUSOLOGICAL EXAMINATIONS
RICKETTSII AND CHLAMYDIA
Viruses are isolate on live cells only:
COLORATION METHODS cell cultures,
- Giemsa, embryonated eggs,
- Romanovski, laboratory animals
- Machiavello,
- Stamp. CELLULAR CULTURES
- primary : fresh cells, chicken fibroblasts, kidney cells,
CULTIVATION MEDIA: embryonic tissues or very young animals
- on living cells, - permanent (serial): the most commonly used tumor
- embryonated eggs, tissues are epithelial or conjunctival malignant tissues with high
- guinea pigs, rates of multiplication.
- mouse. Example:
HeLa (donor – human uterine sarcoma),
BHK21 (cell line obtained from hamster kidneys).
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VIRUSOLOGICAL EXAMINATIONS
READING AND INTERPRETATION OF RESULTS INOCULATIONS ON EMBRYONATED EGGS
By direct methods involving the observation of morphological or
physiological changes of the cellular substrate by the occurrence of: Embryonated eggs are used for 10 to 12 days incubation old.
1.- either a cytopathic effect WORKING STEPS
1. Mirage of the eggs before inoculation with
2. - either a degenerative effects, which consist of:
- decrease in cell volume the marking of the embryo and the air chamber;
- the appearance of intracytoplasmic vacuoles 2. Inoculation with sterile needles and syringes;
- the appearance of intracytoplasmic granulations, 3. Sealing with paraffin the inoculation shell hole.
- pycnotic nucleus,
- a foamed cytoplasm
3. - either modifications with production of inclusions:
- Intracytoplasmic: Newcastle Disease, Bovine fever, Rabies
- Intranuclear: Equine rhinopneumonia, Avian infectious
Encephalomyelitis, Marek's disease
- Intracytoplasmic and Intranuclear: Distemper disease
4. - Proliferative type changes giant cell type:
Parainfluenza virus
INSOLATION OF VIRUSES ON LABORATORY ANIMALS

INOCULATIONS ON EMBRYONIC EGGS Rabbit, is used to isolate the Aujeszky's disease virus, Mixomatosis
Guinea pig, for - Foot-and-mouth disease
INOCULATION EMBRYONATED EGGS ROUTES: Golden hamsters for - vesicular stomatitis, vesicular exanthema
Mice for- swine influence
Chorioallantoic (herpesviruses), Rat for - vesicular stomatitis
Allantoic (vaccines production), The dog - parvovirus, Distemper disease-morbilivirus, Rubarth
Intraviteline (rickettsi and blue-tongue virus), hepatitis, equine pest
Amniotic fluid (varicella viruses). Cat - feline panleucopenia
Chickens - avian poxvirus, infectious avian bronchitis
Changes may occur at: Duck bells - v. Hepatitis
Embryos, The dove - the avian poxvirus
Egg media,
Chorioallantoic membrane.
Inoculation routes: s.c., i.m., i.p., i.v., i.c.
Inoculated animals are marked and separated from healthy ones. CONFIRMATORY DIAGNOSIS
Their observation is according to the clinical observation sheet
BY IMMUNOLOGICAL
including the paraclinical examinations necessary for the
assessment of the physiological and hematological constants.
RESTRUCTURING
RECORDING
Necropsy should be performed immediately after death under
rigorous aseptic conditions.
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IMMUNE CELULAR SYSTEM: system that Immunological effectors recognize on the one hand a CELLULAR RESPONSE
recognizes what is proper to the body, helping that T lymphocyte is responsible for (these effectors are much more prompt
to preserve its immunological integrity. within 24 to 48 hours) and on the other hand, a HUMORAL IMMUNE RESPONSE
that manifests itself after 7 to 14 days post-aggression, represented by
immunoglobulins and complement.
Immune Cell Components:
granulocyte system (myeloblast, myelocyte,
metamielocyte, neutrophil, basophil, eosinophil),
megakaryocito-thrombocyte,
lymphoplasmocitary system
monocitomacrophagic.
Among them, immunology specialized cells for non-

self interaction: lymphocytes and macrophages.
METHODS OF METHODS OF IMMUNODIAGNOSIS

IMMUNODIAGNOSIS
Methods used in practice on the one hand as diagnostic
confirmation or surveillance methods and on the other as methods of
identifying the antigenic structure of a pathogen.
These reactions are based on the high specificity of the

antigen-antibody couple (for each antigen in nature, superior
organisms can synthesize a specific antibody).
The basics components (or necessary) of serological reactions

tests are: the antigens, the antibodies , and the electrolyte
environment, the immune complex resulting from their union becomes
visible in the presence of sodium and chlorine ions.
The antigen may be:

A serological reaction to be used with diagnostic value must meet two
CORPUSCULAR: bacteria, viruses, parasites (also called characteristics:
AGGLUTINOGENS) SPECIFICITY = ability of a serological response to provide a positive response
MOLECULAR: secretions, toxins, flagella (also called to an infected individual;
PRECIPITOGENS)
SENSITIVITY = the ability of a serological reaction to provide a negative
Antibodies can be found in different liquid media of response to an uninfected individual.
the body, its secretions and excretions such as:
In vitro serological reactions are influenced by a number of factors such as:
blood serum,
temperature, concentration of hydrogen ions, pH, electrolytes presence
milk,
vaginal secretions,
lacrimal secretions. The presence or absence of antibodies in the analyzed samples does not
conclude the presence or absence of the disease under investigation.
Depending on the complementary antigen they are called
AGGLUTININS or
PRECIPITINS.
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The outcome of the serological test should take into account:

moment of investigation compared to the time of the infection (if
immunological restructuring occurred),
the age of the animal (the duration of maintaining maternal antibodies
cattle and horses is 6 months, piglets 3 to 5 months, dogs and cats
about 3 months, in chickens about 2-3 weeks). 1. SERO-AGGLUTINATION REACTIONS
the time of infection during the gestation (the risk of newborns with
immunological tolerance).
SERO-AGGLUTINATION REACTIONS SERO-AGGLUTINATION REACTIONS
- Brucella ANTIGEN colored with pink bengal , positive brucella serum,

QUALITATIVE AND QUANTITATIVE. negative serum, glass blades, sample (blood serum),
- Place a drop of serum to be analyzed, a drop of brucella antigen
colored with pink Bengal and mix for 4 minutes by repeated bending of
the blade (positive reaction = agglutination visible after four minutes).
I. QUALITATIVE REACTIONS
The use of whole blood is possible in this type of reaction, an example
A. QUICK AGGLUTINATION REACTION ON GLASS BLADES being the diagnosis of salmonellosis or brucellosis antibodies.
To identify the components of an antigen (somatic, flagella, capsular,

Quick serological method used in practice to highlight surface) can be made with known antibodies by putting in contact with a
serum antibodies using known antigens (Inactivated bacteria bacterial culture on solid nutrient medium.
cultures)
II. QUANTITATIVE REACTIONS
A. Slow Agglutination Reaction

It is used for the evaluation of serum titers in chronic infections
(salmonella, brucella, mycobacterium, mycoplasma, E.coli).
B. RAPID AGGLUTINATION REACTION IN THE TUBE- WRIGHT TEST
(Ringtest) or ring reaction with milk used in the diagnosis of This involves highlighting agglutination antibodies by the occurrence of
brucellosis. the agglutination (submission of the antigen-antibody complex at the
bottom of the reaction tube) interacting with a specific antigen (to
In a Wassermann tube, place 2 ml of milk and 2-3 drops of brucella detect classes of immunoglobulins like IgG2 and IgM).
antigen (hematoxylin or triphenyltetrazolium chloride) is incubated 30-
120 minutes at 37 degrees Celsius (positive reaction = colored ring at Techniques are varied but have the same principle as Wright's
the milk surface) reaction diagnosis of brucellosis in animals. Minimum antibody
concentration expressed as maximum dilution is the agglutinating
titer of the serum to be investigated.
B. Agglutination-lysis reaction with microscope observation
It is used for the diagnosis of leptospirosis and for the identification

of leptospirosis.
5
13
2. SEROPRECIPITATION
https://youtu.be/IKZ5hD80I1c REACTION
https://youtu.be/oXL1rH11MTg
https://youtu.be/3W67OH3v2lU
SEROPRECIPITATION REACTION 2. GEL PRECIPITATION REACTIONS
IMUNODIFUSION:
There are numerous techniques that differ by the way the antigen is
Vertical: with single or one-dimensional and double or bi-dimensional
produced by the reaction medium used.
variants
Horizontal: single or radial (one-dimensional) and double or two-
1. ASCOLI REACTION, is used for the current diagnosis of ANTRAX, has two
dimensional.
options:
• HOT REACTION (used to extract the antigen from the well-
Application: titration of sera to determine immunological or carrier status,
vascularized organs)
identification of antigenic fractions to establish antigenicity, to reveal
• COLD REACTION OR MACERATION (used to extract skin antigen,
immunological deficiencies.
wool)
In a Wassermann tube, place 1 to 2 ml of antigen and with a Pasteur pipette inserts

Principle: depositing in different environmental points of those two
an equal amount of precipitating serum at the bottom of the tube without mixing the reagents and subsequently, during incubation at 25°C, these diffuse to
media. The reaction is considered positive if after 2 - 3 minutes a ring or disc is each other, and to the site of complex formation immune appears a
formed at the contact area of the two media. macroscopically visible as precipitation line.
IMMUNOELECTROPHORESIS
Serological technique combining a physical method (electrophoresis)

with an immunological method (agar gel immuno-diffusion).
Under the action of electric current, Ag or Ab will

migrate, the fractions of the protein mixture are separated https://youtu.be/Ew15yHuqzmY
with each other according to their molecular weight and
the thickness of the agars layer. Separated fractions are https://youtu.be/Bn-w6P_9TUA
identified by the agar gel immune-diffusion reaction. https://youtu.be/4vgyrT0U7S8
https://youtu.be/zUGikX9ZB9U
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3. NEUTRALIZING REACTIONS
A. HEMAGLUTINATION INHIBITION REACTION
It is based on the fact that antibodies contained in an immune-

derived serum from animals passed through the disease or immunized,
are able to neutralize the hemagglutinating capacity of the viruses.
Principle: lay in contact the serum containing specific Ab with the B. SERONEUTRALIZATION TEST
viral suspension and after a 30 minute incubation at 37 ° C, the mixture are
set in contact with red blood cells 0.5%. It is based on the fact that, Antibodies actively formed by a body,
that has come into contact with an antigenic agent, are able to neutralize the
The reaction can be carried out in two processes. infecting value of the corresponding antigen.
- Alpha - the virus suspension is diluted in serial and mixed with amounts equal test Application: identifying an unknown virus based on standardized
known; immune sera. Or the identification of unrecovered serums using typified
- Beta - serum diluted serially is mixed with constant amounts of virus diluted to a viruses (identifying Antibodies in the blood serum).
known number of hemagglutinating units. Seroneutralization can be performed on cell cultures, embryonic
In both cases, if specific coupling occurs
eggs or laboratory animals.
Hemagglutinating Ab-virus, erythrocytes agglutination
phenomenon no longer occurs.
This reaction allows the determination
of the Ab nature of a serum or strain isolated by
cultivation or from the diseased animals organs .
4. COMPLEMENT REACTIONS
COMPLEMENT FIXATION REACTION

Principle: Highlighting the fixation of Alexine on the Ag-Ab complex with
the help of a hemolytic system revealer.
https://youtu.be/YZ67e-YVdrc
Required Elements:
https://youtu.be/CFCi5Q4rhOU
https://youtu.be/nN8MBU8S4EI Ag + Ab + Alexine (complement) = first immune system
Suspension of sheep red blood cells and hemolytic serum = the second
immune system
RFC = negative - positive / hemolysis ( are not specific for an Ag ).
Applicability:
-infectious diseases in which other serological techniques can not be used,

-identification and titration of complement fixation antibodies present in the
blood to determine the disease state or to identify existing antigens in a https://youtu.be/WNmVTKcEN6w
biological product;
-for rapid and accurate determination of infections produced by viruses
https://youtu.be/DPNnZE4OtCM
(foot-and-mouth disease, avian leucosis), bacteria (leptospirosis), https://youtu.be/vboyla11Lyw
protozoa;
- to identify the structure of some antigens, their type and purity using
https://youtu.be/zbf27pCTjRQ
specific sera. https://youtu.be/1Cho6NPtsVw
https://youtu.be/rxvuwwmGC0U
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15
5. IMMUNEECTRONOMICROSCOPY
IEM-TRAPPING TECHNIQUE: specific absorption of
viruses, from a suspension (sample), on the surface of
a pre-coated grid, with specific antiserum or
Principle: visualization on the electron microscope antibodies- IgG, followed by negative staining, and
the immunoglobulin-virus interaction. electron microscope examination.
The method combines serological identity with Technical Advantages and Applications:
morphological identity, which gives IEM the
highest level of specificity. -both whole virions, and their antigenic components
may be examined: antibodies may also attract specific
Variations: aggregates, from the samples with a low
- specific grip (trapping), concentration, making it possible to examine them;
- clumping, -the specificity of reference antisera can be checked;
- specific antibody coating (decorating). - interpretation of results, is much easier, due to the
presence of a large number of viral particles of the
same type.
6. ANTIGEN-ANTIBODY REACTIONS EVIDENCED BY

LABELED REAGENTS
A. IMMUNOFLUORESCENCE REACTION
It is based on the specificity of Ag-Ab reaction, in which

purified immunoglobuline, are chemically bonded by fluorine-free
amine groups, the most commonly used being fluorescein
isothiocyanate.
Immunoglobulin so marked retain their ability to recognize
their specific antigen, visualizing the marker by microscopy in U.V.
light.
The most common IF tests in practice are:
DIRECT IMMUNOFLUORESCENCE TEST (IFD)

- is used to detect the antigen (infectious agent) by known Ab. R + =
https://youtu.be/Vfc_59z05vA
fluorescence https://youtu.be/pteO6FRWo3g
INDIRECT IMMUNOFLUORESCENCE TEST (IFI) https://youtu.be/GKuDbfKUSdk
- can be used for both Ag and Ab detection
NEUTRALIZATION AND IMMUNOFLUORESCENCE TEST

- is used to highlight specific antibodies (both qualitatively and
quantitatively). R + = lack of fluorescence
Benefits:
- Speed
- Simplicity
- Sensitivity
- Small quantities of reagents.
8
16
B. IMMUNOENZIMATIC REACTIONS - ELISA
ELISA principle: detection of antibodies or antigens through enzymes using

Enzyme Linked Immunosorbent Assay the classic antibody-antigen reaction, the enzymes modifying the color of
Antibodies Detection variant (direct method) their specific substrate.
Antigen Detection (simple sandwich)
Benefits:
ELISA - competition (for classical swine fever virus-CSF) - the -very specific - allows identification of a reagent in very small quantities (Nano
competition to occupy Antigen vCSF sites by Antibodies from the grams);
rabbit anti-vCSF serum. -the reagent processing technique allows a high degree of stability;
-widespread use in qualitative diagnosis and / or quantitative of bacteria,
Presence or absence of Antibodies from the serum to viruses, fungi, parasites or vaccine control.
be investigated highlighted with
rabbit anti – Ig G peroxidase.
7. REACTION OF ENZYMATIC AMPLIFICATION IN VITRO GENES

OR PCR (POLYMERASE CHAIN REACTION)
The PCR technique is based on the cyclic function of a DNA polymerase that is
capable of copying a DNA strand used as a model, producing a complementary
strand by elongation starting from the free 3'OH end of an oligonucleotide
https://youtu.be/My375sC5Qkc primer.
https://youtu.be/p8LbQzxpBGE The PCR technique consists of performing "n" successive amplification cycles
https://youtu.be/zR_xlV5v_f4 during which two primers direct the amplification of the double-stranded DNA
sequence it fits. An amplification cycle is composed of three steps, allowing
https://youtu.be/CWkrQrq0yxQ successive completion of denaturation at 95 ° C, hybridization of the primers at
37 °C and extension of the DNA strand under the action of polymerase.
Theoretically, the amount of DNA between the two primers doubles after each
amplification cycle, with an exponential increase.
The amount of DNA obtained by amplification is sufficient to be visualized

directly in the UV.
The specificity of the reaction lies in the choice of the most specific and
constant amplification sequence of the species, the type of nucleic acid to be
identified and in the correct choice of primers relative to the amplified segment.
CONFIRMATION DIAGNOSIS THROUGH GENOM CHARACTERISTIC

SECTION EVIDENCE
ENZYMATIC AMPLIFICATION OF GENES IN VITRO

Animated model
9
17
BIBLIOGRAPHY
https://youtu.be/IbNwwOPH8Cc
https://www.youtube.com/watch?v=om1vzR1B5JM
https://youtu.be/LrbHPx_unDo
https://www.youtube.com/watch?v=cLXSE4RXtJY https://youtu.be/JPlTtFYd6e4
https://www.youtube.com/watch?v=NKNK1MYvZzA https://youtu.be/My375sC5Qkc
https://youtu.be/IbNwwOPH8Cc https://www.youtube.com/watch?v=iKVscHOvj2c
https://www.youtube.com/watch?v=SRI2DEPNgUI
https://youtu.be/p8LbQzxpBGE
https://youtu.be/zR_xlV5v_f4
https://youtu.be/CWkrQrq0yxQ
https://youtu.be/LrbHPx_unDo https://youtu.be/witrkDDm_aY
https://youtu.be/jWkMSw5ynyY
https://youtu.be/Vfc_59z05vA
https://youtu.be/pteO6FRWo3g
https://youtu.be/JPlTtFYd6e4 https://youtu.be/h6ZFI-WMJR8
https://youtu.be/Cb2g9w-A8bU
https://youtu.be/GKuDbfKUSdk
https://youtu.be/WNmVTKcEN6w
https://youtu.be/EE-jHN_c1Dw https://youtu.be/DPNnZE4OtCM
https://youtu.be/vboyla11Lyw
https://youtu.be/Q9OARuvZFQk
https://youtu.be/zbf27pCTjRQ
https://youtu.be/JPlTtFYd6e4 https://youtu.be/1Cho6NPtsVw
https://youtu.be/QBcxFwp3Byo https://youtu.be/YZ67e-YVdrc
https://www.oie.int/doc/ged/D9468.PDF https://youtu.be/CFCi5Q4rhOU
https://www.youtube.com/watch?v=nKQ5M6rCjmI https://youtu.be/nN8MBU8S4EI
https://youtu.be/Ew15yHuqzmY
https://youtu.be/yWtIHuZRwXM
https://youtu.be/Bn-w6P_9TUA
https://youtu.be/dflOQ8hXUbA https://youtu.be/4vgyrT0U7S8
https://youtu.be/0Rhi6wzT9mQ
https://youtu.be/WaTE0bpPgSk
https://youtu.be/3q0NTqXqviU
https://youtu.be/Ew15yHuqzmY
https://youtu.be/Bn-w6P_9TUA
https://youtu.be/4vgyrT0U7S8
https://youtu.be/zUGikX9ZB9U
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PROPHYLAXIS ( eng., prevention) ( ital. Profilassi) ( portug. Profilaxia) ( rom. Profilaxia)

Prophylaxis can be individual or collective, optional or mandatory. It can be medical,
achieved through vaccines, serums or other chemical compounds, with a role in maintaining
health or medical health. The method can be applied to an infected herd or environment -
defensive prophylaxis, or in an free of disease environment or herd, offensive
prophylaxis.
To be effective, prophylaxis must be based on a good knowledge of the epidemiological
characteristics of the disease.
Any control action, which reduces the sources of infection, (especially the treatment or
PREVENTION AND slaughter of affected animals), has favorable prophylactic consequences, because it reduces
the intensity of exposure of susceptible subjects to the pathogen.
All measures, medical and/or hygienic, intended to prevent the occurrence, spread
and multiplication of cases or outbreaks of a disease.
COMBAT OF Examples of prophylactic methods: vaccination, surveillance of certain diseases, preventive
quarantine when introducing an animal into the herd.
INFECTIOUS DISEASES IN ANIMALS
3 MAJOR STRATEGY ELEMENTS OF 4

PROPHYLAXIS:
OBJECTIVES OF PROPHYLAXIS:
Preventing the occurrence of a disease in animal-free flocks; Depending on the sanitary situation of the flocks, the measures
Applying the necessary measures to limit the spread and eradicate a disease; differ as follows,
Protection of human health, by ensuring the healthiness of products of animal origin. a) In healthy herds, free of the disease, measures will be taken to
prevent its introduction, so PROPHYLAXIS.
b) In contaminated flocks, measures are taken to limit and eradicate
the infectious process, so COMBAT.
Depending on the measures and mode of action, on the
etiological agent, the following are described:
a) Non-specific prophylaxis measures such as: Disinfection,
Disinsection, Deratization.
b) Specific measures, such as immunization or vaccination.
Depending on the targeted target, the measures and mode of
action are oriented towards:
a) animals of a livestock in the form of screening actions, treatments,
immunizations,
b) on the environment in which the animals are raised, microclimate
factors, air currents, temperature and humidity, gases from the
shelter, disinfection.
GENERAL NON-SPECIFIC STRATEGIES

5 6
General (non-specific) Prophylaxis measures include:
The permanent supervision, by predetermined methods, of the Health
of the livestock, as well as of the health of the people who come into
contact with these animals;
Permanent supervision of Environmental Factors, those of animal
INTERVENTION STRATEGIES, respectively health, microclimate, nutrition, as well as stress factors;
Prophylaxis or Combat Measures aimed at controlling the Circulation of animals and products
of animal origin; as well as control of Feed Quality (including
microbiological control);
Primary Strategies, applied before the appearance of an infectious,
pathological process, The application of preventive Quarantine measures,
prophylactic, for all animals newly introduced into a herd,
Secondary Strategies, applied after the detection of the first immunological
restructuring, a farm, a household;
Carrying out periodic Disinfections, Disinsects and Deratizations
Tertiary Strategies, which are applied after the clinical-lesional expression of (D.D.D.);
the infectious process.
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GENERAL NON-SPECIFIC STRATEGIES GENERAL NON-SPECIFIC STRATEGIES

7 8
NON-SPECIFIC COMBAT STRATEGIES: MEASURES USED TO INTERRUPT THE TRANSMISSION OF PATHOGENS,
Disinfection
The non-specific combat measures, are a number of actions, intended to It pursues the removal and destruction of pathogenic agents, or conditionally pathogenic agents from the
skin, and from the various objects in the external environment, using mechanical, physical and chemical
lead at the extinction of the disease, such as: means.
It is prophylactic or routine and in the outbreak.
Reporting the occurrence of the disease, subject to the Mandatory Disinfection in the outbreak can be:
Declaration regime; -continues and must last as long as the sick animal eliminates pathogens.
-final at the end of the quarantine period, with the return to health, immunization, or death of the animal.
The establishment of Quarantine Measures and movement control, 1. The mechanical means of disinfection are: washing, wet wiping of surfaces, mechanical
vacuuming of dust, filtering, airing and artificial ventilation.
depending on the degree of danger to human or animal health, measures 2. The physical methods of disinfection are heat, ultraviolet and gamma radiation.
that can be of degree I, II or III, Heat destroys microorganisms by denaturing their structural proteins. They are used as processes:
flaming, incineration, boiling, and treatment with water vapors.
The application of sanitation measures, respectively liquidation of the Irradiation with ultraviolet radiation is only effective for the disinfection of air and smooth surfaces directly
outbreak, through measures to extract only clinically affected animals, exposed, and located at a maximum of 1.5 meters from the source of radiation, the UV lamp.
This method is used in operating rooms and work tables in laboratories where pathogens are worked
infected carriers, or total elimination (by liquidating the entire herd); with.
3. Chemical disinfectants use different groups of substances, with their advantages and
Measures to destroy corpses, to inactivate and denature products disadvantages.
The application methods are: washing or wiping, immersing objects in disinfectant solution, sprinkling
and byproducts from sick or infected animals; large surfaces, vaporizing in closed spaces.
Among the many disinfectants used, we mention: lime water, sodium hydroxide, formalin, chloramine,
Carrying out Disinfections, Disinsection and Deratization in (strong oxidants, which have limited use, in certain concentrations, in the disinfection of spaces), iodine
and hydrogen peroxide (used to disinfect skin and wounds) .
the outbreak of the disease.
GENERAL NON-SPECIFIC STRATEGIES

9 QUARANTINE 10
MEASURES USED TO INTERRUPT THE TRANSMISSION OF
PATHOGENS, Quarantine means the complex of restrictive measures imposed on
Sterilization the free movement of animals, people, products of animal and vegetable
Pursues the destruction of all microorganisms, pathogenic and non-pathogenic, including spores on
objects with veterinary medical use, such as instruments, equipment in operating rooms, milking origin, feed, utensils, means of transport, and excrement from animals.
machines, waterers and feeders, harnesses.
Pest control-Disinsection, The purpose of the quarantine: to stop the spread of diseases,
It seeks to remove, destroy and prevent the multiplication of vectors, both passive ones, which play a transmissible to animals.
role in the transmission of infectious diseases (flies, cockroaches) and biologically active,
hematophagous ones, which are also an epidemiological source (lices, ticks, mosquitoes, fleas). Depending on the extent of the disease, its dynamics, the transmission
Pest control can be prophylactic (preventive) when its aim is to prevent the development of insects,
through permanent hygienic-sanitary measures, hygiene of care staff, housing and stables, food routes and the herds of endangered, threatened animals, quarantine is
hygiene and sanitization of shelters, sanitation of marshy lands and flooded basements, application of applied to:
screens to windows , the application of active insecticides and repellents, as well as the application of
preventive traps. Households, animal farm-holdings, localities, pastures, communes,
Pest control are performed by physical, chemical and biological methods:
a) physical procedures such as: mechanical removal; dry heat, buckling of metal and wooden objects, groups of communes, counties or on much larger territories-countries.
use of adhesive traps.
b) chemical processes, use insect repellent substances and insecticidal substances that can be In certain diseases, only on some herds or categories of animals and on
ingested and contact. the products and by-products derived from them.
c) biological methods are much more indicated, in farms and involve the use of natural predators of
hematophagous vectors, as well as the use of buffer animals that will collect the vectors in the vicinity
of the shelters.
According to the severity of the evolution of the disease, the ease of
Rodent control, its diffusion and the peculiarities of transmission, the quarantine
Prophylactic measures and/or combat measures, using mechanical means (traps) and chemical measures are graded as severity, in several grades with decreasing
means (frequent by ingestion).
severity, grade I, grade II, and grade III.
FIRST DEGREE QUARANTINE

11 SECOND DEGREE QUARANTINE, 12
It applies in the case of:
Highly diffusible, panzoothic diseases, such as Foot and Mouth Disease.
Case of diseases with an exotic character, with high difusibility. Are applied to epizootics with medium difusibility, such as
In these situations, the most severe measures are taken to prohibit free movement: Classical Swine Fever.
of animals (including species not susceptible to the respective disease),
of people, of products and by-products of animal and plant origin, of vehicles, In such situations, movement restrictions are applied to all
of tools and any objects that may be contaminated. animals, products and people, only in contaminated households.
The animals are kept in sealed shelters.
Contaminated households are under guard survay. In the rest of the contaminated city, confinement in shelters is
Animal droppings, litter and contaminated feed scraps are destroyed daily. applied, only for the affected species.
The entry and exit, of people from the quarantined territories is prohibited, and is approved
only in exceptional situations of force majeure, with the application of special disinfection The introduction and removal of susceptible animals, as well
measures under veterinary sanitary supervision. as the removal of products from the affected species, from the
The supply of territories under quarantine is done by transshipment. quarantined area is prohibited.
In the contaminated locality it is possible to drive exceptionally by car, only under the
conditions provided by the specially designed technical instructions. People from the outbreak can only leave with the application of
Until the situation is stabilized, RELIGIOUS, CULTURAL activities and manifestations, and special disinfection measures, with the approval and under the
WEEKLY FAIRS, are prohibited, events that may contribute to the spread of panzoothic or
supervision of the local-territorial Veterinary Sanitary and Food
epizootic disease..
Safety Authority.
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THIRD DEGREE QUARANTINE QUARANTINE

