Guide for

Laboratory
Diagnosis in Herd
Problem
Investigations
Dr.Kedar Karki
Senior Vet.Officer
Central Vet.Laboratory
Tripureswor
Logical Components of Herd
Problem Investigations:
• The unique strength of herd
investigation is that because the
individuals are grouped into a herd.
• you can compare affected animals,
both clinical and subclinical, with
unaffected animals, in both a cross-
section (at one point in time) and
over time to determine the
differences between both the animals
themselves and the factors affecting
them.
 Goals
• Provide examples of how a variety of
laboratory diagnostic techniques are
used in investigational outbreak
settings.
 Laboratory diagnosis can be
used to:

• identify the agent causing an

outbreak;
• confirm cases in an outbreak;
• link cases to the same outbreak,
even with cases that occur over wide
geographic areas;
• identify the strain or serotype of an
agent involved in an outbreak; and
 Laboratory diagnosis can be
used to:

• Each of these uses for laboratory

diagnostics is illustrated below using an
outbreak example.
• Keep in mind that the list is not exclusive.
Each of these examples may feature
multiple aspects of laboratory diagnosis,
and innumerable other outbreaks could
illustrate the same points.
•
 Laboratory diagnosis can be
used to:

• learn more about the epidemiology

of infectious agents for research
purposes (such as to identify new
modes of transmission, to learn more
about newly described or reemerging
infectious diseases, or to evaluate
prevention measures).
 identify the agent causing an
ongoing or recent outbreak.
• Correctly identifying the agent may
allow more effective prevention.
 identify the agent causing an
ongoing or recent outbreak.
• In 1998-1999, 3 clusters of febrile
encephalitis in Malaysia were reported to
the Malaysian Ministry of Health. (1) By
the end of the outbreak, there had been
more than 200 cases and more than 100
deaths. During the same time period, 9
similar cases were reported in Singapore,
including 1 death.
•
 identify the agent causing an
ongoing or recent outbreak.
• Japanese Encephalitis (JE), a viral
encephalitis transmitted through the
bite of a mosquito, was endemic to
the area.
 identify the agent causing an
ongoing or recent outbreak.
• Investigators initially suspected the
JE virus as the cause of the
outbreak, and some specimens
tested positive for this agent.
However, when nervous system
specimens were grown in tissue
culture, a previously unknown virus
grew.
 Identifying the Agent
Causing an Outbreak
• Cases mostly adult men with swine
contact
– JE not usually associated with swine, so JE less
plausible
• Samples from 13 patients sent to CDC for
testing
– JE identified from only 1 specimen
– Samples then examined under an electron
microscope; structure of similar in shape to a
paramyxovirus
– Additional laboratory tests performed
 Identifying the Agent
Causing an Outbreak
• Virus found to be related to Hendra
virus (first identified in Hendra,
Australia)
– Tissues from deceased patients were
antibody positive
– Antibodies also found in the serum of
some patients
– Virus itself found in tissues of other
patients
 Identifying the Agent
Causing an Outbreak
• Similar investigations performed
among swine to examine
epidemiologic link
– Virus found in the central nervous
system, lung, kidney tissues from swine
at affected farms in Malaysia
– Singapore cases handled swine from
Malaysia
 Identifying the Agent
Causing an Outbreak
• To prevent further infection:
– Transport of swine within Malaysia
banned
– Use of personal protective measures
(gloves, masks, etc.) encouraged for
swine workers
– Importation of swine from Malaysia
prohibited by neighboring countries
 Identifying the Agent
Causing an Outbreak
• Research on epidemiology and
transmission of virus among
swine and humans ongoing
outbreak investigation flow diagram below.
1;A Problem is Detected
• A farm event triggers the investigation.
The event can be dramatic, such as a
series of unexpected deaths or a sudden,
dramatic production declines, or it can be
finally the recognition of a chronic problem
that has been occurring for some time.
• The earlier problems are detected,
generally the more successful the
interventions.
1;A Problem is Detected
• He who detects the problem is
often called on to solve it
The three types of problems
are
• 1;Acute: The problem was
precipitated by a temporally-
associated management or
husbandry error of sufficient
magnitude to be a sole cause of the
problem.
The three types of problems
are
• 2;Additive or Cyclic: The problem was
precipitated by a combination of
management or husbandry errors over
time and the effects of cyclical factors
such as season or production cycle stages
such that the combination was sufficient to
precipitate the problem. Ex: the summer
coliform mastitis outbreak that is
associated with the previous winter
change to sawdust bedding.
The three types of problems
are
• 3:Chronic: The problem was
precipitated by the long action of
management or husbandry errors
that required the passage of time for
before the consequences became of
sufficient magnitude to be
recognized, such as the slow spread
of a contagious mastitis agent or of
Mycobacterium paratuberculosis.
 3
• Ex: the recognition of a Staph.
aureus mastitis problem associated
with the adoption of a less efficacious
teat dipping procedure more than a
year previously.
 2:Establish or Verify the
Pathological and Etiological
Diagnosis
• If the diagnosis is not definitive and if one
is needed, start the process to obtain a
definitive pathologic and etiologic
diagnosis by collecting samples and
submitting these to diagnostic
laboratories. If dead or dying animals are
involved, have complete field necropsies
been done of a sufficient number of
representative animals?
Epidemiologic Triad of
Disease

