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Progressive Control Practitioners’ Network
Conducting an NSP antibody Sero-survey
Progressive Control Practitioners’ Network
NSP sero-survey PC Practitioner Network 2
Conducting an NSP-Antibody Sero-survey
Introduction
Kees van MaanenEuFMD component manager MENA and Laboratory networs
Javad EmamiProvincial Veterinary Officer, West Azarbaijan, Iran
NSP sero-survey PC Practitioner Network 3
Chris BartelsEuFMD component
manager PCP
Wilmot ChikurunheShort-term professional Southern Africa
Objective Field
application
Design Laboratory issues
and result
interpretation
Jenny
MaudTraining ProgrammeManager
Practicing the
Network
Introduction
Practitioners’ Network
NSP sero-surveys- To understand level of infection in and
endemic situation- To understand two-stage sampling- To manage sample size calculations for
two-stage sampling- To develop a questionnaire for putative
risk factors- To manage data collection and validation- To analyse risk factors for spread of
infection
Webinar and pre-recorded presentationsToday’s webinar, prerecorded presentation and a final webinar on 9 February
Exercises or assignments- Assessing laboratory results- Estimating animal
prevalence, epi-unit andwithin epi-unit prevalence
- Confidence intervals
Invited experts- Wilmot Chikurunhe- Javad Emami- Kees van Maanen
Knowledge Bank andJob Aidshttps://eufmd.rvc.ac.uk
NSP sero-survey PC Practitioner Network 4
Discussion forum & tutoring- Publications- Propositions- News-feeds- Agenda
A new tool/job-aid- A checklist to conducting a survey- Template for questionnaire to conduct when
sampling
Programme and Practitioners’ view
Agenda for January –February
2017
NSP sero-survey PC Practitioner Network 5
Week Mon Tuesday Wed Thursday Fri
1 16 17 Online available:
exercises, job-aids,
SOP’s related to
‘Survey design’
18 19 Webinar
Introduction and
Survey design
Chris Bartels
Wilmot Chikurunhe
Javad Emami
20
2 23 24 25 26 27
3 30 31 Online available:
exercises, job-aids,
SOP’s related to
‘lab interpretation’
1 2 Pre-recorded
presentations
Laboratory
interpretation
Kees van Maanen
3
4 6 7 8 9 Closing webinar:
results and
explanation of
exercises
10
Programme and Practitioners’ view
12-monthsprogrammefor the Practitioners’Network
NSP sero-survey PC Practitioner Network 6
Month 01
NSP sero-survey Complementing outbreak
reporting
Month 02
Outbreak
investigation Fit for purpose
Month 03
Economic impact
assessment From clinical disease to lost
dollars
Month 04
Month 05
Risk analysis along
value chain Understanding value chains is
the tool to mitigate widespread transmission of
FMD virus
Month 06
Stakeholder
consultation If people make livestock
move, what do we need to know about these people
Month 07
Post-vaccination
monitoring Testing the vaccine and its
performance as not all vaccines and applications are
the same
Month 08
Month 09
Risk-based
surveillance More information for less
input
Month 10
Training of
trainers Integrating capacity building
with strengthening Veterinary Services
Month 11
Review of the year
and future plans What has the Network
brought you and how did you contribute?
Month 12
Programme and Practitioners’ view
ObjectiveDuring this session, we will discuss
• how sero-surveys support understanding of the epidemiology of FMD and monitoring and evaluation of FMD control plans
• Importance to define objective for the sero-survey (objective) – Prevalence estimation versus detection of disease
– Unit of interest versus unit of sampling
– Study population versus target population
– Inclusion and exclusion criteria
• Principles of survey design such as – Two-stage sampling
– Sample size calculation
– Accounting for random error and prevention of bias
– Collect accurate attribute information to be able to conduct further analysis
• Implementation and organisation of conducting a NSP-Ab sero-survey– Developing materials
– Training of staff
– Pretesting of questionnaire and data recording
– Data management
• Laboratory aspects
NSP sero-survey PC Practitioner Network 7
Programme and Practitioners’ view
What does clinical disease tells us about occurrence of FMD?
Information gain– FMD serotype, strain
– Impact
– Vaccine effectiveness
– Risk factors for disease
NSP sero-survey PC Practitioner Network 8
Objective
What does clinical disease tells us about occurrence of FMD? And thus, what not?