13 14
Implementation of the restrictive, quarantine measures is done by the local
councils, through the anti-epizootic headquarters.
The control of compliance with these measures is the responsibility of the
veterinary health and police personnel, who will draw up the documents for
It applies to diseases that are transmitted mainly through sanctioning the violation of the measures ordered.
direct contact, between susceptible animals. Veterinary sanitary personnel are required, to carry out all technical control
operations, according to official instructions from the higher hierarchically superior
For example RABIES, Tuberculosis, and in diseases Veterinary Sanitary and Food Safety Authority, the National Veterinary Sanitary and
Food Safety Agency.
transmitted through mating such as Brucellosis, Herpesvirus
infections. The owners of animals, as well as the heads of units that slaughter animals,
process, store, transport and utilize products of animal origin, are responsible for
observing and applying all the control measures established by the anti-epizootic
In the case of these diseases, restrictions are applied in the commands, and are obliged to ensure the workforce and other necessary
movement or reproduction, of sick and suspect animals, with conditions for carrying out disinfections, pest control, rodents control, as well as
the containment of animals, in order to carry out mandatory sanitary and veterinary
some facilitations regarding the movement of products derived operations, as well as for the application of other measures required for the
from them. purpose of liquidating the epizootic.
The anti-epizootic headquarters are obliged to periodically analyze the evolution
of the epizootic event, take measures to remove the difficulties and deficiencies
that arise, to establish and apply the necessary additional control measures.
The official veterinarians will keep a clear record on the evolution of the
epizootic, periodically reporting to the Directorate of Veterinary Health and Food
Safety, data related to this, and whenever special changes appear in the evolution
of the epizootic.
QUARANTINE
15 SPECIFIC PROPHYLAXIS 16
First and second degree quarantines are instituted by the decision: STRATEGIES:
The Executive Office of the City Council, based on
Are addressed directly to a Single Pathogenic Entity, to one or more species of
The Veterinary Sanitary Act of Finding and Official Declaration of the animals, depending on the epidemiological particularities of the disease.
The specific prophylaxis measures are represented by vaccination and
disease, act drawn up by,
preventive serum antibodies administration.
The Official Veterinarian, of the zonal veterinary sanitary constituency. Immunization is called active, or preventive vaccination, when following it, after
a period of time, of 7-21 days, in the body of the vaccinated animal,
When the territory to be subjected to quarantine measures exceeds
immunoglobulins are produced, and will assure to the vaccinated body, resistance
the area of a municipality, quarantine is instituted by: to clinical expression and/or infection.
Immunization is passive, when preventive sera are performed, containing
The decision of the County Council, based on the proposal,
already existing antibodies, produced in vitro or in vivo, and administered to
County Veterinary Sanitary Directorate and for food safety. animals at risk of infection.
These antibodies confer a resistance to the disease, to the organisms to which they
Level III quarantine is instituted only on the basis of:
were administered, for a variable period of time, but not being the own antibodies of
The Veterinary Sanitary Act of Finding and Official Declaration of the disease, the organism to which they were administered, they gradually disappear from the
act drawn up by, body of the treated one.
The Official Veterinarian, of the zonal veterinary sanitary constituency.
VACCINE is a biological product, obtained from microbial

ACTIVE IMMUNIZATION 17 antigens (integral microorganisms, toxins, or their fragments,
18
subunits) and which, following the administration (usually by
inoculation, injection) of an organism, triggers the mechanisms
Stimulation - Host immune system by an Ag of active immunization, specific to the respective disease by
producing antibodies and specifically stimulated cells.
The conditions that must be met, by an effective vaccine are:
Ensuring a high degree of protection;

Synthesis of specific Synthesis of memory Cells To possess reduced harmfulness (safety, harmlessness);
Ab- IgM,IgG
Not to generate post-vaccination adverse reactions;
NATURAL INHERITED by direct

To determine the reduction or blocking of morbidity, for
contact with pathogenic agent vaccinated animals herds.
ACTIVE IMUNISATION ARTIFICIAL IDUCTED by PRIN The use of vaccines,
VACCINATION
-live (atenuated) Vaccines can only be used for prophylactic purposes; only for
- inactivated (death) therapeutic purposes (eg staphylococcal vaccination);
- subunitary ( sintesis)
or for prophylactic and therapeutic purposes (tetanus vaccination).
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CLASSIFICATION OF VACCINES, VACCINES with Live PATHOGENS

19 20
Live attenuated or heterotypic vaccine antigens, inoculated subcutaneously, will
disseminate in the body, in the same target tissues as in natural infection, where it
According to the nature of the antigen used in the preparation, will produce an infectious process reduced in intensity, followed by the specific
the vaccines can have bacterial origin, viral origin, or parasitic generation of an immune response, as well as the establishment of strong
origin. immunity , lasting.
Disadvantages:
According to the Morpho functional State of the Antigen used
in the preparation, vaccines are produced from live and Non- Post-vaccination accidents may occur, clinically and pathologically similar to
natural infection;
Pathogenic antigens, from Live and Attenuated antigens, from
Inactivated pathogens and Anatoxines (inactivated toxins), The portage of new pathogens, not negligible, can generate new outbreaks with
infectious potential, or the appearance of new pathogenic strains.
vaccines produced from Purified Immunogenic Fractions
(subunitary vaccines), vaccines with viral vectors are described Live vaccines are used, only in necessary vaccinations for the prevention of
using non-replicating adenoviruses, and RNA messenger certain diseases, which do not have harmless immunization, respectively if the
passage through the disease is achieved with the persistance of some sequelae.
vaccines.
Application procedures:
According to the number of antigens-immunogens, vaccines
Simultaneous Vaccination (sero-vaccination-simultaneous administration of
can be: monovalent (one targeted pathogen), or polyvalent
vaccine and antibodies-globulins),
vaccines (against two or more pathogens or toxins).
Vaccination by unconventional means (drinking water, aerosols, conjunctival or
nasal mucosa),
Vaccination with Live Paraspecific Vaccines (heterotypic-for example, lapinized
vaccine against swine plague).
VACCINES PREPARED FROM LIVE

VACCINES CONSISTING OF PATHOGENS
ATTENUATED GERMS 21 22
INACTIVATE AND ANATOXINS
Microbial strains that have lost their pathogenicity, but have kept their
immunogenic properties;will produce a benign, asymptomatic infection, but Vaccines consisting of microbial suspensions killed, by various processes,
generate solid immunity; which, however, preserve the antigenic structure,
Stimulates both Cellular and Humoral immune response; Inactivation is carried out using physical and chemical agents, such as
Live attenuated viral antigenic strains stimulate the immune response to each of heat, and formalin,
the antigenic, immunogenic components of a virus;
Advantages in active immunization, because it presents a practically non-
The immune response is prompt, the installed immunity is solid and lasting; existent risk of infection, they can also be administered to weakened,
Attenuated vaccines retain the ability of the pathogen to replicate in the debilitated animals (for which live vaccines are contraindicated);
vaccinated organism, and it can spread in the animal population, which makes
Disadvantages: it generates a late immune response, a less stable
it possible to immunize even animals in direct contact, but not physically
vaccinated; immunity, with a shorter duration (which involves repeating the vaccination-
the booster, after 1 to 3 weeks after the first vaccination);
The possibility of industrial preparation and mass application, by easy methods,
in drinking water or aerosols. Vaccines consisting of filtrates of bacterial cultures, containing
Disadvantages: reduced stability over time (must be preserved by lyophilisation);
exotoxins, which after inactivation with formalin and heat, are non-toxic
the existence of a residual pathogenicity is described, which can trigger post- preparations with immunizing power-value, called ANATOXINS;
vaccination reactions, or even the risk of the return of the pathogenicity of the Effective active immunization solution against exotoxins produced by
vaccine strains through mutations.
bacteria. Example for vaccines, produced from inactivated toxins, being
Tetanus.
23 24
VACCINES PREPARED FROM
PURIFIED IMMUNOGENEOUS FRACTIONS, MESSENGER RNA VACCINES
Also called Subunitary Vaccines; Messenger RNA vaccines are a newer concept, but not
Active principles are a series of immunogenic fractions existing in the unknown. Messenger RNA vaccines have previously
wall structure or internal structure of antigens or even microbial been studied for viruses that cause influenza, Zika,
metabolites; rabies, and in immunization against human
The immunogenic fractions are, as a rule, viral glycoproteins, viral cytomegalovirus (CMV).
capsid proteins, proteins of the ciliary/pillar apparatus of bacteria, As soon as information about the virus that produces
capsular polysaccharides. COVID-19 was identified and available, scientists
These vaccines are frequently improved with adjuvants like began designing the spyke protein from the messenger
aluminum hydroxide, oily adjuvants, or other adjuvants to increase RNA vaccine.
the immunizing capacity by slowing absorption from the inoculation In addition to vaccines, scientific research related to
site.
oncological conditions has already used
The induced immunity is comparable to that of classic inactivated messenger RNA technology to stimulate the
vaccines, but with a high degree of specificity, towards the antigenic immune system to recognize specific cancer cells.
component for which they were created.
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VACCINES BASED ON VIRAL VECTORS,
Scientists have been studying the production of viral vectors 25 26

since the 1970s. In addition to being used in vaccine
OTHER VACCINES OBTAINED THROUGH
production, viral vectors have been studied for cancer
treatment and molecular biology research. The platform is MODERN TECHNOLOGIES,
intensively studied, hundreds of scientific studies of vaccines
based on viral vectors have been published worldwide. These
vaccines have been produced against diseases such as
Ebola or other infectious diseases such as Zika, influenza
and HIV.
Vaccines obtained by recombinant DNA technology,
Vaccines are usually monovalent vaccines using non- Synthetic vaccines,
replicating viral vector technology. A viral vector is used,
such as adenovirus, whose DNA is modified to become non- Ribosomal vaccines,
replicative, and a gene sequence is attached to it, which codes
for a specific protein, of the virus against which the vaccine is
Anti-idiotype vaccines.
produced. Basically, the vaccinated organism will produce its
own messenger RNA and then the protein, thus blocking the
attachment of the pathogenic virus.
ADMINISTRATION ways OF VACCINE

27 28
VACCINES, ADMINISTRATION
The route of administration differs, so vaccines can be administered by, Vaccine administration schemes are different, depending on the
-scarification, plucking and then bleaching (for example, in fowl pox), Epidemiological Particularities of the disease, so they can be,
-subcutaneously (most vaccines are administered this way), COMPULSORY vaccinations: in Rabies and Newcastle Disease,
intramuscular (vaccines whose immunogenic properties are improved with MANDATORY vaccinations depending on antecedents, for example:
the help of oily adjuvants), Vaccination against, Contagious Agalaxia of sheep and goats: a) vaccinations in
-intra conjunctival (for example, vaccination, anti-Newcastle in chickens),
units and localities with epizootic antecedents: 1. in sheep and goats, in the middle
of the gestation period; 2. in breeding rams and goats, barren sheep and youth, in
-intranasal vaccines used in cats,
the May-June period; 3. in lambs aged 45 days; b) in the units and localities where
-in aerosols (for example, anti-Newcastle vaccination of chickens in
the disease develops, only the clinically healthy population is vaccinated.
batteries),
REQUIRED vaccinations - examples:
-oral vaccination, in baits or drinking water (rabies vaccine for foxes). Vaccination against, Swine Atrophic Rhinitis Vaccinations are carried out in pigs
from farms and localities where the disease has been diagnosed, at the request of
the owners.
Infectious pleuropneumonia of pigs, Vaccinations are carried out in pigs, in farms
and localities where the disease has been diagnosed.
Egg Drop Syndrome, Vaccinations are carried out in birds in farms where the
disease has been diagnosed and are carried out according to the vaccine leaflet.
29 PASSIVE IMMUNIZATION 30
VACCINATION
ADVANTAGES ADMINISTRATION OF SPECIFIC ANTIBODIES
Protection of animals exposed to the risk of disease; ALREADY SYNTHESIZED.
Limiting the Circulation of the disease;
The possibility of eradicating, in time, a disease-for example, human smallpox;
Reduction of economic losses due to illness.
DISADVANTAGES SHORT TIME

Post-vaccination accidents may occur; PROTECTION PERIOD
The antagonism with certain DIAGNOSTIC techniques of animal diseases,
based on the identification of the immune response.
Naturally newborns, will receive
antibodies from mothers
PASSIVE IMMUNIZATION
Administering synthesized products,
in vivo, or in vitro, named immune or
hyper immune serums
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PASSIVE IMMUNIZATION 31 SPECIFIC COMBAT STRATEGIES: 32
without the immunological involvement of the The specific Combating measures are the measures that are taken in
organism to be protected, through the flocks in which the disease evolves:
transfer of active antibodies, produced by Immunization in the Outbreak, which can be Passive, and can be
another animal organism
achieved through Serumization, respectively the administration of
antibodies, to sick animals.
PASSIVE IMMUNIZATION installed immediately Or Active immunization, carried out in disease outbreaks, is carried
out by Vaccinating only healthy animals without clinical signs of
disease.
short duration of protection, from 3 weeks Medical treatment are recommended, when is possible, when the vital
after administration of heterologous sera, up
or economic prognosis are favorable, and the conditions for which the
to 3 months for homologous ones
therapy is carried out, do not have zoonotic potential, or following the
therapy, the animals that have gone through the disease, do not remain
with irreversible sequelae.
Depending on the pathogens against which they are used, these hyperimmune
sera can be monovalent or polyvalent, prepared in vitro or in vivo.
As a source of antibodies, they can be post-vaccinal or post-convalescent.
POST-VACCINATION ADVERSE REACTIONS
It is considered a post-vaccination adverse reaction, a clinical sign, appearing at most 1 month after
33
vaccination, which may or may not be caused by the vaccine product or the vaccination procedure. Only in the
case of certain vaccines, certain adverse reactions can extend over a longer post-vaccination period.
The incidence of these effects in different species and populations of animals is low, because vaccination is
usually addressed to healthy animal flocks and, often, has a mandatory prophylactic character.
The following post-vaccination adverse reactions may require careful medical supervision, in the next
24 hours, from identification, by the veterinarian who performed the vaccination procedure:
1.Severe local reactions-lymphangitis, lymphadenitis, abscesses at the inoculation site, erythema or swelling,
extended to nearby joints lasting more than 3 days, reactions requiring medical supervision;
2.General reactions due to damage to the nervous system-paralysis following vaccination with vaccines
containing attenuated pathogens, with tropism for components of the nervous system; encephalopathies are
described; encephalitis; meningitis; febrile or afebrile convulsions;
3.Other severe side effects that require medical supervision such as anaphylactic, allergic reactions, toxic-
septic syndrome, collapse, hyperpyrexia, arthralgias, myalgias, accompanied by alteration of the general
condition, adynamia, hyperthermia, or worse that can lead to the death of the vaccinated animal.
Depending on the cause, post-vaccination adverse reactions can be classified into:
1.Vaccine-induced reactions-represented by certain particular reactions of an animal to a certain vaccine
product or vaccine compound, effects that would not occur in the absence of vaccination;
2.Reactions potentiated by the vaccine-which can also occur in other situations in certain susceptible animals,
but which are favored by vaccination;
3.Coincidence reactions-which would have occurred even if the animal had not been vaccinated, without
having a causal relationship with the vaccine product;
4.Reactions associated with the vaccination program-due to production defects, handling, transport or
administration errors, deficiencies in the storage of the vaccine preparation;
5.Post-vaccination reactions of unknown cause-when they do not fit into any of the previous categories.
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Contagious, EPIZOOTICO-PANZOOTIC, acute disease common to

BIONGULATES, expressed by vesicular-aphthous rash (F,M,Ma) on
mucous membranes and skin, benign in adults, fatal in youth.
The virus is grown on Sellers kidney cells; on the OIE list, A no. 010.
ETIOLOGY:
Filterable virus, belongs

FAMILY - PICORNAVIRIDAE,
GENOUS - APHTOVIRUS.
Biology: foot-and-mouth disease virus presents
7 major antigenic types:
-A (1-32), O (1-11), C (1-5) will determine panzootic disease;
FOOT AND MOUTH DISEASE -SAT1(7), SAT2 (3), SAT3 (4) which cause epizootics in South Africa;
Fievre aphteuse, -ASIA1 (3) in Asia.
Febra aftoasă,
Maul und klauenseuche
From the point of view of pathogenicity, there are no differences between the 7
antigenic types, regardless of the serotype with they were infected, the animals will
expressed the same clinical signs.
FMD viruses are good immunogens, each type generates antibodies specific to it,
and DO NOT INDUCE CROSS-IMMUNITY. Molecular epidemiology applied in outbreaks in SAT (South African Territories)
highlighted phenomena of genetic evolution through the appearance of mutant
There are also common viral antigens, but antibodies against them do not provide genes: virus escapa mutants.
complete protection (against each type of viral strain).
The virus shows EPITHELIOTROPISM, which explains the pathogenesis,
The diagnosis is simultaneously addressed to viral types A, O, C, spread in the and the location of the lesions.
European geographical area. The existence of the 64 subtypes does not influence
diagnosis and combat, but their study has a role in the development of The virus is ISOLATED ON CELL CULTURES, belonging to the susceptible
epidemiological studies (it makes it easier to track the circulation of viral strains). lines of guinea pigs, and hamster line BHK21.
Clinically, the disease is accompanied by FEVER SYNDROME AND

VESICULAR ERUPTION.
The virulence (pathogenicity) of strains (increased susceptibility of a species or

race to a strain) is expressed by the minimum infectious dose and diffusibility.
Physico-chemical properties of the foot and mouth disease virus:

Sensitive to formalin, alcohol, and to acid pH, lower than 5, the virus is The virus survives the longest, in the bone marrow, then in the
inactivated. tissues that do not suffer maturation processes , it is most easily destroyed in
the muscle tissue.
It resists in the external environment, it resists in biological materials, thanks to
the external capsid proteins, which protect it against denaturation. The capsid is The usual slaughtering technology involves refrigeration shortly after
an arrangement of 60 protomers in a tightly packed icosahedral structure. Each slaughter and evisceration.
protomer consists of 4 polypeptides known as VP (viral proteins) 1, 2, 3 and 4.
The meat products, which result from the necessary slaughter of animals
Once on feed, litter, water, it lasts up to several months. suffering from foot-and-mouth disease, undergo maturation processes (4ºC)
post-sacrifice, which lead to the survival of the foot-and-mouth disease virus.
In the summer, on the pasture, in the aphthae liquid, it lasts for two weeks.
Meat from febrile animals is not given for human consumption.
In non-acidified milk, butter, non-fermented cream, fresh cheese, withstand:
-two weeks if the products are kept in the refrigerator (2to8 degrees)
-one week at 18to20 degrees Celsius;
In meat, the virus is destroyed under the conditions of its acidification.
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EPIDEMIOLOGY:
RECEPTIVITY:
The main domestic species
DOMESTIC AND WILD BIUNGULATES, in descending order: bulls,
pigs, sheep, goats, buffalo, reindeer, deer, camels, antelopes, pigs,
receptive to F.M.D
elephant.
Spontaneous natural infection: HEDGEHOG, RAT, MOUSE.
Refracting naturally are: HORSES AND BIRDS.
Factors that belong to the host and that influence receptivity, in terms of
the severity of the clinical expression, (different evolution in youth
compared to adults, depending on the species, race, infectious dose,
pathogenicity of the virus strain).
The minimum infectious dose (MID) of different virus strains, causing the
same clinical expression of the disease.
Immune status: animals that have gone through the disease have
protective antibodies against a CERTAIN TYPE OF VIRUS, and the
animals become refractory-resistant to infection with that type ONLY.
TRANSMISSION OF INFECTION can be done by:
-oral route, through the mucous membranes, nasal and conjunctival,
-transcutaneous route in the presence of lesions.
Species receptive to F.M.D Species refractory to F.M.D
Species as natural hosts to F.M.D
EPIDEMIOLOGICAL DYNAMICS: DISEASE WITH PANZOOTIC EVOLUTION

PRIMARY SOURCES OF INFECTION:
Time-Space-it has great diffusibility in time-a few days, and space-countries,
Sick Animals and Their Products, continents: it gives the disease a typical Panzootic character.
Animals that have passed through the disease, which remain carriers and shed
the virus for 3-6 months after passing through the disease. Population-several naturally susceptible hosts-Biongulate .
The virus, once eliminated, through secretions-saliva, liquid from aphthae, aerosols,
Buffaloes and capybaras are possible asymptomatic, serologically negative
carriers. survives in the environment for a sufficient time to meet a receptive host.
There are animate and inanimate passive vectors (animals that do not cause disease,
Infected animals eliminate large amounts of virus during the febrile period, the
pathological material richest in virus, being the APHTA LIQUID = APHTAOUS but passively and actively circulate the virus, such as migratory birds, aerosols, air
LYMPH, (through the opening of the aphthae, its liquid mixes with the saliva and currents-African desert storms, with dust particles that circulate the virus in Europe).
thus, the saliva becomes the medium, and the way of transmission of the virus,
from the sick animal to the healthy one. As a result of the immunity specifically induced by each type of foot-and-
mouth virus, pandemics follow each other at intervals of 3-4 years, in the form of
SOURCES OF SECONDARY INFECTION: feed, water, pasture, air currents.
WAVES! Due to the fact that with the renewal of the population of receptive animals,
the immunized ones leave the herd.
If another virus serotype enters the given population, the wave sequence disappears.
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The introduction of the virus into a territory is due to the introduction of PATHOGENESIS:
contaminated products, or of clinically healthy carriers and shedders.
Initially, the appearance of PRIMARY APHTA is described, at the site of virus
The disease is clearly expressed in cattle, strong in pigs and discrete in sheep.
penetration, followed by the appearance of a transient febrile reaction.
Circulation of the virus from endemic areas to free areas is also done by:
SHEEP, the next host being the PIG, which consumes household waste, greasy Viremia occurs, following which the virus is located at the level of the epithelium of
water, and is a virus-eliminating species, in sufficient quantity, to cause cattle the mucous membranes, glabrous skin, Mouth, Mamary gland and Feet, with the
disease. occurrence of SECONDARY APHTHOUS ERUPTION and febrile syndrome.
In a territory, not all susceptible species get sick at the same time.
In the absence of bacterial complications, the canker sores open spontaneously,
In cattle, foot-and-mouth disease can be confused with other diseases of an
they heal "per primam" after 10 days from the eruption, the animal being clinically
enzootic nature. Lameness as a clinical sign is frequently encountered in sheep.
restored,
GEOGRAPHICAL DISTRIBUTION OF FOOT-AND-MOUTH:
Europe is FREE from Foot and Mouth Disease and no longer practices It remains Carrier and Eliminator.
vaccination; vaccination is no longer practiced in Israel; the last wave of foot-
and-mouth disease was registered in England, a few outbreaks were also
described in Holland, France, Germany, but they were liquidated and now these
countries are free.
In South America-Foot-and-mouth disease is ENDEMIC, vaccination is practiced.
It often happens that a traveler from an outbreak area brings foot-and-mouth
disease virus to disease-free areas. Dust circulates the foot-and-mouth disease
virus along with migratory birds.
CLINICAL SUSPICION:
Vesicular exanthema, which on histological examination indicates

vacuolating hydropic degeneration and colicivation necrosis,
in the Malpighi layer without affecting the basement membrane-which
causes rapid, scar-free healing.
Adult cattle-benign form with

febrile infectious syndrome, dyspnea, tachycardia,
characteristic salivation, gurgling noise,
because the animal is trying to swallow.
In the acute evolution, the decrease in milk production is observed.
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LOCATION OF LESIONS:
ORAL: the clinical expression being the characteristic sialorrhea, due to the
opening of the aphthae in the oral cavity, a fact that stimulates the secretion of
FMD-histopathological suspicion, the salivary glands, because the aphthous lesions are painful.
Hydropic vacuolizing degeneration and colicivation necrosis of the Malpighian
layer without basement membrane damage. FOOT: evolution accompanied by secondary infection, leads to exungulations
(rare in cattle). Lesions in the coronary region, the interdigital groove (canker
sores) and acute canker sores lead to lameness.
MAMMARY GLAND: the skin surface of the udder, with a hairy coating is not
affected, especially the nipple is affected, canker sores on the nipple are
observed during milking which becomes difficult, but it is mandatory to avoid the
production of non-milking teats.
SHEEP:
MALIGNANT, SEPTIC-TOXIC FORM (particular) develops in calves, with After an incubation period of 1 to 8 days, the disease manifests itself
mortality below 30-40%, the cause of death being digestive disorders and by febrile syndrome, inappetence, reduced rumination,
febrile syndrome, tympanism, respiratory disorders and impact on
circulation followed by syncope and asphyxiation. aphthous rash on the tongue, on the limbs,
THE REAL SEPTICO-TOXIC form occurs in infants with polymorphic nipples and rarely on the muzzle.
digestive, respiratory, cardiac, nervous clinical expressions. Salivation is a little significant, because more
Atypical /aborted or occult forms without aphthous eruption. FREQUENTLY, IT'S THE LOCATION
FOOT OF SHEEP.
PIG:
Rash present on nose, limbs, in sows
also affecting the mammary gland, occurs in pregnant sows
abortion, and in lactating females, agalaxia.
Areas with fine skin are more frequently affected
(perineum, scrotum, vulva).
MORPHO PATHOLOGICALLY
CONFIRMATION:
The characteristic aphthous lesion: The pathological material is represented by the liquid from canker sores,
Aphtha has an initially transparent, opalescent, protein-rich content, collected from unopened canker sores.
separated from the exterior by a thin, friable wall. Isolation is done on CELL CULTURES.
Apart from canker sores present externally on the skin and explorable
IDENTIFICATION-SEROLOGICAL, against each foot-and-mouth disease
mucous membranes, there are also canker sores on the mucous virus serotype possibly involved (seroneutralization, ELISA-standardized
membranes of the respiratory tract, digestive tract, and prestomach (will methods in all EU countries, OIE affiliated, the result can be compared
cause diarrhea, tympanism). between laboratories).
In calves, cardiac degeneration of the hyalinosis type occurs, as well as
The handling of suspected Foot-and-Mouth Fever samples is done
degeneration of the striated muscles, which takes on the appearance of under special conditions, which prevent the dissemination of the causative
boiled muscle, a lesion called-TIGER HEART. agent, and only certain laboratories meet the biosecurity conditions.
Samples collected without risk of leakage, damage, are placed in the
Biongulate with: aphthous eruption, lameness and fever is: box, then in containers with formalin and removed from the fire, by
Suspicion of foot-and-mouth disease! transshipment to the laboratory.
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Diagnostic
Differential diagnosis
Vesicular-type diseases:
Vesicular stomatitis, Vesicular stomatitis
Virus Antibodies Swine vesicular disease,
Bovine viral diarrhea
Bovine papular stomatitis
Vesicular rash of the pig. Mucous membrane disease (leg injuries)
Infectious bovine rhinotracheitis
ELISA Virus neutralisation Rinderpest
Based on ulcerative lesions in
Bluetongue (leg injuries)
the mouth or muzzle: Plague of Small Ruminants
Virus Agar gel BVD/MD Chemical irritants
isolation/cel.cult. immunodifusion Blue Tongue and other orbiviruses. Swine vesicular disease
Vesicular exanthema of the pig.
IBR
PCR* Rinderpest
Treatment, Control, and Prevention of Foot-and-Mouth Disease in Animals

