A double antibody sandwich ELISA developed by ID-DLO, Lelystad to detect Corynebocterium pseudotuberculosis infection was used on 329 sheep from four pedigree Suffolk flocks in which clinical cases of caseous lymphadenitis (CLA) had occurred. At subsequent necropsy, typical CLA lesions were seen in 133 sheep, and the diagnosis was confirmed on culture. Lesions were most commonly seen in lungs (n = 46), parotid lymph nodes (n = 44), prescapular lymph nodes (n = 38) and mediastinal lymph nodes (n = 31). The sensitivity of the ELISA test for detecting culture-positive sheep was 0.88, while the specificity of the test was 0.55. The antibody ELISA detected 87.5 per cent of sheep that had CLA lesions restricted to internal organs only. It was concluded that the ELISA test has a valuable role in detecting sheep with both clinical and subclinical CLA

Baird, Graham J.

Fee, Seán A.

Kamp, Elbarte M.

King, David C.

Malone, Frank (Frank E.)

Murdock, Fiona E. A.

O'Reilly, Kath M.

English

PubMed

Frank E Malone

Seán A Fee

Elbarte M Kamp

David C King

Graham J Baird

Kath M O'Reilly

Fiona EA Murdock

Springer - Publisher Connector

19
Peer reviewed volume 59 (1) : January 2006    
 irish veterinary Journal 
introduction
Caseous lymphadenitis (CLA) is caused by the bacterium 
Corynebacterium pseudotuberculosis. Although it can affect a number 
of domestic species and man, it is as a disease of sheep that CLA is 
most important. C. pseudotuberculosis causes chronic abscessation 
in lymph nodes (LN) and viscera. The disease is present in many 
countries throughout the world and was first recorded in sheep 
in Northern Ireland in 1999, when the source of the infection was 
traced to imported Scottish sheep. CLA is a notifiable disease in both 
Northern Ireland and the Republic of Ireland, but restrictions have 
been removed in Great Britain since 1991.
Baird (2003) listed a number of factors that make the control of CLA 
difficult. CLA may be detected clinically only in superficial LN, whereas 
in a proportion of affected sheep the lesions may be present only 
in visceral LN or organs such as the lungs. CLA has a relatively long 
incubation period, of between two and four months, before it may be 
detected clinically. In addition, C. pseudotuberculosis may survive for 
long periods in the environment, thus providing a source of infection 
in the absence of clinical cases. 
Dercksen et al. (2000) evaluated a double antibody sandwich ELISA 
test for the detection of CLA in healthy sheep from CLA-free flocks 
and in sheep with culture-confirmed CLA. They found that this ELISA 
test had a specificity of 99±1% and a sensitivity of 79±5%. The present 
study was designed to evaluate its use in four naturally infected flocks.
Materials and methods 
Flock histories 
The study was undertaken in 329 sheep (63 male and 266 female) 
from four flocks, which were depopulated in 2001 and 2002 because 
of widespread CLA infection.
Flock A 
CLA was first detected on this farm in July 2001 during a clinical 
examination of the flock that had been prompted by a backward trace 
of contacts from flock B. The flockowner reported seeing similar 
lesions in his sheep since December of the previous year, but did 
not realise that the cause was CLA. The source of the infection was 
considered to be two rams that were purchased from Scotland in 
August 2000. Subsequently, this flock was depopulated completely in 
batches until July 2002.
peer reviewed
A serological investigation of caseous lymphadenitis in four flocks of sheep
Frank e. Malone1, Seán A. Fee1, elbarte M. Kamp2*, david C. King1, Graham J. Baird3, Kath M. O’reilly4 and Fiona e.A. 
Murdock5
1 Department of Agriculture and Rural Development for Northern Ireland, Veterinary Sciences Division, 43 Beltany Road, Omagh, County 
 Tyrone, Northern Ireland, BT78 5NF
2 Institute of Animal Science and Health (ID-Lelystad), PO Box 65, 8200 AB Lelystad, The Netherlands
 * Deceased.
3 Scottish Agricultural College, Veterinary Science Division, Greycrook, St. Boswells, Melrose, Scotland,  TD6 0EQ
4 Ecology and Epidemiology Group, Biological Sciences, University of Warwick, Coventry, England, CV4 7AL
5 Department of Agriculture and Rural Development for Northern Ireland, Veterinary Service, Dundonald House, Upper Newtownards Road, 
 Belfast, Northern Ireland BT4 3SB
A double antibody sandwich ELISA developed by ID-DLO, Lelystad to detect Corynebacterium pseudotuberculosis infection was used 
on 329 sheep from four pedigree Suffolk flocks in which clinical cases of caseous lymphadenitis (CLA) had occurred. At subsequent 
necropsy, typical CLA lesions were seen in 133 sheep, and the diagnosis was confirmed on culture. Lesions were most commonly 
seen in lungs (n=46), parotid lymph nodes (n=44), prescapular lymph nodes (n=38) and mediastinal lymph nodes (n=31). The 
sensitivity of the ELISA test for detecting culture-positive sheep was 0.88, while the specificity of the test was 0.55. The antibody 
ELISA detected 87.5 per cent of sheep that had CLA lesions restricted to internal organs only. It was concluded that the ELISA test 
has a valuable role in detecting sheep with both clinical and subclinical CLA.
Irish Veterinary Journal
Volume 59 (1) 19-21, 2005
Correspondence:
Frank Malone
Department of Agriculture and Rural Development for Northern 
Ireland, Veterinary Sciences Division, 43 Beltany Road, Omagh, 
County Tyrone, Northern Ireland, BT78 5NF
Tel: +44 (0) 28 822 43337;Fax: +44 (0) 28 822 44228; 
Email: frank.malone@dardni.gov.uk
        Keywords:  Sheep, Caseous lymphadenitis, Serology, Corynebacterium pseudotuberculosis
20
Figure 1: Corynebacterium pseudotuberculosis abscessation in parotid lymph node.
Flock B 
CLA was first detected on this farm by a clinical examination of the 
flock in June 2001. The most probable source of infection was a ram 
purchased from farm A in late October 2000. Subsequently this flock 
was depopulated completely in batches until October 2001.
Flock C 
CLA was first detected on this farm by a clinical examination of the 
flock in August 2001. The source of the infection was considered to 
be a ewe, purchased in November 1999, that had originated from the 
same flock in Scotland that was considered to have infected flock A. 
Subsequently, this flock was depopulated completely in batches until 
February 2002.
Flock D 
CLA was first detected on this farm by a clinical examination of 
the flock in August 2001. The source of infection was traced to 
five ewes purchased from flock A in December 2000. Forty-one in-
contact sheep were depopulated in batches until November 2001. 
On repeated clinical examinations, the remaining sheep, which were 
separately managed, were consistently found to be negative for CLA.
Serology
All sheep were blood sampled for the ELISA test prior to euthanasia 
and postmortem examination. Optical density (OD) readings for the 
ELISA had been determined from experience of using the test in 
The Netherlands (Daan Dercksen, personal communication). An OD 
value above 400 was considered positive, values between 200 and 
400 were considered ‘low positive’, those between150 and 200 were 
inconclusive and those less than 150 were negative. A non-specific 
reaction was identified when the OD with the negative antigen was 
more than 50% of the maximum OD of the positive control.
Postmortem examination 
All sheep were necropsied using a standard protocol. An external 
examination of the carcase was carried out, noting any sinus 
discharges or subcutaneous swellings. The superficial and visceral 
lymph nodes, lungs and liver were then examined for abscessation. 
Suspect lesions were cultured for C. pseudotuberculosis (Cowan and 
Steel, 1974).
results
On postmortem examination, 133 of 329 sheep had lesions typical of 
CLA (Figures 1 and 2); these were confirmed on culture (Table 1). 
The lesions were present in either single or multiple sites (Table 2: 
221 sites in 133 sheep). Lesions in superficial lymph nodes (LN) were 
most frequently present in the parotid LN (n=44) and the prescapular 
LN (n=38). Internal lesions were most frequently present in the lungs 
(n=46) and mediastinal LN (n=31). 
Table 1: Prevalence of caseous lymphadenitis (CLA) lesions in the four flocks 
of sheep
Lesions were present in internal organs only in 32 (24.1 per cent) 
of the 133 sheep (Table 3); of these 28 (87.5 per cent) were ELISA-
positive.
Three hundred and eight sheep were defined as either ELISA-positive 
or ELISA-negative, 18 sheep gave inconclusive tests and three gave 
non-specific results. When inconclusive and non-specific results 
were treated as negative, the overall sensitivity of the ELISA for 
detecting CLA lesions was 0.88 and the specificity was 0.55 (Table 4). 
Sensitivity varied in the different flocks from 0.83 to 1.0 and specificity 
varied from 0.31 to 0.87 (Table 5).
Table 2: Distribution of caseous lymphadenitis (CLA) lesions in 133 sheep
Flock No.of sheep  No.of sheep Prevalence of 
 examined culture-positive CLA in   
  for CLA flocks (%) 
 