Information gain
– FMD serotype, strain
– Impact
– Vaccine effectiveness
– Risk factors for disease
NSP sero-survey PC Practitioner Network 9
• Lack of information – Origin of infection
– Extent of spread of infection
• temporal, spatial, cross species
– Risk factors for transmission of infection
Objective
Outbreak reporting
– being lucky to see
Sero-survey: actively
searching the unseen
NSP sero-survey PC Practitioner Network 10
Objective
Question
What percentage of occurrence of FMD in livestock do you think is reported to the Veterinary Services?
1. Between 1 – 5 per cent
2. Between 6 and 10 per cent
3. Between 11 and 20 per cent
4. Between 21 and 40 per cent
5. More than 40 per cent
NSP sero-survey PC Practitioner Network 11
Objective
W. Azerbaijan serosurvey
80.2% of epi-units had at least 5 calves with a high titre (>75% inhibition)
18% observed clinical signs in their stock in the previous 12 months (questionnaire)
NSP sero-survey PC Practitioner Network 12
Incidence85.71 - 100.0080.71 - 85.7171.43 - 80.7166.67 - 71.4350.00 - 66.67
2011 Incidence FMD serosurvey
Incidence28.57 - 40.0027.27 - 28.5719.81 - 27.2716.67 - 19.818.57 - 16.670.00 - 8.57
Incidence FMD reports (sample)
Incidence13.77 - 32.769.97 - 13.779.18 - 9.977.81 - 9.187.37 - 7.811.52 - 7.37
16 mo prior to survey
Incidence FMD reports
District-level FMD Incidence:
Serosurvey and clinical signs (survey and official reports)
Objective
What are potential reasons for differences between serological and reported data?
NSP sero-survey PC Practitioner Network 13
Objective
What are potential reasons for differences between serological and reported data?
May be due to:1. Under-reporting
• More likely in sheep
• Non aware of reporting
• Non-confident of consequences of reporting
2. Subclinical infection• FMD signs observed by farmers on:
– 18% NSP+ epi-units in W. Azerbaijan
• May vary by FMD strain, species infected, vaccination status
3. Reflection of previous year’s cases?• Importance of limiting serosurvey to young animals
(ideally 4-12 months, possibly more practical to include 6-18/24 months)
NSP sero-survey PC Practitioner Network 14
Objective
Both approaches contribute to assessing and monitoring FMD risk and FMD control programmes
NSP serologyMeasures FMD infection
Less biased (with careful design!)
– Unit of interest, unit of sampling
Combine with questionnaire to give more information about risk factors
Resource intensive
Survey can be combined with post-vaccination monitoring (SP-Ab testing)
NSP sero-survey PC Practitioner Network 15
Objective
Kees van MaanenEuFMD component manager MENA and Laboratory networs
Javad EmamiProvincial Veterinary Officer, West Azarbaijan, Iran
NSP sero-survey PC Practitioner Network 17
Chris BartelsEuFMD component
manager PCP
Wilmot ChikurunheShort-term professional Southern Africa
Objective Field
application
Design Laboratory issues
and result
interpretation
Jenny
MaudTraining ProgrammeManager
Practicing the
Network
Introduction
Step 1 – What and why?
Define objective
Define focus
Prevalence estimate
Detection of disease/Disease
freedom
Comparison of prevalences
Identify target
population
Unit of interest
Unit of sampling
(primary and secondary)
Define diagnostic
tests
Antibody vsantigen
Test characteristics (Sens, Spec)
NSP sero-survey PC Practitioner Network 18
Design
Population
Research question
Sampling strategy
Sample size
Sample
measurements
Analysis
NSP sero-survey PC Practitioner Network 19
Design
Most common research questions
NSP sero-survey PC Practitioner Network 20
Prevalence Estimation
e.g prevalence of FMD infectionDifference in prevalence
e.g. between regions, countries,
case and control group
Detection of disease e.g. presence of disease
at a predefined level or
above?
Freedom of disease
e.g. confidence that disease
is absent
Design
Surveys
Determine what question is being asked... and how best to answer it
Examples of questions?– Is there FMD in my country?
