D5674.PDF

Conf. OIE 2000, 263-270
SWINE VESICULAR DISEASE:
INCIDENCE, PATHOGENICITY, EPIDEMIOLOGY, DIAGNOSIS, ECONOMIC IMPACT
Kris De Clercq
Veterinary and Agrochemical Research Centre, Groeselenberg 99, B-1180 Brussels (Ukkel), Belgium.
Original: English
Summary: Swine vesicular disease (SVD) is a contagious viral disease of pigs classified as a List A
disease of the Office international des Epizooties (OIE). SVD may be indistinguishable from foot and
mouth disease (FMD). Differential diagnosis between FMD, vesicular stomatitis and SVD poses no
problem anymore in the laboratory. Strains of SVDV vary in virulence, and the disease may be
subclinical, mild or severe. SVD is a pen disease that spreads by direct contact between pigs or indirectly
by contact with contaminated faeces and by swill feeding. The resistance of the virus to inactivation may
lead to recrudescence. The disease flamed up again in 1992 in the Netherlands and spread to several
other European countries. SVD is still present in Italy, particular in an endemic way in the south of the
country. The costs of control measures and trade restrictions after a SVD outbreak can be very high.
Singleton reactors cause a lot of trouble in serological testing and therefore in international trade. The
questionnaire sent to OIE Member Countries revealed that 80% of the countries are convinced that SVD
should be maintained on the OIE List A. However, some countries added the remark that the OIE Lists
should be reviewed. Seventy-percent think that SVDV will spread across Europe after removal of SVD
from List A. The majority also predict that at such time FMD identification in the field would be delayed.
Therefore, (clinical) surveillance programmes should be maintained. The majority of the Member
Countries consider a herd as an outbreak of SVD only when SVDV is isolated. Stamping-out of the whole
herd is then implemented, whether clinical signs are present or not. A protection/surveillance zone must
be established and 60% think that serosurveillance should be implemented for at least a year. The
economic consequences of a SVD outbreak for the pig sector are considered to be very high. Half of the
Member Countries think that these consequences are higher than necessary.
1. INTRODUCTION
Swine vesicular disease (SVD) is a contagious viral disease of pigs classified as a List A disease of the Office
international des Epizooties (OIE). SVD may be indistinguishable from foot and mouth disease (FMD) and is thus
important in the differential diagnosis of the latter disease.
Swine vesicular disease virus (SVDV) is a member of the enterovirus genus of the family Picornaviridae. SVDV is
very resistant to environmental factors and many commonly used disinfectants (14), a fact of considerable
epidemiological significance. SVDV can retain infectivity in contaminated faeces for more than four months. The virus
is resistant to fermentation and smoking processes. The virus remains infectious for months in carcasses and processed
meat (ham, dried sausages, processed intestinal casings, etc.).
2. INCIDENCE
In Europe, the disease was first observed in Italy in 1966 (25). It spread to several other European countries beginning
in the 1970s (1971-72: Austria, Bulgaria, Italy, Great Britain and Poland) and lasted until the beginning of the 1980s.
After that period, outbreaks were sporadic. The disease was almost forgotten until it flamed up again in 1992 in the
Netherlands. Once again it spread to several other European countries such as Belgium, Portugal and Spain. In that
period it became clear that SVD was also present in Italy, particular in an endemic way in the south of the country. In
Europe there are two genetically and antigenically distinct variants (4). The year of the last appearance of the disease is
presented in Table 1. As shown the virus has also been identified in samples from some Asian countries or provinces.
Some SVD viruses found in Europe were closely related to the Asian SVD viruses (27).
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Table 1: Year of the last appearance of
swine vesicular disease outbreaks. Based on Dekker (2000)
Europe
Italy
Portugal
Netherlands
Belgium
Spain
France
Germany
Romania
United Kingdom
Austria
Greece
Malta
Ukraine
Russia
Switzerland
Poland
Bulgaria
Year of last
appearance
2000
1995
1994
1993
1993
1983
1985
1985
1982
1979
1979
1978
1977
1975
1975
1972
1971
Outside Europe
Taipei China
Lebanon
Bolivia
Hong Kong
Laos
Macau
Nicaragua
Korea
Japan
Year of last
appearance
1998
1992
1991
1991
1991
1989
1986
1980
1975
Recently only Taipei China and Italy have reported outbreaks of SVD, but the virus is probably present in other parts of
the world. Because SVD generally does not cause serious problems, the disease is most likely under-reported. FMD
outbreaks in Italy (1993) and Taipei China (1997) coincided with increased reports of SVD diagnosis. This suggests
that farmers, who are familiar with the symptoms of SVD, do not report the disease until they think it might be FMD
(8).