A  98   35 35.7   
B  95   37 38.9   
C 100   55 55.0   
D  36    6 16.7  
All four flocks 329 133 40.4
Figure 2: 
Corynebacterium 
pseudotuberculosis 
abscessation in sheep 
lungs.
Lungs 46 (35%) 
Parotid LN 44 (33%) 
Prescapular LN 38 (29%) 
Mediastinal LN 31 (23%) 
Retropharyngeal LN 11 (8%) 
Prefemoral LN 10 (8%)
Bronchial LN 9 (7%)
Submandibular LN 8 (6%)
Liver  5 (4%) 
Supermammary LN 4 (3%)
Mesenteric LN 4 (3%)
Other sites 11 (8%)
Site Number (%) 
  of sheep 
 with CLA 
 lesions
Site Number (%) 
  of sheep 
 with CLA 
 lesions
volume 59 (1) :  January, 2006    Peer reviewed
irish veterinary Journal 
LN: lymph node.
21
Peer reviewed volume 59 (1) : January 2006    
 irish veterinary Journal 
Table 3: Distribution of caseous lymphadenitis (CLA) lesions in 133 sheep
Table 4: Sensitivity and specificity of the ELISA test 
for caseous lympadenitis (CLA)
Table 5: Sensitivity and specificity of ELISA test in individual flocks
discussion
Brown and Olander (1987) reported that the introduction of a single 
CLA-abscessed animal into a naïve flock results in a high incidence 
of abscesses within two to three years. In the present study CLA 
abscesses (Table 1) were detected in 133 animals in these four flocks 
between seven and 21 months after the introduction of CLA-affected 
sheep (flock A: 11 months; flock B: seven months; flock C: 21 months; 
flock D: eight months). Intensive husbandry methods in these flocks 
may have contributed to the relatively rapid spread of infection.
The present study demonstrated that CLA lesions were present only 
in the viscera (lungs and visceral LN) in approximately 24 per cent of 
clinical cases. Consequently, clinical examination alone will not detect 
all infected sheep in a flock. Forty-six of the 133 CLA-positive sheep 
had lung abscesses. These lesions may lead to more rapid spread 
of the disease when sheep are closely confined, as for shearing and 
routine treatments, or when housed.
In this study, the ELISA test was relatively sensitive at detecting CLA-
infected sheep. However, the specificity of the test in detecting animals 
with lesions was low (Table 3). This contrasts with the experience of 
Dercksen et al (2000), who found that the ELISA test had a specificity 
of 99±1% and a sensitivity of 79±5%. In their study, the sensitivity and 
specificity were determined in sera from sheep with clinical abscesses 
from which C. pseudotuberculosis was isolated and in sera from healthy 
sheep in flocks without a record of CLA. The present study differs in 
that both CLA-positive and CLA-negative sheep were drawn from 
infected flocks. 
Dercksen et al (2000) suggested that cross-reactions may occur due 
to infections with other bacteria (for example, other Corynebacterium 
species, Listeria monocytogenes and Mycobacterium avium subspecies 
paratuberculosis). Evidence of these bacterial infections was not found 
in the four flocks examined. Batey (1986) suggested that many CLA 
lesions undergo resolution during the early stages of development. In 
the present study, it is more likely that seropositive, culture-negative 
sheep resulted from sheep in these flocks being exposed to infection, 
but either subsequently not developing the disease or eliminating 
the infection. The wide variation in specificity between flocks may be 
attributed to differences in time exposed to C. pseudotuberculosis.
Twenty-eight of the 32 sheep with internal lesions only were ELISA-
positive. These results indicate that ELISA testing could play a valuable 
role in CLA control by supplementing clinical detection of superficial 
lesions. It is anticipated that such ELISAs will become commercially 
available in the near future.
references
Baird, G.J. (2003). Current perspectives on caseous lymphadenitis. In 
Practice 25: 62-68.
Batey, r.G. (1986). Pathogenesis of caseous lymphadenitis in sheep 
and goats. Australian Veterinary Journal 63: 269-272.
Brown, C.C. and Olander, H.J. (1987). Caseous lymphadenitis of 
goats and sheep: a review. Veterinary Bulletin 57: 1-12.
Cowan, S.T. and Steel, K.J. (1974). Cowan and Steel’s Manual for the 
Identification of Medical Bacteria. Second Edition. Cambridge: Cambridge 
University Press.
dercksen, d.P., Brinkhof, J.M.A., dekker-Norren, T., van 
Maanen, K., Bode, C.F., Baird, G. and Kamp, e.M. (2000). A 
comparison of four serological tests for the diagnosis of caseous 
lymphadenitis in sheep and goats. Veterinary Microbiology 75: 167-175. 
n
Flock A B C d All 4 flocks (%) 
 
Number of  
sheep with  
CLA lesions 35 37 55 6 133 (40.4%) 
 
Number of  
sheep with  
superficial  
LN and  
cutaneous  
lesions only 13 16 37 3 69 (21.0%) 
 
Number of  
sheep with  
internal  
lesions only  
(lungs and  
visceral LN) 12  11     8  1  32 (9.7%)  
Number of  
sheep with  
both internal  
and external  
CLA lesions 10 10 10 2 32 (9.7%)  
Total number  
of sheep  
examined 98 95 100 36 329  
LN: lymph node; CLA: caseous lymphadenitis.
Flock Sensitivity Specificity 
Flock A (n=98) 0.83 0.59  
Flock B (n=95) 0.86 0.53  
Flock C (n=100) 0.91 0.31  
Flock D (n=36) 1.00 0.87  
All 4 flocks (n=329) 0.88 0.55  
CLA lesion positive negative total 
Detected 117  16 133 Sensitivity 0.88 
 
