An evaluation of the comparative tuberculin skin test for detecting tuberculosis in farmed deer

A comparative cervical skin test using 1.0 mg/ml bovine purified protein derivative and 0.5 mg/ml avian purified protein derivative was evaluated as a method for detecting tuberculosis in farmed deer. A positive comparative cervical skin test reaction was defined as a bovine response with a 2 mm or greater increase in skin thickness which was greater or equal to the avian response. Estimates of the sensitivity of the comparative cervical skin test were obtained from a series of experiments conducted on 60 deer intratracheally inoculated with Mycobacterium bovis. Prior tuberculin skin testing was found to suppress the skin reactivity to a subsequent comparative cervical skin test. This effect was most pronounced at short intervals of 3-7 days, but could still be measured 60 days after the previous test. When the test interval was greater than 60 days, the sensitivity of the comparative cervical skin test was 91.4%. The specificity of the comparative cervical skin test was 98.7% when 1157 deer from 17 uninfected herds with a history of nonspecific skin test reactions were examined. There was no statistical difference in the mean skin thickness increases of three groups of infected animals tested with 2 mg/ml, 0.2 mg/ml and 0.02 mg/ml of bovine purified protein derivative respectively.     

The effect of Johne's vaccination on tuberculin testing in farmed red deer (Cervus elaphus)

AIM: To assess the degree of interference with bovine tuberculin testing in farmed red deer that vaccination of young deer with an oil-adjuvanted vs aqueous formulation of live attenuated Mycobacterium paratuberculosis Strain 316F vaccines would be likely to cause, and to compare immunological responses between vaccine formulations.

METHODS: Five-month-old red deer (n=45) were randomly allocated to three treatment groups of 15 animals, which received either no vaccine, a single 2-ml dose of an oil-adjuvanted formulation or two 2-ml doses, 6 weeks apart, of an aqueous formulation of live attenuated M. paratuberculosis Strain 316F vaccine injected subcutaneously (S/C) in the neck (Control, Oil-adjuvant Ptb, and Aqueous Ptb groups, respectively). Injection-site reactions were described and measured on Weeks 3, 6 and 9. Animals were weighed and lymphocyte transformation tests (LTT) and antibody enzyme-linked immunosorbent assays (ELISA) using avian, bovine and Johnin tuberculin were conducted on blood samples collected at Weeks 0, 6, 12, 15, 24, 27, 36 and 39. A bovine mid-cervical skin test (MCT) was applied at Week 12, and comparative cervical skin tests (CCTs) at Weeks 24 and 36. At Week 42, the animals were slaughtered at a commercial deer slaughter premises and subjected to rigorous meat inspection.

RESULTS: Two animals were eliminated at the start of the trial due to a positive cross-reaction with bovine tuberculin in the initial LTT. Almost all animals reacted to the MCT at Week 12, with mean skin thicknesses of 3.9, 2.9 and 1.0 mm for the Oil- adjuvant Ptb, Aqueous Ptb and Control groups, respectively. When the CCT was conducted at Week 24, 2/15 Oil-adjuvant Ptb, 2/14 Aqueous Ptb and 1/14 Control animals were classified as CCT-positive to bovine tuberculin. By Week 36, all animals were CCT-negative. The Oil-adjuvant Ptb vaccination resulted in high persistent levels of antibody that reacted with bovine tuberculin, compared with negligible levels in the Aqueous Ptb group.

Overall, a single dose of the Oil-adjuvant Ptb vaccine in deer stimulated a vigorous, cross-reactive immune response, evidenced by high LTT, skin-test and antibody reactions to bovine tuberculin, with both cell-mediated and humoral characteristics. By comparison, two doses of the Aqueous Ptb vaccine produced less cross-reactivity and a bias towards a cell-mediated response. The Oil-adjuvant Ptb vaccine resulted in moderate injection- site lesions that were quite persistent, whereas the Aqueous Ptb vaccine resulted in smaller nodules that regressed more quickly.

CONCLUSIONS: Vaccination of farmed deer with an oil-adjuvanted Johne's vaccine has the potential to cause significant interference with routine tuberculin skin testing. The cross-reactivity should decline with time and the CCT should be able to clear MCT-positives, but there is a risk of false-positives to the blood test for tuberculosis (BTB), due to high persistent levels of antibody. The CCT could be used as a primary skin test in vaccinated deer on some farms.

The Aqueous Ptb caused fewer problems with skin testing and produced significantly less bovine antibody than the Oil-adjuvant Ptb, but stimulated persistent cell-mediated immune responses that may provide some protection against Johne's disease.     

