Lab 3.1: Detection of Disease Flashcards
sensitivity and specificity def
- Sensitivity: represents the likelihood that an animal with a disease will have a positive test result
- Specificity: represents the likelihood that an animal without a disease will have a negative test result
what are primary and secondary cell lines for virus isolation in cell culture?
I. Primary cell line: prepared directly from animal tissue tissues e.g. kidney, lung, spleen - no sub-cultivation
II. Continuous cell line: these type of cells are transformed for indefinite sub-cultivation. Most frequently used in diagnostic virology
what are the visual signs of a cytopathic effect in cell culture?
Cytopathic effect (CPE)
-Rounding and clumping of cells
-Vacuolation
-Fusion of cells; Syncytia or giant cells
how does immunofluorescence work?
antibody with indicator attaches to viral antigen, then visualized
how does ELISA work?
ELISA: Enzyme - linked Immunosorbent Assay for antigen detection
capture antibody on test sheet attaches to target antigen, and then enzyme-linked detection antibody sweeps over
what is the most versatile, rapid, sensitive and specific test for virus ID?
- Polymerase chain reaction (PCR) and its variations
virus neutralization test
-known amount of virus is added to each well
If enough antibodies are present, they will neutralize the virus and the cells will NOT become infected and NO cytopathic effects will be appreciated
- Which one of the following is CORRECT about diagnostic virology?
a) A rapid test is preferable to a sensitive test?
b) Virus visualization is more important than detecting its nucleic acid
c) Sequencing is not routinely performed for diagnostic testing?
d) Virus isolation is routinely done for diagnosis of animal infections
c) Sequencing is not routinely performed for diagnostic testing?
- Which one of the following is NOT CORRECT about diagnostic virology?
a) Nasal discharge is almost always important sample for all virus infections?
b) Viruses are not alive outside of the infected tissue so storage temperature during shipment WILL NOT affect the diagnostic result
c) There is no need to use personal protective equipment during sample collection since the type of virus is not yet known
d) Diagnostic tests are not that important for disease control and prevention
e) All of the above
f) All except B
e) All of the above
general approaches to treatment of CPV
- Intravenous fluids (balanced electrolyte solution)
- Antibiotics to prevent secondary bacterial infection
- Antiemetics (persistent, severe vomiting)
prognosis of CPV?
- 68-92% of puppies administered appropriate supportive care will survive, having developed long-term (potential life-long) immunity against the virus.
- Recovery time: Approximately 1 week after surviving the first 3-4 days of illness
what to expect in post mortem for CPV
Small intestine:
* Severe necrotic and hemorrhagic enteritis
* Necrosis in the Peyer’s patch
Lymph nodes (intestinal, mesenteric):
* Multifocal hemorrhage
Thymus:
* Severe diffuse hemorrhage (not pictured)
Vaccination strategies for Parvovirus
Inactivated and modified-live vaccine at 6–8, 10–12, and 14–16 weeks
- Booster administered 1 year later and then 3 year later
- Usually given as a combination CORE vaccine→ DAPP (Distemper, Adenovirus, Parvovirus, Parainfluenza) or DHPP (Distemper, Hepatitis, Parvovirus, Parainfluenza)
- Inactivated vaccines for pregnant bitches (reduced risk of abortion)
why might vaccines fail?
- Faulty vaccine - improper storage or administration
- State of immune system at time of vaccination
○ Immature immune system and/or dog is unhealthy at time of vaccination - Breed Differences!
○ Rottweilers and Dobermans are more susceptible to developing parvovirus enteritis despite vaccination! - Maternal immunity - maternal antibodies block the live attenuated vaccine challenge
● Ideal timing of vaccination: when the maternal antibodies levels will be low enough to no longer protect the dog
when is a puppy most vulnerable to CPV and why?
Puppy is most vulnerable to CPV-2 if infected between 8 to 16 weeks old
-period between maternal antibody protection and vaccine protection
when is parvovirus shed and how?
Virus is shed in the feces of infected dogs within 4–5 days of exposure (often before clinical signs develop), throughout the period of illness, and for ~10 days after clinical recovery
how do we kill parvo in the environment?
Killing parvovirus (small naked DNA virus) requires
strong disinfectants:
● Bleach (sodium hypochlorite)
● Potassium peroxymonosulfate
● Hydrogen peroxide
- Quaternary ammonium (soap) disinfectants do not kill parvovirus
Marek’s disease lifecycle
Mdv invades lung air space and infects epithelial cells
> MDV infects cells and spreads to the feather follicales to replicate
>birds shed MDV perticles in skin dander
>birds inhale MDV particles from dust in the environment
how do we vaccinate for Marek’s disease?
In OVO! We can vaccinate 18-day-old embryos! The chick has the best possible start when it hatches and
better disease resistance from day one
Manual subcutaneous vaccination is susceptible to human errors, while in ovo vaccination systems deliver the right dose in the right location
Why do animals get diseases if they have been vaccinated against?
- Particularly virulent strain of virus
Vaccine failure?
* Wrong administration route
* Not administered in correct quantity
* Poor preservation (e.g. vaccine got too hot/cold/old)
- Non-responder to the vaccine / immune status?
- Maternal antibody interference?
- Vaccine does not prevent infection, but prevents disease!
What viruses cause erosive/ulcerative mouth lesions? (6)
Bovine viral diarrhea virus (Flavivirus)
Foot and mouth disease (Picornavirus)
Infectious bovine rhinotracheitis (Alphaherpesvirus) Malignant catarrhal fever (Gammaherpesvirus)
Rinderpest (Paramyxovirus)
Vesicular stomatitis (Rhabdovirus)
*Progressing severe watery bloody diarrhea in young animals
*Oral mucosal ulceration
*Poor-doing calves (scruffiness, poor coordination, small) *Distal limb erosions
=>what diseases fit these symptoms?
*Clinical presentation could fit with mucosal disease OR severe, acute BVDV
How do you test a dairy herd for BVDV?
200 mL of milk from a bulk tank can be pooled from up to 400 animals
If the test is negative: you know every animal who contributed to the bulk tank is negative
If the test is positive: milk should be retested in 3 weeks to rule out acute infection
If the bulk milk sample is positive a second time: you need to evaluate animals individually
How can we control and prevent BVDV?
Identification and elimination of persistently infected animals in a herd > also need to test any replacement animals!
Vaccination:
* At least ten different brands of BVDV vaccines in North America
* Ideally you would want to choose a vaccine that works against both BVDV 1 and 2