Not detected  88 108 196 Specificity 0.55 
 
Total 205 124 329  


A serological investigation of caseous lymphadenitis in four flocks of sheep

Abstract A double antibody sandwich ELISA developed by ID-DLO, Lelystad to detect Corynebacterium pseudotuberculosis infection was used on 329 sheep from four pedigree Suffolk flocks in which clinical cases of caseous lymphadenitis (CLA) had occurred. At subsequent necropsy, typical CLA lesions were seen in 133 sheep, and the diagnosis was confirmed on culture. Lesions were most commonly seen in lungs (n = 46), parotid lymph nodes (n = 44), prescapular lymph nodes (n = 38) and mediastinal lymph nodes (n = 31). The sensitivity of the ELISA test for detecting culture-positive sheep was 0.88, while the specificity of the test was 0.55. The antibody ELISA detected 87.5 per cent of sheep that had CLA lesions restricted to internal organs only. It was concluded that the ELISA test has a valuable role in detecting sheep with both clinical and subclinical CLA.</p

Malone Frank E

Fee Seán A

Kamp Elbarte M

King David C

Baird Graham J

O'Reilly Kath M

Murdock Fiona EA

Directory of Open Access Journals

Irish Veterinary Journal

A double antibody sandwich ELISA developed by ID-DLO, Lelystad to detect Corynebacterium pseudotuberculosis infection was used on 329 sheep from four pedigree Suffolk flocks in which clinical cases of caseous lymphadenitis (CLA) had occurred. At subsequent necropsy, typical CLA lesions were seen in 133 sheep, and the diagnosis was confirmed on culture. Lesions were most commonly seen in lungs (n = 46), parotid lymph nodes (n = 44), prescapular lymph nodes (n = 38) and mediastinal lymph nodes (n = 31). The sensitivity of the ELISA test for detecting culture-positive sheep was 0.88, while the specificity of the test was 0.55. The antibody ELISA detected 87.5 per cent of sheep that had CLA lesions restricted to internal organs only. It was concluded that the ELISA test has a valuable role in detecting sheep with both clinical and subclinical CLA

