The role of Corynebacterium pseudotuberculosis lung lesions in the transmission of this bacterium to other sheep

SUMMARY: :Five groups of 5 shorn and 5 unshorn caseous lymphadenitis (CLA)-free Merino wether weaners were each placed in feedlot pens with 6 Merino ewes, 2 or more of which had CLA lung lesions but no discharging superficial lesions. The sheep were kept together for 5 months.
Twenty-eight per cent of the shorn weaners and 20% of the unshorn weaners developed antibodies to Corynebacterium pseudotuberculosis. At slaughter, 8% of the shorn weaners and 12% of the unshorn weaners had CLA lesions in either lungs, lymph nodes or both. In the absence of contact with CLA-infected ewes, a control group of 5 shorn and 5 unshorn weaners failed to develop antibodies to C. pseudotuberculosis or CLA lesions in the same period. This showed that sheep with CLA abscesses in the lungs but no discharging superficial abscesses were a source of C. pseudotuberculosis infection to other sheep. Aust Vet J. 64: 261–263     

Development and validation of an ELISA to detect antibodies to Corynebacterium pseudotuberculosis in ovine sera

Several enzyme-linked immunosorbent assays (ELISAs) have been developed for the detection of antibodies to Corynebacterium pseudotuberculosis, the causative agent of caseous lymphadenitis (CLA). However, none are commercially available in the UK. It was therefore necessary to develop a new, economic ELISA for use in a research project studying the epidemiology of CLA in UK sheep. The ELISA with its diagnostic qualities is presented. The ELISA was developed using sonicated C. pseudotuberculosis and optimised to detect total antibody or IgG class antibody in serum. Receiver operating characteristic (ROC) curves were obtained and the area under the ROC curve was used to compare the sensitivity and specificity of the two ELISAs. Both versions of the ELISA were evaluated on a panel of 150 positive reference sera and 103 negative reference sera. Using the test at 100% specificity, the sensitivity of detection of total antibody was 71% (95% confidence interval 63-78%), and the sensitivity of detection of IgG antibody to C. pseudotuberculosis was 83% (76-89%), which compares favourably with other reported ELISA tests for CLA in sheep. The sensitivity of the IgG antibody assay may be higher because of the greater affinity of IgG class antibodies compared with the IgM antibodies also detected by the total antibody ELISA. The results of ROC analysis indicated that the IgG isotype ELISA was more accurate than the total antibody ELISA. The efficiency of the test was greatest when serum samples were run in a dilution series than when any single serum dilution was used. The ELISA is considered to be suitable for application in field studies of CLA in UK sheep.     

Comparison of a maedi-visna virus CA-TM fusion protein ELISA with other assays for detecting sheep infected with North American ovine lentivirus strains.

A maedi-visna virus CA-TM fusion protein ELISA (MVV ELISA) was evaluated for the detection of antibody in sheep infected with North American ovine lentivirus (OvLV). The results of the MVV ELISA were compared with other assays for OvLV antibody and with viral infection in an intensively studied group of 38 sheep with a high prevalence of OvLV infection and disease. The sensitivity, specificity, and concordance of assays for OvLV antibody (MVV ELISA, indirect ELISA, Western blot, and AGID), virus (virus isolation, PCR, antigen ELISA), and OvLV-induced disease in each animal were compared with OvLV infection status as defined by a positive result in two or more of the assays. Five sheep met the criteria for absence of OvLV infection. The sensitivity of the MVV ELISA in detecting OvLV infected sheep was 64%, whereas the sensitivity of the other three tests for antibody ranged from 85 to 94%. All the antibody assays were 100% specific in this group of animals. Of the assays for virus, the PCR test had the highest sensitivity and the best concordance with OvLV infection, but it also had the lowest specificity of any of the virus or antibody assays. Among the antibody tests, the concordance of the MVV ELISA compared most favorably with the AGID test for detecting OvLV-infected sheep. Analysis of serum samples from 28 lambs experimentally-infected with one of three North American strains of OvLV suggested that there were no significant strain differences detectable by antibody assay. Twenty virus-inoculated lambs were positive by both the MVV ELISA and the AGID test, five lambs were MVV ELISA negative and AGID test positive, and three lambs were MVV ELISA positive and AGID test negative. No pre-inoculation samples were positive by either assay. In a longitudinal study involving seven lambs, antibodies to OvLV were detected by AGID 3-5 weeks post-inoculation, but were not detected by MVV ELISA until 5-10 weeks post-inoculation. Among 128 naturally and experimentally-infected sheep that were seropositive in the AGID test, the overall sensitivity of the MVV ELISA was higher in the naturally infected sheep (84%) than in the experimentally infected sheep (69%). The data indicated that the MVV ELISA represents a less sensitive, but specific alternative for the detection of OvLV antibodies.     