In regions that are normally FMD-free, control of the disease is typically attempted by culling all animals on infected premises, and
FMD is curable spontaneously; during clinical expression, animals are animal movement controls are imposed to reduce the risk of virus spread
In both normally FMD-free regions and endemic areas, vaccination around outbreaks may be used to limit the spread of the disease
low-cellulose feed, housing hygiene is considered essential. No treatments for infected animals are available
The OIE classifies countries and regions as: FMD-free without vaccination; FMD-free with vaccination; suspended FMD-free status
with or without vaccination; and unrecognized (OIE Terrestrial Animal Health Code, 2019).
Suspicion of foot-and-mouth disease requires the establishment of The current global status of FMD distribution shows geographic areas where FMD prevalence has been high over long periods of
time. They are commonly located in economically challenged countries where veterinary services and resources are inadequate to
control or eradicate FMD.
TOTAL RESTRICTION OF CIRCULATION IN AND OUT OF THE Combined use of trade and movement restrictions of animals and animal products has not completely prevented introductions of FMD
into FMD-free areas. These virus incursions into countries or regions where FMD is not enzootic are usually controlled by culling of all
OUTBREAK. infected and susceptible animals in infected herds, strict restriction of animal and vehicle movement around infected premises, proper
carcass disposal, and environmental disinfection, without the use of vaccines.
Inactivated virus vaccines protect for only 4–6 months against the specific serotype(s) contained in the vaccine. Billions of doses are
used each year and protect animals from clinical illness but not viral persistence in the pharyngeal region, and thus vaccinated
In the outbreak area; it is announced BY NOTIFICATION-veterinary animals can be carriers of infectious virus. Additionally, it is difficult to distinguish infected animals from vaccinated animals unless
purified vaccines are used. Therefore, vaccination is used more in enzootic countries to protect production animals, particularly high-
autorities, the City Hall and the Police. yielding dairy cattle, from clinical illness because slaughter of all at-risk individuals may be economically unfeasible and can cause
food shortages.
Rapid disease reporting is essential to control an FMD outbreak in nonendemic countries. During an outbreak, tracing is done through
epidemiologic inquiries to help identify the source of disease introduction. Sequencing of viruses can also identify the source of
closely related viruses. When mass culling is performed, infected carcasses must be disposed of via incineration, burial, or rendering
on or close to the infected premises. Scavengers and rodents should be prevented or killed to prevent mechanical dissemination of
virus. Buildings should be cleaned with a mild acid or alkaline disinfectant and fumigation, and people that have come into contact
with virus must decontaminate their clothing and avoid contact with susceptible animals for a period of time.
In some regions, FMD persistence in wildlife populations, such as the wild African buffalo, can make the prospect of FMD eradication
very difficult. Control measures, such as fencing of wildlife reserves to prevent contact with domestic livestock, have helped limit the
spread of virus in certain areas. A twice-yearly vaccination buffer zone in livestock near endemic wildlife reserves may additionally
help reduce outbreaks. A progressive control pathway (PCP) has been developed by FAO and adopted by OIE to enable countries to
improve their own FMD control so that the global disease situation will improve.
There is no specific treatment for FMD, but supportive care may be allowed in countries where FMD is endemic.
FMD as zoonothic disease. Foot-and-mouth disease - lesions in Cows
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Foot-and-mouth disease – lesions in Pigs Foot-and-mouth disease – lesions in other species
Bibliografie
https://youtu.be/c0b232noJHg
https://youtu.be/eOzG8ksVUs8
https://youtu.be/Znygl3V4pdI
https://youtu.be/MKf-aMgb-y0
https://youtu.be/V1HbJ8xlQXY
https://youtu.be/dykW8UiaMTU
https://youtu.be/Yn8CC-A7NRA
https://youtu.be/faodAbhtFa0
https://youtu.be/zcZ-86M2kAk
https://youtu.be/fP2mpsd5qPs
https://youtu.be/My3fzEgiBRw
https://www.woah.org/app/uploads/2021/09
/foot-and-mouth-disease.pdf
https://www.woah.org/fileadmin/Home/eng/
Animal_Health_in_the_World/docs/pdf/2.0
1.05_FMD.pdf
https://www.msdvetmanual.com/generalize
d-conditions/foot-and-mouth-disease/foot-
and-mouth-disease-in-animals
DVM PhD Dragos Cobzariu 2022 33
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Etiology:
POXVIRUS
the biggest viruses,
complex structure.
MAMMALS Dense core,

POXVIRUS biconave,
containing double stranded DNA, 200genes
INFECTIONS
Etiology: Etiology:
Smallpox is an infectious disease of viral origin, highly contagious and Schematic representation of
epidemic, caused by a poxvirus. several categories of viruses
The word smallpox comes from the Latin variola, -ae (which means
"small pustule", with the influence of the word varius, "varied,
variegated, spotted, speckled"). Indeed, smallpox is characterized in a
way by a "speckling of pustules". Smallpox was responsible until the
eighteenth century for tens of thousands of deaths per year in Europe
alone.
Smallpox was totally eradicated in 1981, thanks to a campaign by the
World Health Organization (WHO) combining mass vaccination
Thanks to special coloring techniques (Loffler, Morozov,
campaigns, as early as 1958, with a "surveillance and containment Romanovski, Giemsa), due to their large size, viral
particles loaded with dye, become visible under an optical
strategy", implemented in from 1967. In the 21st century, only samples microscope they were given different names for
of this virus were kept for research purposes by laboratories authorized intracitoplasmatic inclusions:
- PASCHEN'S corpuscles in human smallpox , cattle pox
by the WHO. - BORREL'S corpuscles in sheep pox.
By intracytoplasmatic multiplication,
the elementary corpuscles give rise to larger oxyphilic conglomerates,
Etiology:
called INCLUSIONS, easily stainable by the Giemsa method. They were called:
- Buist inclusions in cow pox,
- Guarnieri inclusions in human, sheep and rabbit smallpox.
- Bollinger inclusions in avian smallpox. POXVIRUS have a tropism for tissue:
At the top of the up arrow in the image you see a cell with these
RED INCLUSIONS Ectodermal, Endodermal or Mesodermal.
Variola viruses have a pronounced tropism

for ectodermal tissues, but can also
multiply in endodermal or mesodermal
tissues.
This explains the multiple locations in the
skin cells and organs mucosae.
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POX VIRUS OF MAMMALS

Etiology:
Etiology:
POXVIRUSES have: DNA virus, epitheliotrope
1.COMMON ORIGIN- SIMILAR host specificity
Fam. Poxviridae, subfam. Chordopoxvirinae
Morphological and Biological Genus – Orthopoxvirus
2.DIFFERENCES - Cowpox = cow, man, horse
Immunologycal and Pathologycal. - Camelpox = camel, man
- Smallpox = cow, man,
Although it is accepted that mammalian - Vaccinia virus = pig, cow, horse, man
pox viruses have
a common origin with similar morphological - Capripoxvirus = sheep, goat
and biological properties, - Suipoxvirus = pig
they still differ in terms of immunology and
pathology. - Leporipoxvirus = rabbit disese-Mixomatosis
● Avipoxviruses have been found in 23 orders of birds and at least 270 wild and domestic bird
species.
● Cetaceanpox virus ○ Harbour porpoises (Phocoena phocoena) ○ Long-finned pilot whales
Etiology:
(Globicephala melaena) ○ Striped dolphins (Stenella coeruleoalba) ○ White-beaked dolphins
(Lagenorhynchus albirostris)
● Contagious ecthyma (CE) ○ Bighorn sheep (Ovis canadensis) ○ Chamois (Rupicapra rupicapra) and
Southern chamois (R. pyrenaica) ○ Domestic goats (Capra aegagrus hircus) ○ Domestic sheep (Ovis
aries) ○ Ibex (Capra ibex) ○ Mountain goats (Oreamnos americanus) ○ Oxen (Ovibos moschatus) ○
Reindeer (Rangifer tarandus).
● Cowpox virus ○ In Europe, reservoir hosts are thought to be bank voles (Microtus agrestis) and field Human smallpox vaccine, from a cow strain
voles (Clethrionomys glareolus) ○ Brown rats (Rattus norvegicus) ○ Cheetahs (Acinonyx jubatus) ○
Common shrews (Sorex araneus) ○ Domestic felines (Felis domesticus) ○ Domestic cattle (Bos taurus
and B. indicus) ○ Eurasian lynx (Lynx lynx) and Iberian lynx (L. pardinus) ○ Great gerbils (Rhombomys
of smallpox virus called
opimus) ○ House mice (Mus musculus) ○ Water buffalo (Bubalus bubalis) ○ Wild cats (Felis silvestris)
○ Wood mice (Apodemus sylvaticus) ○ Yellow susliks (Citellus fulvus) VIRUS VACCINIA(latin “vaca” = cow=vache)
● Hare fibroma virus ○ European brown hares (Lepus europaeus)
● Rabbit fibroma virus (RFV) ○ Eastern cottontail rabbits (Sylvilagus floridanus) ○ European rabbits
(Oryctolagus cuniculus)
● Sealpox virus ○ Californian sea lions (Zalophus californicus) ○ Grey seals (Halichoerus grypus) ○
Harbour seals (Phoca vitulina)
● Squirrelpox virus (SQPV) ○ Grey squirrels (Sciurus carolinensis) and red squirrels (Sciurus vulgaris)
The grey squirrel is thought to be the reservoir host and rarely presents with clinical disease.
● Swinepox virus ○ Domestic (Sus scrofa domesticus) and wild swine (S. scrofa)
Etiology: Etiology:
RESISTANCE:
Remarkable resistance
Within AVIAN POXVIRUSES, physical and chemical factors
20 min at 56 °C,
there are several species, 5 min at 80 °C
ANTIGENICALLY AND PATHOGENICALLY DISTINCT 2 min at 100 °C.
LOW TEMPERATURES-dry environment keep alive smallpox virus
up to 2 years by refrigeration,
more than 3 years by freezing
Within the avianpox viruses, there are several species,
distinct from the antigenetic and pathogenic point of more than 4 years by freeze-drying.
view: chicken, dovecote, turkey, canary, quail, starling, At ambient temperatures,
sparrow, psittacine. 2 months on the pasture, and on sheep's wool.
Each species of bird is infected with a specific virus 6 months in shelters 2 months It is very sensitive to the direct action of solar
Viruses confer cross-immunity, a property used for the radiation-UV
production of vaccines.
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Etiology:
All COMMON DISINFECTANTS, (except GENERAL FEATURES:

chloride and alcohol) 1% formalin, sodium Eruptive diseases that manifest themselves as:
hydroxide, 3% phenol, 0.1% potassium - EXANTHEM (on the skin),
- ENANTHEM (on mucosa membranes);
permanganate, hydrogen peroxide,
DESTROY SMALLPOX viruses within minutes. Lesion evolves differently, depending on the species:
- macula, papule, vesicle, pustule, crust (human, cow, horse, goat),
- macula, papule pustule, crust (sheep, pig),
- the evolution is benign, except in the sheep where
the localization occurs also in the internal organs-nodules;
- in pig evolves like in humans;
PASSAGE THROUGH THE DISEASE IMMUNIZES SUSTAINABLY.
DIAGNOSIS of SUSPICION PATHOGENESIS

Epidemiological suspicion:
After entering the body (often in the skin or mucous membranes),
Receptivity: all mammals and birds, including humans, are receptive the virus multiplies in the epithelial cells and forms the VARIOLIC
(each species is infected with a specific virus);
Lesion:
Favorable factors : young age, dry season, external parasites, vitamin- Macula - hyperemia and the appearance of reddish areas of
mineral deficiencies, overcrowding. varying sizes;
Sources of infection: Papule - the serous infiltration of the deep layers of the epidermis
-Primary: animals with lesions (fluid from vesicles, pustules, crusts); which clinically corresponds to the appearance of some slightly
-Secondary: contaminated environmental elements- viral envelope protruding, delimited round rays surrounded by a hyperemic
resistance. area;
Vesicle - the balloon degeneration of hyperplasia cells which
Contamination routes: causes the appearance of vesicles containing a clear, serous,
- respiratory, limbic (lymphatic);
- digestive, Pustule - the result of leukocyte invasion of the vesicle;
- cutaneous (transcutaneous);
Crust - gradual dehydration of tissues;
Dynamics: Sporadic - Enzootic. De crusting - detachment and gradual elimination of crusts
PATHOGENESIS
Mucous or Skin can be first penetration
way virus multiplies - a primary lesion,
3 to 5 days after infection,
but escapes observation.
Inhalation is less effective
Lymph-macrophage + blood.
First viremia + fever.
Skin, mucous membranes, internal organs
Virus multiplies, second viremia
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PATHOGENESIS
The smallpox rash is characterized by the Macula, small red hyperemic spots appear on
progressive development of the rash on the the skin.
skin, with the following stages: Protruded and well-defined, yellowish-white, a
macula, few millimeters in diameter.
papule, Papule instead of macules, formed by
vesicle - blisters hyperplasia of polyhedral cells in the mucous
pustule, body of Malpighi and a serous intercellular
infiltration of the deep layers of the epidermis.
crust and de crusting.
The wall - is not an effective barrier against

• As a result of bloating degeneration of bacteria injected through, before exploding the
hyperplastic cells and necrosis, the content, taking on a purulent appearance with
papules turn into bladder blisters, clearer bacteria, leukocytes, and epithelial cells, the
than the rest of the skin, and contain a Pustule stage.
light yellowish serum, will form Vesicle.
Pustules - break, releasing drying contents, Cattle and Goats- Horses

forming Crust - epithelialization. De crusting
M, P, V, P, Cr, De Cr
PECULIARITIES — OTHER SPECIES.
Healing Without traces, (pig)
Sheep, cell proliferation in the papule phase is
With persistent scars: strong, giving rise to Nodules.
Germ layer-affected complications occurred, Sheep and pigs, poorly expressed vesicular
Eg human smallpox (black vomiting) or stage (witout Vesicles)
sheep.
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Birds bloating degeneration and blistering

are poorly expressed and keratinization of CLINICAL AND LESIONAL
the cell wall occurs, due to which cell lysis
SUSPICION :
can no longer occur.
Incubation period 2 weeks;
Vesicula and Pustula are missing,
Cattle, horse, goat, are passing through ALL STAGES;
Papulo-Crusty lesions.
Sheep and pigs without VESICLES;
Birds without vesicles and pustules, but NODULES appear.
CATTLE
Discrete HYPERTHERMIA,
HYPOGALAXY AND AGALAXY,
MACULA AND PAPULE on the udder and nipples, delimited by the
normal area, by an edge of erythema.
From infection 3 to 4 weeks pass to reepithelization.
There may be lesions - on neck,
- inner thigh,
- scrotum,
- perineal area.
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HORSE: PIG:
Vaccinia-virus-smallpox (Cowpox virus) cattle, buffalo, horse 1.Vaccinia Virus
Virus – Horsepox - extinct
2. Swine pox virus
Intracytoplasmatique oxyphilic
inclusion
Enanthema - bucal,
- peribucal,
- nasal,
- conjunctiva,
- genital;
-hypersalivation;
-abortion;
Exanthema-chitis (local inflammation).
Oral enantema, peri-buccal, nasal, conjunctival mucosa, genital
mucosa; hyper salivation; abortion;
Rash at the level of chitis (local inflammation)-exanthema.
PIG:
SHEEP
Evolution are benigne, affection with lesions in different stages of
evolution; 1. TYPICAL FORM: - febrile syndrome (41 - 42 ° C),
Spreading of the disease is favored by external parasites, like respiratory catarrh, cutaneous sensitivity up to
Hematopinus suis. 1-2 days when small pox eruptions of
exanthema type occur or; abortion;
Healing is spontaneous with overlapping portage diseases.
2. ATYPICAL FORMS:
- Benigne (stone smallpox )= inflammation

purulent hemorrhagic mucous membrane of
the head);
- Malignant: evolves as a enanthema of the respiratory and

digestive system (black spell), nodules, ulcers (internal
gangrenous smallpox - translucent nodes containing
necrotic or tumoral contents in the liver, spleen, kidneys and
serous infiltration in subcutaneous connective tissue).
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CONFIRMATION DIAGNOSIS
1.ISOLATION AND VIRUS IDENTIFICATION:
→ pathological materials: ASEPTIC TRITURATED CRUST

inoculated on corioalantoidian membrane -embryonated eggs 9-11
days old which will cause the formation of NODULES after 5-7
days; inoculation on receptive cell cultures is followed by the
appearance of the CYTOPATHIC EFFECT after 4 to 14 days and the
development of intracytoplasmic oxyphilic inclusions.
PCR
The Antibodies are identified by

indirect immunofluorescence.
2.HISTOPATHOLOGICAL EXAMINATION of the skin with lesions

1. Poxvirus located in the cytoplasm of
in mammals and birds: degenerate cells; ME50,000X (A) and
112,500X (B).
Proliferation of polyhedric cells from the Malpighi mucosal layer
is observed with hypertrophy and balloon
dystrophy in epithelial and mucosal cell
cytoplasm and specific granular inclusions:
→ Borrel corpuscles - sheep; 2. Poxvirus located in a alveolar macrophage -

cytoplasmic eosinophilic inclusions, 100X.
→ Buist corpuscles - cattle;
→ Guarnieri corpusculii - rabbit;
→ Bolinger corpuscles - bird;
→ Paschen corpuscles - man.

3. lung - poxvirus infection: bronchiole (right), blood
vessels (left), necrotic pulmonary parenchyma, 20x.
Poxvirus – celule infectate, model
tridimensional
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DIFFERENTIAL DIAGNOSIS
SANITARY PROPHYLAXIS
The clinical signs of severe sheep pox and goat pox are
highly characteristic. However, in their mild form they can be If culling is not possible, isolation of infected herds and sick
confused with parapoxvirus causing orf or urticaria from animals for at least 45 days after recovery
multiple insect bites. Slaughtering of infected herd if possible
Contagious ecthyma (contagious pustular dermatitis or orf) Proper disposal of cadavers and products - burning or
Insect bites burial is often used
Bluetongue Stringent cleaning and disinfection of farms and equipment
Peste des petits ruminants Quarantine of new animals before introduction into herds
Photosensitisation Animal and vehicle movement controls within infected
Dermatophilosis areas
Parasitic pneumonia Vaccination may be considered when the disease has
spread more widely, or at the importer's request two weeks
Caseous lymphadenitis
prior to delivery.
DVM PhD Dragos Cobzariu 2022 43 • DVM PhD Dragos Cobzariu 2022 44
Live and Inactivated vaccines have been used for the SURVEILLANCE IN SHEEP AND GOAT POX:
control of capripox. All strains of capripoxvirus so far
examined share a major neutralisation site and will cross ► Clinical and anatomopathological surveillance of susceptible species
protect. ► Prophylactic quarantine for transit animals;
There are several attenuated virus vaccines delivered by
► Serological surveillance by virus neutralization or
subcutaneous or intradermal route; conferred immunity lasts immunofluorescence:
up to 2 years. - for the detection of cross-border contamination;
- 1% sheep or goats in zone I,
Inactivated vaccines give, at best, only short-term immunity
- 0.5% sheep or goats in zone II;
Currently, no recombinant vaccines for capripoxviruses are - in animals imported from countries free from disease but with
commercially available. However, a new generation of epidemiological risk;
- at the request of the veterinary administrations of countries
capripox vaccines is being developed that uses the importing receptive animals.
capripoxvirus genome as a vector for the genes of other - If necessary, countermeasures are applied: notification, serological
ruminant pathogens, for instance genes of Rinderpest and examination;
- ROMANIA IS FREE of ovine pox infections, does not vaccinate and
Peste of Small ruminants (PPR) viruses. does not produce vaccines, does not import susceptible animals from
countries that do not have the status of free country;
- Establishment of surveillance zones is done by ANSVSA with IDSA
DVM PhD Dragos Cobzariu 2022 45 approval. DVM PhD Dragos Cobzariu 2022 46
THE RISKS OF POX VIRUSES TO THE PUBLIC HEALTH.
Sealpox virus infection has been reported in humans.

In Canada, two gray seal keepers developed "milking nodules" on their fingers after
contact with gray seals infected with a specific poxvirus.
There are reports of cats, rodents and dairy cows transmitting cowpox to humans.
Pet rabbits can become infected with Leporipox virus if they come into contact with wild
rabbits or hares.
Humans, especially animal handlers and veterinarians, should use appropriate protective
methods to prevent infection.
Smallpox viruses can be transmitted to humans, if they come into contact with an
infected animal or one that has been recently vaccinated; immunosuppressed people are
especially at risk.
People who handle sheep's wool or the skins of small ruminants can contract the
disease; people who are involved in slaughtering small ruminants for religious, cultural or
consumption purposes are also at risk.
Zoos, where many species of domestic and wild animals coexist, are potential sources of
poxvirus infection for visitors.
https://youtu.be/oMhpb8SPe_8
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Bibliographic resources
• https://youtu.be/OBsBtuh-wAk
• https://youtu.be/FXhMs-BjgQ8
• https://youtu.be/oMhpb8SPe_8
• https://youtu.be/2fba-TZuYPg
• https://youtu.be/gmADx6qcJzA
• https://youtu.be/DgBQtrG_7EE
• https://www.woah.org/app/uploads/2021/05/pox-viruses-other-than-those-
listed-by-the-oieinfection-with.pdf
• https://www.woah.org/fileadmin/Home/fr/Health_standards/tahm/3.07.12_S_
POX_G_POX.pdf
• https://www.cdc.gov/poxvirus/diseases.html
• https://www.woah.org/fileadmin/Home/eng/Health_standards/tahm/3.04.12_
LSD.pdf
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ETIOLOGY:
DNA virus, epitheliotrope, host specificity,

Family Poxviridae, subfamily Chordopoxvirinae and Entomopoxvirinae-insects.
Genus - Avipoxvirus - Fowlpox = chickens

- pigeonpox = doves cross-immunity!
- turkeypox = turkeys
- qailpox = quails
- canarypox = canaries
- psittacinepox = parakeets
FOWL POX - sparrowpox = sparrows
Fowlpox, It is a natural pathogen for chickens, guinea fowl, peacocks, Turkeypox virus-pathogenic for turkeys. In pigeons, it causes lesions on the
pheasants. skin, accompanied by the immunity against the Pigeon pox virus.
It is not pathogenic for the pigeons, (it immunizes against Pigeon pox), can be Turkey and Pigeon viruses are antigenically related.
adapted to the pigeon, by successive passages on the pigeon.
It is not pathogenic for turkeys (it immunize against the Turkey pox). However, the cross-immunity is not symmetrical, the pigeon is strongly
Pigeonpox, It is not pathogenic for chickens, turkeys, canaries, it is only immunized with the turkey virus, but the turkey is only partially immunized with
pathogenic for columbiformes (sandgrouse, the pigeons, doves together with the pigeon virus.
their extinct relatives, the dodo and solitaire).
Canarypox - affects canaries, sparrows.
It produces typical lesions of smallpox by experimental inoculation and
immunizes against chicken virus, but also against turkey virus. Qailpox - affects only quail.
Sparrowpox - pathogen only for sparrows.
CULTIVATION AND RESISTANCE

Avian pox viruses are cultivated on
PATHOGENESIS
Poultry cell cultures and embryonated eggs of 9-11 days old, on the The virus replicates at the gate of entry, spreads throughout the body, produces viremia
chorioallantoic membrane, where Bollinger inclusions are highlighted. and spreads to organs, where it causes the characteristic lesions.
Poultry lesion has two different aspects.
The appearance of the lesions in the bird is NODULAR, white-gray, opaque, The smallpox NODULE at the SKIN level, has initially the appearance of a hard
wart-like excrescences, nodules develop on areas without feathers, and papilloma, the size of a lens or a pea, grayish-white in color is described.
mucous. Nodule formation is achieved by hyperplasia of epithelial cells and cornification of the
ectoplasm of hyperplastic cells.
Immunogenicity-avian pox viruses are immunogenic-and confer cross- Ballooning degeneration, of hyperplastic cells, is reduced, at the level of epithelial cells,
immunity. reduced cytoplasm, Bollinger corpuscles, cell nucleus and micro vesicles are observed.
The serous exudate is removed from the surface of the nodule and the presence of a
High resistance to darkness, and in dry scaly crusts, epithelial cells for 2 to 6
DRY CRUST is noted.
years. Vesicle and Pustule stages are not present.
In humid environments at 60ºC it lasts 8 minutes, at 100ºC between two and Diphteroid membranes on MUCOSA-after the papule phase (nodule) and ballooning
degeneration of hyperplastic cells, with the appearance of microscopic vesicles, there is
5 minutes, at 38ºC up to 14 days.
superinfection of lesions with common bacterial flora, accompanied by fibrin production
At room temperature, in crusty powder up to 15 months. and necrotic inflammation in the form of foci, resulting
Usual antiseptics destroy avian poxviruses quickly, there is the phenomenon DIPHTEROID MEMBRANES, made up of cellular detritus, bacteria and fibrin.
The membranes are DIFFICULT TO REMOVE, leave the mucosa bleeding, and
of resistance to some of them. regenerate, causing difficulty in breathing and swallowing.
Acidic pH destroys pox viruses.
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DIAGNOSIS by CLINICAL suspicion,

Eruptive disease, 3 clinical forms being described,
- Smallpox or EXANTHEMA (approximately 8% of cases),
- Diphtheria or ENANTHEMA (2% of cases),
- MIXED DIPHTHERIOVARIOLIC FORM (90% of cases).
The lesions evolution in birds is incomplete, MACLE, PAPULE, CRUST (M,P,C)
SMALLPOX (CUTANEOUS) FORM: NODULES on the crest, chins, pericloacal,
corner of the beak, eyelids, limbs are described.
Healing occurs in 3 weeks, passing through the disease immunizes permanently.
The vital prognosis is favorable. (Important for ornamental birds as pets.)
The DIPHTHERIC FORM (on mucous membranes): are described
-fibrinous, pseudomembranous deposits, adherent to mucous membranes (conjunctival,
oral, esophageal, pharyngeal/laryngeal).
-deposits that, by detaching, leave the mucous membranes exposed, with areas of bleeding.
-relapses occur by restoring fibrinous deposits,
-dysphagia, breathing problems, foreign body sensation (repeated swallowing),
-laryngeal localization of the deposits, leads to death by suffocation;
The vital prognosis is critical.
THE MIXED DIPHTHERIOVARIOLIC FORM, characterized by
-the simultaneous evolution of smallpox and diphtheric forms on the same bird.
The vital prognosis in this form being critical.
Virusurile din subfam. Poxvirinae, genul Avipoxvirus - și implicațiile lor patologice
CLINICAL in TURKEYS
2 weeks incubation;
It is described Cutaneous form,
VARIOLIC EXANTHEMA
Nodules are described at the level of the
caruncles, cutis of the legs, pericloacal,
eyelids, healing takes place in 3-4 weeks;
Passing through disease immunizes
lastingly.
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CLINICAL in TURKEYS
CLINICAL in TURKEYS MIXED DIPHTHEROVARIOLIC
DIPHTHERIC form: ENANTHEMA, FORM, are frequent in turkeys.
characterized by the appearance of
Adherent fibrino-necrotic DEPOSITS,
which, by detaching, leave
MUCOSA bleeding, and are recovering;
-Dysphagia, respiratory disorders,
-Diphtheric deposits of membranes on
mucous membranes:
conjunctiva,
inside of the beak,
esophageal,
-Death by asphyxiation or starvation.
PIGEON CLINICAL SUSPICION

Pigeon pox virus;
Young pigeons, especially the chicks, are very
Clinical evolution:
sensitive, and the evolution of the disease is serious,
with important losses.
Over acute-severe septicaemia, respiratory
Adults make easy shapes;
distress, death within 12 to 24 hours.
The way of transmission is respiratory, digestive, or through skin Acute-fever, anorexia, dyspnoea, weight loss,
lesions; mucosal lesions, death in 6-8 days.
The severity of the disease is given by the state of maintenance: Subacute-with slow evolution, lasting 1-4 weeks,
A good condition determines a clinically evolving, mild skin form; skin rashes, with mucosal or mixed lesions.
The poor state of maintenance causes malignant, enanthema
forms.
CUTANEOUS LOCALIZATION-NODULES are

described, at the level of the eyelids, around the nostrils,
the corner of the beak flaps, around the cloacal opening,
metatarsals, the inner face of the wings.
NODULES in the form of confluent warts, crusts, which in
the distance leave scars.
General condition unchanged.
LOCALIZATION ON THE MUCOUS MEMBRANES-
CONGESTED mucous membranes, covered with false
membranes, deposits, foul-smelling, adherent to the
mucous membranes, which, when detached, leave the
area bleeding, even ulcerated.
-Localized (localized)-corners of beak valves, tongue,
palatal arch, laryngeal, tracheal, conjunctival, nasal
mucous membranes.
-Bad general condition.
MIXED LOCALIZATION is also described in pigeons,
fatal.
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LESIONS Virus isolation and identification,
Papules, Pustules, Scabs which are actually yellowish white Collected pathological materials:-sterilized triturated crusts,
NODULES, on the skin without feathers; Inoculated on the chorioallantoidian membrane of 9-11 days old,
Healing occurs in 2, 3 weeks. embryonated egg, with formation of NODULES after 5-7 days.
DIPHTHEROID MEMBRANES are observed, Inoculation on receptive cell cultures, with the appearance of the
on anterior mucous membranes, respiratory and digestive. CYTOPATHIC EFFECT, after 4-14 days and the development of
intracytoplasmic OXIFILIC inclusions.
IDENTIFICATION OF AVIPOXVIRUSES IS PERFORMED THROUGH:
Immunofluorescence.
Polymerase chain reaction (PCR).
Frequently used quantitative tests:
Transmission electron microscopy (TEM).
Viral culture inoculated onto the chorioallantoic membrane of chick embryos
or avian cell cultures.
Viral neutralization tests.
Enzyme-linked immunosorbent assays for Pox virus, Antibodies (ELISA) are
available.
DIAGNOSTICUL DE CONFIRMARE
Histopathological examination of the skin with lesions, nodules.