Malone, Frank E

Fee, Seán A

Kamp, Elbarte M

King, David C

Baird, Graham J

O'Reilly, Kath M

Murdock, Fiona Ea

LSHTM Research Online

Malone, FE; Fee, SA; Kamp, EM; King, DC; Baird, GJ; O’Reilly,KM; Murdock, FE (2006) A serological investigation of caseous lym-phadenitis in four flocks of sheep. Irish veterinary journal, 59 (1).pp. 19-21. ISSN 0368-0762 DOI: https://doi.org/10.1186/2046-0481-59-1-19Downloaded from: http://researchonline.lshtm.ac.uk/4646841/DOI: 10.1186/2046-0481-59-1-19Usage GuidelinesPlease refer to usage guidelines at http://researchonline.lshtm.ac.uk/policies.html or alterna-tively contact researchonline@lshtm.ac.uk.Available under license: Copyright the author(s)19Peer reviewed volume 59 (1) : January 2006     irish veterinary Journal introductionCaseous lymphadenitis (CLA) is caused by the bacterium Corynebacterium pseudotuberculosis. Although it can affect a number of domestic species and man, it is as a disease of sheep that CLA is most important. C. pseudotuberculosis causes chronic abscessation in lymph nodes (LN) and viscera. The disease is present in many countries throughout the world and was first recorded in sheep in Northern Ireland in 1999, when the source of the infection was traced to imported Scottish sheep. CLA is a notifiable disease in both Northern Ireland and the Republic of Ireland, but restrictions have been removed in Great Britain since 1991.Baird (2003) listed a number of factors that make the control of CLA difficult. CLA may be detected clinically only in superficial LN, whereas in a proportion of affected sheep the lesions may be present only in visceral LN or organs such as the lungs. CLA has a relatively long incubation period, of between two and four months, before it may be detected clinically. In addition, C. pseudotuberculosis may survive for long periods in the environment, thus providing a source of infection in the absence of clinical cases. Dercksen et al. (2000) evaluated a double antibody sandwich ELISA test for the detection of CLA in healthy sheep from CLA-free flocks and in sheep with culture-confirmed CLA. They found that this ELISA test had a specificity of 99±1% and a sensitivity of 79±5%. The present study was designed to evaluate its use in four naturally infected flocks.Materials and methods Flock histories The study was undertaken in 329 sheep (63 male and 266 female) from four flocks, which were depopulated in 2001 and 2002 because of widespread CLA infection.Flock A CLA was first detected on this farm in July 2001 during a clinical examination of the flock that had been prompted by a backward trace of contacts from flock B. The flockowner reported seeing similar lesions in his sheep since December of the previous year, but did not realise that the cause was CLA. The source of the infection was considered to be two rams that were purchased from Scotland in August 2000. Subsequently, this flock was depopulated completely in batches until July 2002.peer reviewedA serological investigation of caseous lymphadenitis in four flocks of sheepFrank e. Malone1, Seán A. Fee1, elbarte M. Kamp2*, david C. King1, Graham J. Baird3, Kath M. O’reilly4 and Fiona e.A. Murdock51 Department of Agriculture and Rural Development for Northern Ireland, Veterinary Sciences Division, 43 Beltany Road, Omagh, County  Tyrone, Northern Ireland, BT78 5NF2 Institute of Animal Science and Health (ID-Lelystad), PO Box 65, 8200 AB Lelystad, The Netherlands * Deceased.3 Scottish Agricultural College, Veterinary Science Division, Greycrook, St. Boswells, Melrose, Scotland,  TD6 0EQ4 Ecology and Epidemiology Group, Biological Sciences, University of Warwick, Coventry, England, CV4 7AL5 Department of Agriculture and Rural Development for Northern Ireland, Veterinary Service, Dundonald House, Upper Newtownards Road,  Belfast, Northern Ireland BT4 3SBA double antibody sandwich ELISA developed by ID-DLO, Lelystad to detect Corynebacterium pseudotuberculosis infection was used on 329 sheep from four pedigree Suffolk flocks in which clinical cases of caseous lymphadenitis (CLA) had occurred. At subsequent necropsy, typical CLA lesions were seen in 133 sheep, and the diagnosis was confirmed on culture. Lesions were most commonly seen in lungs (n=46), parotid lymph nodes (n=44), prescapular lymph nodes (n=38) and mediastinal lymph nodes (n=31). The sensitivity of the ELISA test for detecting culture-positive sheep was 0.88, while the specificity of the test was 0.55. The antibody ELISA detected 87.5 per cent of sheep that had CLA lesions restricted to internal organs only. It was concluded that the ELISA test has a valuable role in detecting sheep with both clinical and subclinical CLA.Irish Veterinary JournalVolume 59 (1) 19-21, 2005Correspondence:Frank MaloneDepartment of Agriculture and Rural Development for Northern Ireland, Veterinary Sciences Division, 43 Beltany Road, Omagh, County Tyrone, Northern Ireland, BT78 5NFTel: +44 (0) 28 822 43337;Fax: +44 (0) 28 822 44228; Email: frank.malone@dardni.gov.uk        Keywords:  Sheep, Caseous lymphadenitis, Serology, Corynebacterium pseudotuberculosis20Figure 1: Corynebacterium pseudotuberculosis abscessation in parotid lymph node.Flock B CLA was first detected on this farm by a clinical examination of the flock in June 2001. The most probable source of infection was a ram purchased from farm A in late October 2000. Subsequently this flock was depopulated completely in batches until October 2001.Flock C CLA was first detected on this farm by a clinical examination of the flock in August 2001. The source of the infection was considered to be a ewe, purchased in November 1999, that had originated from the same flock in Scotland that was considered to have infected flock A. Subsequently, this flock was depopulated completely in batches until February 2002.Flock D CLA was first detected on this farm by a clinical examination of the flock in August 2001. The source of infection was traced to five ewes purchased from flock A in December 2000. Forty-one in-contact sheep were depopulated in batches until November 2001. On repeated clinical examinations, the remaining sheep, which were separately managed, were consistently found to be negative for CLA.SerologyAll sheep were blood sampled for the ELISA test prior to euthanasia and postmortem examination. Optical density (OD) readings for the ELISA had been determined from experience of using the test in The Netherlands (Daan Dercksen, personal communication). An OD value above 400 was considered positive, values between 200 and 400 were considered ‘low positive’, those between150 and 200 were inconclusive and those less than 150 were negative. A non-specific reaction was identified when the OD with the negative antigen was more than 50% of the maximum OD of the positive