Rapid detection of Corynebacterium pseudotuberculosis in clinical samples from sheep

Corynebacterium pseudotuberculosis, a Gram-positive bacterium is the causative agent of caseous lymphadenitis (CLA), a chronic disease of sheep, goats and other warm blooded animals. In the present study, a total of 1,080 sheep reared under semi-intensive system on organized farms situated in the semi arid tropical region of Rajasthan, India, was clinically examined. Pus samples from superficial lymph nodes of 25 (2.31 %) adult sheep showing clinical lesions similar to CLA were collected for laboratory analyses. On the basis of morphological, cultural and biochemical characteristics 12 (48 %) bacterial isolates from pus identified it as C. pseudotuberculosis. A polymerase chain reaction (PCR) assay targeting Putative oligopeptide/dipeptide ABC transporter, nicotinamide adenine dinucleotide phosphate (NADP) oxidoreductase coenzyme F420-dependent and proline iminopeptidase (PIP) genes of C. pseudotuberculosis was developed that showed 14 pus samples as positive. All C. pseudotuberculosis isolates were also found positive for these genes in the PCR. The specificity of the PCR products was confirmed by sequencing of the amplified products that showed 98–100 % homology with published sequences available in the NCBI database. The present study shows the incidence of CLA as 2.31 %, 1.1 % and 1.29 % based on clinical, bacterial culture and direct pus PCR assay, respectively. The PCR assay was rapid, specific and as significant as bacterial culture in detecting bacteria directly in the clinical pus samples. The PCR assay developed in the study can be applied for the diagnosis and control of CLA. Furthermore, the assay can also be applied to detect C. pseudotuberculosis in various clinical samples.     

CORYNEBACTERIUM PSEUDOTUBERCULOSIS INFECTION OF SHEEP: ROLE OF SKIN LESIONS AND DIPPING FLUIDS

Caseous lymphadenitis (CLA) infection was established experimentally in a total of 30, 2-tooth merino wethers. In the first experiment a broth culture of the causative organism, Corynebacterum pseudotuberculosis was placed directly on to a freshly shorn area on the right shoulder of 21 sheep after pretreatment with a defatting agent, water or 4 different commercial sheep dips. Within 6–13 days focal skin lesions appeared on 7 sheep and these were followed by abscessation of the right prescapular lymph node. Lung lesions of CLA developed in 6 sheep; 5 of which had skin lesions. In the second experiment C. Pseudotuberculosis broth culture was applied over the right shoulder region of 14 sheep, 24 hours post shearing. The 7 animals had only 1 application of the culture and the remaining 7 had 4 applications on consecutive days. A total of 9 of the 14 sheep developed skin lesions and subsequent CLA in the right prescapular lymph nod but there was no significant difference between the 2 methods of application. As part of the second experiment a broth culture of C. pseudotuberculosis was added to a commercial arsenical sheep dip and 2 hours later sprayed on to the right side of sheep shorn 24 hours or 2 weeks previously. Within 6–13 days all 14 sheep treated in this way developed skin lesions from which C. pseudotuberculiosis was recovered in dense pure growth. Healing time of skin lesions in all affected sheep ranged from 17–72 days post inoculation. A serological test to detect the presence of C. pseudotuberculosis antitoxin was used at regular intervals on all experimental sheep. The earliest positive reaction in an affected sheep was recorded 4 weeks post inoculation. In 23 known affected sheep tested at 35 weeks post inoculation, 10 were positive to the Zaki test. No false positive reactors were recorded. The localisation of focal lung lesions in the interstitial tissue suggests that pulmonary abscessation caused by C. pseudotuberculosis develops via the haematogenous route.     