Proliferation of polyhedral cells in the Malpighian mucous layer with
hypertrophy and ballooning dystrophy in the cytoplasm of epithelial and
mucous cells with specific granular Bolinger inclusions are identified.
For remember:
→ inclusions Borrel – sheep;
→ inclusions Buist – cow;
→ inclusions Guarnieri – rabbit;
→ inclusions Paschen – man.
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Pigeon pox lesions,

Fowl pox lesions on the crest Eosinophilic inclusions:
Eosinophilic inclusions:
Bollinger corpuscles in
Bollinger corpuscles in
hyperplastic cells of the
hyperplastic cells of the mucous layer
mucous layer
Quail pox Sparrow pox

Oral membrane: intracytoplasmic inclusions in hyperplastic epithelial cells. Skin: eosinophilic inclusions in hyperplastic keratinocytes
Chicken Fowl pox: ME from skin. Viral particles in Bolinger corpuscles

Viral particles in the cytoplasm
The viral particle at the time of budding
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Pigeon pox: the pigeon virus is morphologically similar to the chicken pox
Mature viral particles and envelope fragments are observed in the Bolinger corpuscles
Quail pox: the virus is smaller and
narrower than chicken pox.
It is serologically distinct from the
chicken one.
PROFILAXIE:
Sparrowpox: mature viral particles
within corpuscles
SPECIFIC PROPHYLAXIS - VACCINATION
In birds, prophylactic vaccination - with live VACCINES: SPECIFIC PROPHYLAXIS:

Columbary avian vaccine(COL),
Will vaccinate:
- live varieties of Pigeon pox virus (heterologous vaccine) lyophilized, replacement youth,
- are administrated at the age of 8 to 10 weeks of life(first vaccination), breeding hens,
- control of efficiency: in 7-8 days after vaccination will observe nodules on the skin.
- immunity in 21 days after vaccination, it lasts about 6 months. hens exploited for consumption eggs,
Avian Fowl pox vaccine (GAL), flocks of pigeons (the current year's flocks),
partridges and quails.
- Fowl pox- a chicken virus strain (homologue vaccine), lyophilized,
- is administrated at 4 weeks after the COL vaccine, In localities with an epidemiological history, in the last 12 months,
- application through the Stick method, in the humero-radio-ulnar skin fold, see the movie. according to the vaccination scheme and the manufacturers'
- control of efficiency: 2 to 7 days at the 2 sites of inoculation, are seen nodules.
- immunity in 21 days after vaccination, lasts for 1 year. instructions, for each individual species.
Avian Turkey vaccine for the pigeons,
For pigeons, vaccination is recommended
- is a Turkey pox strain (heterologous vaccine), lyophilized,
- is administrated at the minimum age of 6 weeks, at least 14 days after vaccination against Paramyxovirus.
- control of efficiency: in 7-8 days will identified nodules on the skin.
- immunity in 21 days after vaccination, lasts around 8 months.
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PROPHYLAXIS, PIGEONS
General measures to ensure optimal hygiene conditions in shelters, aviaries;
providing complete vitamin rations (vitamins A and C);
Prophylactic quarantine; the young pigeons are housed in separate
compartments, isolated from the adults.
Periodic disinfection, pest control. Breeding in separate aviaries of different
species of birds.
Specific measures, vaccination,
The vaccine against pigeon pox is a strain of turkey, Turkey pox.
Pluck the feathers from the leg, 10 to 20 feathers from the anterior-outer part of
the , then swab the area with a cotton swab soaked in vaccine.
A local reaction will be identified, through the formation of nodules, 5-7 days after
vaccination. If no such reaction occurs, the pigeon will be revaccinated.
Immunity sets in after 21 to 28 days, and lasts 6 to 8 months..
https://youtu.be/nYCI-fBpK20
COMBAT MEASURES
Official declaration of the outbreak of Avian Pox,
Quarantine of the 3rd degree,
Necessary vaccinations of healthy birds in the outbreak. • https://youtu.be/ABjcVSYgeg4
Isolation and treatment of sick birds (those with diphtheric form on the mucous • https://youtu.be/_Njy7R4ngoo
membranes): • https://youtu.be/5DiW_MLbaM8
-daily elimination of pseudo membranes • https://youtu.be/lDEgg8L13qY
-bleaching with glycerinated iodine tincture, • https://youtu.be/3gz7bGDLO_s
-methyl blue 1%, • https://youtu.be/lDExk_FbCyE
-potassium permanganate 0.1%, • https://youtu.be/tOv-IvFO4Ps
-avoiding secondary bacterial infections by administering antibiotics by general • https://youtu.be/p91dBoauGLY
way. • https://www.msdvetmanual.com/poultry/fowlpox/fowlpox-in-chickens-and-
-protection of epithelia with supplements containing vitamins A, C, and the turkeys
AD3E complex • https://www.woah.org/app/uploads/2021/05/pox-viruses-other-than-those-
listed-by-the-oieinfection-with.pdf
The declaration of the extinction of the disease will be made after 21 days after
the last case of the disease, and 14 days after the necessary vaccination.
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CLASSICAL SWINE FEVER PPC/CSF
AFRICAN SWINE FEVER PPA/ASF

( Montgomery Disease ) CLASSICAL
SWINE FEVER PPC/CSF
DVM PhD DRAGOS COBZARIU 2022
CLASSICAL SWINE FEVER Situation of PPC - Europe 2005 - 2011

“Infectious-contagious swine-specific infection caused by ARN virus,
Flaviviridae family, the PESTIVIRUS genus, characterized by clinical, and
lesional polymorphism (cutaneous, digestive, respiratory, nervous),
with high morbidity and mortality, being considered to be the most damaging
infectious disease specific in pigs”
CLASSICAL SWINE FEVER in the world-2011
Absence des données sur la maladie
Jamais rapportée
Pas rapportée dans la période respective
Suspectée, mais pas confirmée
Confirmée, sans signes cliniques
Confirmée, avec signes cliniques
Confirmată dar limitată la anumite zone
Pig population density in Europe and Romania at the Situation of PPC - world 2020
end of 2010
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Etiology:
CLASSICAL SWINE FEVER in the world RNA virus within the Flaviviridae family, Pestivirus genus
September 2020 Closely related to ruminant pestiviruses causing Bovine Virus Diarrhea(BVD) and
Border Disease.
Not related to African swine fever virus (Asfarviridae, Asfivirus, DNA virus).
Resist - at pH 3.0 - 11.0,
- to heat (up to 56 degrees C)
- freezing, smoking and brining
- in the cold and humid environment (in the absence of U.V.)
 Sensitive to phenol and caustic desinfectants-NaOH
 Cultivated on cell cultures (line PK 15) (no cytopathic effect) –
 Exception: Strain Pav, CAP-cytopathic effect is characterized by the appearance
of cytoplasmic granulation, progressive cytolysis.
Identification by INDIRECT IMMUNOFLUORESCENCE.
 VARIABLE PATHOGENICITY ( EMLED)
- Extremely virulent strains - Serious epizooties;
- Moderately virulent strains - Subacute classical forms;
- Low virulence strains - responsible for Persistent infections;
- Experimentally Diminished virulence strains (modified Vaccine strains)
Antigenicity - Unique Antigenic
Immunogenicity: determines the formation of Protective Neutralizing Antibodies.
FLAVIVIRUS-Pathogenesis
https://youtu.be/3LhWuaTRCME
EPIDEMIOLOGCAL SUSPICION
RECEPTIVITY
- domestic and wild pigs no exception of age, race or sex.
- non-transmissible in humans.
PRIMARY
- infected pigs in all evolutionary phases
(viral excretion may take place some 24 hours after contamination),
- all excretions and secretions from live pigs,
- products and by-products
- their bodies-carcases
! Meat and fresh pork products or preserved from infected pigs can
spread the disease at very long distance (transcontinental)
SECONDARY
- waste water from washing the infected meat
- feed, litter, contaminated vehicles
- non-receptive animals (dogs, cats, birds, rats, etc.)
- iatrogenic (syringe needle, surgical instrument).
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TRANSMISSION PATH:
Vertical :
Semen of infected boars, chronic infections of mothers.
Some Experimentally Diminished Virulence Strains, used to produce the
vaccine, may infect fetuses. Most transplacentary infected piglets, are
VIRUS-CARRERS will elimine viruses through IMMUNOTOLERANT, the inherited colostrale immunity, will delay the
ALL SECRETIONS (oronasal, lacrimal), by appearance of clinical signs, in those piglets, but are strong eliminators of
viruses.
manure, but also through semen that is Horizontally:
Direct contact, by respiratory and digestive ways;
sometimes highly contaminated. Indirect contact with contaminated media (contaminated food residues) -
Iatrogenic - vaccinations, blood harvesting, surgeries,
By air, low possibility of transmitting the virus. Hematophagous insects,
Man, domestic and wild, non-receptive animals, vehicles.
DYNAMICS OF DISEASE - EPIZOOTIC OR ENZOOTIC evolution
Dynamics are influenced by
-virulence of the viral strain,
- the farming system (household, or intensive).
Occurs regardless of season, are worse in youth pigs.
CLINICAL SUSPICION OF DIAGNOSIS

TYPICAL FORMS
OVERACUTE, ATIPICAL FORMS
Sudden onset without prodromal signs, with fever associated with a dull state
and death in 24-48 hours (no skin lesions - "white fever")
ACUTE REPRODUCTIVE DISORDERS
Fever (41°C), anorexia, lethargy, multifocal hyperaemia+ bleeding skin lesions, - abortions, dead pigs, mummified or with malformations at parturition;
conjunctivitis, skin cyanosis, especially of the extremities.
NEONATAL PATHOLOGY
Passive constipation, alternating with diarrhea, occasional vomiting, dyspnea, cough;
ataxia, paresis and seizures;
- at parturition obtaining the IMMUNOTOLERANT PIGLETS, affected by
Death, in 5 - 15 days after, the appearance of clinical signs; mortality in young piglets will persistent infections, the clinical symptoms and death may occur at the age of
reach 100%. 3-4 weeks;
SUBACUTE AND CHRONIC, - faty pigs and young piglets for breeding: are not-growing, have increased
Are identified three periods: body temperature, underdevelopment.
PERIOD I - lasts 10 to 15 days affecting the general state and acute-like symptoms, but INAPPROPRIATE FORMS
attenuated.
Period II – characterized by remission. - possible circulation of the virus in the flock (in faty pigs ) with
Period III - caused by bacterial over infections, associated with changes in general the possible occurrence of clinical cases under favorable
health condition manifested by respiratory and digestive symptoms. conditions.
(pneumo-enterites often with Salmonella spp)
Pigs are losing weight, and die within 1 to 3 months.
ANATOMOPHOPATHOLOGICAL SUSPICION CLASSICAL SWINE FEVER - ACUTE FORM

TYPICAL-EVOLUTION
Hemorrhagic or congestive lesions - lymph nodes (hypertrophy with
congestive or hemorrhagic areas in the cortical or wholly hemorrhagic),
kidneys (hemorrhagic picking "turkey egg„ appearence like), spleen
(marginal infarctions, sometimes hematoma), bladder (visible bleeding on
mucous membranes), tonsils (hypertrophied and hemorrhagic), skin, lung,
digestive tract.
Ulcers - over the entire surface of the digestive tract especially in the
ileocecal valve, colon and cecum (flat, non-perforated, fibrin-coated are
present in subacute or chronic forms).
Bacteria lesions caused by secondary bacterial complications.
ATYPICAL
Various and non-specific lesions.
HEMATOLOGICAL
Leukopenia.
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CLASSICAL SWINE FEVER- ACUTE FORM

CLASSICAL SWINE FEVER
- CRONIC FORM
CLASSICAL SWINE FEVER- LESIONS

– TABLOU LEZIONAL
- CRONIC FORM

CLASSICAL SWINE FEVER-CONGENITAL INFECTION
- LESIONS
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CLASSICAL SWINE FEVER - ACUTE FORM CLASSICAL SWINE FEVER - ACUTE FORM
The renal cortex contains

several patches and infarcts
surrounded by bleeding. The cortex contains There are
several patches numerous cortical
and infarcts dilated petechiall
surrounded by lessions (look like
bleeding. a turkey egg).
There are several infarcts, dark red along the edges of the spleen.
CLASSICAL SWINE FEVER- CRONIC FORM
The renal cortex

contains the
The renal cortex diffused petechial.
contains several patches (hydro nephrosis)
and infarctions and also bleeding.
surrounded by bleeding.
Lymphatic retropharyngeal
node significantly enlarged and The mucosa is red and contains several discrete
haemorrhagic, tonsils contain ("buttons"), ulcers, surrounded by hemorrhagic areas.
multiple hemorrhages
CLASSICAL SWINE FEVER- lesions CLASSICAL SWINE FEVER- lesions
The epiglottis and palatal tonsils

contain multiple outbreaks of
necrosis
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CLASSICAL SWINE FEVER- lesions
CLASSICAL SWINE FEVER-

lesions
CLASSICAL SWINE FEVER- lesions
• https://youtu.be/Csxa3lk_tSI
EPIDEMIO-CLINIC DIAGNOSIS
African swine fever (indistinguishable clinico-pathologically. It is
SUSPICION
essential to send samples for laboratory confirmation.)
Newly introduced animals or the use of non-sterilized slaughterhouse
waste; • Septicemias: erysipelas, eperythrozoonosis, salmonellosis,
Contagious disease affecting pigs of all ages associated with obvious streptococcosis, pasteurellosis, actinobacillosis, and Haemophilus
hyperthermia, general symptoms, local skin, digestive, respiratory and parasuis infections.
various nervous symptoms associated with high mortality rates in 5-10 • Hemorrhage: porcine dermatitis and nephropathy syndrome,
days. haemolytic disease of the newborn, coumarin poisoning,
Pronounced hemorrhagic lesions, especially in lymph nodes, kidneys, thrombocytopenic purpura.
spleen, bladder and tonsils. (In case of doubt, more bodies will be • Weaning: post weaning multisystemic wasting syndrome,
necropsied). enterotoxicosis, swine dysentery, campylobacteriosis
The occurrence of abortions, high neonatal mortality. • Abortions: Aujeszky’s disease (pseudorabies virus),
DIFFERENTIAL encephalomyocarditis virus infection, porcine reproductive and
Clinical not possible with African swine fever respiratory syndrome, parvovirus infections.
Difficult with other diseases with various etiologies likely to cause similar
• Nervous signs: viral encephalomyelitis, salt poisoning
hemorrhagic lesions (salt intoxication, colibacillosis, pasteurellosis,
Aujeszky's disease, see next). • Congenital infection with ruminant pestiviruses: Bovine virus diarrhea,
Border disease
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DIFFERENTIAL DIAGNOSIS PPC-CSF

Streptococcal infection
Swine influenza
Salmonellosis
LABORATORY DIAGNOSIS-CONFIRMATION CONFIRMATION METHODS-Ag

Virus or antigenic Identification
Reverse Transcription polymerase chain
Choice method for detecting early infections in herds is,
collecting whole blood and tissues from multiple febrile or reaction or real time RT-PCR.
recently dead animals. Virus isolation in cell culture, and virus detection
SAMPLES
by immunofluorescence or
• Tonsils
• Lymph nodes (pharyngeal, mesenteric) Immunoperoxidase-(ICC-IF/IP)
• Spleen Confirmatory identification with monoclonal
• Kidney antibodies. (MAB)
• Distal ileum
Immunofluorescence test on cryostat sections of
• Blood on EDTA or Heparin (live cases)
organs from affected pigs. (ICS)
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SEROLOGICAL TESTS- ANTIBODIES

Antibodies are present in body fluids, only during the third
week of illness: submit serum samples of convalescent
pigs, from suspected herds, if more than 3 weeks have
elapsed, since suspected contact was done.
Serum from sows with suspected congenitally infection
litters. Antibodies persist for life in recorded pigs.
The following methods, are used for serological diagnosis
or surveillance, and are also tests recommended by the
OIE, for screening for international trade:
• NEUTRALISATION PEROXIDASE LINKED ASSAY
• FLUORESCENT ANTIBODY VIRUS NEUTRALISATION
• ELISA Diagnosis by Immunofluorescence (IFD)
Isolation of classical swine fever virus

and cultivation on PK15
Detection of antigens in
lymphoid organs of pigs
infected by
immunohistochemistry.
Tonsils-hyperplastic
lymphoid follicles and
abundant
immunoreactivity in the
epithelium.
Brain. Perivascular collar in non-
Brain. Non-purulent
purulent meningoencephalitis
meningoencephalitis extended to
meningees as a result of
inflammatory lymphocyte
infiltration
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PREVENTION AND CONTROL
No treatment. Affected pigs must be slaughtered

and the carcasses buried or incinerated.
SANITARY PROPHYLAXIS
• Effective communication between veterinary
authorities, veterinary practitioners and pig
farmers
• An effective disease reporting system must be
implemented.
• Strict import measures policy for live pigs, pig
semen, fresh and cured pig meat.
SPECIFIC PROPHYLAXIS
SANITARY PROPHYLAXIS Countries free of disease, VACCINATION IS NOT PRACTICED, are
performed epidemiological surveillance by POST-INFECTION
DETECTION OF ANTIBODIES.
• Quarantine of pigs before admission into herd.
Countries where VACCINATION IS PRACTICED,
• Efficient sterilization (or prohibition) of waste food fed - Diagnostic is aimed on highlighting the virus,
to pigs. - Detection of serum antibodies, are used for evaluation of the
immunization -herd
• Efficient control of rendering plants. Prophylactic quarantine of newly introduced pigs
• Structured serological surveillance targeted to Required sacrifices in specially slaughterhouses.
Sterilization of debris from canteens, restaurants, slaughterhouses, hospitals
breeding sows and boars. Correct destruction of the corpses
• Effective pig identification and recording system. Vaccination according to the technical plan - live (attenuated on rabbit) ,
• Effective hygiene measures protecting domestic pigs vaccines 2 times per year, intramuscular.
- Minimum age of vaccination - 2 months
from contact with wild boar. - Only healthy animals
- Filing vaccinations ( can be performed monthly)
SURVEILLANCE OF WILD PIGS:

Serological surveillance - sampling of serum
or thoraco-abdominal fluid from all hunted
pigs aged between 6 months and 2 years • https://youtu.be/lTv8Uc7TVN0
(pairs of samples for virusology) examined
by: ELISA, virus neutralization.
Virusological surveillance - RT-PCR test for
viral genome detection on samples taken
from: wild pigs found, all pigs hunted - IFD
test.
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AFRICAN
SWINE FEEVER
ASF/PPA
https://youtu.be/uWS2Q_0VOjg ( Montgomery Disease)
AFRICAN SWINE FEEVER

Importance - economically speaking!
Etiology: - DNA virus - fam. ASFARVIRIDAE
• https://youtu.be/9eOV5o7qP9Q (African Swine Fever And Related Viruses)
- gen Asfivirus
- tropism: for monocites and macrofages (SRE)
Resistance:
- at 4 ° C → 24 months
- lyophilization → 7 years, frozen→ 6 years,
Sensitivity:
- Na OH 2%, organic solvents, halogens
Dinamyque: Enzootic / Epizootic
Receptivity: Incubation: 3-5 days

- domestic pigs (≈ age, breed, sex)
- wild boars
Sources of infection: Clinical diagnosis: Sp.Ac. , Ac. , Sb.ac. , Cr.
- primary: sick pigs, convalescent (1 year)
warthogs and bush pigs (natural carriers) Pre-symptomatic hyperthermia (41-42 ° C)
- secondary: fod, water, food scraps, objects. Symptoms RDN
Route of infection:
- Respiratory – tachypnea, dispnea
- direct (direct contact),
- indirect - by food products, secondary sources, - Digestive - vomiting, constipation / diarrhea with blood
- by vectors: ticks, lices. - Nervous - hyperexcitation, lateral decubitus (film!)
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Anatomopathological diagnosis: CONFIRMATION DIAGNOSIS:
Hemorrhagic diathesis-generalized, - Virusological - cell cultures (pig leukocytes),

Exanthema with cyanosis and hemorrhages, - Hemadsorption / Inhibition of hemadsorption,
Bloody exudation (weeping, pericardium, peritoneum), - Immunofluorescence,
Splenomegaly (2-4 x), - PCR,
Kidneys-hemorrhagic (with blood clots), - Histopathological,
Lymph nodes-hemorrhagic (aspects of hematomas). - Biological test (piglets),
- Serological: RFC, ELISA,

CONFIRMATION DIAGNOSIS: PPA(ASF)
PROPHYLAXIS:
Non-specific general concerns:
-restrictions on imports from endemic areas,
-preventive quarantine measures,
-measures to maintain biosecurity.
-Disinfection, Disinsection and Deratization.(DDD)
Specific Prophylaxis is ineffective, (although there are
several types of vaccines under study, they are not practiced)
COMBAT:
Quarantinable disease, the outbreak is cleaned up by total
liquidation of the herd, disinfection, disinsection and
deratization are carried out.
Repopulation takes place after 6 months, from the lifting of

quarantine measures (30 days after the last case and the final Regions affected by African swine fever in Romania
disinfection). domestic pigs 2017 - 2018
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Map of locations -last outbreaks

Romania 2020-OIE
Evolution of the African swine fever epidemic in domestic

pigs (red) and wild boars (purple) in Eastern Europe
between 01.01.2018- 23.08.2018
Evolution of African swine fever in the world between 1.01.2018 -

22.09.2018 to domestic pigs (circles) and wild boars (triangles)
Global situation ASF

2020 - 2022 domestic and wild pigs .
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https://youtu.be/4VqbK6YfjPY
https://youtu.be/0ykP5HxP_wQ
Bibliographic Resources GOODBYE! THANKS!

CSF
https://www.researchgate.net/figure/Map-of-countries-with-the-OIE-official-free-status-for-Classical-swine-fever-
2021_fig2_353426765
https://www.woah.org/fileadmin/Home/eng/Health_standards/tahc/2011/en_chapitre_1.15.2.pdf
https://www.woah.org/en/disease/classical-swine-fever/
https://www.msdvetmanual.com/generalized-conditions/classical-swine-fever/classical-swine-fever
https://www.woah.org/fileadmin/Home/eng/Health_standards/tahm/3.08.03_CSF.pdf
https://www.oie.int/fileadmin/Home/fr/Health_standards/tahm/3.08.03_CSF.pdf
https://youtu.be/5J65jG9g-ew
https://youtu.be/Je9241JArrQ
https://youtu.be/4Ry2z3T-Rfo
https://youtu.be/ejqVCKUsHmk
https://youtu.be/fHEBbISDv84
https://youtu.be/sQTs9Al47Zs
ASF
https://www.msdvetmanual.com/generalized-conditions/african-swine-fever/african-swine-fever
https://www.woah.org/en/disease/african-swine-fever/
https://www.woah.org/app/uploads/2022/05/asf-report11.pdf
https://www.oie.int/fileadmin/Home/fr/Health_standards/tahm/3.08.01_ASF.pdf
https://youtu.be/zb-9tdJPxf8
https://youtu.be/Tj1qg8NaCMA
https://youtu.be/4Ej3dyACu9A
https://youtu.be/gQLkacWl0kA
https://youtu.be/g0BYrjnNMC0
https://youtu.be/0ykP5HxP_wQ
https://rr-asia.woah.org/wp-content/uploads/2020/03/asf_vittorio_guberti_part_2.pdf
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NEWCASTLE DISEASE
Big problem in countries, where birds are
growing in the households system, than in
countries where the intensive system is
almost exclusively, practiced.
Epizootic, but it tends to Panzootic.