– How much FMD is in my country?• What is the NSP sero-prevalence?
• What is the incidence of clinical FMD?
– Is the level of FMD higher in one area than another?
– What are risk factors for FMD? (must define factors of interest)
– Is vaccination effective?• What is the level of sero-conversion after vaccination?
• Are vaccinated animals less likely to develop disease?
– What is socio-economic impact of FMD on dairy farms?• Measure the decrease in milk production associated with FMD
• Quantify the economic loss due to an outbreak of FMD
NSP sero-survey PC Practitioner Network 21
Design
Unit of interest (unit of analysis): what are you counting?
NSP sero-survey PC Practitioner Network 22Control AI Vaccination AI & ND Vaccination
Treatment Blocks
Kulon Progo District
v
v v
v v
v
v
v
v
v
Individual animal
Nr animals sick/total nr animals
Pens
Nr Infected pens/total nr pens
Farms
Households/villages
Districts
Design
Calculate sample size
Assumptions Primary and secondary
sampling unit
Define sampling procedure
Random, Convenience, Selective,
Risk-based
Single or multiple stage
Selection procedure
Develop collection
instruments
Sampling materials
Questionnaire Recording
Pre-test and instruct
Supervision EnumeratorsLivestock owners
Define objective
Define focus
Identify target
population
Define diagnostic
tests
Step 2 – How to conduct?
NSP sero-survey PC Practitioner Network 23
Design
NSP sero-survey PC Practitioner Network 24
The concept of a sample
Measuring whole population - census
Design
Target Population
NSP sero-survey PC Practitioner Network 25
Sample
Measure and infer
results to whole
population
Design
Single-stage and 2-stage sampling?
• Based on number of animals involved
• Based on management practices and observed clustering of animals
• In most cases never a simple, single stage random sampling because the animals numbers are almost always very high and exist in some form of clustering
• Most surveys fall in the 2-stage sampling category
• Rarely do surveys involve more than 2 stages
NSP sero-survey PC Practitioner Network 26
Design
Two stage sampling
Primary sampling units (PSU)
– Herds, villages, flocks
– First level of clustering of
individual animals
– Often the unit of our primary
interest for estimation of
prevalence as FMD control
aims at epi-unit level
(livestock owner, village)
more than on individual
animal
– Bulk milk testing may help
to simplify the design
Secondary sampling unit (SSU)
– Animals, individuals
– Units required to provide
information about the
primary unit (it is just not
possible to sample the herd,
village directly to define it
infected or not)
– Thus, to qualitatively define
the disease / infection status
of the herd, village, flock
referring to detection of
‘disease’
NSP sero-survey PC Practitioner Network 27
Design
First Stage Sampling
NSP sero-survey PC Practitioner Network 28
Animal
clusters
(as the sampling frame)
Design
Second Stage Sampling (Selection of animals)
NSP sero-survey PC Practitioner Network 29
Second stage sample
Sampling frame? - Each of the epi units in the first stage sample
Design
Have you worked with a 2-stage sampling approach?
Yes
No
NSP sero-survey PC Practitioner Network 30
Design
Sample size determination at both stages
Use of software eg Winepiscope
NSP sero-survey PC Practitioner Network 31
You need:
•Design prevalence -
a hypothetical value that
is given for disease
presence in a herd or
population when planning
a survey. Herd level and
within herd
•Level of confidence
•Population size
Design
Sample surveys (bias)
NSP sero-survey PC Practitioner Network 32
There will be always a difference between the true value of the parameter (most of the times unknown) and its estimate
This error can be broadly classified into:
Random error • Sample size
Systematic error
• Selection
• Measurement
• Information
BIAS
Design
Questionnaire
When sampling livestock, it is important to collect additional information to understand association between lab result and factors (attributes)
– At animal level• Age
• Purpose
• Origin (born where sampled or introduced
newly)
• Vaccination status (if known at individual level)
– At household level• Common grazing, common drinking
• Herd structure, presence of other livestock
species
• Trading activities of owner/family
– At village level
• History of FMD
• Vaccination history
• Location near animal market, main road,
nature reserves
HOUSEHOLD – ask owner- QUESTIONNAIRE IDENTIFICATION - VV/GG/01-
X- COORDINATE ……………
Y-COORDINATE ……………
1 Are there (or have there been in the last 12 months)
sheep or goats in this household? (3)
0 No
1 Yes
2 Where did ruminants go for drinking in the last 12
months? (more than one answer possible)