The use of pestivirus antigen ELISA currently available for the detection of hairy shaker disease/border disease virus in sheep

AIMS: To estimate the number of cases of scrapie that would occur in sheep of different prion protein (PrP) genotypes if scrapie was to become established in New Zealand, and to compare the performance of two commercially available, rapid ELISA kits using ovine retro-pharyngeal lymph nodes (RLN) from non-infected and infected sheep of different PrP genotypes.

METHODS: Using published data on the distribution of PrP genotypes within the New Zealand sheep flock and the prevalence of cases of scrapie in these genotypes in the United Kingdom, the annual expected number of cases of scrapie per genotype was estimated, should scrapie become established in New Zealand, assuming a total population of 28 million sheep. A non-infected panel of RLN was collected from 737 sheep from New Zealand that had been culled, found in extremis or died. Brain stem samples were also collected from 131 of these sheep. A second panel of infected samples comprised 218 and 117 RLN from confirmed scrapie cases that had originated in Europe and the United States of America, respectively. All samples were screened using two commercial, rapid, transmissible spongiform encephalopathy ELISA kits: Bio-Rad TeSeE ELISA (ELISA-BR), and IDEXX HerdChek BSE-Scrapie AG Test (ELISA-ID).

RESULTS: If scrapie became established in New Zealand, an estimated 596 cases would occur per year; of these 234 (39%) and 271 (46%) would be in sheep carrying ARQ/ARQ and ARQ/VRQ PrP genotypes, respectively. For the non-infected samples from New Zealand the diagnostic specificity of both ELISA kits was 100%. When considering all infected samples, the diagnostic sensitivity was 70.4 (95% CI=65.3–75.3)% for ELISA-BR and 91.6 (95% CI=88.2–94.4)% for ELISA-ID. For the ARQ/ARQ genotype (n=195), sensitivity was 66.2% for ELISA-BR and 90.8% for ELISA-ID, and for the ARQ/VRQ genotype (n=107), sensitivity was 81.3% for ELISA-BR and 98.1% for ELISA-ID.

CONCLUSIONS: In this study, the ELISA-ID kit demonstrated a higher diagnostic sensitivity for detecting scrapie in samples of RLN from sheep carrying scrapie-susceptible PrP genotypes than the ELISA-BR kit at comparable diagnostic specificity.

CLINICAL RELEVANCE: The diagnostic performance of the ELISA-ID kit using ovine RLN merits the consideration of including this assay in the national scrapie surveillance programme in New Zealand.     

Evidence of decreased concanavalin A induced suppressor cell activity in the peripheral blood and pulmonary lymph nodes of sheep with ovine progressive pneumonia

Concanavalin A (Con A) induced suppressor cell activity was evaluated in a group of ovine progressive pneumonia virus-antibody positive sheep (OPPV+). Decreased levels of suppressor activity were observed in the peripheral blood and regional pulmonary lymph nodes of animals with clinical and pathological evidence of ovine progressive pneumonia (OPP) when compared to animals with no lesions and animals with caseous lymphadenitis involving the lungs and pulmonary lymph nodes. Decreased levels of Con A stimulated lymphocyte proliferation was also observed in the peripheral blood and iliac lymph nodes of sheep with OPP. Sheep with OPP were found to be hypogammaglobulinemic. Sera from OPPV+ sheep had no effect on T and B cell mitogen stimulated responses of lymphocytes from sheep seronegative for antibodies to ovine progressive pneumonia virus (OPPV-) when compared to normal sheep serum.     