NEWCASTLE
DISEASE
Economic and health importance

• Decrease in egg production.
• Costs of immunoprophylactic measures,

and the losses due to the restrictive
measures, imposed by the control plan.
• Can be transmitted to humans, producing

conjunctivitis or a flu syndrome, and in
children nervous symptoms. https://youtu.be/jkNxmTrrZSk
ETIOLOGY
Family Paramyxoviridae, Avulavirus Genus, ARN
Avian paramyxovirus 1 (VPA 1).
Avian paramyxovirus type 2to9, or paramixoviruses 2to9 (VPA 2-9), do not

cause Newcastle disease.
Differentiation by hemagglutination or inhibition of hemagglutination tests.
Pleomorphic virus, wrapped in a lipoprotein-type envelope, the surface of

which is finding a large number of spicules, the hemagglutinin sites.
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https://youtu.be/NnzqDRU9Nug
NEWCASTLE DISEASE
In concentrated state, the virus also has RNA virus - avian paramyxovirus 1 (APMV-1 or VPA1) have 5 patotypes
• velogenic viscerotrop
hemolytic action, on chicken erythrocytes. • velogenic neurotrop
• mesogenic
• lentogenic or respiratory
• asymptomatic enteric: initial replication in the intestine -
The virus is grown on 7-12 day old avirulence infection
Rarely, membership of a certain patotype is clear
embryonated eggs, or various cell cultures
Inactivated at 56°C, after 3 hours, or 60°C, in 30 minutes.
of either avian origin, such as chicken Inactivated by acid pH ≤ 2.
Sensitive to ether; formalin-inactivated, phenols and oxidizing agents (e.g., Virkon);
embryo fibroblasts, or mammalian origin, chlorhexidine, sodium hypochlorite (6%)
such as HeLa, BHK 21. Resistant to the environment (frozen 180 days, lyophilized for 3 years, in sheltered for
2 months and in death animals 1 month)
- It produces embryonic mortality (24-100 h p.i.), cytopathic and
haemagglutinating effect (for red blood bird)
- antigenicity - homogeneous;
- immunogenicity - protective titer, Ab at 21 days p.i. (minimum titre at 7 days)
• Velogenic strains are wild pathogenic strains that cause

disease by natural infection. Are described 4 types of clinical evolution depending on the
type of pathogenicity of the virus involved in the infectious
process:
• Mesogenic strains - medium pathogenic viruses that can Velogenic strains,
be isolated from natural infection or which have been - Doyle type (1927) - all ages – Disorders : general, digestive,
artificially created in laboratories by attenuating the respiratory and nervous.
pathogenicity of some velogenic strains - Beach type (1942) - chicks, youth - Disorders : general,
respiratory and nervous.
Mesogenic strains,
• Lentogenic strains - lacking pathogenicity are found as in - Beaudette type (1946) - chicks, youth -Disorders: respiratory
nature from where they have been isolated or have been and nervous.
artificially created in laboratories by attenuating the adults – respiratory disorders.
pathogenicity of some velogenic and mesogenic strains Lentogenic strains,
- Hitchner type (1948) - chicks - respiratory disorders.
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Epidemiologic suspicion diagnostic Sparrows, will have variable susceptibility; some species do not
Hosts: express clinical signs of disease, but excrete the virus, while others
Numerous species of domestic and wild birds: develop a serious illness.
Hens are very susceptible to disease; turkeys do not tend to develop severe
signs;
Cases of fatal disease, have also been reported in crows, and
Birds of hunting (pheasants, partridges, quail and guinea fowl) and psittacids (true ravens (genus Corvus).
parrots) have variable susceptibility; cacatuidale (nymphs) are susceptible; Acute form of ND, was also recorded in penguins (Pelecaniformes
order).
Wild birds and water birds.
Some isolated viral VPA1 strains, from certain genotypes, can cause infections in
all bird species that populate a particular habitat at the same time. PALMIPEDS ARE CONSIDERED RESISTANT SPECIES.
The disease was observed in ostriches, and it is known that pigeons are
susceptible.
Morbidity and mortality rates vary, depending on the host species
Rapier birds are usually resistant to ND, except the bearded eagle (Gypaetus and the viral strain.
barbatus), the haliaeetus albicilla (Haliaeetus albicilla), a fisherman eagle
(Pandion heliaetus) and several species of hawk.
NEWCASTLE DISEASE
Epidemiology CLINICAL SUSPICION DIAGNOSIS:
Incubation 2-15 days (5-6 days); in some species it may exceed 20 days.
Sources of infection
Primary:
- diseased/contaminated birds by excretions and secretions. • General disorders : inappetence, horiplumation,
- their bodies and carcasses. • Digestive disorders : indigestion, diarrhea (possibly with blood),
- contaminated eggs. • Respiratory disorders: dyspnea, sneezing,
Secondary: water, feed, bedding, inventory items, contaminated means of • Nervous disorders: convulsions, myoclonus, paresis, paralysis, death.
transport, vectors: hematophagous insects it is not clear the role of flies as
mechanical vectors.
Vaccinated birds, although immune, may be asymptomatic Clinical evolution: - supraacute (≤24h),
- acute (2-6 days),
carriers of velogenic strains on mucous membranes, where the - subacute (7-14 days),
virus can be multiplied, being protected from circulating - chronic (up to 30-45 days with secondary infection),
antibodys of carrier. Secondary bacterial infections: Escherichia, Hemophillus, Mycoplasma (in
chronic forms).
Epidemiology
Epidemiology
• Eggs from infected flocks, may contain the virus
on the shell, or inside the egg
The path of infection:
- horizontal - respiratory, digestive, transcutaneous.
• In the case of attenuated strains, infected
embryos can reach to the hatching age, - vertical – epidemiologically irrelevant (embryonic
resulting infected one day old chicks. mortality).
• Vaccines against infectious bronchitis, avian Dynamics: Explosive epizootic-panzootic

paramixovirus, or other diseases of birds, if were
prepared on infected egg with lentogenic ,
mesogenic and velogenic strains of ND, can be
sources of infection.
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Clinical evolution
In typical forms, evolution can be supra
acute, acute or subacute
Supra acute form, lasting up to one day,

sometimes occurs in chicks and is
manifested by uncharacteristic symptoms:
inappetence, depression, convulsions
Clinical evolution Sometimes bilateral conjunctivitis, edema of

eyelids, epiphosis and corneal opacification
Acute form is the usual form of clinical manifestation. are also noted.
Fever (43-44ºC), inappetence, deep depreciation of
general condition, and stopping of laying.
Sick birds are sleepy, indifferent to what is happening The clinical picture is dominated by either
around them, staying immobile, with rough feathers. digestive symptoms, respiratory or nervous
The body, wings and tail are often left down, symptoms, depending on the pathogen
sometimes supported by the ground. (virulence and tropism) of the infective
strain.
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NEWCASTLE DISEASE
NEWCASTLE DISEASE
Nervous system
disorders
seizures
myoclonus
paresis
Paralysis
Death
TORTICOLIS, PARESIS
TORTICOLIS
NEWCASTLE DISEASE NEWCASTLE DISEASE

Nervous system disorders Nervous system disorders
seizures seizures
myoclonus myoclonus
paresis paresis
Paralysis Paralysis
Death Death
NEWCASTLE DISEASE
Respiratory disorders
Dyspnoea
Sneeze
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Clinical evolution
Subacute form, the symptoms are similar,
fever is less pronounced, nervous disorders
are more severe, the disease last longer
than (7-14 days) and have a lower mortality
rate (30-80%).
Birds that survive, some of them are healing
completely, but others remain with long-
lasting or definitive nervous problems.
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Clinical evolution
ANATOMO-PATHOLOGICAL SUSPICION DIAGNOSIS:
Rare atypical forms, ND is manifested either hemorrhagic collar at the entry into the glandular stomach-
exclusively by respiratory, digestive or nervous proventricule,
symptoms, or only by decreasing egg production. - ingluvial indigestion,
- subcutaneous edema in the head and neck,
Were described forms only manifested by rhino - tonsillitis and hemorrhagico necrotic proctitis,
conjunctivitis and / or edema of the head and neck. - proventriculitis and hemorrhagico-necrotic enteritis,
- hemorrhagico necrotic laryngotracheitis, pulmonary edema /
bloody pneumonia,
A form of ND produced by viscerotrope velogenic
- serous bleeding (bleeding diathesis).
strains manifested only by inflammatory edema
Histopathological examination of nervous system:
comprising the eyelids, conjunctiva, beard, the skin
- neural degeneration, and lymphocytic infiltration.
of the head and finally, eyeball.
ANATOMO-PATHOLOGICAL SUSPICION DIAGNOSIS: ANATOMO-PATHOLOGICAL SUSPICION DIAGNOSIS:
Supra acute forms, especially found at On the oral, pharyngeal and esophageal
chickens, lesions are either missing or mucosa, the mucous papillae are swollen.
discrete, consisting of catarrh hemorrhagic
inflammation of the intestine or respiratory Hemorrhagic lesions, located mainly on the
system tip of the glandular papillae, disseminated
In typical, acute and subacute forms the throughout the proventricular mucosa or
lesions are sometimes characteristic and are organized as hemorrhagic cord or collar, at
mainly due to hemorrhagic infiltration due to the entrance or exit of the proventricular
degeneration, necrosis and vascular rupture. mucosa.
Subacute forms, due to hemorrhagic Inflammatory lesions are formed, with the
infiltration, local necrosis of the cells also appearance of plaques, or diphtheroid
occurs, inflammation becoming hemorrhagic buttons, are difficult to remove, thickened
or necrotic. and visible, even through the transverse wall
Intestinal lesions - hemorrhages with ovoid of the unopened intestine, will have the form
shape, on average sizes of 2-8 mm, few in of violet-red spots.
number, which can be found on the mucosa
of the small intestine, lesions are infiltrate by Hemorrhagic or necrotic inflammation, of
an fibrinogen exudate, and local necrosis lymphoid formations from entrance of cecal
occurs. sacs (cecal tonsils).
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Ovarian hemorrhages, sometimes with Hemoragic collar
follicular breakage and vitelline peritonitis ,

hemorrhages at the base of the heart,
internal sternum, mesentery, and other
serosae.
Central Nervous System, histologically are
described congestions and hemorrhagic
infiltrations of meningeal membranes, lymph
systemic meningoencephalitis, perivascular
Proventriculitis and haemorrhagico-
infiltration, glial proliferation and neuronal necrotic enteritis
degeneration.
Haemorrhagico- Hemorrhagico-
necrotic necrotic Inflammation of the cecum and
tracheitis proventriculitis hemorrhagico-necrotic proctitis
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HISTOPATHOLOGICAL EXAMINATION
Image: Dr Peter Hooper, Australian Animal Health Laboratory (AAHL)
Immunohistochemistry:
Immunohistochemistry: viral particles in
viral particles in cardiac conjunctival mucosa are colored in brown
myocytes of turkeys.
CONFIRMATORY DIAGNOSIS :
Pathogen identification
Samples: live animal: tracheal and cloacal swabs (or
Bird cholera feces); corpses: oro-nasal swabs, lungs, kidneys,
Avian Influence intestines (all with content), spleen, brain, liver and heart,
Infectious avian laryngotracheitis separated or as a fused sample.
Avian diphtheria
Psittacosis (clamidiasis) (psittacine) Isolation on embryonated eggs or cell cultures with confirmation of
mycoplasmosis presence through cytopathic effect.
Infectious Avian Bronchitis Evaluation of haemagglutinating activity + IHA with anti-ND virus,
Antibodies specific serum
Aspergillosis - IFD (direct immunofluorescence)
Management errors: water shortages, ventilation, feeding - ELISA
In companion birds: Pacheco's disease of parrots, salmonellosis, - Molecular methods: RT-PCR, rtRT-PCR, nucleotide sequencing
adenoviroya, and other paramyxoviruses
In cormorants and other wild birds: botulism, avian cholera,
conformation abnormalities
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DIFFERENTIATION OF THE VIRUS

EVALUATION OF PATHOGENICITY –
• Serological techniques,
– 28 MoAb panel for sero grouping, Plaque tests on hen embryo fibroblasts.
– Useful for larger sero groups, limited specificity, Average time for killing embryos.
• Real-time RT-PCR, Intra venous pathogenicity index in 6-week-old chickens.
– L-gene screening, is a real challenge for detecting all
types. Intra cerebral pathogenicity in one day old chicks (ICPI)
- ICPI is the mean per bird per day observed for 8 days in
• Nucleotide sequencing and phylogeny, 10 intra cerebrally inoculated birds with the susceptible
– Safe, high specificity. virus are marked:
– The data are presented as phylogenetic trees. - 0 = normal 1 = ill 2 = dead ICPI = 0 = no sign seen within
– Will provide data on epidemiological interferences. 8 days ICPI = 2 = all birds died within 24 hours.
Serological tests
Samples: blood serum, blood.
• Hem agglutination, inhibition reaction (vaccine virus used
as antigen)
• ELISA
Definition of EU: a bird disease produced by an avian paramyxovirus strain 1

with an intracerebral pathogenicity index (ICPI) in one day chickens more than
0,7. Virus with ICPI greater than 0.7 is considered virulent
PROPHYLAXIS :
PROPHYLAXIS : SPECIFIC
General Non-specific, Performed in flocs of hens, turkeys, pheasants and pigeons.
Strict isolation of outbreaks. With live attenuated vaccines (LaSota lentogenic strains).
Destroying all infected and exposed birds. Mode of administration: - in the conjunctival sac, aerosols, intramuscular,
Judicious washing and disinfection of shelters (prophylactic DDD). subcutaneous.
Appropriate disposal of carcasses. Vaccination schemes:
Control of parasites in flocks. In farms: at 8-10 days, 22-24 days, 40-42 days, 70-72 days, 115-120 days,
Depopulation, and 21 days vacuum health, fallowed by restocking. then from 3 to 3 months.
Avoiding contact with birds, with unknown health status. In households: at 2 months of age, then at 4 to 4 months, in three annual
Control of human movement: sanitary filter / firm fencing / guard. campaigns.
Exploitation on the totally empty principle: a single age on the farm. In farms are performed the antibodies serological control, after 3rd anti-
Newcastle Disease vaccination.
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COMBAT:
Official statement
Quarantine of first degree.
Stamping-out
Lifting quarantine measures, after 30 days from the last case
and the final disinfection.
RESTOCKING
Repopulation is done by introducing sentinel birds (SPF),
holding them for 40 days on the farm, testing for specific anti-
ND antibodies (negative result), and then full repopulation;
Birds in the repopulated shelters, can not leave the holding
before carrying out negative serological tests.
• https://www.oie.int/fileadmin/Home/eng/Animal_Health_in_the_World/docs/pdf/Disease_cards/NE
WCASTLE_DISEASE.pdf
• https://www.cdfa.ca.gov/ahfss/animal_health/pdfs/V_N_D_Photos-EngSpan.pdf
• https://www.cdfa.ca.gov/ahfss/animal_health/pdfs/V_N_D_Veterinarians.pdf
SURVEILLANCE DIAGNOSIS • https://www.cdfa.ca.gov/ahfss/animal_health/newcastle_disease_info.html
• https://www.aphis.usda.gov/aphis/ourfocus/animalhealth/animal-disease-
information/avian/virulent-newcastle/vnd
NCD surveillance is performed according to the Program of Action for the
• https://www.msdvetmanual.com/poultry/newcastle-disease-and-other-paramyxovirus-
Prevention and Control of Animal Diseases through: infections/newcastle-disease-in-poultry
- permanent clinical and anatomo pathological surveillance of • https://www.woah.org/fileadmin/Home/fr/Health_standards/tahm/3.03.14_NEWCASTLE_DIS.pdf
birds • https://www.aphis.usda.gov/animal_health/emergency_management/downloads/sop/sop_nd_e-
e.pdf
- serological surveillance of poultry in poultry holdings, households, • https://youtu.be/LYLAQ3OsqTQ -vaccinare
import. • https://youtu.be/GUuiGHuiPUo -semne clinice
• https://youtu.be/NnzqDRU9Nug - etiologie si morfopatologie
• https://youtu.be/MQuOVZxDtEQ
• https://youtu.be/l_MpPBv2-mQ
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BOVINE ENZOOTIC LEUCOSIS

(LEB, BLV) B-108
EQUINE INFECTIOUS ANEMIA
(AIE, EIA) B-205
(LEB, BLV) B-108
2022
2022 2
BOVINE ENZOOTIC LEUCOSIS BOVINE ENZOOTIC LEUCOSIS
 Horse
DVM PhD DRAGOS COBZARIU DVM PhD DRAGOS COBZARIU
2022 3 2022 4

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https://youtu.be/EGfMRVBDxaY
https://youtu.be/HhhRQ4t95OI
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EPIDEMIOLOGYCAL SUSPICION
Receptivity:
- natural infection: cattle and bubaline - whatever their age
ETIOLOGY : (maximum incidence ≈ 7 - 8 years)
- experimental infection: sheep, goats
Risk factors:
- RNA virus- fam. Retroviridae - age (> 2 years);
- genus Deltaretrovirus, type C (BLV) - breed - red-dressed breeds (Danish Red!)
- Tropism for - B lymphocytes - wrestling breeds - milk breeds - most affected!)
- Stress
- Macrophages lack defence Sources:
- T lymphocites-partially - Primary - infected animals (lymphocytes in blood or milk!)
Minimum quantity of blood for transmission: 0.1 ml!
- Sensiblity: - low in the external environment - Secondary - Surgical instruments
- 30 ”→ 80 ºC - objects contaminated with infected blood
- max. 2 h at changes in pH (<6-> 8) - Vectors - blood-sucking insects (horseflies!)
NOT TRANSMITTED through semen, saliva.

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Route of infection: LEB 2019

- Vertical (transplacental)
(congenital infection in 4 - 8% calves of BLV + cows)
- Calves infected "in utero" from the age of 80 days can be identified;
- Horizontal - Digestive (colostrum / contaminated milk)
- Transcutaneous - blood-sucking insects
- Iatrogenic - punctures / operations
(with contaminated instruments)
- transrectal transmission
(pregnancy control)
- Direct contact (wounds, small lesions)
Dynamics:
Sporadico-Enzoothic with stationary character

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CLINICAL SUSPICION:
LEB 2020 - Long incubation (→ 4-5 weeks) with chronic - sometimes acute course
Enzootic leucosis - leukemic phase - lymphoid!

- phase of persistent lymphocitosis
- tumoral phase
- myeloid phase (rare)
Sporadic leucosis - Juvenile (young cattle up to 2 years)
- Thymic (cattle between 6 - 30 months)
- Dermal - rare (cattle between 6 months - 4 years)
- Paraclinical example (concluding in the cytemic-leukemic phase)

- Hematological - leukocytosis (100,000 leucocytes / mm3, of which 95% lymphocites)
- Cytomorphologic - lymphoblastosis, myeloblastosis

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CLINICAL STAGES OF BOVINE LEUCOSIS BOVINE ENZOOTIC LEUCOSIS

(according to Dr. Barna, 1994)
Stage Lesions Duration
I. Asymptomatic carrier Leukocytosis, lymphocitosis, eosynophilia, 3-6
of virus transient monocytosis. months
Precipitating antibody against BLV.
II. Lymphadenopathy Unilateral or bilateral hypertrophy of explorable 1–2
lymph nodes (including those difficult to years CLINICAL STAGES
palpate transcutaneously), seroconversion
III. Leukemic Alteration of the dominant blood-hematological 3-6 Average incubation 4 - 6 weeks (3 - 9).
hemograme on the lymphoid series. months
IV. Generalized Generalized lymphosarcoma-type tumors in 2-3 Clinical expression after 2 - 5 years.
lymphosarcoma tissues and organs. months
+ Digestive - tumors; - esophagus - strictures. Chronic evolution (over acute / acute, rare-youths).
+ Circulator - heart failure.
+ Genital ap.- infertility, metritis, vaginitis, tumors.
+ Urinary tract - dysuria, hematuria. Death at the end, remissions during evolution.
+ Nervous sist. - excitement, paresis, paralysis
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STAGE I
Asymptomatic carrier for 3 - 6 months; STAGE II
- Leukocytosis, lymphocitosis, eosynophilia, transient monocytosis;
Hypertrophy lnn deep-interns and explorable lnn.
- are mobile, non-painful and hard (reticulosarcoma)
Nuclear - are soft (lymphosarcoma);
Athypia
Cytoplasmatic
} → Prolymphoblasts, Lymphoblasts, Rieder cells - Apathy, Anemia, Loss of appetite - Progressive evolution
Antibodies-Ab appear after 3 - 6 weeks

Precipitating antibodies → sero + ═> CONFIRMING diseased animal
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STAGE III
STAGE IV
Lymphoid leukemia - Malignant Lymphocytic Lymphoma
- Tumors lnn. + Viscera
- Diagnostic keys: Bendixen, Goetze, Tolle, CEE
Key of the European Economic Community (EEC) Antibodies precipitating in 70% of infected animals-DIAGNOSTICS
to diagnose bovine leukosis
Leukemic
Age in years Negative Exam Values Positive LEUCOSES
-limphocites
0–1 11.000 11.000 – 13000 13.000 Aleukemic
1–2 10.000 10.000 – 12.000 12.000
2–3 8.500 8.500 – 10.500 10.500 1.ACUTE: to stem cells; Hemocytoblastic, Paralymphoblastic and
3–4 7.500 7.500 – 9.500 9.500
Lymphoblastic; Reticulosarcoma; Lymphosarcoma; Hodgkin disease,
4–5 6.500 6.500 – 8.500 8.500
5–6 6.000 6.000 – 8.000 8.000
Giantfolliculary lymphoma, Myeloma.
More than 6 5.500 5.500 – 7.500 7.500
years 2. CHRONIC: Monocytic, Lymphocytic, Plasmocytic
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SUSPICION - CONCLUSION:
ANATOMOPATHOLOGICAL SUSPICION:
EPIDEMIOLOGICAL:
DIFFUSE OR / AND NODULAR INFILTRATIONS in: - Race
- hematopoietic organs (marrow, lymphatic nodes) - Age
- Veterinary technologies
- fetal appearance of bone marrow
- Operating system
- viscera (clot, liver, spleen, kidneys, CNS, uterus.)
CLINICAL:
Alopecia, Emaciation- muscle masses - ex. Hematologic: Count / Leukocyte
Formula
Adenopathie +
HISTOPATHOLOGICAL:
Abundance of neoplastic cells (more than 90% lymphoblastic)

in infiltrated tissues
{ - Medulograme
Adenograme
Immunological

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DIAGNOSTIC - CONFIRMATION: DIAGNOSTIC - CONFIRMATION:
-Isolation and Identification of virus

Identification of Virus (forme Integrated Provirus - PCR )
p 10, p 12, p 24
- Evidence of the antigenic components of LEB with MoAb
gp 58 (F, G, H)
-Identification of specific antibodies-Ab: ser, colostrum

- IDGA (3 months after infection) - for infected animals
- ELISA - for uninjured staff
-Cytomorphological (smear or erythrocyte concentrate)

-Medullogram
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PROPHYLAXIS PROPHYLAXIS
- Control of imports → only free animals; •Eliminating the movement of blood from infected animals to naive animals is the cornerstone of prevention
- Non-specific - prophylactic quarantine (minimum 4 months!) protocols. In calves, feeding colostrum from seronegative cows is often advocated. However, most
epidemiologic evidence suggests that the protective effect of colostral antibody outweighs the risk of
- Annual monitoring-surveillance of the LEB; infections, particularly in high prevalence herds. The replacement of whole milk feeding with high-quality milk
replacer may also be considered. Bloody milk should never be fed to calves.
•Cautery or other bloodless methods of dehorning should be used. Equipment used for castration, tattooing,
-Guarantees for breeding animals, seminal material. ear tagging, or implanting should be adequately cleaned and disinfected between animals.
•Transmission can be decreased in adult cattle by changing rectal sleeves in between cows. Artificial
insemination or embryo transfer (using negative recipients) may limit transmission. In beef herds, the use of a
-Free Farms/ Exploatations -Surveillance negative bull may limit transmission, but natural service is an uncommon method of viral transmission unless
breeding is traumatic.
•Additional recommendations include disinfection of equipment that has come in contact with blood or body
Serological by ELISA and / or ID in cattle> 6 months for: tissue. Single use, disposable needles should always be used for blood collection and IM injections. It is
- bulls and cows - when allowed and 2 times / year; preferable to use single-use disposable needles for vaccination, but the risk of transmitting BLV virus via SC
vaccination is low. Handling facilities that become contaminated with blood should be cleaned between
- cows, buffaloes, heifers - once / year; animals. Fly control helps minimize the potential for tabanid-associated transmission. Blood transfusions and
vaccines containing blood, such as those used for babesiosis and anaplasmosis, are particularly potent ways to
- imported cattle - control in prophylactic quarantine; spread the disease, and donors must be carefully screened.
- exported cattle - control 30 days before delivery;
- cattle for fattening - once a year.
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COMBAT MEASURES
There is no treatment for viral infection or for lympho sarcoma in cattle, although
parenteral corticosteroids can transiently decrease the severity of clinical signs.
COMBAT MEASURES Eradication programs have been developed but success has been variable, primarily
- Official declaration of the disease because of the expense and high prevalence of infection among cattle, relative to the
- 3rd degree quarantine economic cost of disease. The most commonly recommended eradication protocol is as
follows:
1) identify infected animals using a serologic test,
1 or more animals infects: 2) cull seropositive animals immediately,
3) retest the herd in 30–60 days,
<15% R + ═> unit în sanitation by extraction. 4) use PCR to test young calves and as a complementary test to clarify test results in
herds with a low prevalence of infection, and
5) repeat testing and cull until the entire herd tests negative. Testing is then repeated
> 15% R+ ═> reorganization by liquidation every 6 mo. The herd is declared free when there have been no positive tests for 2 yr.
Additions to the herd should have two negative tests 30 and 60 days before arrival.
When test and cull programs are economically untenable, test and segregation programs
have been recommended but are rarely implemented. These programs necessitate
running two completely separate operations and require additional resources, including
money, time, and available workforce.

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EQUINE INFECTIOUS
ANEMIA
(AIE, EIA) B-205
https://youtu.be/1lhMkrbOuDY
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EQUINE INFECTIOUS ANEMIA EQUINE INFECTIOUS ANEMIA
Etiology: - RNA virus - fam. Retroviridae

- genus Lentivirus
- Tropism for the blood-forming tissue
and reticulo-endothelial tissue
- Not immunogenic!
- Phenomenon of „DRIFT OF VIRAL ANTIGENS” →
antigenic variants! (p26 - stable!)
- Resistant in the environment:
- 3 months cadavers
- 6 months fibrous feed
- 1 month in water
- 1 hour at 60 ° C
 Horse

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Viral infectious disease specific to solipeds, clinically characterized Sources of infection :
by fever, anemia, cardiovascular disorders, bleeding diathesis and
reticuloendothelial tissue hyperplasia. Primary :
- contaminated animals (with or without clinical signs)
DIAGNOSIS OF SUSPICION
(carry and shed the virus all their lives)
- corpses of contaminated animals Low risck!
EPIDEMIOLOGY
- meat, products and by-products
Receptivity: Solipeds (horse, mule, hedgehog, donkey- i.m.p.m.)
- regardless of race or sex
Secondary:
Favorising factors: - water, fodder, droppings, slurry
- age - more sensitive young horses - dressings, harness (for collective use)
- food and working conditions - vectors – HEMATOPHAGOUS insects High risck!
- geoclimatic particularities (heat and humidity, forests) (horseflies, flies, mosquitoes, ticks,)
(hot and humid regions → blood-sucking insects).

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AIE 2019
Transmission routes:
Horizontaly
- Transcutaneous
- Bites of blood-sucking insects
- Friction wounds (loose harness, grooming)
- Iatrogenic - non-sterilized instruments (needles, scalpel, probes)
- digestive
- Direct contact possible, but not epidemiologically significant!
Verticaly - transplacentary
- Favorable conditions: long cohabitation + large population- vectors
Dynamics: Enzootic - recently infected herds.

Sporadic - herds with old infections.
(seasonal exacerbations → hot and humid months)

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AIE 2020

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CLINICAL SUSPICION
Incubation 4-6 weeks.
1st moderate access, with quick remission
BIOLOGICAL MODIFICATIONS (paraclinical)
ACUTE or SUB-ACUTE
- Intermittent fever 41oC, ↓ ↑ 1oC / hour - Leukopenia (neutropenia, lymphopenia)
- Anorexia, depression, debilitation, ataxia
- Delicate edemas: abdomen, foreskin, limbs, petechial lesions - Trombocytopenia
on the mucous membranes
- Cardiac changes: increased intensity of heart sounds, tachiycardia,
- Erytrhopenia (normocytic, normochromic)
arrhythmias, serosanguineous nasal discharges.
- Splenomegaly
Abortions. - Decrease total serum protein, maintaining the ratio between fractions
CRONIC
- Clinical remissions with RELAPSES at 2-3 week intervals - Early precipitating antibodies → IDGA at 45 days p.i.
→ death the first year → worsening (corticosteroid therapy, stress)
→ clinical remission → CHRONIC CARRIER AND ELIMINATOR - Donkeys - colostral antibodies up to 65-182 days.

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ANATOMO-PATHOLOGICAL SUSPICION CONFIRMATION DIAGNOSIS
- Anemic mucous membranes and hemorrhagic diathesis

- Cardiac dilation and hypertrophy + subepicardial hemorrhages
- Splenomegaly and hepatomegaly (bitten liver)
- Phoetalization islands the marrow long bones (femur,humerus)
- Cachexia and hypertrophy ln.
- Ex. Virusological histopathological and haematological.