3. PATHOGENICITY
Clinical disease after a SVD infection is restricted to pigs. SVDV enters the pig through the mucosa of the digestive
tract or through skin lesions (19, 22). The incubation period of SVD is between two and seven days, and a transient
fever of up to 41°C may occur. Strains of SVDV vary in virulence, and the disease may be subclinical, mild or severe.
The latter is usually only seen when pigs are housed on a concrete floor in humid conditions (14, 16, 18). Pigs
introduced into an environment contaminated with SVDV can already become viraemic after one day and serologically
positive after three to four days (IgM) (9). Vesicles appear principally around the coronary bands of the feet, including
those of the accessory digits. These may affect the whole coronary band, resulting in loss of the hoof. The vesicles may
extend to the skin of the lower limbs. Affected pigs may be lame and off their feed for a few days. Vesicles occasionally
develop on the snout, but seldom on the tongue. In lactating sows, lesions on the udder and teats can also be seen.
Sudden death, due to heart degeneration, which is often observed in young piglets affected with FMD, is not seen in
SVD. Recovery is usually complete within two to three weeks, with the only evidence of infection being a dark,
horizontal line on the hoofs where growth has been temporarily interrupted. Severe clinical signs of SVD are
indistinguishable from FMD, vesicular stomatitis (VS) or vesicular exanthema. Recent European outbreaks of SVD
have been characterised by less severe or no clinical signs, diagnosis frequently being dependent on serology.
4. EPIDEMIOLOGY
Affected pigs may excrete virus from the nose and mouth and in the faeces up to 48 hours before the onset of clinical
signs. Most virus is produced in the first seven days after infection, and production from the nose and mouth will have
stopped after two weeks, but virus may continue to be shed for up to three months in the faeces (1). Airborne spread of
the virus is not significant. The disease is spread by direct contact between pigs or indirectly by contact with
contaminated faeces (in the pig house, in markets or on contaminated vehicles) and by swill feeding (12). Contact with
a SVD-infected environment has been shown to be as infectious as direct contact with SVD infected pigs, causing rapid
spread of the disease (9). Minor skin trauma caused, for example, by walking on concrete or by transport, can have a
significant bearing on the severity of the disease. In many pigs infection is mild enough to go unnoticed by farmers and
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the movement of such infected pigs (through markets and collecting centres) is important in spreading the disease to
new farms.
The resistance of the virus to inactivation not only favours indirect spread by fomites but may also lead to
recrudescence of the disease when farms are restocked after inadequate disinfection. Feeding pigs insufficiently cooked
swill is an important method of dissemination of the disease. Findings in Europe indicating that SVDV is related to
Asian strains show the epidemiological importance of using food from airplanes.
Although infection usually spreads rapidly in groups of pigs in direct contact, spread from pen to pen on a farm is rather
slow (14) and may fade out within the pig farm. SVDV carriers are very uncommon. Only in one experimental study
could SVDV be recovered as late as 126 days (21).
In most List A diseases clinical signs will be evident in case of infection. In those cases, continuous serosurveillance
will not shorten the time between introduction of the agent and detection (High Risk Period). It is therefore not useful as
an early warning system. In the case of SVD, clinical disease may go unnoticed. Continuous serosurveillance might
therefore be effective. However, as in the case of classical swine fever (6, 15), it has been concluded that for SVD the
serosurveillance programme has not contributed to early detection (8).
5. DIAGNOSIS
Since FMD may be suspected in the presence of any vesicular condition in pigs, special precautions must be taken for
the safe packaging of suspected samples (17). Laboratory diagnosis of clinical cases of all the vesicular virus diseases
should be conducted within biosecure facilities. When the presence of SVDV in a holding is suspected, appropriate
samples from representative groups of pigs must be collected for disease confirmation and differential diagnosis.