Peste des petitis ruminants morbillivirus infection in lambs and young goats at Qassim region, Saudi Arabia

An outbreak of peste des petits ruminants (PPR) in lambs and young goats of Najdi breed of sheep and goats occurred during winter 2005 at Qassim region of central Saudi Arabia. The PPR infection was confirmed by demonstration of antibodies against the virus in the serum of clinically-ill young sheep and goats using competitive ELISA test. Clinical examination of infected animals showed fever, salivation, lacrimation, mucopurulent nasal discharge, difficult breathing and diarrhoea. The disease was particularly severe in the goats. Morbidity was about 20% and mortality was less than 3 percent. Autopsy showed necrotic and ulcerative lesions in the mouth, stomach and intestine. Mesenteric lymph nodes were swollen and congested. The lungs were patchy pneumonic mostly at the diaphragmatic and apical lobes. Liver and kidney lesions were seen in goats only and both organs were congested and necrotic. Histopathological examination revealed necrotic inflammation in the gastrointestinal tract. Intracytoplasmic viral inclusions were seen in the enterocytes of goats. Lung sections showed bronchopneumonia and syncytial and giant cells. The bronchial epithelium of goats had intracytoplasmic viral inclusions. Extensive coagulation necrosis, fatty degeneration and presence of intracytoplasmic viral inclusions were seen in hepatocytes and syncytial cells were evident in biliary epithelium of goats. Congestion, coagulation necrosis and syncytial cells were seen in the renal tubular epithelium of goats only. In a survey to determine the magnitude of the outbreak, PPR antibodies were evidenced in 363/996 (36.6%) sheep and 530/962 (55.1%) goats.     

Serum antibody responses in sheep after natural infection with Bacteroides nodosus

Soluble outer membrane protein of Bacteroides nodosus extracted with potassium thiocyanate (KSCN) was employed as antigen in an enzyme linked immunosorbent assay (ELISA) to detect serum antibody in sheep naturally infected with a heterologous serogroup. Serum antibody responses in 55 sheep were monitored for 2 years and maximum levels were directly related to the severity of clinical foot lesions. Serum antibody levels rose 2 weeks after foot lesions developed and declined within several months of resolution of lesions. After the first footrot transmission period, antibody levels persisted significantly (P less than 0.001) longer in sheep that did not become affected in the next transmission period compared with sheep in which footrot recurred. Antibody response did not appear to result in resolution of foot lesions. ELISA using KSCN antigen gave similar results to whole cell ELISA where cells prepared from an homologous serogroup were used as antigen. Both these assays were more sensitive than ELISA in which heterologous whole cell antigen was used. Proteins extracted from the outer membrane of B. nodosus, which are known to be immunogenic in natural infection and common to different serogroups of B. nodosus, appear to be useful antigens for serological investigations of ovine footrot.     

Health of finishing steers: effects on performance, carcass traits, and meat tenderness

The impact of respiratory disease during a 150-d feedlot finishing period on daily gain, carcass traits, and longissimus tenderness was measured using 204 steer calves. Feedlot health status was monitored in two ways. First, clinical signs of respiratory infection were evaluated each day; treatment with antibiotic was based on degree of fever (if rectal temperature exceeded 40 degrees C then calves were treated). Steers that were treated (n = 102) had lower (P<.05) final live weights, ADG, hot carcass weights (HCW), less external and internal fat, and more desirable yield grades. Steers that were treated had a higher prevalence of carcasses that graded U.S. Standard than steers that were never treated. Second, as an alternative index of health status, lungs of all steers were evaluated at the processing plant using a respiratory tract lesion classification system; this health index included presence or absence of preexisting pneumonic lesions in the anterioventral lobes plus activity of the bronchial lymph nodes (inactive vs active). Lung lesions were present in 33% of all lungs and were distributed almost equally between treated (37%) and untreated cattle (29%). Steers with lesions (n = 87) had lower (P<.05) daily gains, lighter HCW, less internal fat, and lower marbling scores than steers without lesions. Compared to steers with lesions but inactive bronchial lymph nodes (n = 78), steers with lung lesions plus active lymph nodes had lower (P<.01) ADG and dressing percentage. Longissimus shear force values for steaks aged 7 d were lower (P = .05) from steers without lung lesions than those for steaks from steers with lung lesions. Overall, morbidity suppressed daily gains and increased the percentage of U.S. Standard carcasses. Compared to health assessment by clinical appraisal (based on elevated body temperature), classification based on respiratory tract lesions at slaughter proved more reliable statistically and, thereby, more predictive of adverse effects of morbidity on production and meat tenderness.     

