Foot-and-mouth disease is a highly contagious viral disease of livestock that causes severe economic losses. Implementing effective surveillance and control programs is important for preventing outbreaks. Rapid testing for antibodies to non-structural proteins allows for quick, on-site screening to identify infected animals, establish infection perimeters, and monitor imported livestock, helping refine Kerala's control program and work towards eradicating the disease.

1. FOOT-AND-MOUTH DISEASE: IMPLEMENTING AN EFFECTIVE DISEASE CONTROL
PROGRAMS AND ROLE OF NSP ANTIBODY TESTING
V I Bishor and Faisal Siyavudeen
ubio Biotechnology Systems Pvt. Ltd
Technology Incubation Center, Kinfra Hitech Park,
Kalamassery, Kerala, India.
Foot and Mouth disease: Overview
Foot and Mouth Disease (FMD) is a severe, highly contagious viral disease of livestock
with significant economic impact. The disease causes severe production losses and while
the majority of affected animals recover, the disease often leaves them weakened and
debilitated, with severe reduction in milk production. In a susceptible population,
morbidity approaches 100% and intensively reared animals are more susceptible to the
disease than traditional breeds. While the disease is rarely fatal in adult animals, there is
high mortality in young animals due to myocarditis or by lack of milk when the dam is
infected by the disease. Because of the huge economic impact, FMD is enlisted in the
World Organisation for Animal Health (OIE) Terrestrial Animal Health Code, 2009, and
world-wide guidelines are in place for detection and control of the disease.
FMD virus is found in all excretions and secretions from an infected animal. The virus
may be present in milk and semen for up to 4 days before the animal shows clinical
signs of disease. Infected animals notably breathe out a large amount of aerosolized
virus, which can infect other animals via the respiratory or oral routes. All these factors
make the virus spread very fast among populations. Animals that have recovered from
infection also serve as carriers of the virus.
The key elements the strategy for dealing with FMD are early detection, warning, and
prevention. These include continuous monitoring of FMD occurrence and prevalence,
and characterisation of FMD viruses. Protection of FMD- free areas should be enhanced
with stringent controls and surveillance on import and cross-border animal movement.
The recommended control measures include controlling introduction of new animals to
existing stock, monitoring and immediate reporting of illness and sound bio-security
practices at farm level.
According to OIE, A good FMD surveillance system should be under the responsibility of
the Government Veterinary Authority, as is done in Kerala today. A procedure should be
in place for rapid collection and transport of samples, and subsequent testing and
diagnosis. This would ensure that disease occurrence is detected as soon as possible,
and quarantine measures initiated quickly to avoid outbreaks. The surveillance systems
should also include an early warning system throughout the production, marketing and
processing chain for reporting suspicious cases. Farmers and workers who have day-
today contact with livestock, veterinarians and other technical staff should report
promptly any suspicion of FMD. They should be supported directly or indirectly (e.g.
through private veterinarians or veterinary para-professionals) through government
information programmes and the Veterinary Authority. All suspect cases of FMD should