Preferred samples are epithelium and vesicular fluid. Blood samples from the suspected pigs and in-contact pigs must
be taken for serological testing. Faecal samples from these pigs and from the floor of their pen are necessary.
Clarified suspensions of samples must be inoculated onto sensitive cell cultures, such as IBRS-2 cells, SK6 or PK-15, to
isolate and grow the virus. If a cytopathic effect (CPE) develops, the supernatant fluid must be used in an ELISA
(enzyme-linked immunosorbent assay) for virus identification. A blind passage is performed on negative cultures. In the
absence of CPE after a further blind passage, the sample can be declared negative for the presence of live virus.
An indirect sandwich ELISA or a monoclonal antibody-based ELISA may be used for SVD antigen detection and for
differentiating FMD, VS and SVD. This is performed directly on the clarified sample or after the virus has grown on
cell culture.
Nucleid acid recognition methods can be used to detect SVD viral genome using PCR, to differentiate between FMD,
VS and SVD, and to establish relationships between isolates of SVDV by nucleotide sequencing. The PCR is rapid and
sufficiently sensitive. Where sub-clinical infection is suspected, or when samples are collected after the resolution of
clinical disease or when processing faecal samples, enhanced RT-PCR technique (5, 11, 20) provides a detection system
as sensitive and considerably more rapid than multiple passages on tissue culture. The immune-PCR (11) technique was
extensively validated and proved more sensitive than virus isolation, particularly for faecal samples.
FMD-VS-SVD differential diagnosis no longer poses a problem in the laboratory.
Serosurveillance can be carried out on a non-discriminatory basis in the absence of SVD or on holdings in the
protection/surveillance zone and on in-contact animals after an outbreak. Blood samples must be collected for
serological tests.
The virus neutralisation test (VNT) (10, 13) is the reference test to detect antibodies against SVDV. It has the
disadvantage that it takes two to three days to complete, requires cell culture facilities and is performed with live virus.
Antibodies can also be detected by a monoclonal antibody-based competitive ELISA (3). An indirect trapping ELISA
using isotype-specific monoclonal antibodies to detect swine IgM or IgG specific for swine vesicular disease virus is
helpful in assessing the time of infection in the pig or in the infected premise.
Singleton reactors cause a lot of trouble in serological testing and therefore in international trade. A singleton reactor
(SR) is a single seropositive pig in a holding, which yields a positive result in serological tests for SVD. The pig has,
however, no history of contact with SVDV. There is no evidence of spread of infection to in-contact pigs from this pig.
The prevalence of SR is approximately 1 per 1000 pigs. The number of SR decreases considerably when the serological
profile is determined by combining the results of the three serological tests mentioned (7). Alternatively, the pig must
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be tested a second time together with neighbouring pigs. The SR phenomenon is most probably not caused by another
enterovirus but by a purely aspecific IgM reaction.
6. ECONOMIC IMPACT
If no stamping-out is implemented, economic loss associated with a severe case of SVD on the farm level would be due
to loss of weight and delay in finishing fattening pigs. The cost due to secondary bacterial infections and their treatment
should also be taken into account. This can be difficult in a system of industrialisation with low profit per capita. Loss
of suckling piglets can occur if the udder of the lactating sow is affected.
Mild and subclinical cases are often not noticed by the pig holders or veterinarians. A large number of pigs might
become infected and the disease may spread to other farms and collecting centres. These forms cause little direct
economic loss. As the virus is quite resistant to disinfectants, it is very difficult to get rid of it. The disease, however,
can turn into the severe form through environmental factors or change in the virulence of the virus.
Severe clinical signs of SVD are indistinguishable from FMD, vesicular stomatitis or vesicular exanthema. Therefore, it
is generally accepted that SVD could not be tolerated in countries free from these diseases. For this reason, SVD was
placed on the OIE List A. SVD is therefore a notifiable disease. In case of an outbreak in the European Union (EU),
SVD is strictly controlled by stamping-out and restricting livestock movement. Several cases of recurring infection after
stamping-out and disinfection have been reported.
Pig holdings found to be serologically positive (virus isolation negative) during serosurveillance and with an
epidemiological link to a SVD outbreak are treated in the same way in the EU as an SVD outbreak with positive virus
isolation.
The use of inactivated SVD vaccines has been studied (23, 24), but no data have yet been published about their use in
the field.