0 At the house
1 Within village
2 Around village
3 In nearby village
3 Where were ruminants fed in the last 12 months?
(more than one answer possible)
0 At the house
1 Within village
2 Around village
3 In nearby village
4 Were ruminants introduced/purchased in last 12
months? (more than 1 answer is possible)
0 No
1 Yes, from this village
2 Yes, from this municipality
3 Yes, from this district
4 Yes, from this governorate
5 Yes, from –governorate-
5 For what purpose are ruminants raised?
(more than 1 answer is possible)
1 Milk production
2 Fattening
3 Other
6 How did you treat the manure in the last 12 months?
(more than 1 answer is possible)
0 Keep for himself
1 Lend to his neighbors in the
same village
2 Sold out the villages
7 Did you buy/introduce manure from other farms in
the last 12 months?
0 No
1 Yes
8 Have you sold any ruminants in the last 12 months? 0 Yes
1 No
NSP sero-survey PC Practitioner Network 33
SAMPLE SHEET QUESTIONNAIRE IDENTIFICATION - VV/GG-
X- COORDINATE ……………
Y-COORDINATE ……………
Serial number Age in
months
Sex
0 = male
1 = female
Species
0 = cattle
1 = buffalo
Born at owner
0 = yes
1 = no, in village
2 = no, in municipality
3 = no, in district
4 = no, in different
governorate
Owner Coding for
household
questionnaire
VV/GG/01/01
VV/GG/01
VV/GG/01/02
VV/GG/02/03 VV/GG/02(1)
VV/GG/02/04(2)
VV/GG/03/05 VV/GG/03
VV/GG/03/06
VV/GG/04/07 VV/GG/04
VV/GG/04/08
VV/GG/05/09 VV/GG/05
VV/GG/05/010
VV/GG/06/011 VV/GG/06
VV/GG/06/012
VV/GG/07/013 VV/GG/07
VV/GG/07/014
VV/GG/07/015
VILLAGE – ask local veterinarian - QUESTIONNAIRE IDENTIFICATION - VV/GG -
X- COORDINATE ……………
Y-COORDINATE ……………
1 Is there a ruminant market in this village? 0 No
(Only one answer possible) 1 No, but in same municipality
2 No, but in same district
3 Yes
2 Have there been clinical signs for FMD in the last 12
months? (Only one answer possible)
0 No
1 No, but in same municipality
2 No, but in same district
3 Yes
3 Was there vaccination against FMD in the last 12
months? (Only one answer possible)
0 No
1 Yes, 1 time, - month -
2 Yes, 2 times, - month -
4a What is the estimated number of cattle in this village? Cattle …………….
4b What is the estimated number of buffaloes in this village? Buffaloes …………….
4c What is the estimated number of sheep in this village? Sheep …………….
4d What is the estimated number of goats in this village? Goats …………….
IDENTIFICATION -Fill out yourself- QUESTIONNAIRE IDENTIFICATION - VV/GG -
X- COORDINATE ……………
Y-COORDINATE ……………
Name of owner ………………………… Village ……………………..
Municipality ………………………… District ……………………..
Governorate …………………………
Location of local clinic 0 Same village
1 Different village
1 What are geocoordinates (decimal degrees) of the
village?
X …………………………..
Y …………………………..
Design
Available on our PC Practitioner site areSupporting documentation and publications
NSP sero-survey PC Practitioner Network 34
Design
Kees van MaanenEuFMD component manager MENA and Laboratory networs
Javad EmamiProvincial Veterinary Officer, West Azarbaijan, Iran
NSP sero-survey PC Practitioner Network 36
Chris BartelsEuFMD component
manager PCP
Wilmot ChikurunheShort-term professional Southern Africa
Objective Field
application
Design Laboratory issues
and result
interpretation
Jenny
MaudTraining ProgrammeManager
Practicing the
Network
Introduction
Calculate sample size
Define sampling
procedure
Develop collection
instruments
Pretest
Define objective
Define focus
Identify target
population
Define diagnostic
tests
Conduct survey
Monitor implementation
Support service
Collect and enter
data
Data flow field to central
Check for completeness and accuracy
Validate and
analyse
Descriptive Analytical
Interpret Follow-up actionsFeedback to participants
Step 3 – Do it and learn!