2. be investigated immediately. Where suspicion cannot be resolved by epidemiological
and clinical investigation, samples should be taken and tested immediately. The
surveillance program should also implement regular and frequent clinical inspection and
serological testing of high-risk groups of animals, such as those adjacent to an FMD
infected area.
Foot and Mouth Disease Control in Kerala
Kerala’s Animal Disease Control Project (ADCP) is lauded as the model for the entire
country. The vaccination program conducted as part of ADCP has led to significant
reduction in the occurrence of FMD in the state. Continuous monitoring and re-
vaccinations have kept the disease away from the state for years altogether. However,
we have seen increased incidence of FMD outbreaks in recent years and this points to
new influential factors that have come into play. These factors include:
1. Increased movement of slaughter animals into the state across the border without
any control.
2. Introduction of new dairy animals from infected areas into the state, especially as
part of government programs targeted at ensuring increase of livestock population.
3. Lack of continuous disease monitoring among livestock populations in the state that
slows down outbreak detection, emergency response and epidemic prevention.
The FMD control program has to be refined by incorporating control measures that
consider these new factors, so that we sustain and improve the FMD disease status
across the state.
FMD Disease control in Kerala: Proposed measures.
The following measures are to be implemented to refine the FMD disease control
program in Kerala.
1. Clinical Surveillance
2. Migration Control and Monitoring*
3. Control of cross-border livestock trade
4. Early detection measures and epidemic control
Clinical surveillance
Even though we have implemented a successful routine vaccination program, we do not
have a continuous monitoring system to identify virus circulation or silent carriers in the
population. A major outbreak is often followed by a series of smaller outbreaks caused
by previously infected carriers, and lack of identification of such carriers makes epidemic
control less effective. To mitigate these risks, a separate monitoring program should be
put in place in addition to the vaccination efficacy screening. Such a program would
involve:
1. Regular random sampling of serum from across the state
2. Testing the sample for the presence of infection
3. Identification of infected carriers or new infections
4. Quarantine and control measures including emergency vaccination
Migration Control and Monitoring
The State Government provides financial incentives for purchase of new livestock
animals under various schemes. However, these incentives have been utilized more for
intra-state trade, without any increase in livestock population. To ensure that financial

3. incentives result in increased livestock populations, the government has started insisting
that such incentives will be available only for import of animals from other states. While
this is a welcome development from a demographical point of view, it has a serious
negative impact on disease control programs. Most areas adjoining our state have
higher incidence of FMD with less developed disease control systems, there is a high
chance that FMD carriers may be imported through inter-state purchase of new dairy
animals. A program where every new dairy animal imported into the state under
government schemes has to be tested and certified free of FMD before the purchase is
made has to be considered.
Control of Cross-Border livestock trade.
The number of slaughter animals imported into the state through our border check-
posts is huge. These animals do not mingle with local livestock, and they spend just a
few hours in the state before they are slaughtered, which makes the risk of infections
from these animals lesser in a general sense. However, this does not hold for FMD, as
the disease is so contagious that even animals that live near the road through which
infected animals are transported are at risk of infection.
As the number of slaughter animals imported is large, the cumulative risk is very high. In
fact, there is strong suspicion that the recent outbreaks at various places in Kerala were
caused by infected slaughter animals. A screening program may be implemented at
border check-posts to screen and exclude infected animals. Ideally, every animal should
be checked for infection and the physical or financial responsibility to dispose of the
infected animal should be fixed on the importer. The tagging system at the check-posts
should be extended to retain test data and associate it with individual animals. A good
vigilance program that randomly screens animals at slaughter-houses can then ensure
accountability and assure efficacy of the screening program by cross-checking results
from the check-post.
Early detection measures and epidemic control
Emergency response is a critical part of epidemic control. Today, treatment is focused
on areas where outbreaks are reported, and treatment often follows the spreading
outbreak. However, the most difficult aspect of an epidemic is its fast spread which
makes control problematic, and more attention must be given to containment. By the
time an FMD infection is detected in an animal, the disease would be incubating in all
the animals in direct contact with the infected animal. The most important step in FMD
epidemic control is to quickly establish an infection perimeter. Today, this perimeter is
established empirically. We propose that all susceptible animals in infected and
adjoining areas and areas with high risk of contagion be screened to identify early
infections whenever an epidemic is reported. This data should be used to establish the
infection perimeter quickly and accurately. This perimeter should then be used to
implement quarantine measures in the infected area, and emergency vaccination should
be performed around the perimeter to block spread, as is done today. Of course, this
should be followed by intensive supportive therapy for infected animals, thereby
containing the outbreak quickly. Once the epidemic is over, all the animals in the area
should be screened again for the presence of carriers so that follow-up outbreaks do not
occur.