Host

Agent Environment
 The Natural History of
Disease

Outcome
Healthy Cure
Control
Disease Symptoms Seek Diagnosis Treatment
Onset Care Disability
Death
 3:Establish a Case
Definition
• Establish a case definition, precise as
reasonable, to exclude those cases that
are due to endemic background problems.
• You may wish to establish a case definition
with different degrees of certainty (e.g.,
certainly affected, possibly affected,
possibly unaffected, certainly unaffected).
Remember the iceberg principle and
the spectrum of disease.
 and Magnitude of the
Problem with Objective
Data
• Obtain objective data to
document and verify the
magnitude of the problem; do
not rely only on the memories
and perception of management
and employee .
• Remember: More mistakes are
made from not looking than from
not knowing!
 and Magnitude of the
Problem with Objective
Data
• A:Establish the Timing of the
problem (the temporal pattern -
When?)
• B: Establish the Place of the
problem (the spatial pattern -
Where?)
• C:Establish the Demographics of
affected vs. non-affected animals
(Who?)
5:Assemble and Analyze
the Data
• For an outbreak, establish an
epidemic curve.
• For endemic problems, plot risk over
time by cohort group.
• Examine the effects of other factors
that vary over time
 6:Generate Hypotheses
(Differential Diagnoses)
about Key Determinants
• Key determinants are those risk
factors causing the problem that
can be modified on this premises.
7:Test Your Hypotheses
• Make predictions of the form "if
this cause is present, then this
finding should be present".
 8:Design Interventions
and/or Prospective Studies
• Generate action items that are
compatible with the specific facilities,
economic limitations and
management scheme of the
premises.
 9:Report Your Findings
• "The faintest pen is stronger than
the strongest mind" and that
"Success has many fathers but
failure is an orphan." If your
recommendations are successful,
because of the passage of time you
will not likely get the credit due you
unless they were documented.
 10:Monitor Results of
Interventions
• Develop a monitoring scheme to
provide early warning of the
problem. If the herd doesn't have a
good production accounting system
to monitor changes in production but
one is warranted, propose one.
Investigation Execution:
• 1:Record the physical layout of
premises.
• 2:Record the current numbers of
animals in each pen or area and
the intended maximum capacity
of these areas.
• 3:Record the animal "calendar"
intended by management for the
relevant animals
 Summary
• tests can be used to:
– Solve outbreak investigations
– Identify agents
– Investigate remaining questions about
infectious diseases
• Laboratory diagnostic techniques are an
integral part of public health surveillance,
investigation, and research
• Understanding the basics of how these
tests work will improve your conduct of
outbreak investigations