control.Postmortem examination All sheep were necropsied using a standard protocol. An external examination of the carcase was carried out, noting any sinus discharges or subcutaneous swellings. The superficial and visceral lymph nodes, lungs and liver were then examined for abscessation. Suspect lesions were cultured for C. pseudotuberculosis (Cowan and Steel, 1974).resultsOn postmortem examination, 133 of 329 sheep had lesions typical of CLA (Figures 1 and 2); these were confirmed on culture (Table 1). The lesions were present in either single or multiple sites (Table 2: 221 sites in 133 sheep). Lesions in superficial lymph nodes (LN) were most frequently present in the parotid LN (n=44) and the prescapular LN (n=38). Internal lesions were most frequently present in the lungs (n=46) and mediastinal LN (n=31). Table 1: Prevalence of caseous lymphadenitis (CLA) lesions in the four flocks of sheepLesions were present in internal organs only in 32 (24.1 per cent) of the 133 sheep (Table 3); of these 28 (87.5 per cent) were ELISA-positive.Three hundred and eight sheep were defined as either ELISA-positive or ELISA-negative, 18 sheep gave inconclusive tests and three gave non-specific results. When inconclusive and non-specific results were treated as negative, the overall sensitivity of the ELISA for detecting CLA lesions was 0.88 and the specificity was 0.55 (Table 4). Sensitivity varied in the different flocks from 0.83 to 1.0 and specificity varied from 0.31 to 0.87 (Table 5).Table 2: Distribution of caseous lymphadenitis (CLA) lesions in 133 sheepFlock No.of sheep  No.of sheep Prevalence of  examined culture-positive CLA in     for CLA flocks (%)  A  98   35 35.7   B  95   37 38.9   C 100   55 55.0   D  36    6 16.7  All four flocks 329 133 40.4Figure 2: Corynebacterium pseudotuberculosis abscessation in sheep lungs.Lungs 46 (35%) Parotid LN 44 (33%) Prescapular LN 38 (29%) Mediastinal LN 31 (23%) Retropharyngeal LN 11 (8%) Prefemoral LN 10 (8%)Bronchial LN 9 (7%)Submandibular LN 8 (6%)Liver  5 (4%) Supermammary LN 4 (3%)Mesenteric LN 4 (3%)Other sites 11 (8%)Site Number (%)   of sheep  with CLA  lesionsSite Number (%)   of sheep  with CLA  lesionsvolume 59 (1) :  January, 2006    Peer reviewedirish veterinary Journal LN: lymph node.21Peer reviewed volume 59 (1) : January 2006     irish veterinary Journal Table 3: Distribution of caseous lymphadenitis (CLA) lesions in 133 sheepTable 4: Sensitivity and specificity of the ELISA test for caseous lympadenitis (CLA)Table 5: Sensitivity and specificity of ELISA test in individual flocksdiscussionBrown and Olander (1987) reported that the introduction of a single CLA-abscessed animal into a naïve flock results in a high incidence of abscesses within two to three years. In the present study CLA abscesses (Table 1) were detected in 133 animals in these four flocks between seven and 21 months after the introduction of CLA-affected sheep (flock A: 11 months; flock B: seven months; flock C: 21 months; flock D: eight months). Intensive husbandry methods in these flocks may have contributed to the relatively rapid spread of infection.The present study demonstrated that CLA lesions were present only in the viscera (lungs and visceral LN) in approximately 24 per cent of clinical cases. Consequently, clinical examination alone will not detect all infected sheep in a flock. Forty-six of the 133 CLA-positive sheep had lung abscesses. These lesions may lead to more rapid spread of the disease when sheep are closely confined, as for shearing and routine treatments, or when housed.In this study, the ELISA test was relatively sensitive at detecting CLA-infected sheep. However, the specificity of the test in detecting animals with lesions was low (Table 3). This contrasts with the experience of Dercksen et al (2000), who found that the ELISA test had a specificity of 99±1% and a sensitivity of 79±5%. In their study, the sensitivity and specificity were determined in sera from sheep with clinical abscesses from which C. pseudotuberculosis was isolated and in sera from healthy sheep in flocks without a record of CLA. The present study differs in that both CLA-positive and CLA-negative sheep were drawn from infected flocks. Dercksen et al (2000) suggested that cross-reactions may occur due to infections with other bacteria (for example, other Corynebacterium species, Listeria monocytogenes and Mycobacterium avium subspecies paratuberculosis). Evidence of these bacterial infections was not found in the four flocks examined. Batey (1986) suggested that many CLA lesions undergo resolution during the early stages of development. In the present study, it is more likely that seropositive, culture-negative sheep resulted from sheep in these flocks being exposed to infection, but either subsequently not developing the disease or eliminating the infection. The wide variation in specificity between flocks may be attributed to differences in time exposed to C. pseudotuberculosis.Twenty-eight of the 32 sheep with internal lesions only were ELISA-positive. These results indicate that ELISA testing could play a valuable role in CLA control by supplementing clinical detection of superficial lesions. It is anticipated that such ELISAs will become commercially available in the near future.referencesBaird, G.J. (2003). Current perspectives on caseous lymphadenitis. In Practice 25: 62-68.Batey, r.G. (1986). Pathogenesis of caseous lymphadenitis in sheep and goats. Australian Veterinary Journal 63: 269-272.Brown, C.C. and Olander, H.J. (1987). Caseous lymphadenitis of goats and sheep: a review. Veterinary Bulletin 57: 1-12.Cowan, S.T. and Steel, K.J. (1974). Cowan and Steel’s Manual for the Identification of Medical Bacteria. Second Edition. Cambridge: Cambridge University Press.dercksen, d.P., Brinkhof, J.M.A., dekker-Norren, T., van Maanen, K., Bode, C.F., Baird, G. and Kamp, e.M. (2000). A comparison of four serological tests for the diagnosis of caseous lymphadenitis in sheep and goats. Veterinary Microbiology 75: 167-175. nFlock A B C d All 4 flocks (%)  Number of  sheep with  CLA lesions 35 37 55 6 133 (40.4%)  Number of  sheep with  superficial  LN and  cutaneous  lesions only 13 16 37 3 69 (21.0%)  Number of  sheep with  internal  lesions only  (lungs and  visceral LN) 12  11     8  1  32 (9.7%)  Number of  sheep with  both internal  and external  CLA lesions 10 10 10 2 32 (9.7%)  Total number  of sheep  examined 98 95 100 36 329  LN: lymph node; CLA: caseous lymphadenitis.Flock Sensitivity Specificity Flock A (n=98) 0.83 0.59  Flock B (n=95) 0.86 0.53  Flock C (n=100) 0.91 0.31  Flock D (n=36) 1.00 0.87  All 4 flocks (n=329) 0.88 0.55  CLA lesion positive negative total Detected 117  16 133 Sensitivity 0.88  Not detected  88 108 196 Specificity 0.55  Total 205 124 329  