4. FMD control measures: implementing Challenges.
While the control measures proposed above can lead to vast improvements in our FMD
control program, there are specific challenges that must be overcome before they can
be effectively implemented. A clinical surveillance scheme that aims to detect silent
FMD carriers cannot rely on clinical examination, and would require laboratory
diagnosis. Today, the basic laboratory facility for FMD diagnosis is available only at one
centre in Kerala – the CDIO/IAH&VB labs at Palode. It is not practical for daily random
samples to be sent to a single center within the state. The sample management problem
cannot be solved even if district-wide diagnosis facilities are established, as considerable
manpower would be required for serum transport and lab analysis. This option would
also entail high capital expenditure in the form of additional equipment for serum
handling, storage and lab analysis.
Migration control and monitoring programs that require ‘FMD-free’ certification of dairy
animals before purchase would need a testing method, which would allow the
veterinary doctor to test the animal at the point of purchase. It is not possible for the
doctor to collect the serum of the animal to be purchased, then send it to a lab for
testing and close the deal after the result is available, as the turnaround time will be in
days. A mobile lab that accompanies the doctor is one option, but the cost is prohibitive.
Similar challenges also exist for check-post screening. A typical animal carriage vehicle
will spend 30-45 minutes at the check-post for animal tagging and clearance. It is
impossible to screen all the animals in the carriage within this time using conventional
methods. Serum collection and off-site lab diagnosis is not an option here, as infected
animals would have carried the infection into the state by the time the results are
available.
In the case of disease screening to establish infection perimeters, quick results are very
important as the disease spreads by the minute. A 2-3 day delay in obtaining results
would make the test data almost useless as the disease perimeter would have morphed
and expanded by that time.
In all these measures, it is important to make sure that vaccinated, non-infected animals
are not judged as infected by looking at the presence of FMD antibodies in their serum,
and that vaccinated silent carriers are identified. Screening of antibodies of Non
structural protein is a simple way of differentiating the infected and vaccinated animals.
NSP antibodies can be monitored by Enzyme Linked Immunosorbent assay technique. A
recently introduced lateral flow assay test for NSP antibodies offers much easier and
portable way of screening these antibodies in bovine serum sample.
Rapid FMD NSP Testing
All the FMD control measures outlined above involve fast, reliable, de-centralized
screening of the disease. The implementation challenges outlined in the previous
section can be used to derive a set of requirements for the ideal diagnostic technique to
be used in the FMD disease screening. They are:
1. Quick results should available in just a few minutes.
2. Should require no instrumentation, instruments should be portable if present.
3. Should be easy to use.
4. Should allow decentralized implementation (should not require serum transport)

5. 5. Should require no, or low, capital expenditure.
6. Should be able to execute the test and read result in the field itself.
7. Should not require refrigerated storage.
8. Should differentiate between vaccinated and infected animals.
The only diagnostic technique available today that satisfies these requirements for FMD
diagnosis is the FMD Non-structural-protein (NSP) rapid lateral flow assay. Various FMD
control measures can be effectively implemented by using this kind of assay for
screening.
Assay Overview and Usage
Rapid FMD NSP test (available as ‘ubio quickVET FMD NSP’) uses recombinant FMD NSP
antigens and monoclonal antibodies to detect the disease. Antibodies to NSP antigen
are present only at the time of infection, and these antibodies are not type-specific.
Hence, the presence of these antibodies indicates the presence of FMD infection,
irrespective of the virus type. FMD NSP rapid test is designed so that even a technician
with limited expertise can perform the test easily. The veterinarian or the technician
draws blood from the animal using a syringe or a vacutainer, optionally coated with a
clot-activator for faster serum separation. The syringe is kept till the serum separates,
and then, 3 drops of serum are added to the sample hole on the test card using a
dropper. The results can be read in just 5-10 minutes. After the test is complete and
result recorded, the used kits should be disposed of according to standard disposal
procedure followed for clinical specimens in veterinary hospitals and labs.
Conclusion
Kerala’s FMD control program can be refined and disease outbreaks can be prevented
by implementing effective screening programs. These programs require easy, rapid tests
which can be executed on site, without access to lab facilities. FMD NSP lateral flow
assay provides a perfect tool that would allow us to implement such screening programs
Efficiently thus making the state FMD free.
Address for correspondence
Dr. V I Bishor. Phone: +91 9744122269, e mail: bishor@ubio.in