Utility of 2 Immunological Tests for Antemortem Diagnosis of Equine Protozoal Myeloencephalitis (Sarcocystis neurona Infection) in Naturally Occurring Cases

Background: Antemortem diagnosis of equine protozoal myeloencephalitis (EPM) is challenging. Limited information is available regarding a commercial test (surface antigen 1 [SAG‐1] ELISA). Performance of another commercial test (indirect fluorescent antibody test [IFAT]) using samples from an independent group has not been well described. Hypothesis/Objectives: The primary goal was to evaluate the SAG‐1 ELISA and IFAT using naturally occurring EPM cases. A secondary goal was to obtain more information regarding clinical presentation. Animals: Hospital cases were admitted over 20 months and classified into 4 groups. Confirmed positive cases (n = 9) had asymmetric or multifocal neurologic deficits or both and postmortem lesions consistent with EPM. Confirmed negative cases (n = 17) had variable clinical signs and postmortem lesions consistent with another neurologic disease (or no lesions). Suspected positive cases (n = 10) had asymmetric or multifocal deficits or both, marked improvement after treatment for EPM, and other likely diseases excluded. Suspected negative cases (n = 29) had orthopedic disease and no neurologic deficits. Methods: Results of immunological testing (SAG‐1 ELISA and IFAT on serum or cerebrospinal fluid [CSF] or both), neurologic examinations, CSF analyses, and postmortem examinations were analyzed retrospectively. Results: SAG‐1 ELISA sensitivity was 12.5% (95% CI, 1.6–38.4) and specificity was 97.1% (95% CI, 84.7–99.9) using serum. IFAT sensitivity was 94.4% (95% CI, 72.7–99.9) and specificity was 85.2% (95% CI, 66.3–95.8) using serum; sensitivity was 92.3% (95% CI, 64.0–99.8) and specificity was 89.7% (95% CI, 72.7–97.8) using CSF. Conclusions and Clinical Importance: Low sensitivity of the SAG‐1 ELISA limited its usefulness for antemortem diagnosis of EPM in this patient population.     

Sensitivity and specificity of two serological tests for the detection of ovine paratuberculosis

OBJECTIVE: To determine the sensitivity and specificity of an absorbed ELISA and an AGID test for the detection of clinical and subclinical paratuberculosis in sheep. DESIGN: By testing a panel of sera from 1257 Australian Merino and crossbred sheep greater than 1 year of age, of which 1137 sheep were not infected with Mycobacterium avium subsp paratuberculosis and 120 sheep had paratuberculosis. PROCEDURE: Sera were collected from 457 sheep in Victoria and 800 sheep in Western Australia. Presence of M a paratuberculosis infection in Victorian sheep was determined by histological examination of intestinal tissues, whereas sheep from Western Australia were presumed to be free of Johne's disease. The ability of an absorbed ELISA to discriminate between infected and uninfected sheep was described by test sensitivity and specificity, the distribution of ELISA OD, and the area under a receiver operating characteristic curve. RESULTS: The absorbed ELISA had a specificity of 98.2 to 99.5% (CI) and a sensitivity of 35 to 54% (CI). In sheep from infected flocks in Victoria, the AGID test had a specificity of 99 to 100% (CI) and a sensitivity of 38 to 56% (CI). The sensitivity of serological tests was higher in sheep with a body condition representative of the lower quintile of their flock of origin. CONCLUSION: The AGID test and absorbed ELISA are useful tests for the detection of ovine paratuberculosis. Although the tests had a similar accuracy, they detected different subpopulations of infected sheep with only moderate overlap. The AGID test had a higher specificity than the absorbed ELISA.     