NSP sero-survey PC Practitioner Network 37
Field application
IRANIAN NSP-SEROSURVEY TO ESTIMATE
FMD INFECTION LEVEL WHERE NON-
PURIFIED FMD VACCINE IS USED
J Emami, M McLaws, A Emami, R Ghaffari,
N Sedghi, N Rasouli, C Bartels
Field application
Implementing an
NSP-Ab Sero-survey:
Experiences from West Azerbaijan province of Iran
NSP sero-survey PC Practitioner Network 39
Field application
NSP sero-survey PC Practitioner Network 40
Country
Neighboring countries
Size
Human population: 80 million
Small scale farming : 21 million
of people
Dairy farms
Beef farms
Province
Neighboring countries
FMD focus area of
3-years project (EuFMD)
FMD control program
Iran West Azerbaijan
Field application
Objectives
1. To estimate prevalence of epi-units with indication of recent FMD infection
2. To identify putative risk factors
3. To use results in
targeted FMD control
4. To monitor impact of FMD
control program over time
NSP sero-survey PC Practitioner Network 41
Field application
Animal
Infected with FMD virus
SP antibodies
NSP antibodies
Vaccinated with purified vaccine
SP antibodies only
Vaccinated with
impure Vaccine
SP antibodies
NSP antibodies?
Background: FMD structural proteins (SP) and non-structural proteins (NSP)
42
Field application
ThereforeNSP serosurvey is an accepted way to measure
previous FMD infection, even in vaccinated populations, providing the vaccine is purified However, the vaccine used in Iran is locally
produced and impure. How does this affect the interpretation of NSP sero-survey results?
43
Field application
Materials & Methods I
2-stage sampling
Assumptions: 1. 35% epi units infected (+/- 5%, 95% confidence)
300 epi-units sampled randomly from a complete list from 4 production type
– 200 villages, 40 dairy farms, 30 beef farms,
30 farms with cattle and small ruminants
2. Within epi-unit: 10% animals infected
30 head of livestock within each epi-unit sampled
Sample youngstock (6-24 mo), NSP Ab is reflection of recent infection(0-2 vaccinations)
NSP sero-survey PC Practitioner Network 44
Field application
Materials & Methods II
All 17 districts
Questionnaire survey
– Animal
– Household (villages)
– Epi-unit characteristics
Provincial laboratory with trained staff, use PrioCheck NSP-antibody ELISA
NSP sero-survey PC Practitioner Network 45
Field application
Preparing for field
First Step
- Provincial manager of sero-survey selected and trained in Iran Veterinary Organization(IVO) under FAO- EuFMD Epidemiologists to implement sero-survey
-Preparing a preliminary questionnaire from previous experiences of FAO and EuFMD in different countries (specially Egypt) for Iran
-Sharing preliminary questionnaire with provincial experienced veterinarians and improve it many times
-Sharing and pretest and improve preliminary questionnaire with farmers
-Prepare second questionnaire that was ready to use in the field in pretest district(Salmas) in north part of province
-NSP sero-survey PC Practitioner Network 46
Field application
Pretest(Questionnaire and Sampling)
Salmas district
NSP sero-survey PC Practitioner Network 48
Field application
Questionnaire– Data – 3 Level – Animal level
Animal level Informationمورد خونگیری داممشخصات
districtpoldashtنام شهرستان کد واحد
code
3216012
9
epunitنام واحد اپیدمیولوژیک
nameoroj mohamad
row numberردیف
_ نام دامدار
farmer
name
شماره گوش
گاو در
صورت وجود
ear tag
number (if
exist)
) سن دام به ماه
ط خونگیری فق
تا 6از دامهای
ماهه انجام 24
age in( شود
months
نر )جنس دام
= ماده / 0=
1)sex(ma
le=0 /
1=female
)
سن دام هنگام
از شیر گیری
ageبه ماه
at
weaning
in months
آیا دام در این روستا یا دامداری
و بلی 0=خیر )متولد شده است؟
، بلی همین شهرستان1=همین واحد
, (3=، شهرستانهای دیگر 2=
yes No=0 in this epi-unit
yes=1, in this district =2 ,
other districts = 3
)نوع استفاده از دام
گوشتی/ 0=شیری
=1 )purpose
(dairy=0/beef=
1)
آیا به تب برفکی در یک
ت؟ سال گذشته مبتلا شده اس
( 0= خیر /1= بلی )
Clinical Signs of
FMD observed
(Y=1/N=0, if yes:
when?