- Ex. Serological - ELSA, (Ac- after 30-40 days P.I.)
- COGINS-immunodiffusion in gellose
- PCR
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EQUINE INFECTIOUS ANEMIA PROPHILAXY:
-Prophylactic quarantine;
Recommendations for the equidae import of Veterinary Authorities of importing Strict control of the movements of equines;
countries should require the presentation of an INTERNATIONAL VETERINARY Biosecurity measures in equine farms;
CERTIFICATE attesting to: Serological monitoring by ID and / or ELISA - horses over 6 months
Histological and anatomo-pathological surveillance of dead equines in quarantine, or
1) the animals showed no clinical sign of infectious equine anemia on the day of with suspicion of disease.
shipment, nor during the previous 48 hours -Clinical and serological monitoring of healthy solipeds
2) no case of equine infectious anemia has been associated with the premises in
FIGHT AGAINST DISEASE:
which the animals have been kept during the three months prior to shipment
Incurable disease, ineffective treatment
3) that in the event of permanent importation, the animals have been tested for - 3rd degree quarantine officially declared
infectious anemia in equines by means of a diagnostic test carried out on blood - Isolation of the suspected of infection from the rest;
samples taken during the 30 days prior to shipment, the result of which was negative, - Pest Control and deworming;
or - Monitoring of equine marketing - live animal fairs;
4) that, in the event of temporary importation, the animals have been tested for - Elimination of sick equines, within 10 days after receipt of the analysis report with a
infectious anemia of equines by means of a diagnostic test carried out on blood positive result
samples taken during the 90 days prior to loading, the result of which was negative. - End of quarantine 6 months after the last case
Romanian counties at high epidemiological risk for AIE: Sălaj , Satu Mare ,
Sibiu , Maramureş , Mureş , Bihor , Bistriţa , Arad
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https://youtu.be/60YO4eZiv3s
2022 45
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• ^ Cromer D, Grimm AJ, Schlub TE, Mak J, Davenport MP (January 2016). "Estimating the in-vivo HIV template switching and recombination rate". AIDS. 30 (2): 185–
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^ Jolly C (March 2011). "Cell-to-cell transmission of retroviruses: Innate immunity and interferon-induced restriction factors". Virology. 411 (2): 251–9. doi:10.1016/j.virol.2010.12.031. PMC 3053447. PMID 21247613.
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^ Aiewsakun P, Katzourakis A (January 2017). "Marine origin of retroviruses in the early Palaeozoic Era". Nature Communications. 8: 13954. Bibcode:2017NatCo...813954A. doi:10.1038/ncomms13954. PMC 5512871. PMID 28071651.
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^ Desport M, ed. (2010). Lentiviruses and Macrophages: Molecular and Cellular Interactions. Caister Academic. ISBN 978-1-904455-60-8.
https://youtu.be/hxAYxvPdQFY • ^ Ross, S. R. (2018). Cellular Immune Responses to Retroviruses. In Retrovirus-Cell Interactions (pp. 401–420). Elsevier. https://doi.org/10.1016/B978-0-12-811185-7.00011-X
• ^ Burrell, C. J., Howard, C. R., & Murphy, F. A. (2017). Retroviruses. In Fenner and White’s Medical Virology (pp. 317–344). Elsevier. https://doi.org/10.1016/b978-0-12-375156-0.00023-0
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^ Lauber C, Seitz S, Mattei S, Suh A, Beck J, Herstein J, et al. (September 2017). "Deciphering the Origin and Evolution of Hepatitis B Viruses by Means of a Family of Non-enveloped Fish Viruses". Cell Host & Microbe. 22 (3): 387–
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^ Krupovic M, Blomberg J, Coffin JM, Dasgupta I, Fan H, Geering AD, et al. (June 2018). "Ortervirales: New Virus Order Unifying Five Families of Reverse-Transcribing Viruses". Journal of Virology. 92 (12). doi:10.1128/JVI.00515-
18. PMC 5974489. PMID 29618642.
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^ Fact-checking Judy Mikovits, the controversial virologist attacking Anthony Fauci in a viral conspiracy video, By Martin Enserink, Jon Cohen, May 8, 2020, accessed June 17, 2022, science.org website.
^ Neil, Stuart J.D.; Campbell, Edward M. (2020). "Fake Science: XMRV, COVID-19, and the Toxic Legacy of Dr. Judy Mikovits". AIDS Research and Human Retroviruses. 36 (7): 545–
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^ Virus Conspiracists Elevate a New Champion, by Davey Alba, May 9, 2020, nytimes.com
• ^ Rutherford GW, Sangani PR, Kennedy GE (2003). "Three- or four- versus two-drug antiretroviral maintenance regimens for HIV infection". The Cochrane Database of Systematic Reviews (4):
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MAREK DISEASE(MD-NDV) MAREK DISEASE(MD-NDV)

ENZOOTIC PARALYSIS,
AVIAN LEUCOSIS NEUROLYMPHOMATOSIS,
ACUTE LEUCOSIS,
2022 SKIN LEUCOSIS.
Infectious and contagious disease, specific to hens,

expressed by lymphocytic infiltration of peripheral nerves
and tumors in different organs.
Aladar Marek
ETIOLOGIE:
Gallid herpesvirus 2, (GaHV-2) genus Mardivirus, a DNA virus that induces
tumor transformation.
In infected organisms, the herpesvirus → integrated form, causes CYTOLYSIS
(which determines the proliferation of the free cell, not associated with the
genome, present in the skin - plumifer follicle, sub-epidermal structures -
hence it is eliminated with keratinization and exfoliation).
HERPESVIRUS - ELIMINATED THROUGHOUT LIFE- INFECTED BIRD.
Herpesvirus in environment: envelope, protected by the proteins of the
dequamation product against radiation and temperature.
Dissemination through atmospheric air with fine particles and dust, in
• https://youtu.be/b8n86DSJdPQ shelters.
Dr. DVM PhD Dragos Cobzariu 3 Dr. DVM PhD Dragos Cobzariu 4
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In the order of Herpesvirales and the family of Herpesviridae, the Gallid herpesvirus 2, (GaHV-2), Serotypes and Pathotipes of MDV
herpesviruses of birds, grouped together in the subfamily Alphaherpesvirinae,
infect many avian species (chickens, turkeys, ducks, pigeons, parrots, eagles,
storks, falcons, cranes , hake, cormorants, penguins, owls).
The main avian herpesviruses are:
Oncogenic Marek's disease (Gallid herpesvirus 2, genus Mardivirus),
Infectious laryngotracheitis (Gallid herpesvirus 1, genus Iltovirus),
Virus enteritis or duck plague (Anatid Herpesvirus), not classified, but close to
genera Mardivirus.
Varicellovirus and Simplexvirus in the same subfamily.
These Alphaherpesvirinae can cause significant economic and ecological
losses. Other herpesviruses can infect pigeons (Columbid herpesvirus 1),
parrots (Psittacid herpesvirus 1 responsible for Pacheco's disease) and other
species of companion or wild birds.
Ord. Herpesvirales ,
In most shelters and aviaries where birds are raised, the infection is widespread. Virtually all birds become infected in the first weeks of
Fam. Herpesviridae, life, although this can sometimes be delayed by strict biosecurity measures. Due to the high prevalence of viruses, Gallid herpesvirus 2,
Subfam. Alphaherpesvirinae serotype 1, which presents variable pathogenicity, from mild to highly pathogenic, and serotypes 2 and 3, non-oncogenic, in the
environment of birds, they can be infected with more than one specific herpes strain of MDV. There is some evidence to suggest that
Gallid herpesvirus 2, with aging, the frequency of non-oncogenic, low-virulence or non-pathogenic virus isolates increases.
Unlike virulent strains of Marek's disease virus, which are highly contagious, turkey herpesviruses are not easily transmitted to chickens
(GaHV-2) genus Mardivirus, (although they are easily transmitted among turkeys, their natural hosts). Attenuated strains of Marek's disease virus vary greatly in
transmissibility.
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PATHOGENESIS OF MAREK DISEASE IN POULTRY

Currently, four phases of infection with Marek's disease in vivo are recognized:
• Early cytolytic infection (productive-restrictive),latent infection
• Second phase of cytolytic, productive-restrictive infection coincident with permanent
immunosuppression
• Proliferative phase, involving nonproductively infected lymphoid cells that may or may
not progress to the point of lymphoma formation.
• Productive infection may occur transiently in B lymphocytes within a few days after
infection with virulent Marek disease virus strains and is characterized by antigen
production, which leads to cell death.
Because few if any virions are produced, this has also been termed a restrictive-
productive infection.
Productive infection also occurs in the feather follicle epithelium, in which enveloped
virions are produced.
Latent infection of activated T cells is responsible for the longterm carrier state.
No antigens are expressed, but virus can be recovered from such lymphocytes by
cocultivation with susceptible cells in tissue cultures.
Some T cells, latently infected with oncogenic Marek disease virus strains, undergo
neoplastic transformation. These transformed cells, provided they escape the immune
• https://youtu.be/fH1zS7hlW54
system of the host, may multiply to form characteristic lymphoid neoplasms.
DVM PhD DRAGOS COBZARIU 7 Dr. DVM PhD Dragos Cobzariu 8
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EPIDEMIOLOGYCAL SUSPICION
Sources of infection:
Receptivity: INFECTED BIRDS WITH OR WITHOUT SIGNS OF DISEASE, OR VACCINES!
Hens - maximum receptivity: 1 DAY-OLD POULT; receptivity declines with
advancing age; may establish resistance against the development of Vaccination does not protect against infection with the wild-type
lymphomatous infiltrations. virus, it does protect against the development of tumors; its eradication
is not possible; vaccination cannot be stopped).
Risk factors: The virus: ubiquitous in the workforce; there are birds that do not have
- Sex, tumors, but excrete the wild virus by desquamation;
- Age,
- Stress, Usually the shell of the contaminated egg is wet and sticky by the
- Subdivisions, surfactant of the genital tractus, the dust from the shelter is placed on
- Air currents, its surface, along with the virus, causing continuous contamination
The genetic structure of the host (resistance to Marek disease is codified (eggs are always taken and disinfected); when the chickens come out of
by the dominant B21 allele; heterozygotes that have the allele are resistant the egg, they are automatically contaminated.
to Marek disease).
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CLINICAL SUSPICION
The clinical evolutionary dynamics is determined by the single or associated infection of different herpesvirus pathotypes, which
possess different levels of virulence, as well as by the performance and subsequent immunization, following prophylactic actions,
respectively vaccination of the chicks on the first day of life.
A) CLASSIC FORMS OF MAREK'S DISEASE:
Clinical signs of Marek's disease usually appear around the age of 3-7 weeks, and the peak of clinical symptoms occurs
between 10-30 weeks, but difficult to suspect clinically.
Transmission: Birds with visceral tumors, are adynamic, will presente laziness, prostration and are often cachectic before death, which may
occur after another 3 weeks.
Inhalation (respiratory tract). NEURAL FORM-early also called Neurolymphomatosis,
Nodular lymphocytic infiltration of peripheral nerves, leading to paralysis, is characteristic of Marek disease.
Birds are affected by lymphocytic infiltration, frequently asymmetric, or unilateral of the peripheral nerves, belonging to the sciatic
Dynamics: and brachial plexuses, infiltration leading to asymmetric partial paralysis, of the bird's wings and legs and/or death when the
vagus nerve is affected.
ENZOOTIC-explosive, relative to the age at which the infection appeared. GaHV-2 can also infect the brain, leading to transient paralysis or persistent neurological disease.
OCULAR FORM -late
Chickens contaminated on the 1st day of life, excrete the virus after 2 Blindness is observed clinically, it is due to lymphocytic infiltration of the optic nerve. Lymphocytic infiltrations at the level of the
iris, are easily observed clinically, detecting changes in the color and shape of the iris slit.
weeks, in non-vaccinated herds. CUTANEOUS FORM improperly named, “cutaneous leucosis”
The cutaneous form, also known in the past as "cutaneous leucosis“, is represented by the
location at the level of the plumifer follicles.
Nodular lesions at this level may involve several scattered follicles, or the lesions may converge, a local inflammatory process is
identified by the presence of a reddish coloration of the cutis in the areas with lesions.
VISCERAL FORM
Visceral tumors are the most common lesions, which give uncharacteristic organ symptoms.
Different localizations can be observed, often associated and in several possible combinations.
Tumors are mainly located in the liver, spleen, gonads, kidneys, heart, glandular stomach - proventricule, and rarely in the
Bursa of Fabricius.
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B) ACUTE FORM. LESIONAL SUSPICION

Begins at the age of 7-16 weeks,
Evolution of 2-5 days, MACROSCOPIC:
Death preceded by paralysis; TUMORS (in classic forms and in adult birds) of the reproductive system
Discrete paralysis, pallor of the ridges. (ovary, testes), in the kidneys, liver, skin, lung, heart, muscles, peripheral
High morbidity rate: nerves.
- 90% in laying hens and
- 8-10% in 7-week-old broilers. Table of PLEOMORPHIC LESIONS - lymphocyte T infiltrations will give,
Skin tumors, too. thickening uniform or nodular-like (bead) of perpherical nerves, will affect the
Paralysis , soon followed by death. brachial plexus, sciatic nerve, cervical nerves.
C) OVERACUTE FORM Peripheral nerves infiltration - histological examination in acute forms, too.
Produced by hypervirulent herpesvirus strains,
Mortality rate: 100% in the first 4 weeks of life. Differential diagnosis
There are no nervous signs, but kidney tumors may appear. Avian Leukosis (where HEPATOMEGALY AND SPLENOMEGALY are
The over acute or acute forms completely compromise the non observed and their tumors, the onset of elevated B lymphocites).
vaccinated birds floks.
Marek's Disease - swelling in the liver, spleen, other tissues and organs
(ovary, testis, heart, lung).
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Normal poultry eye (left); poultry eye with

Marek's disease (right);
Marek's disease - neoplastic infiltration

in the iris, with changes in the pupil.
Marek's disease - ocular form: normal eye

(left), diseased eye (right)
Marek's disease - paralysis of the limbs and
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neck
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Marek's disease - skin lesions
Marek's disease. Characteristic posture for paralysis of a hen legs
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Marek's disease -
swelling of peripheral
nerves, sciatic plexus
Marek's disease - skin lesions
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DIFFERENTIAL DIAGNOSIS:
Avian infectious nephritis,
Avian infectious bursitis,
Dysmetabolies that cause kidney damages

Marek's disease - swollen, tumorized spleen and liver.
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MICROSCOPIC DIAGNOSTICS:
CONFIRMATORY DIAGNOSIS: On microscopic examination, Marek disease lymphomas are characterized

by a mixture of pleomorphic lymphocytes. Some of these cells are true
Histological, tumor cells that carry T cell surface antigens and the GaHP-2 antigen are
expressed in all tumors in Marek disease; others are probably host T cells
Virological, reacting against viral and tumor antigens.
A number of 75% of lymphocites are tumorized and have appearance of
Serological. lymphoblasts, plasma cells, mononuclear, polyinuclear lymphocytes T,
eosinoplils.
Marek's disease-MD, the target cell is

T lymphocyte → polymorphic proliferation
Avian leukosis, the target cell is
B lymphocyte → monomorphic proliferation
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HISTOLOGICAL
Marek's disease is characterized by an INFILTRATION OF
MONONUCLEAR CELLS in the peripheral nerves, gonads, CONFIRMATION DIAGNOSIS:
various viscera, iris, muscle and / or skin.
ISOLATION OF THE VIRUS
VIROLOGICAL EXAMINATIONS (into monolayer cultures of
Although peripheral nerve enlargement and visceral chicken kidney cells or duck embryo fibroblasts);
lymphomas are common, no injury is seen consistently. PCR and indirect immunofluorescence with labeled antibodies
Histological examination,
Criteria such as age (4-20 weeks, except in breeders and layers IDENTIFICATION OF ANTIBODIES (over 4 weeks of age)
where the frequency of tumors due to GaHV-2 increases at the Serological examinations, with anti-tumoral serum.
ELISA, AGID, IF.
start of lay), the distribution of lesions and the absence of
tumors due to GaHV-2.
Other viruses such as Avian leukosis (ALV) or
Reticuloendotheliosis virus (REV) should also be considered.
Dr. DVM PhD Dragos Cobzariu 25 DVM PhD DRAGOS COBZARIU 26
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DIAGNOSTICUL DE CONFIRMARE:
Virological / Serological
Virus can be isolated as early as one or two days after inoculation in
chickens, five days after contact /exposure, and then throughout the life of
the bird.
The virus can be obtained from infected samples of heparinized whole
blood, suspensions of lymphocytes, isolated tumor cells, as well as non-
cellular preparations of the skin, feather follicles or the base of the feathers of
infected chickens.
Cultures on chicken kidney cells and duck embryo fibroblasts are usually
used for the isolation of GaHV-2.
Cultures develop typical plaques within 4-14 days.
Identity of serotype determined Immunofluorescence with serotype-

specific monoclonal antibodies.
Currently, the diagnosis of the GaHV-2 pathotype requires in vivo
challenge experiments.
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PROPHYLAXIS SPECIFIC PROPHYLAXIS - MEDICAL
GENERAL NON-SPECIFIC PROPHYLAXIS Vaccination to PROTECT AGAINST CLINICAL EXPRESSION, but not
The use of vaccines should never be an excuse for poor management or preventing the circulation of the viruses;
lack of biosecurity measures. MEDICAL PROPHYLAXIS:
Dander, feathers and litter from infected flocks are contaminated with Vaccination in the hatchery- live, lyophilized vaccines (at - 86 ° C ):
infectious MDV, which can remain infectious for many months. -HVT (3) / VHT of heterologous turkey tulpins, which confers cross-
Removal and appropriate disposal of dead and infected birds, manure and immunity (protects in 80% of cases);
litter along with disinfection of buildings are important aspects of disease -Rispens (1) protects against tumor development
control, especially in view of the possibility of selection for pathogens with -SB1 (2) - reduced patogenity, belonging to serotypes 1 and 2.
increased virulence.
Furthermore, placing chicks in an environment heavily contaminated with 95% protection
virus, before their immune system is matured and developed, can lead to
vaccination failure.
In addition, avoiding multi-age flocks and air management in the farm are Vaccines can be:
recommended. - monovalent;
Strict biosecurity measures, are also necessary to prevent the introduction of - bivalent. (CVI988 / Rispens)
new MDV strains into a farm. Complies with vaccine preservation measures (sensitive: radiation and temperature)

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Misuse of the vaccine is one of the main reasons for the increased mortality from
the GaHV-2 virus.
The most effective and widely used MD vaccines are made from cell cultures and
should be stored at -196 ° C during transport and until thawed before use.
Vaccine ampoules should be thawed quickly in cold water and the vaccine once
thawed and diluted should be kept cold and used within two hours.
In addition additives, such as antibiotics which can damage the vaccine, should be
avoided.
Vaccines introduced since the 1970s have helped limit the economic losses due to
GaHV-2, but because no vaccine provides sterilizing immunity, the virus has
spread in poultry farms around the world. .
Current vaccines protect against current strains, but new strategies will be
needed in the future so that today's solution is not a concern tomorrow.
Vaccination and biosecurity practices should help delay the onset of more virulent
strains of the infectious agent.
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Genetic resistance CONCLUSION
• Marek's disease is a highly contagious neoplastic

Genetic resistance to MD is well documented, resistant chicken lines can
be developed through progeny testing.
disease characterized by T-cell lymphomas in organs,
and nodular infiltrates of peripheral nerves.
Two distinct genes places, with a major role in the control of resistance was • Paralysis is a common clinical sign.
identified. • The presumptive diagnosis is based on the
epidemiological history, clinical signs, and the
The best characterized association is between the histocompatibility identification of tumors in the organs.
complex (MHC) and resistance to MD, associated with the B 21 allele.
• Vaccines will prevent the clinical evolution, not against
the infection, vaccines must be used together with good
biosecurity measures against the disease.

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AVIAN LEUCOSIS-ALV
Lymphoid Leukosis, Large Liver Disease,
Visceral Lymphoma, Lymphocytomatosis,
• https://youtu.be/Q3DsZQor-TM Lymphomatosis
Dr. DVM PhD Dragos Cobzariu 35
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Oncogenic virus Avian Alpharetrovirus type C, family Retroviridae.

Infectious contagious disease→ malignant tumors → affect the hematopoetic
system.
Historical:
Were described (3rd decade of the 20th century) later than Marek's disease,
as „disease of the large liver” (with wasting of muscle masses and cahexia)
With the progress of genetics, some links of dissemination of this infection
have been deciphered.
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ETIOLOGY:
Avian type C oncogenic virus have numerous strains, grouped in relation to

the speed and initiation of the oncogenic process:
-strains with a long latency period → determined by the genetic structure,
-strains with a short latency period → presence in the single-stranded RNA
genome only of the gag, env and pol genes or / and the presence, with it or
replacing one of them, genes responsible for oncogenesis (the myc genes ).
Avian oncogenic virus genome:

-multiple genes; The env → genes determine the enzymatic functions, and the synthesis of
GAG → Ag synthesis of specific group, p27 (protein); - in all type C viruses; specific envelope proteins with a role in the antigenic differentiation between
POL → synthesis of revers transcriptase enzyme. strains, 6 subgroups are described based on envelope differences, and within each
ENV → enzymatic functions; it determines the synthesis of specific envelope subgroup, strains are described that have or do not have oncogenic capabilities,
proteins, playing a role in antigenic differentiation between tulpines; depending on the presence or absence of genes, myc.
GAG AND ENV → synthesis of proteins, induce the synthesis of specific Ab.
MYC → genes responsible for oncogenesis.
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PATHOGENESIS
INFLUENCED BY:
Multiple determinism, dependent on the causative agent, → and the
genetic structure, of the hosts (several independent genes that trigger the
infection; not well known).
Elements that underlie the genetic sensitivity to this infectious disease
= the consequence of the interaction between several independent
genes from different loci.
Pathogenesis:
Exogenous infection (with clinical expression and elimination of virus);
Endogenous infection (the viral genome is included in the host
genome; perpetuated according to Mendelian laws and transmitted to
offspring without direct clinical expression).
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Bird population (gallins reared on farms) → MIXTURE OF SUBJECTS

WITH DIFFERENT IMMUNOLOGICAL STATUS
EPIDEMIOLOGYCAL SUSPICION 1.- chickens without virus, without antibodies-Ab, use↔ genetically
resistant lines;
Sources of infection: 2.- chickens without virus, with Ab, infected horizontally, exogenous;
Primary: they can host the virus in the liver, spleen, temporarily eliminating it in the
-the birds that host the infection carry the virus through the circulation; blood and do not develop tumors; protection provided by the Ab;
-others host it in parenchyma,
-others eliminate it by secretions, feces, egg (congenitally). 3.-10 - 40% viremic chickens without Ab → endogenous infection,
represents an element for congenital transmission;
Dissemination of the infection → succession between the 4.- chickens who have the virus and the Ab, → exogenous cycle of
horizontal transmission (close cohabitation) and congenital (via egg). infection, horizontal elimination of the virus
Infected subjects In congenital transmission (by genes) sex of bird, plays an important role,
-produce Ab-specific targeted genes - are not protective Ab, indicator of female chickens providing transmission.
infection.
-produce Ab directed to the envelope protein gene, with a neutralizing role. The interaction between the virus and the body determines the appearance
of specific and neutralizing antibodies.
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Evolution of tumoral proces:

Avian leukosis refers to several leukaemia-like proliferative diseases caused by the avian leukosis virus (ALV).
ALVs consist of 10 subgroups designated A to J. 6 of the 10 affect chickens. Subgroups A and B are the most
common, especially in egg laying hens, followed by subgroup J which occurs in broilers and egg laying hens.
Risk factors in tumor development are: Subgroup E viruses are not oncogenic, and are widely present in non-commercial domestic chickens.
The general forms of disease associated with avian leukosis virus include:
- the transmission route; Lymphoid leukosis: the most common type of cancer caused by the ALV in chickens. It occurs in chickens four
- the prevalence of the infection; months of age or older. Tumors often develop in the liver, spleen, and bursa of Fabricius. Less commonly, the
kidney, lung, gonad, heart, mesentery, and bone marrow. Tumor growth may be nodular, miliary, diffuse, or a
- sex; combination of these forms. The bursa of Fabricius is usually always involved. Microscopically diagnostic
- the genetic structure of the host - 3 alleles of different bonds compete features are the uniform large lymphocytes (lymphoblasts) that comprise the tumors, the presence of intrafollicular
tumors in the bursa, and the tendency for the tumors in other tissues, such as the liver and spleen, to grow in an
with the susceptibility of the host (sensibility is the dominant place). expansive nodular fashion.
Erythroblastosis (Erythroid leukosis): an intravascular erythroblastic leukemia which can occur in birds
Heterozygotes are susceptible to the tumor process. younger than 4 months old. It can affect chickens as young as five weeks old and in adults. Infected chickens are
often anemic, with muscle hemorrhages and occasionally abdominal hemorrhage and ruptured liver. Myeloid
leukosis:
Resistance to tumoral process is genetically determined, by the Myeloid leukosis, caused by subgroup J, occurs in two, often overlapping forms referred to as Myeloblastosis
(myeloblastic myeloid leukosis) and Myelocytomatosis (myelocytic myeloid leukosis). Myelocytomatosis causes
interaction between the groups deterministic genes in favor of sensibility multiple masses (myelocytomas) on the chicken’s shanks, head, and oral cavity, trachea, and eye. The tumors are
genes. usually nodular and multiple, with a soft, friable consistency and of creamy color. This form of cancer occurs
mainly in adult chickens, but sometimes in birds as young as five weeks of age. Both myeloblastic and myelocytic
forms are often marked by leukaemia, and bone marrow becomes replaced by neoplastic myeloid cells. Avian
Different types of tumors appear after infection with a specific strain of Osteopetrosis: Avian osteopetrosis, also referred to as MARBLE BONE DISEASE, alters the growth and
the oncogenic virus specific for an subset cells of the hematopoietic line, differentiation of osteoblasts, resulting in uniform or irregular diaphyseal or metaphyseal thickening, usually in the
long bones of the legs, and less frequently the wings. A variety of other tumors have been associated with ALV,
leading to leucosis. including hemangiomas, histiocytic sarcomas, nephroblastomas, fibrosarcomas, chondromas, osteomas,
myxomas, adenocarcinomas, mesotheliomas. These tumors can occur alone or with the leukoses.
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CLINICAL SUSPICION LESIONAL SUSPICION:

The disease has no specific expression - it is the consequence of the
development of the tumor process of the nodular type – miliary or diffuse Increased dimensions of the liver and changes of its color, correlating
infiltrative - it compress and lacerates the tissues where it has developed with the proliferated lymphocyte line. Usually, in lymphoid leucosis, the liver
→ appearance of symptoms of insufficiency of the organ. has a clayey yellow appearance, while hemorrhages seen, on its surface or
section.
GENERAL:
Weakening, pallor of the ridge, chins, mucous membranes, discoloration of the Death occurs by hepatic rupture → massive internal bleeding.
skin;
Gigantic spleen, increased 8-10 times, with milky, nodular or uniform
THE LIVER - organ constantly damaged, target of tumors → liver failure and proliferation (infiltration).
disturbances of protein synthesis, assimilation,
Weakening of the organism, cachexia, pallor of the skin and mucous Tumors in other tissues and organs: the intestinal wall, Fabricius bursa
membranes (due to hematopoietic disorders), disruption of biliary function (as (if not fully involute at the time of infection), kidneys.
result profuse diarrhea, treatment resistant, will lead to death), modification of
the biological parameters of the organism.
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Avian leucosis - tumor process - DIFFERENTIAL DIAGNOSIS:

milIiary
or infiltrating diffuse at the hepatic
level in a broiler
Marek disease
Tuberculosis,
Coligranulomatosis,
Different peculiarities of lesions; similar
epidemiological evolution and dynamics; all these
are chronic diseases have the following symptoms:
pallor, inflammation; reduced prevalence.
Salmonellosis.
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CONFIRMATION DIAGNOSTIC
CONCLUSION: LABORATORY TECHNIQUES for identification are indirect, irrelevant and difficult to
use; we can't do therapy.
The limfoid line affected in Avian leucosis: Isolation of virus from cell cultures: fibrosis of embryonic fibroblasts and
resistance of cell culture to infection with Rous sarcoma virus (rapid tumorisation).
B lymphocyte will give imunosuppression.
RIF = resistance inducing factor - relies on the resistance of embryonic fibroblasts
from chickens infected with avian leucosis virus against reinfection with Rous
Erythrocyte line will give anemia,
sarcoma virus. It signifies the induction of resistance factor by the leukemia virus by
(consequence of aplasia of the organs that produce red blood cells and cytolysis carried out by the Rous virus.
lymphocytes).
Liver Damage (change in transaminase levels), leading to hypoproteinemia COMPLEMENT FIXATION = hyper immune sera prepared on the hamster = COFAL
and ascites. (Complement Fixation for Avian Leucosis);
In latent infections: decreased egg production, delayed sexual maturity. ELISA = identification of the specific group protein, of leucosis viruses, p27 (resulting
from the synthesis of the gag gene); it does not detect endogenous infections, but
can detect the virus in egg white (not only in serum) - non-invasive investigation,
specific to leucosis virus.
2022 2022
GENERAL PROFILAXIS :
Eradication of exogenous ALV from flocks
This depends on breaking the vertical transmission cycle of virus from dam to progeny and prevention of re-infection of the
progeny and involves the identification and elimination of hens that shed ALV.
These hens belonging to the infective classes V+A-S+ and V-A+S+ are identified by testing their cloacal or vaginal swabs, or
albumen from their eggs.
The test procedures carried out at different ages and all positives are eliminated to ensure that only clean birds are allowed to
continue in the flocks. (Payne and Venugopal, 2000).
Hatched chicks are reared in isolation in small groups and tested for viremia and ALV antibodies from about 8 weeks of age to
verify freedom from infection. Exclusion of incubation of eggs from the floks with 1-2% disease cases to prevent congenital
transmission.
Hygiene
Good hygiene and biosecurity are very important adjuncts to disease control, particularly for leucosis for which there are no
vaccines and where commercial flocks free from infection are at risk from re-infection.
Good general farm management procedures include isolation of premises, all-in all-out management, cleaning and disinfecting of
premises between crops, use of new litter, safe disposal of old litter, and site security.
Hatchery hygiene is of equal importance. ALV is a fragile virus outside the bird, with a half-life of only a few hours at room
temperatures, and is susceptible to all common disinfectants.
Immunization and Vaccines
Relatively little research has been carried out on vaccine development because it is believed that they would not be effective
against a vertically transmitted virus which induces immunological tolerance. However, the propensity of subgroup J ALV especially
to induce tolerant rather than immune infections following early contact infection has stimulated interest in developing vaccines to
protect chicks against early exposure to ALV-J.
Selection for genetic resistance
Avian leukosis virus - Cell culture (electron microscopy) Two main types of genetic resistance to leucosis have been recognized: genetic resistance to ALV infection and genetic resistance
to development of leucemic tumors.
Resistance to infection depends on the lack of specific ALV receptors on the cell membrane, which interact with viral envelope
glycoprotein and allow infection to occur. The presence or absence of these receptors is under simple genetic control.
Three autosomal loci, tva, tvb and tvc, with dominant susceptibility genes encoding the presence of virus receptors and recessive
resistance genes encoding their absence, control susceptibility to infection by ALV of subgroups A, B and D, and C, respectively.
Poultry breeders can artificially select for the presence of the resistance genes, ar and br, at the tva and tvb loci, and thus develop
strains of chickens resistant to infection by the common A and B subgroup ALV.