The costs of control measures and trade restrictions can be very high. After the SVD outbreak in 1993 in the
Netherlands, the export of pigs was blocked for one month. The cost due to loss of export was estimated to be
approximately USD 18 million (26). To prove freedom of SVD after the outbreak, the Netherlands has adopted a
serosurveillance programme, in which farms are clinically examined and serologically tested three times a year.
Currently, the cost of the SVD serosurveillance is over USD 3.3 million per year (8). Currently, about USD 8 million is
spent yearly on a European programme to monitor SVDV in Italy (based on the 1995 figures EU VI/7260/94-EN
Rev. 1). There is an added cost associated with the destruction of animals (e.g. a 1999 Italian outbreak in the north of
the country resulted in the destruction of 65 000 animals at an approximate cost of USD 7.3 million). In serological
surveys of pig populations, including those of SVD-free countries, a small number of pigs will inevitably give falsepositive reactions (singleton reactors). Consequently, considerable time, expense and effort has been spent in the EU on
the serological aspects of SVD.
The aforementioned considerations have led the State veterinary personnel of some countries to express the opinion that
the economic hardships which a country suffers when it declares SVD are too severe and that the disease should
therefore be deleted from OIE List A. A discussion ensued within the FAO EUFMD Research Group (2), during which
the following points were made for and against removing SVD from the OIE List A.
In favour of removing from OIE List A:
- Considerable effort and cost of repeated serological surveys. However, the economic savings of removing SVD
from List A would depend on whether there was general international acceptance of the change.
- Excessive trade and economic penalties result for countries which report outbreaks, especially related to serological
findings only
- Removal would not necessarily result in the cessation of control programmes against SVD.
Against removing from OIE List A:
- If SVD were allowed to spread there would be a decline in farmer and field veterinarian awareness and possibly an
increased risk of delayed FMD reporting.
- The presence of SVD on List A makes it is easier to justify the cost of SVD control programmes and high
containment laboratories.
7. RESULTS OF THE QUESTIONNAIRE: SUMMARY
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Conf. OIE 2000
The following OIE Member Countries completed the questionnaire (Appendix 1): Albania, Armenia, Austria, Belarus,
Belgium, Croatia, Cyprus, Czech Republic, Estonia, Finland, France, Germany, Hungary, Iceland, Ireland, Israel, Italy,
Kyrgyzstan, Latvia, Lithuania, Luxembourg, Moldavia, Netherlands, Norway, Poland, Portugal, Russia, Slovakia,
Spain, Sweden, Switzerland, Tajikistan, United Kingdom and Uzbekistan. The results are presented in Table 2.
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Conf. OIE 2000
Table 2: Results of some of the questions (indicated with their number) of the questionnaire
expressed in percentage of the number of Member Countries having provided the answer mentioned in the first column
Questions:
1.5.
1.6.
1.7.
swill
1.7
1.7
imported imported
animals products
1.7
1.8. 1.9. 1.10. 1.11. 1.12.
vehicles
1.12
1.12
1.12
2.1.
1 year
3 years
irregular
never
63
3
25
9
85
4.5.
4.6.
4.7.
4.8.
4.9.
94
82
85
15
62
2.2.
2.2
2.2
SVDV
Ab
PCR
84
97
57
2.3.
2.3
2.3
2 days
week
59
41
Answers:
Yes
High
Moderate
Low
Questions:
22
3.3.
63
13
66
22
53
23
23
81
16
3
44
28
28
47
38
16
3.3
3.3
3.3
3.4.
3.5.
74
28
41
31
4.1. 4.2. 4.3. 4.4.
imported exported imported exported
pigs
pigs
product product
84
61
35
3
4.10. 4.11. 4.12. 4.13. 4.14.
4.15.
all
all
herds herds
Answers:
Yes
High
Moderate
Low
74
59
100
79
50
62
94
93
91
71
23
6
63
79
68
82
21
59
19
22
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Epidemiology of SVD
Table 3 gives the number of primary/secondary outbreaks of SVD according to the results of the questionnaire.