WA_ poldasht_1 2404310نورالدین شجاعی
WA_ poldasht_22414300نورالدین شجاعی
WA_ poldasht_32414300نورالدین شجاعی
WA_ poldasht_42414300نورالدین شجاعی
WA_ poldasht_52415300نورالدین شجاعی
WA_ poldasht_61214100محمود آقازاده
WA_ poldasht_71514000محمود آقازاده
WA_ poldasht_81514000محمود آقازاده
WA_ poldasht_91214200محمود آقازاده
WA_ poldasht_102414300یونس آقازاده
WA_ poldasht_111514300یونس آقازاده
WA_ poldasht_121204310یونس آقازاده
NSP sero-survey PC Practitioner Network 49
Field application
Questionnaire– Data – 3 Level – Farmer level
Farmer level informationاطلاعات دامدار Farm /Houshold informationیا خانه ای که خونگیری می شود؟ ( شیری یا گوشتی )مشخصات دامداری
نام
شهرست
ان
Name
of
district
poldasht
کد واحد
epiunit
code
32160129نام واحد اپیدمیولوژیک
epiunit name oroj mohamad
ردیف
row
numb
er
نام دامدار
_
farmer
name
تاریخ نمونه
Dateبرداری
of Sampling
تعداد گاو
و گاومیش
موجود در
زمان
خونگیری
numbe
r of
cattle,
د و تعداد گوسفن
بز موجود در
زمان خونگیری
number
of,
_other
ruminants
کود یک سال گذشته
دامهای خود را چکار
خودم استفاده)کردید؟
، می 0=می کنم
= ، هر دو 1=فروشم
2 )Manure
treatment (Own
use=0 , SELL =1,
Both =2)
خرید و فروش دام های
ماه 12دامداری در
= ا بلی در روست) گذشته
، بلی در میدان دام 1
=، بلی سایر موارد 2=
( 0= ، خیر 3
Trading of
Animals in Last 12
monthes-No=0,
YES in this
village=1, in
animal market=2 ,
yes other places ,
no=3
آیا دام دارای چرای خارج
از خانه یا دامداری است؟
grazed outdoors خیر
1=بلی در همین واحد 0=
ا ، بلی در مرتع مشترک ب
2= روستای های همجوار
، ( 3=، بلی در سایر جاها
No=0,yes in this ep
uint=1 , common
pasture=2 other
villages=3 , other
places = 4
دام معمولا در کجا نگهداشته می
شود؟ همیشه داخل دامداری یا
، گاهی خانه و گاهی 0=خانه
، در 1=در نزدیکی روستا
( 2=مناطق دور از روستا
the animal is always
kept within the
compound (0), within
the epi-unit (1), or has
been outside the epi-
unit (2)
آیا علائم بیماری تب
برفکی در دامداری یا
خانه مورد خونگیری
ماه گذشته دیده 12در
Clinicalشده است؟
FMD signsin last
12 monthes
outbreak seen
(yes=1/no=0)
آیا علاوه بر گاو ،
گوسفند و بز هم در
خانه یا دامداری
نگاهداری می شود؟
0= ، خیر 1= بلی
If there are
sheep/goats in
the same
compound
yes=1 and No
=0
1نورالدین
شجاعی1390/04/0790012200
0
1390/04/076010200محمود آقازاده21
1390/04/0751000200یونس آقازاده31
1390/04/076001200رجابر پاشاپو40
NSP sero-survey PC Practitioner Network 50
Field application
Questionnaire – Data – 3 Level – Epiunit level
Ep unit level informationاطلاعات واحد اپیدمیولوژیک
نام
شهرست
ان
distric
t
name
نام واحد
Epiunit
Name
کد واحد
Epiunit
Code
علائم تب
برفکی در
ه یکسال گذشت
این واحد
گزارش شده
= بلی)است ؟
شته.تاریخ نو
0=خیر/ شود
Clinical
FMD
outbreak
seen
(yes= Dat
e should
inter /no=0)
تاریخ آخرین مایه
در کوبی تب برفکی
Lastواحد ؟
FMD
vaccination
date
ه تاریخ مای
ل کوبی راپ
در آخرین
فاز تب
برفکی؟
Last
Booster
Vaccin
ation
Date
ه تاریخ مایه کوبی ک
قبل از آخرین مایه
کوبی انجام شده
FMDاست؟
vaccination
date before
last
vaccination
Type ofنوع واحد
Epiunit ، 1=روستا )
، 2= گاوداری شیری
گروه گاوداری پرواری
ام و پرواری با شیری تو
، گروه دامداری 3=
د های دارای گاو و گوسفن
vilage=1( 4= وبز
, dairy farm=2 ,
beef farms =3 ,
smal ruminants=4
تعداد گاو و
گاومیش در
زمان
خونگیری
Number of
cattle and
calves
فاصله از
دان نزدیکترین می
دام به کیلومتر
distance to
closest
animal
market (in
km)
ند تعداد گوسف
و بز در زمان
خونگیری
Number
of sheep
and goat
نام نوع
اده واکسن استف
شده در این
تترا ) واحد
، 0= والان
= منووالان
1)type of
vaccine
used
0=Monov
alent=1/T
etravalent
نام شرکت
سازنده
واکسن
استفاده شده
در این واحد
= رازی )
= مریال / 0
1 )
0=Merial
=1/Razi
poldashtoroj
mohamad32160129
در گوسفند دیده
1389/05/17شده1389/12/031389/10/21111225520
0
1
1
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Field application
Combining Field survey data with Lab Reported Data