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GENERAL PROPHYLAXIS:
-Sanitary - prevention of horizontal transmission of infection by

cohabitation;
-Hygienic -use of single-use instruments and routine disinsection to

reduce or cancel transcutaneous transmission;
-Exclusion of incubation of eggs from the floks with 1-2% disease cases
to prevent congenital transmission.
• https://youtu.be/sJuaBL2lf6U
2022 2022
BIBLIOGRAPHY BIBLIOGRAPHY
• http://www.simv.org/sites/default/files/manuel.frv2__0.pdf • Fadly AM; Witter RL, 1998. Oncornaviruses: leukosis/sarcoma and reticuloendotheliosis. In: Glisson JR, Jackwood DJ,
Pearson JE, Reed WM, Swayne DE, eds. A Laboratory Manual for the Isolation and Identification of Avian Pathogens.
• https://www.oie.int/doc/ged/D9316.PDF Kennett Square, PA, USA: American Association of Avian Pathologists, 185-196.
• OIE Handistatus, 2002. World Animal Health Publication and Handistatus II (dataset for 2001). Paris, France: Office
• https://www.woah.org/fileadmin/Home/eng/Health_standards/tahm/3.03.13_MAREK_ International des Epizooties.
DIS.pdf • OIE Handistatus, 2003. World Animal Health Publication and Handistatus II (dataset for 2002). Paris, France: Office
International des Epizooties.
• https://www.msdvetmanual.com/poultry/neoplasms/marek-s-disease-in-poultry# • OIE Handistatus, 2004. World Animal Health Publication and Handistatus II (data set for 2003). Paris, France: Office
• https://www.cabi.org/isc/datasheet/76376 International des Epizooties.
• OIE Handistatus, 2005. World Animal Health Publication and Handistatus II (data set for 2004). Paris, France: Office
• https://youtu.be/b8n86DSJdPQ International des Epizooties.
• https://youtu.be/fH1zS7hlW54 • Payne LN, 1985. Genetics of cell receptors for avian retroviruses. Poultry genetics and breeding. Proceedings of the 18th
Poultry Science Symposium, in association with the 25th British Poultry Breeders Round Table, 1983., 1-16; [Poultry
• https://youtu.be/Q3DsZQor-TM Science Symposium 18]; 103 ref.
• Payne LN, 1992. Biology of avian retroviruses. In: Levy JA, ed. The Retroviridae, Vol. 1. New York, USA: Plenum Press,
• https://youtu.be/ablLNoE2HHU 299-404.
• https://youtu.be/nTt2k298EiY • Payne LN, 1998. HPRS-103: a retrovirus strikes back. The emergence of subgroup J avian leukosis virus. Avian Pathology,
27(Supp 1):S36-S45; 35 ref.
• https://youtu.be/vKyWO3Hrx4w • Payne LN; Fadly AM, 1997. Leukosis/sarcoma group. In: Calnek BW, ed. Diseases of Poultry. Ames, USA: Iowa State
• https://youtu.be/cxAqnsyE3ng University Press, 416-466.
• Payne LN; Venugopal K, 2000. Neoplastic diseases: Marek's disease, avian leukosis and reticuloendotheliosis. In: Diseases
• https://youtu.be/PHgr6s6-iEY of Poultry: World Trade and Public Health Implications. Office International Des Epizooties (OIE) Scientific and Technical
Review, 19(2):544-564.
• https://youtu.be/jzbgwl8oM0A • Venugopal K, 1999. Avian leukosis virus subgroup J: a rapidly evolving group of oncogenic retroviruses. Research in
• https://youtu.be/sJuaBL2lf6U Veterinary Science, 67(2):113-119; 32 ref.
• Weissmahr RN; Schupbach J; Boni J, 1997. Reverse transcriptase activity in chicken embryo fibroblast culture supernatants
• https://youtu.be/PWgxjlA16Vs is associated with particles containing endogenous avian retrovirus EAV-0 RNA. Journal of Virology, 71:3005-3012.
DVM PhD DRAGOS COBZARIU 57 DVM PhD DRAGOS COBZARIU 58
Dr. DVM PhD Dragos Cobzariu 59

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RABIES
2021
• https://youtu.be/MZR0CGKZ6Uc
10/17/2022 DVM PhD Dragos Cobzariu 2
Rabies viruses belong to the genus Lyssavirus (from the Greek lussa,
madness). Determining the sequence of the viral genome encoding the N
protein makes it possible to define 7 genotypes:
Distribution
Génotype Virus espèces concernées Efficacité du vaccin
géographique
Homme, carnivores sauvages

Rage
1 mondiale et domestiques, chauves- oui
classique
souris.
chauves-souris frugivores,
2 Lagos bat Afrique non
chats, chiens.
Homme, musaraignes, chats,

3 Mokola Afrique non
chiens, rongeurs.
Homme, chauves-souris
4 Duvenhage Afrique du Sud non
insectivores.
5 EBL-1 Europe partielle
insectivores.
6 EBL-2 Europe oui
insectivores.
Viruses of the Rhabdoviridae family (from the Greek rhabdos, rod, after the "rectangular"
shape of the virion) are part of the order Mononegavirales Homme, chauves-souris
7 ABL Australie oui
their genome is an unsegmented RNA (Mono) of negative polarity (nega) frugivores et insectivores.
they are enveloped viruses (and therefore fragile viruses)
10/17/2022 DVM PhD Dragos Cobzariu 3 10/17/2022 DVM PhD Dragos Cobzariu 4
Rabies is an infectious disease of viral etiology, affecting all species of

HOMEOTHERMIC ANIMALS, also transmissible to humans. It is
characterized by an acute course of nervous manifestations, expressed
by hyperexcitability and aggressiveness, followed by paralysis and death.
Etiology
Family Rhabdoviridae-genus Lyssavirus-rabies virus
Rhabdoviruses are in the form of a rod (Æ = 80 nm, variable length from 120 to
180 nm) with one flat end and the other rounded, giving them a very characteristic Rhabdovirus multiplication cycle.
"revolver bullet" appearance. There are filamentous forms up to 300 nm. The entire cycle is neuronal-intracytoplasmic.
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Nucleocapsids and virions collect in a fibrous matrix (M) and form

characteristic intracytoplasmic inclusions, observable under an optical
microscope: the BODIES OF NEGRI (named after the Italian physician,
Adelchi Negri, who described this lesion in 1903).
Rabid cattle neuron. Observation of a Negri body under optical

microscopy (a) and electron microscopy (b). • https://youtu.be/Lbex10NVgq4
Rabies-a zoonosis, that is to say an animal disease-to be transmitted

to humans: all WARM-BLOODED animals are susceptible to rabies.
Epidemiology
Receptivity
Domestic and wild mammals, regardless of age, race, sex, state of maintenance,
season and climate.
In particular, birds are affected.
Sources of infection
Disease of natural focus: natural reservoirs of infection.
-domestic or urban reservoirs-domestic animals (dogs, other Canidae)
silvatic or wild reservoir-represented by several wild animals (dominant: the fox
and the wolf).
From humans to animals, the required conditions,
a Virus reservoirs,
A zoonosis can only exist thanks to the permanence of an animal reservoir: in the
case of rabies, the reservoirs of viruses are wild mammals, which harbor the virus for
a very long time.
b Distributors of viruses (vectors),

PRIMARY DISTRIBUTORS, are the reservoirs of the virus: infected wild animals
become excretors of the virus in saliva, and transmit it by bite, either to their
congeners-which maintains the reservoir-or to other animals, which 'they meet.
SECONDARY DISTRIBUTORS, are domestic animals bitten by wild animals,
excretors: dogs, cats, cattle, horses, which have not been vaccinated ...
c Men
In general, the rabies virus will be transmitted to humans accidentally, by
inoculation of virulent saliva, from a rabid, wild or domestic animal: by bite and, Pathophysiology of rabies showing the more or less long CENTRIPETAL
more rarely, by scratching, or by licking a 'a wound, or a mucous membrane. PATHWAY OF INFECTION TO THE BRAIN, then the CENTRIFUGAL
Once declared, rabies is encephalitis, which is always fatal. PATHWAY TO THE SALIVARY GLANDS leading to the secretion of
the virus in saliva.
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Ecology of animal rabies.

1-wild rabies.
Rabies viruses are perpetuated in two major natural cycles:
a. Rage of wild predators. The rabies of predators is present on all
continents. Only a few countries are preserved by their insularity and
draconian sanitary measures at their borders: Great Britain, Japan,
Australia, Pacific Islands. The vectors of the virus vary from country to
country: the red fox in Europe, the raccoon and skunk in North America,
the jackal in Africa, the wolf in the Middle East, the mongoose in South
Africa.
Rabies reservoir-
distributors around
the world
Wild rabies in USA

b. Rabies in Bats: Bats constitute a quarter of the planet's mammals with nearly
In bats-flying mammals-rabies most often presents as a chronic infection: they 1,000 species, including around thirty in Europe. The Pipistrelle
can excrete the virus in saliva and urine for long periods, which makes them represents nearly 2/3 of the bats, living in France: its population is
primary distributors potentially formidable. estimated at several tens of millions of individuals. Small in size-3 to 5
Depending on the species, bats can be infected with different viruses of the cm-it stays in the cracks in the walls.
genus Lyssavirus and the situation varies depending on the continent. Viruses of
genotype 1 (classical rabies virus from which the vaccine is prepared) are only
found in bats of the American continent. Viruses of genotypes 2 and 4 are found in
African bats and genotype 7 in Australia. In Europe, genotypes 5 and 6 are the
only ones found in native bats, which are all insectivores. The species which
seems to be the most affected among the 33 species is the common Serotin. In
France, all species are protected.
Bat rabies is present across much of the globe, including countries free from
terrestrial carnivore rabies such as Great Britain and Australia.
Blood-sucking bats (vampires) in Latin America are responsible for paralytic rabies
in cattle, killing several hundred thousand animals each year.
Bat rabies is uncommon in France: 20 cases (affecting serotin commune) have been
recorded since 1989 (these statistics reflect the degree of vigilance rather than the
real situation). On the left, rabid blood-sucking bats (vampires).
The main populations at risk of exposure are bats (300 to 400 people), especially Right, a vampire bite on a cow.
those who handle animals (about half) and wildlife care workers.
Human contamination
2 ° Urban Rage or "street rage" 1-Cutaneous route-the most frequent (99%).
Healthy skin is an impassable barrier for the rabies virus.
Dogs are the main reservoir and vector of the virus around the world.
Often from a bite from a rabid virus-shedding animal, and more
The WHO estimates that street rabies is responsible for over 99% of human Rarely :
rabies cases and at least 50,000 deaths each year. Stray dogs are the -licking on a fresh wound, excoriated skin,
intermediaries between wild rabies and urban rabies: they transmit rabies -from a scratch (cat) by claws soiled with drool,
to other wild animals, herbivores and unvaccinated domestic carnivores (dogs, -handling of a rabid animal (dead or alive).
cats). Canine rabies is rife in the form of enzootics in economically disadvantaged -licking or the projection of droplets of virulent saliva on the conjunctival, olfactory
areas of Africa, Asia and South America. On the other hand, except accidental or or labial mucous membranes by the excoriated skin.
illegal introduction, street rabies has disappeared from North America, Western 2-Airway-exceptional:
Europe and Japan, because these countries eliminate stray dogs and vaccinate -inhalation of an aerosol of viral particles
domestic animals. -visit a cave and manipulate bats
3-Care for a rabid man-theoretically possible
4-Corneal transplants from a donor incubating rabies
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Rabies endemic areas in the world

Routes of transmission-transmission to animals
Transmission transcutanée-innocultion of virulent saliva by bite or scratch
risk factors-deep bite, close to the central nervous system or strongly inervated.
The saliva of infected dogs includes the virus 10-14 days before the onset of
clinical signs of the disease (in foxes, even 29 days before the onset of clinical
signs of the disease).
The virus is present in the saliva throughout the illness.
Blood, meat, milk, urine, feces-limited virulence, transient, variable.
Infection by contact of the virus with the mucous membranes (conjunctival,
pituitary, respiratory) or the skin with recent lesions.
Transmission through the digestive tract is possible (the fox).
The transplacental route-cited by the literature as a possibility for dogs, cattle and
sconcs
Epidemiological dynamics
Usual-Sporadic
Sometimes-Endemic (areas with high potential for rabies transmission). Enzootic: contagious disease affecting animals in a
region. enzootic is the animal equivalent of endemic
Rabies endemic areas in Europe.
RABIES PHYSIOPATHOLOGY 3-Centrifugal diffusion from the brain

1-Penetration of the virus The virus then spreads to all tissues by a centrifugal route, infecting the salivary
The rabies virus is most often inoculated to its host when bitten by an infected glands but also the eye, hair follicles, pancreas and kidneys
animal:
it multiplies first in muscle cells.
it enters the nervous system by endocytosis at the level of free nerve endings and
neuromuscular junctions.
2-Centripetal invasion of the nervous system
Virions are transported in the axon (by

dynein) to the cell body where the virus
multiplies. Virions that bud from the
infected neuron are released into the
intersynaptic space and infect the next
postsynaptic neuron.
The virus reaches the brain where it
continues to replicate. Maturation of new
virions can take place on the cell surface
and inside the cytoplasm.
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B. State period-the dog becomes aggressive against other animals, people and
Clinical objects.
We describe 2 clinical forms: furious and paralytic. His gaze is fixed, prowling and ferocious.
Amid his attacks of aggression, his gaze is wandering, his face expressionless.
AT THE DOG Dromomania hoarse voice; the bark ends with a long howl
Average incubation: 20-60 days (5 days-6 years-11 years). In some subjects: convergent or divergent strabismus, myosis or mydriasis, unequal
pupils, vomiting or diarrhea with blood
1.The Furious form
Duration of this phase 2-6 days.
A. Early Perioad-often unnoticed:
-changes in behavior: alternating states of depression and nervousness C. Paralytic period-dog cannot swallow, loses voice; his lower jaw is inert; his
hyperesthesia at any excitement (noise, light, touch etc.), dripping saliva runs out; his eyes become still, her 3rd eyelid-prominent.
auditory and visual halucinations The paralysis sets in gradually; paraplegia appears; after a few hours of agony, the
-changes in appetite: present at the beginning, gradually diminished, dog died of asphyxiation.
deterioration of taste sometimes itching instead of the bite Duration of this phase: 1-3 days, rarely 5 days.
thirsty, present or even exaggerated; absence of hydrophobia.
-exaggeration of the reproductive instinct
voice louder, hoarse; short, jerky bark.
Duration of this phase: on average, 1-3 days.
2 THE PARALYTIC FORM

Onset with depression, followed by paralysis of the faringian and masseter At the Fox:
muscles. Radical change in behavior
Swallowing becoming impossible; the mouth remains slightly open; a thin saliva It enters the localities, in the shelters of domestic animals in broad daylight.
flows from her. He becomes soft, lets himself be stroked, grabbed and bound.
Bark hoarse, then aphonia. Without becoming aggressive, it bites animals and people, with a discretion that
Sad, suffering faces, absent gaze. makes superficial wounds unnoticed.
Death occurs after the generalyzed paralysis, 3-4 days later. The animals have no reactions anymore, are staring, are aggressive, have
merry-go-round movements and opisthotonus; they are rolling; then, paralyzes
AT THE CAT set in; the march becomes uncertain. The fox can no longer stand and dies 3-4
Average incubation: 10-15 days (10-260 days) days later.
Usually evolution in furious form
Aggressive cat since the prodromal phase; he meows incessantly, with a shrill
voice, then hoarse; it attacks animals and people, bites and scratches.
Heavily dilated eyelids, sometimes uneven
In the excitement phase convulsions are frequent; cats roll over and may die
suddenly.
In the paralytic phase paralysis of different muscle groups appears, starting with
those of the larynx and masseters.
In this form, the duration of the disease is 3-6 days.
To the humans
To the Bats The completely silent incubation lasts an average of six weeks, used for serovaccination (it
The bat also changes its behavior (difficulty in flying, prostration). Bites may go is important to go faster than the virus).
unnoticed because they are small, painless, and located in areas like the scalp. The duration of incubation is shortened in case of deep or multiple bites of the face and
hands-areas rich in nerve endings-
In children, the incubation period is significantly shorter than in adults.
Because of their size, children are more often bitten on the face.
Conversely, the incubation can be exceptionally long.
statut
Phase symptômes durée statut viral immunol
ogique
multiplicatio
n du virus
60 -
dans le tissu
Incubation asymptomatique 365 0 (1)
musculairep
jours
eu de
virions
peu de
fièvre, nausées, virions dans
2 - 10
Prodromes anorexie, douleur au le SNC et 0 (1)
jours
niveau de la morsure dans le
cerveau
Anticorps
spasmes
dans le
pharyngés, hydroph 2 - 7 titre en
Neurologique 1 sérum et
obie, hyperactivité, jours virions élevé
le SNC
anxiété, dépression
(LCR)
Neurologique 2 Paralysie
coma, arrêtcardiaque,
0 - 14 titre en
Coma hypotension,
jours virions élevé
hypoventilation
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LESIONS
The strategy implemented by the rabies virus is therefore
Macroscopic changes are non-characteristic:
dehydration, weight loss, dirty coat, recent skin wounds, broken teeth, foreign devilish:
bodies in the stomach, venous stasis, weakly coagulated blood, brown.
The cerebrospinal axis is congested, with edema and petechiae. At the bite level, viral multiplication does not produce a cytopathogenic effect
capable of presenting viral antigens to the immune system.
Histopathological:
Characteristic lesion = oxyphilic inclusions (Negri corpuscles) in the citoplasm After entering the nervous system, it escapes, almost completely, the immune
of neurons and their extensions, especially neurons of the horn of Ammon, surveillance of the host. Antibodies do not appear until the terminal stage of rabies.
pyramidal cells of the cerebral cortex and Purkinje cells of the cerebellum.
The presence of inclusions is different in terms of frequency and location, with The multiplication of the virus in the brain, in particular in the limbic system (which
respect to the species and the development of the pathogenetic process. controls emotions and behavior), makes the host aggressive: an essential condition
Non-specific lesions: for its transmission to a new host.
Perivascular and diffuse lymphocytic histiocytic infiltrations
neuronal alterations (hydropic degeneration, chromatolysis, pycnosis, In the nervous system, virions produced by an infected neuron immediately fuse
karyolysis). with neighboring neurons without causing cell destruction.
Nodular infiltrations: accumulations of glial cells with or without neurophagy; the In the salivary glands, virions formed by cells are secreted into saliva along with
same lesions, accompanied by neurophagy, which are found in the mucus, even in the preclinical phase, hence the importance of veterinary forensic
cerebrospinal ganglia are referred to as the Van Gehucten and Nellis nodules. surveillance.
The virus can thus be transmitted before its host dies
• https://youtu.be/2JovOAhpETA • https://youtu.be/KthucjWBCJA
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Specimens
DIAGNOSTIC
After telephone agreement from the recipient, the samples are sent under
refrigeration (4 ° C) in a triple sealed packaging.
Depending on the animal species involved, the following are sent to the laboratory:
The clinical diagnosis of rabies is never a definite diagnosis. -whole animal, if it is a small mammal (marten, ferret, squirrel, etc.),
The only indisputable diagnosis is the biological diagnosis carried out in the laboratory. -entire head for larger animals (dog, cat, fox), detached at the level of the cervical
This diagnosis is the exclusive role of the CNR, which should always be contacted in vertebrae,
the event of suspicion in order to carry out the adequate samples and send them in -only the brain if it is a large herbivore.
optimal conditions, in compliance with the safety rules imposed by the legislation in
force. In general, research focuses on areas particularly
In France, the two official structures authorized by the Ministries of Agriculture and rich in rabies virus: the horn of Ammon located in
Health to carry out the biological diagnosis of rabies are the Institut Pasteur in Paris, the convolution of the hippocampus, the medulla
for any suspected human case and for any animal likely to have transmitted the oblongata, the cerebellum, the cortex and the
disease. rabies in a man (see contact details at the end of the text) and the National salivary glands.
Center for Rabies Studies in Nancy (reference laboratory for animal rabies, out of
contact with humans). For updated techniques see the OIE Manual
All handling of suspect specimens should be performed under P3 type
laboratory containment conditions.
Identification de l'antigène
Identification des anticorps
CONFIRMATION DIAGNOSIS CONFIRMATION

1. Identify antigens, rabies in cells,
Direct immunofluorescence 2.ELISA method (sandwich technique)
On impressions of Ammon's horn, bulb and cortex.
As a reference technique, it can lead to a definite diagnosis Rabies antigens are immunocaptured.
in a few hours, on fresh or well-preserved material. The wells of a microtiter plate are coated
The test is based on the recognition of the rabies antigen by specific antibodies with the antibodies of a rabbit serum
coupled to fluorescein: the fluorescent antibodies will only bind to the antigen, immunized against the antigens of the
revealing the latter when reading the slide with the aid of a fluorescent microscope. core.
Immunofluorescence-specific and rapid test; diagnosis of certainty: The supernatant of the ground tissue is
between 98-100% of cases, in 24 hours. deposited in the wells. Antigen, if present,
is captured by the antibody.
After washing, the anti-rabies antibodies
coupled to the peroxidase are added.
The added substrate undergoes the action
of the enzyme: the appearance of a yellow
color, a sign of the presence of the rabies
antigen in the tissue tested.
The test is simple, rapid (a few hours),
sensitive and specific.
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CONFIRMATION
CONFIRMATION
3. ISOLATION OF THE VIRUS ON CELL CULTURE (cells in continuous lines of
mouse neuroblastoma) is a very sensitive test and allows a rapid diagnosis (less
4.Biological test-inoculation of brain, cerebellum and bulb triturations in
than 24 hours) if the sample has been correctly taken.
suspension of physiological saline solution in 10 mice, nv.ne, 3-4 weeks old,
Isolation confirms the detection of viral antigens and allows the serotype of the
inoculated intracerebrally with 0.015 ml and 0.03 ml, respectively. These mice are
isolated strain to be determined using monoclonal antibodies.
kept under observation for 28 days. Their death within this interval involves the
Viral antigens are searched for in the inoculated cells by the direct
application of confirmatory methods for the identification of the antigen.
immunofluorescence or immunoenzymology tests described.
The technique is faster, more reliable and less dangerous than mouse inoculation
5.Histopathological-done on sections of Ammon's horn, hippocampus,
cerebellum; they are stained by the Mann, Lenz, Giemsa methods, to highlight the
Babeş-Negri corpuscles
The presence of the corpuscles = confirms the rabies
The absence of corpuscles does not exclude rabies.
CONFIRMATION
6. Detection of viral RNA
From saliva or CSF, the viral genome can be detected by gene amplification: RT-
PCR (reverse transcription of viral RNA into DNA amplified by polymerase chain
reaction (Polymerase Chain Reaction).
7. Virus genotyping
After the amplification by RT-PCR, the sequencing of the gene for nucleoprotein
N and glycoprotein G.
8. Antibody assay
Antibody titration makes it possible to verify and assess the degree of immunity
of subjects against rabies, before or after exposure to the risk of contamination
(immunizing threshold:> 0.5 IU per ml).
An immunoenzymatic technique of the ELISA type is used.
The detection of anti-rabies antibodies in the blood is of very limited
interest in the diagnosis of rabies because they appear only late, or to
confirm post-vaccination immunization.
RABIES PROPHYLAXIS AND CONTROL

