D.

FAMILY FILOVIRIDAE
- Name comes from the Latin word: filo = 'threadlike'.
- Filoviruses are a major category of emerging viruses
- Belongs to the order Mononegavirales
- 1967: Marburg/Frankfurt, Germany
- Laboratory workers preparing primary cell cultures from African green
monkeys resulted in an outbreak of a previously unrecognized disease.
- Highly infectious:
31 ases with 7 deaths (some probably survived due the first
therapeutic administration of interferon)
- 1976: 'Ebola disease' outbreak in Zaire & Sudan:
- previously unrecognized hemorrhagic fever
- 500 diagnosed cases, 460 deaths! (92% mortality rate)
- From these two outbreaks, 2 novel viruses (Marburg & Ebola viruses) were
isolated; placed in a new family, the Filoviridae
- This family has strong structural & genetic similarities to both Rhabdoviruses &
the Paramyxoviruses

GENUS FILOVIRUS
● Properties of Genus Filovirus
- pleomorphic, elongated, 80nm diameter x 130-14,000nm long
- sometimes straight, but may be curved or hooked - 'U' or '6' shaped.
- particles are rounded at one end with a distended swelling at the other.
- the core has a striated appearance similar to that of Rhabdoviruses.
- enveloped
- stable when stored at 15oC to 20oC
- sensitivity: a. acid environment at ph 5
b. alkaline environment at ph 8
c. lipid solvent, phenol, formaldehyde, and ß-propiolactone
d. infectivity is reduced after exposure to irradiation (UV and
gamma)

Genome:
- Single stranded, unsegmented, (-)sense RNA, ~19kb
- Size varies with length of particle - optimum infectivity for Marburg
~790nm, Ebola ~970nm.

• Members
1. Ebola Virus
2. Marburg Virus
DISEASES CAUSED BY FILOVIRUS

1. Ebola Hemorrhagic Fever (Ebola HF)

- severe, often fatal disease of humans and non-human primates
- 4 identified serotypes
3 have caused disease in humans
Ebola Zaire
Ebola Sudan
Ebola Ivory Coast
1 has caused disease in non-human primates but not in humans
Ebola Reston

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 - Has a tendency to be spread through airborne particles

● Epizootiology and Pathogenesis of FILOVIRUS

- The clinical manifestations of Ebola virus infection are severe.
- The incubation period varies between four & sixteen days.
- The initial symptoms are:
a. severe frontal & temporal headache
b. generalized aches & pains
c. malaise
- By the second day the victim will have a fever.
- Later symptoms include:
a. watery diarrhea
b. abdominal pain
c. nausea and vomiting
d. dry sore throat
e. anorexia
- By day seven of the symptoms
- patient will have a maculo-papular (small slightly raised spots) rash
- develop thrombocytopenia & hemorrhagic manifestations, particularly in
the gastrointestinal tract, & the lungs, but it can occur from any orifice,
mucous membrane or skin site
- by day twelve the skin starts to peel away from the rash
- Ebola causes lesions in almost every organ, although the liver & spleen are the
most noticeably affected. Both are darkened & enlarged with signs of necrosis.
- The cause of death is normally shock, associated with fluid & blood loss into the
tissues.
- The hemorrhagic & connective tissue complications of the disease are not really
understood, but may be related to the fact that the VP40 protein is
antigenically related to human cell matrix proteins (abdominal aortic aneurism
protein & MFAP-4), leading to autoimmune attack

● Laboratory Tests Used:

1. Antigen captured ELISA
2. IgM ELISA
3. PCR
4. Virus isolation

● Treatment
There is no standard treatment. Patients must receive supportive therapy. This
includes balancing of patients’ fluid and electrolytes, maintaining their oxygen
status and blood pressure, and treating them for any complication.

2. Marburg Disease
- caused by Marburg Disease
- severe fatal viral hemorrhagic fever of humans
- first reported in Marburg, Germany in 1967, among laboratory workers exposed
to African green monkey

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G. FAMILY PARAMYXOVIRIDAE

Paramyxovirus
- enveloped
- pleomorphic
- linear stranded RNA
- nucleocapsid with helical symmetry
3 genera:
1. Morbilliviruses
2. Paramyxoviruses
3. Pneumoviruses

DISEASES CAUSED BY MORBILLIVIRUSES

1. Canine Distemper
- caused by Canine Distemper Virus
- most serious viral disease of dogs
- an acute viral infection with most frequent symptoms as follows:
diphasic fever ocular and nasal discharges
diarrhea hyperkeratosis of footpads
respiratory signs signs of CNS involvement
- Incubation Period is 3-5 days