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Field application
Implementing of survey in the field
Second Step
-A coordination session with all colleagues(provincial and district representatives) in this survey and explanation them all details
-Holding other coordination sessions in every district veterinary office among field groups
-Starting of questionnaire field survey and blood sampling in the same time in different epi-units in pre-defined dates
-All blood samples transferred to district veterinary offices and sera prepared in same day
-All sera labeled and checked very carefully due to hemolysis (often 2-4 more sera sampled)
-All samples (with questionnaires) were sent to provincial veterinary office in 24-48 under cold chain(2-8°C)
-
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Field application
Lab test and combine data
Third Step
-There was a responsible man in provincial veterinary office to take over samples and questionnaires and check them
-All samples sent to laboratory of provincial veterinary office and stored in proper temperature
-All samples were tested with a trained man with NSP test kits(Priocheck)
-All lab results were sent to survey manager and combined with field data and all missed data revised with district representatives.
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Field application
Providing final dataset
Fourth Step
-All epi-units data combined together in every district
-All district data combined together
-One final data(one excel file including 3 level data(epi-unit-farmer-animal) developed
-Final file was sent to FAO-EuFMD epidemiologist
-All data checked many times to delete errors and correct missed data
-Final dataset prepared and analyzed by epidemiologist
-
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Field application
Descriptive results
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• 281 epi units
• 1798 different owners
• 8378 animals were sampled
Po
sitiv
e
Ne
ga
tive
• Manufacturer recommended 50%
inhibition as cut off
• Graph shows distribution of titers
• 53.7% of all samples positive for
NSP-Ab
• 279 epi-units with 1 or more
seropositive cattle (˃99% positive epi-units)
• 6 epi-units with all 30 cattle
seropositive
50 % cut
off as
reference
line
Field application
540 Samples retested at lab in Brescia – ItalyIstituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna
(IZSLER): FAO collaborating laboratory for FMD NSP ELISA
Cedi test
ELISA
IZSLER ELISA
Positive Negative Total
Positive 227 61 288
Negative 18 234 252
Total 245 295 54057
Compared with IZSLER Elisa
• Agreement 85%
• Kappa 0.71
- Rel. Se. 92.6%
- Rel. Sp. 79.3%
- Higher cut off (60-75% inhibition),
higher agreement(90%)
Higher
cut off
Field application
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Field application
Available on our PC Practitioner site areSupporting documentation and publications
Kees van MaanenEuFMD component manager MENA and Laboratory networs
Javad EmamiProvincial Veterinary Officer, West Azarbaijan, Iran
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Chris BartelsEuFMD component
manager PCP
Wilmot ChikurunheShort-term professional Southern Africa
Objective Field
application
Design Laboratory issues
and result
interpretation
Jenny
MaudTraining ProgrammeManager
Practicing the
Network
Introductions
IAH
FMD antibody detectionObjectives
– confirmation of infection, particularly subclinical infection– retrospective diagnosis when epithelium not available e.g. after
resolution of acute infection– import/export– during epidemiological surveys
Screening by ELISA– rapid– Sensitive
Confirmation by VNT– 'Gold standard' for international trade
Laboratory aspects
Which Antibodies can be found in animals that have been in contact with FMDV ??