A serological investigation of caseous lymphadenitis in four flocks of sheep.

A double antibody sandwich ELISA developed by ID-DLO, Lelystad to detect Corynebacterium pseudotuberculosis infection was used on 329 sheep from four pedigree Suffolk flocks in which clinical cases of caseous lymphadenitis (CLA) had occurred. At subsequent necropsy, typical CLA lesions were seen in 133 sheep, and the diagnosis was confirmed on culture. Lesions were most commonly seen in lungs (n=46), parotid lymph nodes (n=44), prescapular lymph nodes (n=38) and mediastinal lymph nodes (n=31). The sensitivity of the ELISA test for detecting culture-positive sheep was 0.88, while the specificity of the test was 0.55. The antibody ELISA detected 87.5 per cent of sheep that had CLA lesions restricted to internal organs only. It was concluded that the ELISA test has a valuable role in detecting sheep with both clinical and subclinical CLA

Malone, F.E.

Fee, S.A.

Kamp, E.M.

King, D.C.

Baird, G.J.

O'Reilly, K.M.

Murdock, F.E.A.

Name not available

Warwick Research Archives Portal Repository

 University of Warwick institutional repository: http://go.warwick.ac.uk/wrap  This paper is made available online in accordance with publisher policies. Please scroll down to view the document itself. Please refer to the repository record for this item and our policy information available from the repository home page for further information.  To see the final version of this paper please visit the publisher’s website. Access to the published version may require a subscription. Author(s):  Frank E Malone, Seán A Fee, Elbarte M Kamp, David C King, Graham J Baird, Kath M O'Reilly and Fiona EA Murdock Article Title: A serological investigation of caseous lymphadenitis in four flocks of sheep Year of publication: 2006 Link to published article: http://dx.doi.org/10.1186/2046-0481-59-1-19 Publisher statement:  None  19Peer reviewed volume 59 (1) : January 2006     irish veterinary Journal introductionCaseous lymphadenitis (CLA) is caused by the bacterium Corynebacterium pseudotuberculosis. Although it can affect a number of domestic species and man, it is as a disease of sheep that CLA is most important. C. pseudotuberculosis causes chronic abscessation in lymph nodes (LN) and viscera. The disease is present in many countries throughout the world and was first recorded in sheep in Northern Ireland in 1999, when the source of the infection was traced to imported Scottish sheep. CLA is a notifiable disease in both Northern Ireland and the Republic of Ireland, but restrictions have been removed in Great Britain since 1991.Baird (2003) listed a number of factors that make the control of CLA difficult. CLA may be detected clinically only in superficial LN, whereas in a proportion of affected sheep the lesions may be present only in visceral LN or organs such as the lungs. CLA has a relatively long incubation period, of between two and four months, before it may be detected clinically. In addition, C. pseudotuberculosis may survive for long periods in the environment, thus providing a source of infection in the absence of clinical cases. Dercksen et al. (2000) evaluated a double antibody sandwich ELISA test for the detection of CLA in healthy sheep from CLA-free flocks and in sheep with culture-confirmed CLA. They found that this ELISA test had a specificity of 99±1% and a sensitivity of 79±5%. The present study was designed to evaluate its use in four naturally infected flocks.Materials and methods Flock histories The study was undertaken in 329 sheep (63 male and 266 female) from four flocks, which were depopulated in 2001 and 2002 because of widespread CLA infection.Flock A CLA was first detected on this farm in July 2001 during a clinical examination of the flock that had been prompted by a backward trace of contacts from flock B. The flockowner reported seeing similar lesions in his sheep since December of the previous year, but did not realise that the cause was CLA. The source of the infection was considered to be two rams that were purchased from Scotland in August 2000. Subsequently, this flock was depopulated completely in batches until July 2002.peer reviewedA serological investigation of caseous lymphadenitis in four flocks of sheepFrank e. Malone1, Seán A. Fee1, elbarte M. Kamp2*, david C. King1, Graham J. Baird3, Kath M. O’reilly4 and Fiona e.A. Murdock51 Department of Agriculture and Rural Development for Northern Ireland, Veterinary Sciences Division, 43 Beltany Road, Omagh, County  Tyrone, Northern Ireland, BT78 5NF2 Institute of Animal Science and Health (ID-Lelystad), PO Box 65, 8200 AB Lelystad, The Netherlands * Deceased.3 Scottish Agricultural College, Veterinary Science Division, Greycrook, St. Boswells, Melrose, Scotland,  TD6 0EQ4 Ecology and Epidemiology Group, Biological Sciences, University of Warwick, Coventry, England, CV4 7AL5 Department of Agriculture and Rural Development for Northern Ireland, Veterinary Service, Dundonald House, Upper Newtownards Road,  Belfast, Northern Ireland BT4 3SBA double antibody sandwich ELISA developed by ID-DLO, Lelystad to detect Corynebacterium pseudotuberculosis infection was used on 329 sheep from four pedigree Suffolk flocks in which clinical cases of caseous lymphadenitis (CLA) had occurred. At subsequent necropsy, typical CLA lesions were seen in 133 sheep, and the diagnosis was confirmed on culture. Lesions were most commonly seen in lungs (n=46), parotid lymph nodes (n=44), prescapular lymph nodes (n=38) and mediastinal lymph nodes (n=31). The sensitivity of the ELISA test for detecting culture-positive sheep was 0.88, while the specificity of the test was 0.55. The antibody ELISA detected 87.5 per cent of sheep that had CLA lesions restricted to internal organs only. It was concluded that the ELISA test has a valuable role in detecting sheep with both clinical and subclinical CLA.Irish Veterinary JournalVolume 59 (1) 19-21, 2005Correspondence:Frank MaloneDepartment of Agriculture and Rural Development for Northern Ireland, Veterinary Sciences Division, 43 Beltany Road, Omagh, County Tyrone, Northern Ireland, BT78 5NFTel: +44 (0) 28 822 43337;Fax: +44 (0) 28 822 44228; Email: frank.malone@dardni.gov.uk        Keywords:  Sheep, Caseous lymphadenitis, Serology, Corynebacterium pseudotuberculosis20Figure 1: Corynebacterium pseudotuberculosis abscessation in parotid lymph node.Flock B CLA was first detected on this farm by a clinical examination of the flock in June 2001. The most probable source of infection was a ram purchased from farm A in late October 2000. Subsequently this flock was depopulated completely in batches until October 2001.Flock C CLA was first detected on this farm by a clinical examination of the flock in August 2001. The source of the infection was considered to be a ewe, purchased in November 1999, that had originated from the same flock in Scotland that was considered to have infected flock A. Subsequently, this flock was depopulated completely in batches until February 2002.Flock D CLA was first detected on this farm by a clinical examination of the flock in August 2001. The source of infection was traced to five ewes purchased from flock A in December 2000. Forty-one in-contact sheep were depopulated in batches until November 2001. On repeated clinical examinations, the remaining sheep, which were separately managed, were consistently