● Properties of the CD Virus

- Variable in shape and size
- With helical symmetry of nucleocapsid
- Viral capsid is composed of 6 major polypeptides
Glycosylated H polypeptide – responsible for the adsorption of CDV to
receptor sites of susceptible cells
Glycosylated F polypeptide – causes fusion of cells
- Resistance: a) optimal ph stability is 7.0.
b) persist longer in cool, shady environments or in serum or tissue
debris
- Susceptibility: They are labile in extreme temperature, pH and several
disinfectants
Viral infectivity is lost at pH 10.5 or above and pH 4.3 or below
a) Inactivated by the following physical means:
UV light
Visible light
Heating at 60oC
b) Inactivated by the following disinfectant:
0.2% Roccal (QAC)
0.75% Phenol solution
 - Isolated and propagated in primary canine and ferret kidney cell cultures
- Worldwide in distribution
- Dog is the main reservoir
- No evidence to produce infection in human
- Transmission in dogs is via aerosol

● Laboratory Diagnosis:
1. Isolation and identification of CDV
specimen: secretions or tissue suspension from dogs
2. Demonstration of CDV antigen in monoclonal cells in the blood or
conjunctival smears by:
a. Immunofluorescent Technique
b. Peroxidase Technique
3. Demonstration of rising IgG titers in paired sera by ELISA

● Treatment and Control

1. Supportive in nature
antibiotics- prevent secondary bacterial infections
electrolyte to restore fluids
anticonvulsants to provide relief of nervous disorders
2. Vaccination – best means of controlling the disease
- modified live or inactivated CD vaccine
Maternal antibodies interfere vaccination
Alternatives for early vaccination:
a. Booster dose at 12 to 14 weeks of age regardless of the time of the
first dose
b. Measles viral vaccine followed by CD viral vaccine some weeks later
measles is antigenically related to CD virus. It confers resistant to
CDV infection in puppies. Upon exposure to CDV, measles is able
to sensitize antibody-producing cells to produce rapid and
pronounced CD neutralizing antibody titer
- On the other hand, measles are not neutralized by antibody
of CDV

2. Phocine Distemper
- infectious disease of seals similar to CD
- caused by Phocine Distemper Virus

3. Rinderpest
- caused by Rinderpest virus
- an acute or subacute viral disease of ruminants particularly cattle
- characterized by fever, lymphopenia, nasal and lacrimal discharges, diarrhea
and necrotic stomatitis
- IP is 3 to 8 days

● Properties of Rinderpest Virus

- similar with CDV
- inactivated by: sunlight within 2 hours

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 UV light rapidly
Lipid solvents
Disinfectants such as phenol or strong alkaline compounds
- adapted to grow in: laboratory animals such as rabbits, mice and guinea pigs
embryonated chicken eggs
- cultivated in primary or continuous cell cultures of bovine, ovine and porcine
origin.
- Transmitted thru direct contact of susceptible animals with secretions and
excretions of infected animals.

● Laboratory Diagnosis
- must differentiate Rinderpest from:
Bovine Viral Diarrhea FMD
Infectious Bovine Rhinotracheitis Bluetongue
Malignant Catarrhal Fever Salmonellosis
Coccidiosis
1. Isolation and identification of Rinderpest virus in susceptible animals and
cell cultures
2. Demonstration of specific antigen in tissues by:
a. FAT
b. Direct Peroxidase Test
3. Demonstration of significant titer rise by ELISA

● Treatment and Control

- no treatment
- can be prevented by:
1. Restricting live animal importation from enzootic areas
2. Slaughtering of affected herds
3. Vaccination for enzootic areas only

DISEASES CAUSED BY PARAMYXOVIRUSES

1. New Castle Disease

- caused by New Castle Disease Virus, the most serious disease-causing avian
paramyxoviruses
- highly infectious disease in chickens
- characterized by respiratory distress, diarrhea and neural signs
- incubation period is 4-11 days
- different strains:
Velogenic strains – most virulent strains
- produce 90% mortality rates
ex: GB, Milano, Herts, Ca-1083 and Largo
Mesogenic strains - less severely pathogenic
- mortality rate is <25%
ex: Roakin, Mukteswan and Haifa

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 Lentogenic strains - almost avirulent strain
- frequently employed as vaccine
ex: B1, La Sota, Ulster, F and V4

● Properties of Newcastle Disease Virus

- 6 major polypeptides; 2 are glycosylated and 4 are not glycosylated
Glycoprotein HN – responsible for hemagglutination and neuraminidase
activities
Glycoprotein F – causes fusion among infected cells