Post-shearing management affects the seroincidence of Corynebacterium pseudotuberculosis infection in sheep flocks

Associations between the incidence of caseous lymphadenitis (CLA) in sheep and post-shearing management and environmental factors were examined in a prospective study. CLA incidence was measured in 126 groups of 1- and 2-year-old sheep in 70 Western Australian flocks selected from the prevalence of cull-for-age ewes with CLA lesions at abattoirs. CLA incidence was assessed using a CLA toxin ELISA. Dichotomous and polychotomous stepwise logistic regression methods were compared in examining the effects of management and environment on CLA incidence. Shower dipping sheep for ectoparasite control after shearing increased the odds of high CLA incidence by five to six times and keeping sheep under cover for 1 h or more after shearing increased the odds of being in high CLA incidence categories three-fold. The seroprevalence of existing CLA infection within each age group affected incidence more than did the overall slaughter-based flock estimate. This suggests that CLA spreads mostly within groups of sheep shorn together.     

Comparison of a commercial H1N1 enzyme-linked immunosorbent assay and hemagglutination inhibition test in detecting serum antibody against swine influenza viruses.

Recently a commercial enzyme-linked immunosorbent assay (ELISA) kit for detecting antibody against H1N1 swine influenza virus (SIV) has been made available to diagnosticians and veterinary practitioners. Because the hemagglutination inhibition (HI) test has been considered the standard test for SIV serology, diagnostic performance of the new ELISA was evaluated using positive (n = 60) and negative (n = 188) serum samples from young pigs with known status of SIV infection and compared with that of the HI test. Both ELISA and HI test identified all negative animals correctly. None of the serum samples (n = 64) from pigs inoculated with H3N2 SIV was positive by ELISA for SIV antibody. The H1N1 SIV antibody detectable by ELISA appears to develop more slowly in comparison with antibody detectable by HI test. Although antibody was detected by HI test in all inoculated animals (n = 20) by day 7 postinoculation (PI), antibody was detected by ELISA in 0%, 75%, and 100% of the inoculated animals on days 7, 14, and 28 PI, respectively. Discrepancy in test results between the 2 serologic tests appeared to be because of differences in antibody isotypes detected by each test. Enzyme-linked immunosorbent assay mainly detected IgG antibody, whereas the HI test detects IgM antibody very efficiently as well as IgG antibody. Collectively, the commercial ELISA is highly specific for antibody to H1N1 SIV but may not identify positive animals at the early stage of infection as effectively as the HI test, particularly when SIV is introduced to a na?ve swine population.     

Immunomorphologic and morphometric changes in pulmonary lymph nodes of sheep with progressive pneumonia

Morphologic, immunohistochemical, and morphometric studies were conducted on the posterior mediastinal lymph nodes of eleven sheep with naturally occurring ovine progressive pneumonia and four apparently healthy sheep with no pulmonary lesions (three seropositive, one seronegative for antibody to ovine progressive pneumonia virus). Compared with lesion-free sheep, sheep with ovine progressive pneumonia had a seven-fold increase in B lymphocyte areas and a 21/2-fold increase in T lymphocyte areas of these lymph nodes. Immunochemistry revealed cytoplasmic immunoglobulin G in scattered cells of germinal centers, medullary cords and interfollicular areas and membrane-associated immunoglobulin G in dendritic cells of germinal centers. Immunoglobulin M staining cells were widely scattered in germinal centers and medullary cords. Although B cell hyperplasia seemed to be the predominant process in lymph nodes of sheep with ovine progressive pneumonia, this was not accompanied by the expected degree of plasmacytosis, morphologically and immunohistochemically. These findings may represent an aberrancy of immunoregulation in ovine progressive pneumonia.     