found to be negative for CLA.SerologyAll sheep were blood sampled for the ELISA test prior to euthanasia and postmortem examination. Optical density (OD) readings for the ELISA had been determined from experience of using the test in The Netherlands (Daan Dercksen, personal communication). An OD value above 400 was considered positive, values between 200 and 400 were considered ‘low positive’, those between150 and 200 were inconclusive and those less than 150 were negative. A non-specific reaction was identified when the OD with the negative antigen was more than 50% of the maximum OD of the positive control.Postmortem examination All sheep were necropsied using a standard protocol. An external examination of the carcase was carried out, noting any sinus discharges or subcutaneous swellings. The superficial and visceral lymph nodes, lungs and liver were then examined for abscessation. Suspect lesions were cultured for C. pseudotuberculosis (Cowan and Steel, 1974).resultsOn postmortem examination, 133 of 329 sheep had lesions typical of CLA (Figures 1 and 2); these were confirmed on culture (Table 1). The lesions were present in either single or multiple sites (Table 2: 221 sites in 133 sheep). Lesions in superficial lymph nodes (LN) were most frequently present in the parotid LN (n=44) and the prescapular LN (n=38). Internal lesions were most frequently present in the lungs (n=46) and mediastinal LN (n=31). Table 1: Prevalence of caseous lymphadenitis (CLA) lesions in the four flocks of sheepLesions were present in internal organs only in 32 (24.1 per cent) of the 133 sheep (Table 3); of these 28 (87.5 per cent) were ELISA-positive.Three hundred and eight sheep were defined as either ELISA-positive or ELISA-negative, 18 sheep gave inconclusive tests and three gave non-specific results. When inconclusive and non-specific results were treated as negative, the overall sensitivity of the ELISA for detecting CLA lesions was 0.88 and the specificity was 0.55 (Table 4). Sensitivity varied in the different flocks from 0.83 to 1.0 and specificity varied from 0.31 to 0.87 (Table 5).Table 2: Distribution of caseous lymphadenitis (CLA) lesions in 133 sheepFlock No.of sheep  No.of sheep Prevalence of  examined culture-positive CLA in     for CLA flocks (%)  A  98   35 35.7   B  95   37 38.9   C 100   55 55.0   D  36    6 16.7  All four flocks 329 133 40.4Figure 2: Corynebacterium pseudotuberculosis abscessation in sheep lungs.Lungs 46 (35%) Parotid LN 44 (33%) Prescapular LN 38 (29%) Mediastinal LN 31 (23%) Retropharyngeal LN 11 (8%) Prefemoral LN 10 (8%)Bronchial LN 9 (7%)Submandibular LN 8 (6%)Liver  5 (4%) Supermammary LN 4 (3%)Mesenteric LN 4 (3%)Other sites 11 (8%)Site Number (%)   of sheep  with CLA  lesionsSite Number (%)   of sheep  with CLA  lesionsvolume 59 (1) :  January, 2006    Peer reviewedirish veterinary Journal LN: lymph node.21Peer reviewed volume 59 (1) : January 2006     irish veterinary Journal Table 3: Distribution of caseous lymphadenitis (CLA) lesions in 133 sheepTable 4: Sensitivity and specificity of the ELISA test for caseous lympadenitis (CLA)Table 5: Sensitivity and specificity of ELISA test in individual flocksdiscussionBrown and Olander (1987) reported that the introduction of a single CLA-abscessed animal into a naïve flock results in a high incidence of abscesses within two to three years. In the present study CLA abscesses (Table 1) were detected in 133 animals in these four flocks between seven and 21 months after the introduction of CLA-affected sheep (flock A: 11 months; flock B: seven months; flock C: 21 months; flock D: eight months). Intensive husbandry methods in these flocks may have contributed to the relatively rapid spread of infection.The present study demonstrated that CLA lesions were present only in the viscera (lungs and visceral LN) in approximately 24 per cent of clinical cases. Consequently, clinical examination alone will not detect all infected sheep in a flock. Forty-six of the 133 CLA-positive sheep had lung abscesses. These lesions may lead to more rapid spread of the disease when sheep are closely confined, as for shearing and routine treatments, or when housed.In this study, the ELISA test was relatively sensitive at detecting CLA-infected sheep. However, the specificity of the test in detecting animals with lesions was low (Table 3). This contrasts with the experience of Dercksen et al (2000), who found that the ELISA test had a specificity of 99±1% and a sensitivity of 79±5%. In their study, the sensitivity and specificity were determined in sera from sheep with clinical abscesses from which C. pseudotuberculosis was isolated and in sera from healthy sheep in flocks without a record of CLA. The present study differs in that both CLA-positive and CLA-negative sheep were drawn from infected flocks. Dercksen et al (2000) suggested that cross-reactions may occur due to infections with other bacteria (for example, other Corynebacterium species, Listeria monocytogenes and Mycobacterium avium subspecies paratuberculosis). Evidence of these bacterial infections was not found in the four flocks examined. Batey (1986) suggested that many CLA lesions undergo resolution during the early stages of development. In the present study, it is more likely that seropositive, culture-negative sheep resulted from sheep in these flocks being exposed to infection, but either subsequently not developing the disease or eliminating the infection. The wide variation in specificity between flocks may be attributed to differences in time exposed to C. pseudotuberculosis.Twenty-eight of the 32 sheep with internal lesions only were ELISA-positive. These results indicate that ELISA testing could play a valuable role in CLA control by supplementing clinical detection of superficial lesions. It is anticipated that such ELISAs will become commercially available in the near future.referencesBaird, G.J. (2003). Current perspectives on caseous lymphadenitis. In Practice 25: 62-68.Batey, r.G. (1986). Pathogenesis of caseous lymphadenitis in sheep and goats. Australian Veterinary Journal 63: 269-272.Brown, C.C. and Olander, H.J. (1987). Caseous lymphadenitis of goats and sheep: a review. Veterinary Bulletin 57: 1-12.Cowan, S.T. and Steel, K.J. (1974). Cowan and Steel’s Manual for the Identification of Medical Bacteria. Second Edition. Cambridge: Cambridge University Press.dercksen, d.P., Brinkhof, J.M.A., dekker-Norren, T., van Maanen, K., Bode, C.F., Baird, G. and Kamp, e.M. (2000). A comparison of four serological tests for the diagnosis of caseous lymphadenitis in sheep and goats. Veterinary Microbiology 75: 167-175. nFlock A B C d All 4 flocks (%)  Number of  sheep with  CLA lesions 35 37 55 6 133 (40.4%)  Number of  sheep with  superficial  LN and  cutaneous  lesions only 13 16 37 3 69 (21.0%)  Number of  sheep with  internal  lesions only  (lungs and  visceral LN) 12  11     8  1  32 (9.7%)  Number of  sheep with  both internal  and external  CLA lesions 10 10 10 2 32 (9.7%)  Total number  of sheep  examined 98 95 100 36 329  LN: lymph node; CLA: caseous lymphadenitis.Flock Sensitivity Specificity Flock A (n=98) 0.83 0.59  Flock B (n=95) 0.86 0.53  Flock C (n=100) 0.91 0.31  Flock D (n=36) 1.00 0.87  All 4 flocks (n=329) 0.88 0.55  CLA lesion positive negative total Detected 117  16 133 Sensitivity 0.88  Not detected  88 108 196 Specificity 0.55  Total 205 124 329  