- Inactivated in varying degrees by physical and chemical agents as:

Heat Phenol
UV light Lysol
Ph Detergents
Oxidation process Butylated hydroxytoluene
- Survives in: contaminated areas and eggshells for several weeks
fresh eggs for many months
frozen carcass for few years
- Can be propagated in:
a. chorioallantoic cavities of 10-12 day-old embryonated chicken eggs
b. cell cultures: primary chick embryo kidney
primary chick fibroblast
baby hamster kidney cells
- Transmitted: most commonly thru aerosol via respiratory tract
lies can also transmit the virus
- Different forms:
1. Doyle’s form – caused by velogenic strain involving visceral organs
2. Beach’s form – caused by velogenic strain involving CNS
3. Beaudette’s form – caused by mesogenic strain involving respiratory and
nervous systems
4. Hitchner’s form – caused by lentogenic strain which is mild and inapparent

● Laboratory Diagnosis
1. Isolation and identification
specimens: respiratory exudates
tissue suspension (spleen, lung and brain)
substrates: embryonated hen’s egg
cell cultures
2. Demonstration of NCDV antigen in affected tissues or cell cultures by
immunofluorescence
3. Demonstration of rising NCDV antigen titers by HI, Neutralization or
ELISA

● Treatment and Control

1. Sanitation
2. Vaccination with:
a. live lentogenic strains – thru drinking water and aerosol
b. live mesogenic strains – at 4 weeks old via wing web, feather follicle or

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 intramuscularly
c. inactivated oil emulsion vaccines - parenteral route

2. Parainfluenza
- caused by parainfluenza viruses
- associated with upper respiratory infections in humans and animals

a. Infectious Tracheobronchitis/ Kennel Cough in Dogs

- caused by Parainfluenza type-2 virus
- characterized by sudden onset of mild fever, slight to copious nasal
discharge with harsh and non-productive cough
- results in more severe clinical disease when dually infected with Bordetella
bronchiseptica

● Properties of PI-2 Virus

- unstable; rapidly inactivated at ph 3.0 and at 37oC and above
- one strain causes laryngotracheobronchitis in children
- diagnosed by isolation and identification as well as serology

● Treatment and Control

1. antibiotics - counter B. bronchiseptica
2. expectorants containing codeine
3. isolation of infected dogs
4. vaccination with combined CD or Canine Hepatitis or B. bronchiseptica

b. Shipping Fever
- caused by Parainfluenza Type-3 Virus (PI-3)
- affects cattle
- one of the clinical disease syndromes in: Bovine Respiratory Disease Complex
- characterized by:high fever (41.5oC or above)
conjunctivitis
respiratory distress
mucopurulent rhinitis
pneumonia
- other contributors to the pathogenesis of Shipping Fever are:
1. Bovine Diarrhea Virus
2. Infectious Bovine Rhinotracheitis Virus
3. Bovine Respiratory Syncytial Virus
4. Pasteurella multocida
5. Pasteurella hemolytica
6. Mycoplasma spp

● Properties of PI-3
- inactivated at pH 3.0 and at 55oC for 30 minutes
- propagated in cell cultures of bovine, porcine, equine and human origin
- its pathogenic role in shipping fever of cattle can only be assumed in the
presence of other viruses and bacteria

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 - target cells are:
a. epithelial cells of respiratory tract
b. type 2 alveolar cells

● Laboratory Diagnosis:
1. Isolation
2. Serology

● Treatment and Control

1. Anti-prostaglandin compound: e.g. flunixin and meglumine to treat
pneumonia
2. Vaccination –usually combined with other agents causing shipping fever
protects calves but with questionable protection in adult
cattle
3. Isolation of infected animals
4. Good management practice

GENUS PNEUMOVIRUS
- Members of this genus are antigenically related
- Members lack neuraminidase

DISEASES CAUSED BY PNEUMOVIRUSES

1. Bovine Respiratory Syncytial Viral Infection

- caused by Bovine Respiratory Syncytial Virus
- an acute pneumonia in calves
- diagnosed by:
a. direct staining of nasopharyngeal smears with immunofluorescent antibody
to BRSV
b. ELISA of serum antibody to BRSV
- prevented by vaccination
- no treatment