The effect of Johne's vaccination on tuberculin testing in farmed red deer (Cervus elaphus)

AIM: To assess the degree of interference with bovine tuberculin testing in farmed red deer that vaccination of young deer with an oil-adjuvanted vs aqueous formulation of live attenuated Mycobacterium paratuberculosis Strain 316F vaccines would be likely to cause, and to compare immunological responses between vaccine formulations. METHODS: Five-month-old red deer (n = 45) were randomly allocated to three treatment groups of 15 animals, which received either no vaccine, a single 2-ml dose of an oil-adjuvanted formulation or two 2-ml doses, 6 weeks apart, of an aqueous formulation of live attenuated M. paratuberculosis Strain 316F vaccine injected subcutaneously (S/C) in the neck (Control, Oil-adjuvant Ptb, and Aqueous Ptb groups, respectively). Injection- site reactions were described and measured on Weeks 3, 6 and 9. Animals were weighed and lymphocyte transformation tests (LTT) and antibody enzyme-linked immunosorbent assays (ELISA) using avian, bovine and Johnin tuberculin were conducted on blood samples collected at Weeks 0, 6, 12, 15, 24, 27, 36 and 39. A bovine mid-cervical skin test (MCT) was applied at Week 12, and comparative cervical skin tests (CCTs) at Weeks 24 and 36. At Week 42, the animals were slaughtered at a commercial deer slaughter premises and subjected to rigorous meat inspection. RESULTS: Two animals were eliminated at the start of the trial due to a positive cross-reaction with bovine tuberculin in the initial LTT. Almost all animals reacted to the MCT at Week 12, with mean skin thicknesses of 3.9, 2.9 and 1.0 mm for the Oil-adjuvant Ptb, Aqueous Ptb and Control groups, respectively. When the CCT was conducted at Week 24, 2/15 Oil-adjuvant Ptb, 2/14 Aqueous Ptb and 1/14 Control animals were classified as CCT-positive to bovine tuberculin. By Week 36, all animals were CCT-negative. The Oil-adjuvant Ptb vaccination resulted in high persistent levels of antibody that reacted with bovine tuberculin, compared with negligible levels in the Aqueous Ptb group. Overall, a single dose of the Oil-adjuvant Ptb vaccine in deer stimulated a vigorous, cross-reactive immune response, evidenced by high LTT, skin-test and antibody reactions to bovine tuberculin, with both cell-mediated and humoral characteristics. By comparison, two doses of the Aqueous Ptb vaccine produced less cross-reactivity and a bias towards a cell-mediated response. The Oil-adjuvant Ptb vaccine resulted in moderate injection site lesions that were quite persistent, whereas the Aqueous Ptb vaccine resulted in smaller nodules that regressed more quickly. CONCLUSIONS: Vaccination of farmed deer with an oil-adjuvanted Johne's vaccine has the potential to cause significant interference with routine tuberculin skin testing. The cross-reactivity should decline with time and the CCT should be able to clear MCT-positives, but there is a risk of false-positives to the blood test for tuberculosis (BTB), due to high persistent levels of antibody. The CCT could be used as a primary skin test in vaccinated deer on some farms. The Aqueous Ptb caused fewer problems with skin testing and produced significantly less bovine antibody than the Oil-adjuvant Ptb, but stimulated persistent cell-mediated immune responses that may provide some protection against Johne's disease.     

Blood biochemical reference values for farmed red deer

The activities and concentrations of 30 whole blood and plasma constituents including enzymes, substrates, electrolytes and vitamins were determined in samples from 91 clinically healthy farmed red deer (Cervus elaphus) hinds ranging in age from one to 15 years. The hinds were randomly selected from a herd maintained on a mixture of reseeded and permanent hill pasture at an experimental research station in north east Scotland. The results were analysed statistically and the calculated means, standard errors and tolerance intervals provide a basis for diagnostic clinical chemistry in red deer. The data are compared with data from earlier reports on blood biochemistry in deer and other livestock species.     

Type II ostertagiosis in farmed red deer

A mild form of type II ostertagiosis was recorded in a group of 12, 22-month-old red deer stags, slaughtered in an abattoir in April 1989. The animals appeared clinically normal but they were light and in rather poor condition. Mixed ostertagid infections were present in very variable numbers. In the seven most heavily infected animals, 70 per cent of the worms were in the early fourth stage and assumed to be hypobiotic. In some animals the worms were maturing and in these there were lesions similar to those associated with bovine ostertagiosis.     

Serological responses of farmed red deer to a vaccine against Leptospira interrogans serovars hardjo and pomona

Six adult, female, red deer were vaccinated with a hardjo-pomona vaccine followed by a second vaccination four weeks later and thereafter, at yearly intervals for two years. Serological responses were determined at intervals using a standard microscopic agglutination test. Increased responses to hardjo were observed in two deer which were seropositive to this serovar before vaccination compared to those which were initially seronegative. Some initially seronegative deer developed titres to hardjo and pomona ranging from 1:24 to 1:48 following the first dose of vaccine, and all deer produced a serological response to both serovars ranging from 124 to 1:96 following the second vaccination except one animal which failed to produce a pomona titre of 124 or greater. Responses to annual revaccinations were more variable both within and between years. Most deer produced titres which ranged from 1:24 to 1:96 for three to live months, although some deer failed to seroconvert following the annual revaccinations. Peak responses were similar to those recorded in cattle following an identical vaccination programme with the same vaccine but titres fell to undetectable levels after three to live months whereas titres reported in cattle generally persisted for at least 12 months.     