Parameter estimation and simulations of a mathematical model of Corynebacterium pseudotuberculosis transmission in sheep

Caseous lymphadenitis (CLA) is an infectious disease of sheep caused by Corynebacterium pseudotuberculosis. It is prevalent in most sheep producing countries and was introduced into the UK sheep population in 1991. The pathogen invades the host through epithelium and forms an abscess in the local draining lymph node. Typically, disease presents as clinical, with overt (externally visible) swollen lymph nodes (the parotid, submandibular, prefemoral, prescapular, popliteal or mammary) or sub-clinical, with abscesses in the lungs and associated thoracic (bronchial and mediastinal) lymph nodes.

We present a mathematical model in which disease is categorised as overt and/or respiratory (sub-clinical), using the above groupings. In both situations sheep may be infected and may or may not be infectious. In the model, overt abscesses may resolve and respiratory abscesses are considered to be present for life. Using the location of the abscesses, three routes of transmission are postulated: overt to overt, respiratory to overt and respiratory to respiratory. Data from four naturally infected flocks were used to describe populations of sheep with epidemic CLA and to estimate transmission coefficients for each of the postulated transmission routes. The infection process parameters were derived from literature where possible. Parameters were estimated using maximum likelihood methods and compared to the data using a multinomial distribution.

The distribution of abscesses in the flocks was similar to endemic data reported in other studies. In the model most infected sheep developed abscesses, and approximately 36% of sheep with overt abscesses recovered from infection. The average time for respiratory abscesses to become infectious was 41 days. In these data, overt to overt transmission was the most frequent route of transmission since it had the highest coefficient in the model compared with respiratory to overt and respiratory to respiratory transmission. Transmission coefficients specific for each flock significantly (P < 0.05) improved the model fit to the data.

In simulations using values of best-fitting parameter combinations, the proportion of sheep infected was between 0.39 and 0.60 at equilibrium. This is the first mathematical model of C. pseudotuberculosis infection, the parameter estimates indicate that aspects of the infection process could be utilised to design control strategies.     

Ovine interleukin-2: partial purification and assay in normal sheep and sheep with ovine progressive pneumonia

Interleukin-2 (IL-2), a T cell derived lymphokine, acts in nonspecific hormone-like fashion to maintain proliferation of activated lymphocytes in vitro and is believed to play a key role in cell-mediated immune function in vivo. The parameters of induction and assay of factors with IL-2 activity were examined in a group of clinically normal sheep seronegative for antibodies to ovine progressive pneumonia virus (OPPV-). Supernatants from cultures of Concanavalin A (Con A) stimulated mononuclear leukocytes (ML) derived from peripheral blood and lymph nodes contained factors with the capacity to maintain continued proliferation in Con A stimulated lymphoblasts. This activity was localized by gel chromatography to fractions containing proteins of 17,000-20,000 daltons. In a group of sheep seropositive for antibodies to OPPV (OPPV+), decreased levels of IL-2 activity were found in ML culture supernatants derived from the posterior mediastinal lymph nodes of sheep with clinical and pathological evidence of OPP, when compared to OPPV+ sheep with no lesions and sheep with visceral caseous lymphadenitis. This decrease in IL-2 activity appeared not to be associated directly with levels of prostaglandin E2 in these supernatants. These findings may correlate with virus induced alterations in cell mediated immune function in lymphoproliferative lesions of OPP.     

Surveys on Coxiella burnetii infections in Swedish cattle,sheep, goats and moose

Background

Q fever is a zoonotic disease caused by the bacterium Coxiella burnetii. Prevalence data in ruminant species are important to support risk assessments regarding public and animal health. The aim was to investigate the presence of or exposure to C. burnetii in cattle, sheep, goats and moose, and to compare two enzyme-linked immunosorbent assays (ELISAs). National surveys of antibodies against C. burnetii were performed for dairy cattle (n=1537), dairy goats (n=58) and sheep (n=518). Bovine samples consisted of bulk milk, caprine of pooled milk, and ovine of pooled serum. Antibodies were investigated in moose samples (n=99) from three regions. A one-year regional cattle bulk milk survey was performed on the Isle of Gotland (n=119, four occasions). Cattle, sheep and goat samples were analysed with indirect ELISA and moose samples with complement fixation test. For the sheep, goat, and parts of the cattle survey, samples were run in parallel by ELISAs based on antigens from infected ruminants and ticks. Bulk milk samples from the regional cattle survey and vaginal swabs from a subset of the sheep herds (n=80) were analysed for the agent by polymerase chain reaction. Spatial clustering was investigated in the national cattle survey.