Which information do they provide us ?
Which tests fit for different purposes ?
FMD virus structure : mosaic of many different antigensStructural Proteins = SP Non Structural Proteins = NSP
FMD Non-structural and Structural Proteins
Laboratory aspects
O A C Asia 1 SAT 1 SAT 2 SAT 3
anti-SP Antibodies serotype-specificSeven different assays, one for each FMD virus type
anti-NSP antibodies identical for 7 serotypesO / A / C / Asia 1 / SAT 1 / SAT 2 / SAT 3NSP tests A unique assay for all FMD virus types
Laboratory aspects
NSP antibody detection ELISAs1. Test principles
• Commercially available test kits
• Indirect ELISAs versus blocking versus competition ELISAs
• Advantages and disadvantages
2. Test characteristics• Test validation
• Technical information manufacturer
• Independent/peer reviewed information
• Sensitivity (in vaccinated and in non vaccinated animals)
• Specificity (in vaccinated and in non vaccinated animals)
• Repeatability and reproducibility
• Positive and negative predictive values
3. Interpretation of test results, validity of data, pitfalls
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Laboratory aspects
Available on our PC Practitioners’ site areExercises
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Exercises available
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NSP Sero-survey
Exercise 1
Estimating individual animal prevalence, herd prevalence and within-
herd prevalence.
In the datasheet “NSP sero-survey_exercise 1” you find the result of
5000 samples collected as part of the NSP sero-survey explained by Dr
Javad Emami, Urmia, Iran.
In this dataset we have copied information on
The questions to answer are the following:
1) What is the estimated animal prevalence, and the 95%
confidence interval?
Available on our PC Practitioners’ site areExercises
Exercises available
In summary
During this session, we discussed
• NSP sero-survey complements information about outbreak notifications
• It allows for assessing the extent of virus circulation and putative risk factors for infection
• A proper design makes inference of results possible• use in developing a Risk-based strategy
• tool to monitor the impact of such strategy
• Implementation and organisation• Develop procedures in consultation with people involved in particular with
fieldstaff responsible for bleeding and questionnaire as it will not be
possible to repeat it
• Consider entry of data be performed centrally to safeguard consistency
• Perform data validation thoroughly and double check with enumerators
• Laboratory aspects
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Summary
Kees van MaanenEuFMD component manager MENA and Laboratory networs
Javad EmamiProvincial Veterinary Officer, West Azarbaijan, Iran
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Chris BartelsEuFMD component
manager PCP
Wilmot ChikurunheShort-term professional Southern Africa
Objective Field
application
Design Laboratory issues
and result
interpretation
Jenny
MaudTraining ProgrammeManager
Practicing the
Network
NSP sero-survey PC Practitioner Network 71
Getting started
https://eufmd.rvc.ac.uk
Look out for new
features!
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Getting started
Track your progress with course completion
Complete your profile
Discussion forum:• Ask and answer questions• Notification of new
resources
Select training and resources that meet your needs
Online quizzes allow us to track your progress
Receive recognition for the training you undertake
Each network activity is associated with credits, 1 credit = 1 hour’s training time
Recognition for number of credits accumulated
– Certificate
– Bronze, Silver and Gold levels
Getting started
To get started:
E-mail [email protected] with:
Your full name
Location
Brief background
OR visit https://eufmd.rvc.ac.uk and follow links from the homepage.
Share this opportunity with your colleagues
NSP sero-survey PC Practitioner Network 74