2. Feline Respiratory Syncytial Viral Infection

- very mild; not a clinical disease
- not included in new classification

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E. FAMILY PICORNAVIRIDAE

Picornavirus

- comprise a large family of small RNA viruses that have great

significance in human and veterinary medicine
- 5 genera:
1. Aphthovirus
2. Enterovirus
3. Cardiovirus
4. Rhinovirus
5. Hepatovirus – no veterinary significance
- cause hepatitis A in human
● General Family Characteristics
- exhibit a cubic symmetry
- diameter: 22 to 30 nm
- capsid is icosahedral, composed of 60 subunits
- the viral genome is composed of a single piece of single stranded RNA
- non-enveloped

GENUS APHTHOVIRUS
- commonly referred to as “Foot and Mouth Disease (FMD) Virus
- causes one of the most important diseases of food animals
- economic losses include disease-associated losses and the interference of
international animal movement

DISEASE CAUSED BY APHTOVIRUS

Foot and Mouth Disease

- caused by Foot and Mouth Disease Virus
- infects wild and domestic CLOVEN-HOOVED animals
- most dramatic in cattle and swine
- sheep and goats are not severely affected

● Character of the Disease

- fever, depression, excessive salivation, lameness and formation of
vesicular-type lesions on the mucous membranes of the:
a. mouth – tongue, dental pad and gum
b. skin of the muzzle
c. inter-digital spaces
d. udder and teat
 e. coronary band
f. also in pharynx, trachea, larynx, esophagus, rumen and
muscles of heart
- lesions in the mouth result in:
a. reduced consumption
b. weight loss
c. emaciation
- secondary bacterial infection is responsible for the large part of FMD-
associated damage by complicating foot, respiratory and udder infections
- great economic loss is due to extensive morbidity; losses are due to
decreased productivity and protracted convalescence

● Properties of FMD Virus

- 7 serotypes: O, A, C, Sat1, Sat 2, Sat 3, and Asia 1
- serotypes A and O serve as genus prototype
- Resistance:
a. survives for extended periods in animal secretion and products
b. remains viable for 3 months in frozen meat
c. remains viable for 2 months in ham, bacon and sausage
d. for longer periods in lymph and bone marrow
e. resists inactivation by lipid solvents
f. tolerates ph 6.6 - 10
- Susceptibility:
a. inactivated by heating above 50oC
b. acid sensitive (<ph 5.0)
c. alkaline sensitive (>ph 11)
d. 1% NaOH2 – recommended to disinfect premises following
FMD outbreaks
● Epizootiology and Pathogenesis
- complex epizootiology- there’s an interplay of: viral strain, animal host
and environment
- transmitted by: direct contact
aerosols
fomites
possibly arthropod vector
- virus can be recovered from: vaginal discharge semen
nasal discharge milk
membranes of aborted fetus feces
saliva tears
urine
- Respiratory route is the predominant route of FMD infection.
- Initial replication occurs in the pharynx
- Malignant form occurs when the virus infects heart muscles, causing
degenerative changes and necrosis. It is characterized by longitudinal
yellow-whitish streaks representing myocardial necrosis, referred to as
“Tiger Heart”, where death occurs early in infection, especially in young
animals.

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 ● Immunity
- duration of immunity is longer (about 1 year) in cattle than in swine

● Diagnosis
- must be differentiated from: VS (Vesicular Stomatitis)
SVD (Swine Vesicular Disease)
VES (Vesicular Exanthema of Swine)
1. Isolation in cell cultures and laboratory animals
2. Electron Microscopy – rapid diagnosis
- physicochemical characterization
3. Serologic tests- identifies antibody obtained from previous infection,
not from previous vaccination:
a. AGID – Agar Gel Immunodiffusion
b. Recombinant FMDV nonstructural proteins

● Treatment and Control

- control is very difficult due to:
a. highly contagious in nature
b. multiple host range
c. viral stability
d. multiple antigenic types and subtypes
e. short-term immunity
- Proper animal husbandry practices reduce losses and secondary bacterial
infection:
a. impose strict import regulations on animals, animal
products and potential viral-contaminated fomites from
FMD + countries
b. slaughter and quarantine in affected areas
c. vaccination - to avoid additional viral strains since cross-
protective immunity between strains is not
guaranteed
- must be done one to three times a year since
immunity is short term
i. inactivated vaccines of tissue origin – most common
ii. live attenuated vaccines - questionable

GENUS ENTEROVIRUS
- thermo-labile – infectivity is destroyed quickly at 50oC
- insensitive to detergent treatment
- stable at low ph – can survive at ph 3

DISEASES CAUSED BY ENTEROVIRUS

1. Porcine Enteroviral Infections

- 11 serotypes
serotype 1 –Teschen and Talfan Disease Viruses
- Swine Enterovirus prototype