Health prophylaxis
Objective-liquidation of the rabies virus reservoir.
FIGHT AGAINST RABIES
Strict control of the movement of dogs (leash + muzzle) and cats
Reduction in dog population
Official declaration and 3rd degree quarantine.
Capture and destruction of animals without an owner (dogs and cats)
Rabid animals / suspected rabies / carnivores suspected of contamination will be killed
Systematic preventive vaccination of all dogs.
and destroyed after taking samples for laboratory diagnosis.
In wild animals:
Exception:
Control of the fox and wolf population
1) solipeds, ruminants and pigs suspected of rabies can be placed under sanitary and veterinary
Vaccination campaigns with baits
observation, in conditions of complete isolation and safety; they will be kept alive to deny or
Medical prophylaxis / Immunoprophylaxis
confirm rabies;
Vaccines
2) dogs suspected of contamination, special use or high value dogs
-Vaccines with modified live virus / inactivated vaccines.
-Dosage-strictly according to the manufacturer's instructions
Must be placed under veterinary observation for 10 days carnivores suspected of rabies
Vaccines with modified virus
and contamination, having bitten or scratched or having otherwise come into contact with other
Adapted on a poultry embrion; tulpines Flury LEP
animals or people, as well as solipeds, ruminants and pigs suspected of having entered in
Suitable for cell cultures: canine cell line, pig cells, bovine kidneys, hamster cell lines.
contact with persons in conditions which could have resulted in the contamination of the latter.
Inactivated vaccines
If rabies is not declared during this time, animals suspected of rabies exit into the observation.
Vaccine virus replicated on the brain of adult animals, the brain of newborn animals, cell
As for the carnivores suspected of contamination, they will be killed, because they can
cultures or embryonated eggs.
fall diseases later.
Vaccine virus inactivates by physical (temperature, ultraviolet) or chemical (phenol, beta-
The 10-day veterinary health observation must also apply to animals without signs of
propriolactone) methods.
rabies or with a history (considered healthy) having bitten or scratched people or
Post-vaccination accidents-the case of brain vaccines
animals.
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When cases of rabies appear in domestic production animals, those with clinical signs of
RABIES VETERINARY HEALTH SURVEILLANCE ACTIONS (B-058)
disease are first killed; then the other animals will be vaccinated against rabies if they
have not been for the last 6 months and will be kept under veterinary health surveillance
1. Surveillance by direct immunofluorescence and, if so, by histological, virusological
for 3 months.
examinations and identification of the viral genome in domestic and wild carnivores, as well
as in other domestic mammals which have died or had to be sacrificed and have shown
When domestic or wild carnivores, rabies or suspected of rabies, animals with signs of
symptoms. nervous.
bites are (even if vaccinated against rabies previously) have entered a herd of solipeds,
ruminants or pigs, they will be sacrificed, under health surveillance. veterinarian, in
2. Mandatory clinical surveillance of carnivores having bitten or scratched people and
maximum 6 days after the date of the bite.
animals, within 14 days after the date of the bite or scratch.
Their heads, spine and the bite area are confiscated.
The rest of the animals in which no bite marks are seen will be vaccinated against rabies,
3. Remove the heads of all suspect animals (having bitten, scratched, or having been bitten or
if they have not been vaccinated in the past 6 months; small ruminants and pigs will be
scratched).
quarantined and under surveillance for 3 months; large ruminants and solipeds will be
quarantined and under surveillance for 6 months.
4. Killing of wild animals with changed behavior (rabies suspects) with respect for legal
Meat, milk and any other product of rabid animals or suspected of rabies will be
provisions in terms of protection and welfare of animals and taking tests to make a diagnosis.
destroyed.
The places where sick or suspected rabid animals have remained or have been killed, as well
5. Active surveillance of rabies in foxes, to determine the efficacy of the vaccine by
as objects in contact with them, will be disinfected.
determining the vaccine marker and laboratory examination of 8 foxes hunted / km2.
The extinction of the disease and the lifting of the quarantine measures take place 3
months after the last case of death or killing due to rabies and if in the respective locality
6. Screening and identification of STRAINS of rabies virus, by molecular biology
there are no more animals in quarantine or under veterinary health observation for this
investigations.
disease.
1. The notification of the disease is made according to the forecasts of the MAA VETERINARY HEALTH SURVEILLANCE ACTIONS FOR ZOONOSES (DISEASES
Order, for the approval of the veterinary sanitary standard on the announcement, TRANSMISSIBLE FROM LIVING ANIMALS TO HUMANS)
declaration and notification of certain communicable diseases of animals. In the case of rabies: anatomical, pathological and laboratory monitoring in receptive species.
2. The tests (the heads of the dead or sacrificed animals) taken for the Specific sanitary and veterinary prophylaxis for rabies:
virusological examination (including the biological test) and histological are sent to 1. Oral vaccination of foxes, according to the specific prophylaxis program approved by
LSVSAJ and / or, as the case may be, to IDSA, with the respect of sanitary ANSVSA
conditions. mandatory veterinarians on their removal, packaging, identification and 2. Vaccination of dogs and cats over 3 months old once a year, between October and
shipping. December, with additional vaccinations.
3. Vaccination, if applicable, of animals that have come into contact with animals suspected of
3. Brain tests from domestic and wild ruminants with negative diagnosis of
infection.
rabies will be histologically examined for EST, Aujeszky, listeriosis-diag. Diff. 4. Oral vaccination of foxes, twice a year, in April-May and October-November.
4. The collection of wild animals suspected of disease is done whenever 5. Expenses on the purchase of vaccine baits are incurred by ANSVSA.
necessary, according to the provisions of the common protocol between ANSVSA, 6. Expenses on the distribution of vaccine bait are incurred by the owners of the hunting lots.
AGVPS, RNP-ROMSILVA and approved as an emergency by MAPDR. 7. The effectiveness of oral vaccination of foxes is verified by tests taken from vaccinated
5. Investigations by authorized and accredited PCR-reference laboratory animals (hunted foxes).
6. In farms with diagnoses of rabies, restrictions on the movement of animals 8. The value of the vaccine and the labor to vaccinate city dogs and cats is incurred by their
are imposed, in accordance with the provisions of the legislation in force. owner.
9. The value of the vaccine and the labor to vaccinate village dogs and cats is committed by
ANSVSA.
10. The countervalue of the vaccine and labor for rabies vaccines when needed is incurred by
ANSVSA.
Links
https://www.oie.int/doc/ged/D11042.PDF
Rhabdoviridae.pdf (gla.ac.uk)
Thank You! Questions!??
https://www.oie.int/fileadmin/Home/fr/Health_standards/tahm/3.01.17_RABIES.pdf
Rhabdoviridae - Rhabdoviridae - Negative-sense RNA Viruses - ICTV (ictvonline.org)
ICTV (ictvonline.org)
https://www.pasteur.fr/fr/sante-publique/cnr/les-cnr/rage
http://www.microbes-edu.org/etudiant/rhabdoviridae.html
BEH 2005 n°24-25 « Santé des voyageurs et recommandations sanitaires » et n° 29-30 « Calendrier vaccinal 2005 »
« La rage : état des lieux en 2004 et persistance des risques en France » SPECTRA BIOLOGIE, par Hélène PEIGUE-LAFEUILLE et Hervé BOURHY, n°143 – Janvier Février 2005
« La rage humaine en France en 2004 : état des lieux et prise en charge » par H. PEIGUE-LAFEUILLE et al., Médecine et maladies Infectieuses 34 (2004) 551-560 accessible
sur www.sciencedirect.com
Traité de Virologie Médicale, par JM HURAUX, H PEIGUE-LAFEUILLE, JC NICOLAS, H AGUT, 2003, éd. ESTEM-AUF
Site de l’OMS www.who.int/en/
Site de l’Organisation Mondiale de la Santé Animale www.oie.int/
Site de l’Institut de Veille Sanitaire www.invs.sante.fr
Site de l’Agence Française de Sécurité sanitaire des Aliments www.afssa.fr (publications puis éditions)
Site du ministère de l’agriculture www.agriculture.gouv.fr
Site de l’Union nationale des centres de soins pour la faune sauvage www.chez.com/uncs
Site du ministère de la santé www.sante.gouv
Site de la société Française pour l’étude et la protection des mammifères www.sfepm.org
https://youtu.be/MZR0CGKZ6Uc
https://youtu.be/2JovOAhpETA
https://youtu.be/KthucjWBCJA
https://youtu.be/Lbex10NVgq4
10/17/2022 DVM PhD Dragos Cobzariu 53
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99
Aujeszky
Disease 2022
Herpesvirosis common to several species of

animals, sporadic-enzootic, with an acute
evolution
LETHAL-as encephalomyelitis
BENIGN-as part of the porcine respiratory complex
ETIOLOGY
herpesvirus 1 Herpesviridae,
Alphaherpesvirinae, Varicellovirus.
icosahedral symmetry
nucleocapsid,
double-stranded DNA
8 proteins.
structural proteins, (gE, gB, gC,
gD, gI, gG).
non-structural proteins thymidinekinase (TK).
gB, gC and gD appear to play a major role in the induction of immunity
TK, in pathogenicity.
grown on the chorioallantoic membrane of the chick embryo primary
cell cultures or a wide range of cell lines PK15, IBR-S2 and BHK 21
cytopathic-ECPP effect,
1
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100
eosinophilic plaques, and intranuclear inclusions

antigenic unique strains
differences in pathogenicity tropism, neurotropes
polyorganotropes
VARIABILITY
variants or mutants passages on
laboratory animals chicken embryos certain cell
cultures by deletion
vaccine strains.
variably successful
Rabbit, is most susceptible 72 hours, with scratch lesions
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101
Virus resistance quite high. EPIDEMIOLOGY

retains its infectivity for 2 to 3 years
low temperatures dry fodder-up
Receptivity:
pork;
dog, cat, cow, sheep, goat, horse;
rat, boar, ferret, fox.
100°C-immediately Favoring elements:

AGE: more severe are clinical forms in
young animals
Sources of infection: WAYS OF TRANSMISSION:

Primary:
Sick, convalescent and cured animals. PIG: horizontally and vertically
Pigs eliminate the virus through nasal, ocular, bronchial, genital Contamination is frequent → by the respiratory route, by inhaling contaminated
secretions, milk and urine. particles (dust, aerosols) and by the digestive route, by ingesting infected breast
Saliva becomes virulent only if mixed with BRONCHIAL SECRETIONS. milk or transplacentally.
Seminal material becomes virulent if contaminated by foreskin lesions
THE FAECES ARE NOT VIRULENT. OTHER SPECIES:
-carnivores and rodents → contamination through the digestive tract-
Because of cadavers, products from sick, convalescent or cured pigs, the
consumption unheated feed from infected pigs.
pig becomes the main reservoir for maintaining the virus in an areal -cattle, horses and rodents → cutaneous route also mentioned.
(because the virus persists for a long period in the organisms after -transmission by insects → not proven.
recovery).
Vaccinated pigs that have come into contact with the virus become DYNAMICS:
potential virus shedders, without showing clinical signs of disease.
Secondary: fodder, water, shelter, objects, contaminated vehicles. -In pigs, the disease evolves as an ENDEMIC focus, without the tendency of its
The virus can be spread, also by air masses, but only over a distance of a spread outside the focus, having a stationary character.
few kilometres. -Aujeszky's disease is SPORADIC in other species of susceptible animals.
CLINICAL DIAGNOSIS
Incubation: 3-6 days.

PORK
-Piglets born infected: lack of appetite, depression, recumbency, chills, muscle convulsions,
aphonia, death;
-Piglets born infected after birth: hyperthermia, depression, lack of appetite, foaming at the
mouth, convulsions, aphonia, rapid breathing, vomiting, diarrhea, death within a few hours;
-Piglets from 2-12 weeks: after a period with only febrile syndrome → manifestations of
encephalomyelitis: agitation of the skin hyperesthesia, ataxic walking, dromomania, walking
in carousels, abnormal head positions, sideways falls with pedaling and teeth grinding, knocks
surrounding obstacles (decreased visual acuity), epileptiform seizures lasting 5-10 minutes,
which are repeated more and more often as the disease progresses; the disease lasts 1-3
days → death or torticollis if the piglet survives, slowing of growth.
-Piglets older than 3 months and adults: BENIGN DEVELOPMENT, hyperthermia (41°C),
depression, inappetence, throwing up of mucous serum, then mucous and purulent, sneezing,
coughing, dyspnea, “sitting dog” position, weakness; signs remit in 6-10 days;
mortality rate: 3-5%.
-Pregnant sows: abortion, premature births, vomiting diarrhea; infection in the first month of
gestation: embryonic resorption, with estrus;
-Incidence rate-reproductive disorders in pregnant sows: 20%.
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102
Aujeszky's disease-skin lesions in the area

of the snout Aujeszky's disease– mumified Aujeszky's disease-nervous symptoms
fœtus
Aujeszky's disease-aborted fetuses Aujeszky's disease-mortality in piglets and a cat
CATTLE, SHEEP, GOATS
FEBRILE SYNDROM
NERVOUS SYNDROM: muscle tremors, foaming salivation, aphonia,
pruritus-gradually accentuated → scratching, even self-mutilation.
Frequent pruritus: on the head, limbs, mammary gland, genitals and at
the base of the tail.
Paralysis and paralysis,
Phenomena of hyperexcitability and abortions.
Death from exhaustion 1-3 days later
Aujeszky's disease-self-
harm (scratching) in the
head area in goats
Rabbit is most susceptible-dies in 72 hours

with scratch lesions at site of inoculation-
bioprobe diagnostic method
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HORSE
FEBRILE SYNDROME followed by: NERVOUS SYNDROM
restlessness, dromomania, unsteady gait, abnormal attitudes,
paralysis and death 1 to 4 days later.
pruritus-very rarely.
DOG
ANATOMO-PATHOLOGICAL DIAGNOSIS
NERVOUS SYNDROM
onset with depression and inappetence, then worry, anxiety, barking without reason,
cutaneous hyperesthesia-HC PORK
after a maximum of 12 hours → pruritus, which can lead to self-harm; sometimes, we Lesions according the clinical form.
observe aggression-against other animals, against humans -Infant piglets: congestion, meningitis, pinpoint haemorrhages in the nervous
Sometimes paralysis, coma and death appear after 18 to 26 hours. substance, kidneys and gastric mucosa, mile necrosis in the liver, kidneys and lungs.
2 atypical clinical forms of Aujeszky's disease in dogs: -Abortions and piglets born dead: subcutaneous edema, exudates in large cavities
Sudden Death and Gastro-Intestinal Form. and necrotic foci similar to those seen in infants
-Fat and adults: ulcerations covered by white-yellowish pseudomembranous
CAT
NERVOUS SYNDROM deposits, necrotic foci and small abscesses in the tonsillar, epiglottic and posterior
abrupt onset with depression and inappetence, weeping mewing, followed by abundant part of the palate; pulmonary edema, serohemorrhagic pneumonia, acute myocarditis,
salivation, swallowing disorders, change in the voice to aphonia, rapid pulse, abdominal pain, hemorrhagic gastroenteritis, hemorrhagic placentitis.
vomiting, skin hyperesthesia (common at the base of the tail and at the lumbar level). -Animals with pruritus: depilated skin, oedematous, with erosions and deep
30% of cases → pruritus in the head and neck. wounds.
mydriasis, anisocoria, hyperexcitability and even aggressiveness
death occurred in 12 to 36 hours, preceded, for a short period, by paresis and paralysis.
Aujeszky's disease-rhnitis
Aujeszky's disease-yellowish-
white necrotic foci in the spleen Aujeszky's disease-congestive lung damage
HISTOPATOLOGICAL DIAGNOSIS
perivascular lymphocyte sleeves,
neuronophagy,
neuron degradation,
presence of spheroidal, elongated, semilunar or other forms of acidophilic or
basophilic intranuclear inclusions in the ganglion and glial cells, as well as
around the foci of necrosis of the various organs, called Hurst inclusions.
Aujeszky's disease-milliary necrosis in the liver
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104
DIAGNOSIS CONFIRMATION of Aujeszky's disease
Isolation of virus-antigen
Pathological materials: brain, spinal cord, tonsils, lungs, runny nose secretions, runts.
Types of cell lines or primary cell cultures are used → often used: porcine kidney
cell line PK 15 (under the inverted microscope: appearance of the characteristic
cytopathic effect 24-72 hours after inoculation).
If this cytopathic effect does not appear after the first passage, a second “blind”
passage is performed; only if one does not see and cytopathic after this, one gives
the negative result (absence of the Aujeszky disease virus).
Identification, using a method, with high specificity and sensitivity:

-serum neutralization reaction, SN
-immunofluorescence, IF
Aujeszky's disease-Histopathological diagnosis-left: acidophilic inclusions in -ELISA
the nucleus of the epithelial cells of the tonsillar crypts; right: acidophilic and -PCR (RT-PCR).
basophilic inclusions in the nucleus of neurons.
2. Demonstration of Immunological Restructuring-Antibodies Ab

methods APPLIED ONLY IN PIGS (this species can survive, long enough to develop an
immune response).
Specific antibodies → appear in infected pigs, 7 days after infection.
Serological methods used:
- Latex-Agglutination → retrospective diagnosis.
- ELISA
- Virus neutralization, NV
Latex-Agglutination-simple and quick test (10 minutes)- certificate infection

6 to 7 days after infection.
ELISA-differentiation of Ab. post-infections, and Ab. post-vaccination after use of deleted

Aujeszky's disease-ME-left: Porcine herpes virus in porcine kidney cell vaccines, with the demonstration of antibodies 7 to 8 days after infection.
culture PK 15, viral capsid in nucleus (N), immature virions in citoplasm;
right: viral particles (mature virus) outside the cells
Virus neutralization-less sensitive than the first 2 tests presented, identifying antibodies 8-
10 days after infection.
PROPHYLAXIS
General measures:
PROGNOSIS Purchase of pigs only from free units, prophylactic quarantine,
Periodic examinations, periodic extermination of the rats.
ADULT PIGS-FAVORABLE Judicious sterilization of slaughterhouse and culinary waste used in animal feed,
Other categories of pigs and animal species-UNFAVORABLE Prohibition of access or remains of pig carcasses on farms.
For carnivores: the administration of meat only after heat treatment is the most
effective prophylactic measure
TREATMENT
Specific measures:
For pigs with respiratory disorders-symptomatic treatment, combined with Used only for pigs-VACCINATION
antibiotherapy to prevent the appearance of additional bacterial infections. Live Attenuated, Inactivated or Genetically Modified Vaccines (viral mutants by
deletion, without gp E, which allows differentiation of Ab by ELISA the infectious
Other categories of pigs and species-no treatment. antibodies- of -vaccine antibodies).
Many vaccines are available for sale. The companies that produce them make
available different vaccination schedules, ensuring protection against this disease
throughout the period of exploitation of the animals.
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105
PORCILIS® BEGONIA is a live vaccine, against Aujeszky's disease, for pigs in

the form, of a vial containing, a creamy white freeze-dried pill ,accompanied by a vial
of diluent, (Diluvac Forte) for reconstitution. PSEUDORABIVAC, live, freeze-dried vaccine against Aujeszky's disease-it is a freeze-
This product is a live, labeled, attenuated and freeze-dried vaccine, against dried suspension of Aujeszky's disease virus, tipe A-8/94, deleted by glycoprotein gp1,
Aujeszky's disease. Each dose, contains at least 106.0 TCID50, of Aujeszky Begonia with a minimum titer of 104.5 DICP50; accompanied by an adjuvant (aluminum
virus. hydroxide).
The diluent, is an injectable aqueous solution, with immunostimulating properties. The It is used preventively and when needed for pig herds diagnosed with infection with
Begonia strain, does not produce glycoprotein I (g I-), and thymidine kinase (tk-), so Aujeszky's disease virus.
animals infected with wild-type virus, will be serologically different, from those In infected efectives, vaccination is done like this:
vaccinated with Porcilis Begonia. Reconstitute the vaccine, with Diluvac Forte diluent. at the mother-3 times a year; in young pigs: at 8, 12 and 16 weeks. In the cohorts
Each animal, is vaccinated intramuscularly, at the back of the neck, behind the ear without disease or without clinical manifestation: cohort-mother vaccinated twice a
with 2 ml of vaccine. year, in trimesters II and IV; young-at 55 and 75 days. The vaccine dose: 2 ml,
Immunization program whatever the category of pigs; subcutaneous after the ear. The vials with reconstituted
Fatty pigs: first vaccination: from 14 weeks. For piglets without maternal antibodies, a vaccine should be shaken prior to inoculation. The instruments used to administer the
single vaccination, is sufficient. However, most animals have maternal antibodies, vaccine will be sterilized only by boiling. Animals with nutritional and metabolic
requiring a second vaccination, 2 weeks later. diseases, parasitic or produced by pathogenic germs may have an inappropriate
Breeding pigs: first vaccination, at 14 weeks with a 2 week booster. It continues with, immune response.
3 vaccinations per year, at intervals of 4 months. Withdrawal period for slaughter: 6 weeks after the date of vaccination.
Sick animals are not vaccinated.
PSEUDORABIVACOL, inactivated, oily vaccine against Aujeszky's disease-it is an

inactivated and freeze-dried suspension of Aujeszky's disease virus, type A-8/94,
obtained from cell cultures; the minimum titer: 107.0 DICP50/ml, with diluent.
Pseudorabivacol is used to prevent Aujesky's disease. COMBAT MEASURES
The vaccine can be used on all categories of pigs, regardless of their age.
The 1st dose must be followed by a 2nd, 3 weeks later. Illness officially declared; quarantine measures, 3rd degree.
Sick animals-isolated immediately, treated or sacrificed; after sterilization-they
The maintenance immunity of these animals is done by vaccines twice a year. The
are given away, or be eaten.
instruments used to administer the vaccine will be sterilized by boiling. The vials with
diluent for Pseudorabivacol are taken out of the refrigeration space, in the room at
-Infant, sick piglets-killed.
temperatures of 15-20 degrees Celsius, 2-3 hours before their use, to ensure their -Corpses and waste, after the sacrifice: destroyed or processed to fodder flour.
fluidity. The vial of Pseudorabivacol must be shaken, after reconstitution by diluting, from -Restriction of traffic, for animals.
the beginning and until the end of the vaccination, to homogenize the product. Animals -Periodic disinfection and repeated deratting.
with nutritional and metabolic diseases, parasitic or produced by pathogenic germs may -Vaccination requires- for healthy animals from contaminated farms.
have an inappropriate immune response. -This disease in pigs is considered extinct and the quarantine measures are
Accidental inoculation with the vaccine of operating personnel, in any area of the body, lifted 3 months after the last case of death, cut or recovery, if the necessary
leads to emergency medical consultation and monitoring. vaccination and final disinfection have been carried out.
Waiting period for slaughtering animals: 6 weeks after the date of vaccination.
NATIONAL MONITORING, CONTROL AND ERADICATION PROGRAM

MONITORING
Planned inspection for numbers of pigs without infection: in units supposed to quarantine new
animals.
SURVEILLANCE PRÉCISIONS TECHNIQUES PRÉCISIONS EXÉCUTION
Serological (ELISA gE): Inspection planifiée pour les effectifs
For 5% of breeding animals of authorized pig holdings, which do not vaccinate or vaccinate de porcins sans infection: dans les
with marker vaccine-once a year. unités censées mettre en
If there is a suspicion of disease, morphopathological and virusological examinations are quarantaine les animaux nouveaux.
Sérologique (ELISA gE): Les épreuves de sang sont recoltés
carried out and, depending on the case, the viral genome is identified by PCR a) Pour 5% des animaux de des porcines non-vaccinés ou
at the NRL in the IDSA laboratory. reproduction des exploitations vaccinés avec duvaccin marker.
TECHNICAL DETAILS autorisées de porcines, qui ne
vaccinnent pas ou vaccinnent avec
Blood tests are collected from non-vaccinated pigs or pigs vaccinated with marker vaccine.
du vaccin marker- 1 fois/an.
Ruminant brain tests with Aujeszky's disease symptoms will also be examined for transmissible S’il y a suspicion de maladie, on fait Les épreuves de cerveaux des Les épreuves de cerveaux des
spongiform encephalopathies. des examens morphopathologiques, ruminants avec symptomatologie de ruminants avec symptomatologie de
EXECUTION DETAILS virusologiques et, selon le cas, on maladie d’Aujeszky seront maladie d’Aujeszky seront
identifie le gènome viral par PCR au examinées aussi pour des examinées aussi pour la maladie d’
Ruminant brain tests with Aujeszky's disease symptoms will also be examined for Aujeszky's LNR au sein de l’IDSA. encéphalopathies spongiformes Aujeszky (mét. HEA, mét. Mann) et
disease (met. HEA, met. Mann) and for transmissible spongiform encephalopathies (HE transmissibles. pentru encéphalopathies
method, rapid test) at the NRL within the IDSA Institute of Diagnostics and Animal Health spongiformes transmissibles
(méthode HE, teste rapide ) au LNR
au sein de l’IDSA
7
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106
https://youtu.be/uvRAsbninoY
Pseudorabies. In The Merk Veterinary Manual. Eighth Edition. 964 – 966

Aujeszky’s Disease. OIE. Terrestrial Manual. http://www.OIE int.
https://youtu.be/5AXHEAsttic
https://youtu.be/uvRAsbninoY
https://youtu.be/fLkvB-RTxeo
https://youtu.be/ShZzLtDzr74
https://youtu.be/GrPX39YE3qI
https://youtu.be/3KbeHhNlKwQ

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PRACTICAL WORK GUIDE IN

INFECTIOUS DISEASES AND

DVM PhD DRAGOS COBZARIU

Practical Work Guide in

Coordinator: Head of works Dr. Dragoș Cobzariu

© 2022 All rights to this edition are reserved.

The practical work in the Discipline of Infectious Diseases and

Notifications of labor protection, fire prevention and extinguishing

PURPOSE OF PRACTICAL WORKS:

Infectious disease-definition DIAGNOSIS OF INFECTIOUS DISEASES

Within the SUSPICION-ASSUMPTION-mandatory diagnosis, three large groups of EPIDEMIOLOGICAL DIAGNOSIS

• Other data necessary for the DEVELOPMENT OF

EPIDEMIOLOGICAL INQUIRY EPIDEMIOLOGICAL INQUIRY

 Species and animals categories affected,

- Carcasses of dead animals Mobile supports: water, air (aerosols), vehicles,

LIVING RECEPTIVE ORGANISMS

A vaccinated flock can be infected in a silent manner, the

LIVING RECEPTIVE ORGANISMS CARCASSES OF DEAD ANIMALS

Destruction the carcasses of the dead animals is

-clinical expression, If additional examinations are required, all necessary

PRODUCTS OF ANIMAL ORIGIN PRODUCTS OF ANIMAL ORIGIN

RECEPTIVITY OF ORGANISMS TRANSMISSION WAYS

Measures prior to clinical examination, -Inspection,

CLINICAL OBSERVATION SHEET ANATOMOPATHOLOGICAL EXAMINATION

characteristic lesions for an organ (points to a group of diseases),

to be executed immediately after death or slaughter

EXTERNAL EXAMINATION OF THE CORPSE:

The external appearance (cadaveric changes,

INTERNAL EXAMINATION OF THE CORPSE:

ANATOMOPATHOLOGICAL EXAMINATION IN BIRDS

Samples are kept in a refrigerator (2 to 8 degrees Celsius) until shipping.

Newcastle Disease - birds:

Birds pasteurelosis CHOLERA

DVM PhD Dragoș Cobzariu 2022 39

 Isolation and identification of the CAUSATIVE AGENT,

ISOLATION AND IDENTIFICATION OF THE CAUSATIVE ISOLATION AND IDENTIFICATION OF THE

INSULATION AND IDENTIFICATION OF THE CAUSATIVE

It assumes seeding on culture media. 2. BACTERIOLOGICAL EXAMINATION

Cultivation conditions and Cultural characters.

THE OBJECTIVES OF THE EXAMINATION OF CULTURAL

For bacterial species with antigenically differentiated populations, the

Sensitivity to certain antibiotics.

INSOLATION OF VIRUSES ON LABORATORY ANIMALS

Inoculation routes: s.c., i.m., i.p., i.v., i.c.

Among them, immunology specialized cells for non-

METHODS OF METHODS OF IMMUNODIAGNOSIS

These reactions are based on the high specificity of the

The basics components (or necessary) of serological reactions

The antigen may be:

The outcome of the serological test should take into account:

SERO-AGGLUTINATION REACTIONS SERO-AGGLUTINATION REACTIONS

- Brucella ANTIGEN colored with pink bengal , positive brucella serum,

To identify the components of an antigen (somatic, flagella, capsular,

II. QUANTITATIVE REACTIONS

A. Slow Agglutination Reaction

B. Agglutination-lysis reaction with microscope observation

It is used for the diagnosis of leptospirosis and for the identification

SEROPRECIPITATION REACTION 2. GEL PRECIPITATION REACTIONS

In a Wassermann tube, place 1 to 2 ml of antigen and with a Pasteur pipette inserts

Serological technique combining a physical method (electrophoresis)

Under the action of electric current, Ag or Ab will

It is based on the fact that antibodies contained in an immune-

COMPLEMENT FIXATION REACTION

RFC = negative - positive / hemolysis ( are not specific for an Ag ).

-infectious diseases in which other serological techniques can not be used,

6. ANTIGEN-ANTIBODY REACTIONS EVIDENCED BY

It is based on the specificity of Ag-Ab reaction, in which