Suspicions of SVD are infrequent and the majority of the Member Countries consider a herd has suffered an outbreak
only if SVDV is isolated. There is a moderate risk of spread of SVDV in Europe but the majority of Member Countries
think that SVD will become endemic if stamping-out is not implemented. The main risk of the spread of SVD is by
importing live animals subclinically infected with SVDV. Swill feeding, import of animal products and transport
vehicles are seen as high risks by half of the Member Countries. Swill feeding is still practised in many cases. Cleaning
vehicles is an effective preventive measure if well conducted. In most Member Countries the SVD survey is included in
the general animal survey programme.
Table 3: Number of primary/secondary
outbreaks of SVD according to the results of the questionnaire
Belgium
France
Germany
Italy
Netherlands
Poland
Portugal
Spain
Switzerland
United Kingdom
1960-69
1970-79
1980-89
1990-99
0
0
0
0
0
0
1/5
3/0
179/1/1
2/0
1/0
71/375
0
4/0
4/0
46/0
0
0
11/75
2/0
0
0
92/28
2/7
1/0
2/0
0
0
Laboratory support
Almost all Member Countries have a laboratory or an agreement with a laboratory from another Member Country and
therefore have the possibility of performing SVD diagnosis and differentiating SVD from FMD (50% of the
laboratories can do it within two days).
The pig sector
All Member Countries import pig products. Most Member Countries also import live pigs and export pig products.
Sixty-percent export live pigs. The economic consequences of a SVD outbreak for the pig sector are considered to be
very high. Half of the Member Countries think that these consequences are higher than necessary.
Control of SVD
Most Member Countries use the classical swine fever contingency plan. The majority of the Member Countries
implement stamping-out of the whole herd in the event of SVDV isolation, with or without clinical signs. Sixty-percent
are convinced that this is acceptable to the public. A protection/surveillance zone must be established and 60% think
that serosurveillance should be implemented for at least a year.
Eighty-percent of the Member Countries answered that SVD should be maintained on the OIE List A. However, some
countries added that the OIE Lists should be reviewed (together with SVD). Seventy-percent are convinced that the
SVDV will spread across Europe after the removal of SVD from List A. The majority also thinks that then FMD
identification in the field would be delayed. Therefore, (clinical) surveillance programmes should be maintained.
Vaccination against SVDV would not be implemented.
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REFERENCES
1.
Anonymous (1996). Manual of standards for diagnostic tests and vaccines, 3rd edition, Paris, France: Office
International des Epizooties, 21-27.
2.
Anonymous (1999). Report of the Session of the FAO Research Group of the Standing Technical Committee of
the European Commission for the Control of Foot and Mouth Disease, 29 September - 1 October, Paris, France,
p. 11-12.
3.
Brocchi E., Berlinzani A., Gamba D. & De Simone F. (1995). Development of two novel monoclonal antibodybased ELISAs for the detection of antibodies and the identification of swine isotypes against swine vesicular
disease virus. J. Virol. Methods, 52, 155-167.
4.
Brocchi E., Zhang G., Knowles N.J., Wilsden G., McCawley J.W., Marquardt O., Ohlinger V.F. & De Simone F.
(1997). Molecular epidemiology of recent outbreaks of swine vesicular disease: two genetically and
antigenically distinct variants in Europe, 1987-1994. Epidemiol. Infect., 118 (1).
5.
Callens M. & De Clercq K. (1999). Highly sensitive detection of swine vesicular disease virus based on a single
tube RT-PCR system and DIG-ELISA detection. J Virol Methods, 77, 87-99.
6.
Crauwels A.P, Nielen M., Stegeman J.A., Elbers A.R., Dijkhuizen A.A. & Tielen M.J. (1999). The effectiveness
of routine serological surveillance: case study of the 1997 epidemic of classical swine fever in the Netherlands.
Rev. Sci. Tech. Off. Int. Epi., 18(3): 627-37.
7.
De Clercq K. (1998). Reduction of singleton reactors against swine vesicular disease virus by a combination of
virus neutralisation test, monoclonal antibody-based competitive ELISA and isotype specific ELISA. J. Virol.
Methods, 70, 7-18.
8.
Dekker A. (2000). Pathogenesis, diagnosis and epizootiology of swine vesicular disease. PhD thesis, IDLelystad.
9.
Dekker A., Moonen P., de Boer-Luijtz E.A. & Terpstra C. (1995). Pathogenesis of swine vesicular disease after
exposure of pigs to an infected environment. Vet. Microbiology, 45, 243-250.