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 serotype 9 – Swine Vesicular Disease Virus

a. Teschen Disease
- produce mild or severe form of polio-encephalomyelitis
- mild forms are referred to as: Talfan disease
Benign enzootic paresis
Polioencephalitis
b. Swine Vesicular Disease
- resembles: FMD
VS (Vesicular Stomatitis)
VES (Vesicular Exanthema of Swine)
- characterized by:
a. fever, weight loss, and lameness
b. development of vesicular lesions in:
i. feet (coronary bands, soles and inter-digital areas)
ii. tongue
iii. nares
iv. lips
v. skin
a. ulceration – occur in the metacarpal and metatarsal
areas
b. sometimes shedding of hoof

● Diagnosis
1. Identification of virus or viral antigen in lesions by FA staining
in electron microscopy
2. Viral isolation via:
a. primary porcine cell cultures
b. suckling mice
3. Serology
Nested RT-PCR – can detect viral RNA in clinical specimens
- more sensitive than viral isolation on cell
cultures
- distinguishes SVD from FMD, VS & VES

2. Bovine Enteroviral Infection

- 7 serotypes
- uncertain role in disease

3. Duck Hepatitis
- contagious and highly fatal disease of young ducklings
- caused by Duck Hepatitis Virus

● Properties of Duck Hepatitis Virus

- resistant to: lipid solvent
acidic ph
trypsin
- thermostable and survive for long periods in fecal materials

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 - cultivated in chicken embryonated eggs or duck embryonated egg thru
allantoic inoculation

● Epizootiology and Pathogenesis

- Brown rats are potential carriers
- Transmitted directly or indirectly (thru contaminated feces). No vertical
transmission
- Not well-defined pathogenesis
- Primary gross lesion – enlarged and hemorrhagic liver

● Diagnosis
1. Suggested by rapid spread and per acute nature of the disease
2. Pathognomonic hemorrhagic lesions in the liver of ducklings
3. FA staining- definitive diagnosis
Source of specimen:
Necropsy tissues
Inoculated avian embryo
Note: serology has not been useful in DHV diagnosis

● Treatment and Control

- No treatment
- Controlled thru proper sanitation and administration of immune serum
- Prevented by vaccination

4. Avian Encephalomyelitis (AE)

- Caused by AEV – Avian Encephalomyelitis Virus
- A disease in young chickens (1-2 weeks old). Also infects turkeys,
Japanese quail and pheasants
- Characterized by dull expression in eyes, progressive ataxia and tremors
- Death is due to lack of food and water, as well as trampling by other
chicks.
- Propagated in: 1. Chick embryos thru the yolk sac route of inoculation
2. Cell cultures- neurological cells, embryo fibroblast,
brain cell and pancreatic cells
● Epizootiology and Pathogenesis
- transmitted directly or indirectly (thru viral contaminated feces)
- both vertical and horizontal transmission
- oral infection is the usual route of entry
- from the GIT, virus infects other visceral organs and CNS

● Diagnosis
- must be differentiated from: NCD
Ricketts
1. Histopathology – provide presumptive diagnosis
2. Demonstration of Disease to inoculated embryo
3. Identification of viral antigen
4. Serology

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 ● Treatment and Control
- no treatment
- supportive care – proper husbandry
- prevented by vaccination

GENUS CARDIOVIRUS

DISEASE CAUSED BY CARDIOVIRUS

Encephalomyocarditis (EMC)
- natural disease of rodents
- young pig is the only domestic animals affected

● Properties of Cardiovirus
Encephalomyocarditis (EMC) virus- only one serotype recognized
\ 4 strains: 1. Mengo virus 3. Columbia SK virus
2. Maus Elberfeld virus 4. MM virus
● Epizootiology
- lacks pig to pig transmission
- infection in pigs is initiated from rodent sources
- Gross lesion:
a. spleen is devoid of blood
b. myocardial lesions include white longitudinal tracts of
necrosis, zone of lymphoid infiltration and calcification
● Diagnosis
1. Viral Isolation
2. PCR
* since immune response require about 5 days and may not
develop prior to death, the virus cannot be tested with serum

GENUS RHINO VIRUS

- insensitive to acid (<5-6 ph)
- resistance to ether
- thermo-stable
- has tropism to respiratory tract
- includes: 1. human rhinovirus
2. bovine rhinovirus – uncertain significance as a pathogen
3. equine rhinovirus – can infect multiple species such as:
guinea pigs, rabbits, monkeys, and
human

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