GJ Baird

RG Batey

CC Brown

ST Cowan

DP Dercksen

Crossref

NARCIS 

Malone, FE

Fee, SA

Kamp, EM

King, DC

Baird, GJ

O'Reilly, KM

Murdock, FEA

Spiral - Imperial College Digital Repository

19Peer reviewed volume 59 (1) : January 2006     irish veterinary Journal introductionCaseous lymphadenitis (CLA) is caused by the bacterium Corynebacterium pseudotuberculosis. Although it can affect a number of domestic species and man, it is as a disease of sheep that CLA is most important. C. pseudotuberculosis causes chronic abscessation in lymph nodes (LN) and viscera. The disease is present in many countries throughout the world and was first recorded in sheep in Northern Ireland in 1999, when the source of the infection was traced to imported Scottish sheep. CLA is a notifiable disease in both Northern Ireland and the Republic of Ireland, but restrictions have been removed in Great Britain since 1991.Baird (2003) listed a number of factors that make the control of CLA difficult. CLA may be detected clinically only in superficial LN, whereas in a proportion of affected sheep the lesions may be present only in visceral LN or organs such as the lungs. CLA has a relatively long incubation period, of between two and four months, before it may be detected clinically. In addition, C. pseudotuberculosis may survive for long periods in the environment, thus providing a source of infection in the absence of clinical cases. Dercksen et al. (2000) evaluated a double antibody sandwich ELISA test for the detection of CLA in healthy sheep from CLA-free flocks and in sheep with culture-confirmed CLA. They found that this ELISA test had a specificity of 99±1% and a sensitivity of 79±5%. The present study was designed to evaluate its use in four naturally infected flocks.Materials and methods Flock histories The study was undertaken in 329 sheep (63 male and 266 female) from four flocks, which were depopulated in 2001 and 2002 because of widespread CLA infection.Flock A CLA was first detected on this farm in July 2001 during a clinical examination of the flock that had been prompted by a backward trace of contacts from flock B. The flockowner reported seeing similar lesions in his sheep since December of the previous year, but did not realise that the cause was CLA. The source of the infection was considered to be two rams that were purchased from Scotland in August 2000. Subsequently, this flock was depopulated completely in batches until July 2002.peer reviewedA serological investigation of caseous lymphadenitis in four flocks of sheepFrank e. Malone1, Seán A. Fee1, elbarte M. Kamp2*, david C. King1, Graham J. Baird3, Kath M. O’reilly4 and Fiona e.A. Murdock51 Department of Agriculture and Rural Development for Northern Ireland, Veterinary Sciences Division, 43 Beltany Road, Omagh, County  Tyrone, Northern Ireland, BT78 5NF2 Institute of Animal Science and Health (ID-Lelystad), PO Box 65, 8200 AB Lelystad, The Netherlands * Deceased.3 Scottish Agricultural College, Veterinary Science Division, Greycrook, St. Boswells, Melrose, Scotland,  TD6 0EQ4 Ecology and Epidemiology Group, Biological Sciences, University of Warwick, Coventry, England, CV4 7AL5 Department of Agriculture and Rural Development for Northern Ireland, Veterinary Service, Dundonald House, Upper Newtownards Road,  Belfast, Northern Ireland BT4 3SBA double antibody sandwich ELISA developed by ID-DLO, Lelystad to detect Corynebacterium pseudotuberculosis infection was used on 329 sheep from four pedigree Suffolk flocks in which clinical cases of caseous lymphadenitis (CLA) had occurred. At subsequent necropsy, typical CLA lesions were seen in 133 sheep, and the diagnosis was confirmed on culture. Lesions were most commonly seen in lungs (n=46), parotid lymph nodes (n=44), prescapular lymph nodes (n=38) and mediastinal lymph nodes (n=31). The sensitivity of the ELISA test for detecting culture-positive sheep was 0.88, while the specificity of the test was 0.55. The antibody ELISA detected 87.5 per cent of sheep that had CLA lesions restricted to internal organs only. It was concluded that the ELISA test has a valuable role in detecting sheep with both clinical and subclinical CLA.Irish Veterinary JournalVolume 59 (1) 19-21, 2005Correspondence:Frank MaloneDepartment of Agriculture and Rural Development for Northern Ireland, Veterinary Sciences Division, 43 Beltany Road, Omagh, County Tyrone, Northern Ireland, BT78 5NFTel: +44 (0) 28 822 43337;Fax: +44 (0) 28 822 44228; Email: frank.malone@dardni.gov.uk        Keywords:  Sheep, Caseous lymphadenitis, Serology, Corynebacterium pseudotuberculosis20Figure 1: Corynebacterium pseudotuberculosis abscessation in parotid lymph node.Flock B CLA was first detected on this farm by a clinical examination of the flock in June 2001. The most probable source of infection was a ram purchased from farm A in late October 2000. Subsequently this flock was depopulated completely in batches until October 2001.Flock C CLA was first detected on this farm by a clinical examination of the flock in August 2001. The source of the infection was considered to be a ewe, purchased in November 1999, that had originated from the same flock in Scotland that was considered to have infected flock A. Subsequently, this flock was depopulated completely in batches until February 2002.Flock D CLA was first detected on this farm by a clinical examination of the flock in August 2001. The source of infection was traced to five ewes purchased from flock A in December 2000. Forty-one in-contact sheep were depopulated in batches until November 2001. On repeated clinical examinations, the remaining sheep, which were separately managed, were consistently found to be negative for CLA.SerologyAll sheep were blood sampled for the ELISA test prior to euthanasia and postmortem examination. Optical density (OD) readings for the ELISA had been determined from experience of using the test in The Netherlands (Daan Dercksen, personal communication). An OD value above 400 was considered positive, values between 200 and 400 were considered ‘low positive’, those between150 and 200 were inconclusive and those less than 150 were negative. A non-specific reaction was identified when the OD with the negative antigen was more than 50% of the maximum OD of the positive control.Postmortem examination All sheep were necropsied using a standard protocol. An external examination of the carcase was carried out, noting any sinus discharges or subcutaneous swellings. The superficial and visceral lymph nodes, lungs and liver were then examined for abscessation. Suspect lesions were cultured for C. pseudotuberculosis (Cowan and Steel, 1974).resultsOn postmortem examination, 133 of 329 sheep had lesions typical of CLA (Figures 1 and 2); these were confirmed on culture (Table 1). The lesions were present in either single or multiple sites (Table 2: 221 sites in 133 sheep). Lesions in superficial lymph nodes (LN) were most frequently present in the parotid LN (n=44) and the prescapular LN (n=38). Internal lesions were most frequently present in the lungs (n=46) and mediastinal LN (n=31). Table 1: Prevalence of caseous lymphadenitis (CLA) lesions in the four flocks of sheepLesions were present in internal organs only in 32 (24.1 per cent) of the 133 sheep (Table 3); of these 28 (87.5 per cent) were ELISA-positive.Three hundred and eight sheep were defined as either ELISA-positive or ELISA-negative, 18 sheep gave inconclusive tests and three gave non-specific results. When inconclusive and non-specific results were treated as negative, the overall sensitivity of the ELISA for detecting CLA lesions was 0.88 and the specificity was 0.55 (Table 4). Sensitivity varied in the different flocks from 0.83 to 1.0 and specificity varied from 0.31 to 0.87 (Table 5).Table 2: Distribution of caseous lymphadenitis (CLA) lesions in 133 sheepFlock No.of sheep  No.of sheep Prevalence of  examined culture-positive CLA in     for CLA flocks (%)  A  98   35 35.7   B  95   37 38.9   C 100   55 55.0   D  36    6 16.7  All four flocks 329 133 40.4Figure 2: Corynebacterium pseudotuberculosis abscessation in sheep lungs.Lungs 46 (35%) Parotid LN 44 (33%) Prescapular LN 38 (29%) Mediastinal LN 31 (23%) Retropharyngeal LN 11 (8%) Prefemoral LN 10 (8%)Bronchial LN 9 (7%)Submandibular LN 8 (6%)Liver  5 (4%) Supermammary LN 4 (3%)Mesenteric LN 4 (3%)Other sites 11 (8%)Site Number (%)   of sheep  with CLA  lesionsSite Number (%)   of sheep  with CLA  lesionsvolume 59 (1) :  January, 2006    Peer reviewedirish veterinary Journal LN: lymph node.21Peer reviewed volume 59 (1) : January 2006     irish veterinary Journal Table 3: Distribution of caseous lymphadenitis (CLA) lesions in 133 sheepTable 4: Sensitivity and specificity of the ELISA test for caseous lympadenitis (CLA)Table 5: Sensitivity and specificity of ELISA test in individual flocksdiscussionBrown and Olander (1987) reported that the introduction of a single CLA-abscessed animal into a naïve flock results in a high incidence of abscesses within two to three years. In the present study CLA abscesses (Table 1) were detected in 133 animals in these four flocks between seven and 21 months after the introduction of CLA-affected sheep (flock A: 11 months; flock B: seven months; flock C: 21 months; flock D: eight months). Intensive husbandry methods in these flocks may have contributed to the relatively rapid spread of infection.The present study demonstrated that CLA lesions were present only in the viscera (lungs and visceral LN) in approximately 24 per cent of clinical cases. Consequently, clinical examination alone will not detect all infected sheep in a flock. Forty-six of the 133 CLA-positive sheep had lung abscesses. These lesions may lead to more rapid spread of the disease when sheep are closely confined, as for shearing and routine treatments, or when housed.In this study, the ELISA test was relatively sensitive at detecting CLA-infected sheep. However, the specificity of the test in detecting animals with lesions was low (Table 3). This contrasts with the experience of Dercksen et al (2000), who found that the ELISA test had a specificity of 99±1% and a sensitivity of 79±5%. In their study, the sensitivity and specificity were determined in sera from sheep with clinical abscesses from which C. pseudotuberculosis was isolated and in sera from healthy sheep in flocks without a record of CLA. The present study differs in that both CLA-positive and CLA-negative sheep were drawn from infected flocks. Dercksen et al (2000) suggested that cross-reactions may occur due to infections with other bacteria (for example, other Corynebacterium species, Listeria monocytogenes and Mycobacterium avium subspecies paratuberculosis). Evidence of these bacterial infections was not found in the four flocks examined. Batey (1986) suggested that many CLA lesions undergo resolution during the early stages of development. In the present study, it is more likely that seropositive, culture-negative sheep resulted from sheep in these flocks being exposed to infection, but either subsequently not developing the disease or eliminating the infection. The wide variation in specificity between flocks may be attributed to differences in time exposed to C. pseudotuberculosis.Twenty-eight of the 32 sheep with internal lesions only were ELISA-positive. These results indicate that ELISA testing could play a valuable role in CLA control by supplementing clinical detection of superficial lesions. It is anticipated that such ELISAs will become commercially available in the near future.referencesBaird, G.J. (2003). Current perspectives on caseous lymphadenitis. In Practice 25: 62-68.Batey, r.G. (1986). Pathogenesis of caseous lymphadenitis in sheep and goats. Australian Veterinary Journal 63: 269-272.Brown, C.C. and Olander, H.J. (1987). Caseous lymphadenitis of goats and sheep: a review. Veterinary Bulletin 57: 1-12.Cowan, S.T. and Steel, K.J. (1974). Cowan and Steel’s Manual for the Identification of Medical Bacteria. Second Edition. Cambridge: Cambridge University Press.dercksen, d.P., Brinkhof, J.M.A., dekker-Norren, T., van Maanen, K., Bode, C.F., Baird, G. and Kamp, e.M. (2000). A comparison of four serological tests for the diagnosis of caseous lymphadenitis in sheep and goats. Veterinary Microbiology 75: 167-175. nFlock A B C d All 4 flocks (%)  Number of  sheep with  CLA lesions 35 37 55 6 133 (40.4%)  Number of  sheep with  superficial  LN and  cutaneous  lesions only 13 16 37 3 69 (21.0%)  Number of  sheep with  internal  lesions only  (lungs and  visceral LN) 12  11     8  1  32 (9.7%)  Number of  sheep with  both internal  and external  CLA lesions 10 10 10 2 32 (9.7%)  Total number  of sheep  examined 98 95 100 36 329  LN: lymph node; CLA: caseous lymphadenitis.Flock Sensitivity Specificity Flock A (n=98) 0.83 0.59  Flock B (n=95) 0.86 0.53  Flock C (n=100) 0.91 0.31  Flock D (n=36) 1.00 0.87  All 4 flocks (n=329) 0.88 0.55  CLA lesion positive negative total Detected 117  16 133 Sensitivity 0.88  Not detected  88 108 196 Specificity 0.55  Total 205 124 329  

http://doaj.org/search?source=%7B%22query%22%3A%7B%22bool%22%3A%7B%22must%22%3A%5B%7B%22term%22%3A%7B%22id%22%3A%224a99c075249046a499f84ef693541cd8%22%7D%7D%5D%7D%7D%7D

A serological investigation of caseous lymphadenitis in four flocks of sheep

Abstract

Similar works

Full text

Available Versions