10.
Donaldson A.I., Ferris N.P., Knowles N.J., & Barnett I.T.R. (1983). Comparative studies of United Kingdom
isolates of swine vesicular disease virus. Res. Vet. Sci., 35, 295-300.
11.
Fallacara F., Pacciarini M., Bugnetti M., Berlinzani A. & Brocchi E. (2000). Detection of swine vesicular disease
virus in faeces samples by immune PCR assay. Proceedings of the Fifth Congres of the European Society of
Veterinary Virology, pp. 173-174.
12.
Geering W.A., Forman A.J. & Nunn M.J. (1995). Exotic Diseases of Animals: a field guide for Australian
veterinarians. Australian Government Publishing Service, Canberra.
13.
Golding, S.M., Hedger, R.S., Talbot, P. & Watson, J. (1976). Radial immuno-diffusion and seroneutralization
techniques for the assay of antibodies to swine vesicular disease. Res. Vet. Sci., 20, 142-147.
14.
Hedger R.S. & Mann J.A. (1989) Swine vesicular disease virus. In: Virus infections of porcines. Ed. Pensaert,
M.B., Elsevier Science Amsterdam, the Netherlands, pp. 241-250.
15.
Horst S. (1999). Economic aspects of diagnostic tests related to contagious animal diseases. Proceedings of the
Dutch Society for Veterinary Epidemiology and Economics, 12, 15-18.
16.
Kanno T., Mackay D., Inoue T., Wilsden G., Yamakawa M., Yamazoe R., Yamaguchi S., Shirai J., Kitching P.
& Murakami, Y. (1999). Mapping the genetic determinants of pathogenicity and plaque phenotype in swine
vesicular disease virus. J. Virology, 73 (4), 2710-6.
17.
Kitching R.P. & Donaldson A.I. (1987). Collection and transportation of specimens for vesicular virus
investigation. Rev. sci. tech. Off. int. Epiz., 6, 263-272.
- 271 -
Conf. OIE 2000
18.
Kodama M., Ogawa T., Saito T., Tokuda G. & Sasahara J. (1980a). Swine vesicular disease viruses isolated
from healthy pigs in non-epizootic period. I. Isolation and identification. Natl Inst Anim Health Q (Tokyo), 20, 110.
19.
Lai S.S., McKercher P.D., Moore D.M. & Gillespie J.H. (1979). Pathogenesis of swine vesicular disease in pigs.
American Journal of Veterinary Research, 40, 463-468.
20.
Lin F., Mackay D.K. & Knowles N.J. (1997). Detection of swine vesicular disease virus RNA by reverse
transcription-polymerase chain reaction. J Virol Methods, 65, 111-21.
21.
Lin F., Mackay D.K. & Knowles N.J. (1998). The persistence of swine vesicular disease virus infection in pigs.
Epidemiol Infect, 121, 459-72.
22.
Mann J.A. & Hutchings G.H. (1980). Swine vesicular disease: pathways of infection. Journal of Hygiene, 84,
355-363.
23.
Mitev G., Tekerlekov P., Dilovsky M., Ognianov D. & Nikolova E. (1978). Experiments on the preparation and
testing of associated vaccine against foot and mouth disease and vesicular disease in swine. Archiv für
Experimetelle Veterinarmedizin, 32, 29-33.
24.
Mowat G.N., Prince M.J., Spier R.E. & Staple R.F. (1974). Preliminary studies on the development of swine
vesicular disease vaccine. Archiv für die Gesamte Virusforschung, 44, 350-360.
25.
Nardelli L., Lodetti E., Gualandi G.L., Burrows R., Goodridge D., Brown F. & Cartwright B. (1968). A foot and
mouth disease syndrome in pigs caused by an enterovirus. Nature, 219, 21-22.
26.
Terpstra C., Dekker A., Reek F.H. & Chenard G. (1993). Swine vesicular disease: threat or challenge for the
Dutch pig industry? Lelystad, The Netherlands : Annual Report of the Central Veterinary Institute (CDI-DLO),
1993 : 15-18.
27.
Zhang G., Haydon D.T., Knowles N.J. & McCauley J.W. (1999). Molecular evolution of swine vesicular disease
virus. J. Gen. Virol., 80, 639-51.
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