Results

The prevalence of antibodies in dairy herds was 8.2% with large regional differences. High risk clusters were identified in the southern regions. The prevalence among dairy herds on the Isle of Gotland varied from 55.9% to 64.6% and 46.4% to 58.9.0% for antibodies and agent, respectively, overall agreement between agent and antibodies was 85.2%. The prevalence of antibodies in sheep was 0.6%, the agent was not detected the vaginal swabs. Antibodies were not detected in goats or moose, although parts of the moose samples were collected in an area with high prevalence in cattle. The overall agreement between the two ELISAs was 90.4%.

Conclusions

The prevalence of antibodies against C. burnetii in dairy cattle in Sweden shows large regional differences. The results suggest that C. burnetii is a rare pathogen among Swedish moose, dairy goat and sheep. ELISAs based on ruminant and tick antigen performed in a similar manner under Swedish conditions.     

Brucellae through the food chain: the role of sheep, goats and springbok (Antidorcus marsupialis) as sources of human infections in Namibia

A confirmed case of human brucellosis motivated an investigation into the potential source of infection in Namibia. Since domestic animals are principal sources of Brucella infection in humans, 1692 serum samples were screened from sheep, goats and cattle from 4 presumably at-risk farms and 900 springbok (Antidorcas marsupialis) serum samples from 29 mixed farming units for Brucella antibodies by the Rose-Bengal test (RBT) and positive cases confirmed by complement fixation test (CFT). To assess the prevalence of human brucellosis, 137 abattoir employees were tested for Brucella antibodies using the standard tube agglutination test (STAT) and by enzyme linked immunosorbent assay (ELISA). Cattle and sheep from all 4 farms were negative by RBT and CFT but 2 of the 4 farms (Ba and C) had 26/42 and 12/285 seropositive goats, respectively. Post mortem examination of seropositive goats revealed no gross pathological lesions typical of brucellosis except enlarged mesenteric and iliac lymph nodes seen in a single buck. Culture for brucellae from organs of seropositive animals was negative. None of the wildlife sera tested positive by either RBT or CFT. Interviews revealed that besides the case that prompted the investigation, a family and another person from other farms with confirmed brucellosis shared a common history of consumption of unpasteurised goat milk, home-made goat cheese and coffee with raw milk and prior contact with goats, suggesting goats as the likely source of infection. All 137 abattoir employees tested negative by STAT, but 3 were positive by ELISA. The 3 abattoir workers were clinically normal and lacked historical connections with clinical cases. Although goats are often associated with B. melitensis, these studies could not explicitly implicate this species owing to cross-reactivity with B. abortus, which can also infect goats. Nevertheless, these data reinforce the need for a better National Control Programme for brucellosis in Namibia.     

Balantidiasis in the gastric lymph nodes of Barbary sheep (Ammotragus lervia): an incidental finding

A 4-year-old female Barbary sheep (Ammotragus lervia) was found dead in the Gwangju Uchi Park Zoo. The animal had previously exhibited weakness and lethargy, but no signs of diarrhea. The carcass was emaciated upon presentation. The main gross lesion was characterized by severe serous atrophy of the fat tissues of the coronary and left ventricular grooves, resulting in the transformation of the fat to a gelatinous material. The rumen was fully distended with food, while the abomasum evidenced mucosal corrugation with slight congestion. Microscopic examination revealed the presence of Balantidium coli trophozoites within the lymphatic ducts of the gastric lymph node and the abdominal submucosa. On rare occasions, these organisms may invade extra-intestinal organs, in this case the gastric lymph nodes and abomasum.