Osteochondrosis associated with copper deficiency in young farmed red deer and wapiti x red deer hybrids

Osteochondrosis is reported in association with copper deficiency in young red deer and wapiti x red deer hybrids on eight deer farms throughout New Zealand. On two farms, more than 30% of fawns were affected. Affected animals were lame, often had one or more swollen joints, and in some cases had an abnormal “bunny-hopping” gait or “cow-hocked” stance. Lesions were most common in the carpal, tarsal, stifle and hip joints, and were usually bilateral. Defects in articular cartilage ranged from loose flaps to complete separation with exposure of subchondral bone and the presence of cartilage fragments within the joint space. In advanced cases, the joints had features of degenerative arthropathy. Bilateral epiphyseolysis of the femoral head was observed in some severely lame deer. All deer with osteochondrosis had low serum and/or liver copper concentrations.     

Osteochondrosis associated with copper deficiency in young farmed red deer and wapiti x red deer hybrids

Osteochondrosis is reported in association with copper deficiency in young red deer and wapiti X red deer hybrids on eight deer farms throughout New Zealand. On two farms, more than 30% of fawns were affected. Affected animals were lame, often had one or more swollen joints, and in some cases had an abnormal "bunny-hopping" gait or "cow-hocked" stance. Lesions were most common in the carpal, tarsal, stifle and hip joints, and were usually bilateral. Defects in articular cartilage ranged from loose flaps to complete separation with exposure of subchondral bone and the presence of cartilage fragments within the joint space. In advanced cases, the joints had features of degenerative arthropathy. Bilateral epiphyseolysis of the femoral head was observed in some severely lame deer. All deer with osteochondrosis had low serum and/or liver copper concentrations.     

Risk factors for dystocia in farmed red deer (Cervus elaphus).

OBJECTIVE: To analyse data from a study of deer herd health and productivity to identify potential risk factors for the occurrence of dystocia. DESIGN: A 2-year longitudinal observational study of 14 red deer farms was carried out in New Zealand. About 2700 hinds were monitored each year for individual characteristics including body condition score (BCS), and reproductive performance. Grazing and calving management practices were recorded. During calving, farmers recorded calving difficulties (dystocia) that needed human assistance or caused the death of the hind. Risk factors potentially affecting the occurrence of dystocia were identified and analysed by multivariable statistical techniques. RESULTS: The percentage of hinds with dystocia was 0.52% (95% CI 0.32 to 0.78). Hinds with a post-winter BCS over 3.5 were 2.7 times as likely to experience dystocia as hinds scoring 3.5 or less. Given the same BCS class, bodyweight gain in late pregnancy was associated with an increased risk of dystocia. Grazing steep paddocks in the last trimester of pregnancy decreased the risk of dystocia in hinds with a BCS over 3.5. CONCLUSIONS: The risk of dystocia on red deer farms may be reduced if key individual animal, management and environmental factors are optimal.     

Dictyocaulus infection in farmed red deer (Cervus elaphus)

Post mortem examination of red deer calves on a deer farm situated on hill ground in north-east Scotland revealed infection by a lungworm morphologically similar to Dictyocaulus viviparus. Trials were conducted to monitor the natural development of D viviparus infection in red deer, to investigate the value of a commercial lungworm vaccine and to evaluate methods of treating clinical cases. The findings indicate that the syndrome may be less apparent in red deer than it is in cattle, protection might be gained by vaccination and that housing and medication provide useful therapy. The extent of clinical disease is likely to depend on the general health, bodily condition and nutritional status of the animals versus the weight of infection acquired from the pasture. However, various factors can affect both sides in this confrontation.     

Real-time ultrasonography for pregnancy diagnosis and estimation of fetal age in farmed red deer

Twenty-nine farmed red deer hinds two years of age or older were observed during mating in April, and the dates of oestrus and, or, mating were recorded. From May 28 to August 13, at approximately weekly intervals, the deer's uteri and associated structures were examined by rectal ultrasound using a 5 MHz transducer while the deer were held standing in a restraining device. The uterine diameter, amniotic sac diameters, crown-rump length, head length, nose length, chest depth, chest width and placentome base-apex and width were measured for each scan. Quadratic regression equations were computed for each measurement with age, using data from hinds with known mating dates, and the chronological sequence of appearance of a range of uterine and fetal characteristics was recorded. There were significant quadratic regressions. (P less than 0.001) for each of the measurements. The accuracy of pregnancy detection to 117 days after conception was 99.3 per cent and to 132 days after conception 97.4 per cent. The confirmation of non-pregnancy was 96.5 per cent accurate. The regressions of each measurement by age should provide an accurate method for the estimation of fetal age in red deer.     

Intra-uterine transmission of paratuberculosis (Johne's disease) in farmed red deer

AIM: To determine whether intra-uterine transmission of paratuberculosis (Johne's disease) occurs in farmed red deer (Cervus elaphus) in New Zealand. METHODS: On four different farms, nine late-stage pregnant hinds with Johne's disease were slaughtered and samples were taken from them and their 10 fetuses. Samples of the hepatic, ileocaecal and mesenteric lymph nodes and the posterior ileum were collected from the hinds. The lung, liver, spleen, jejunum and ileum from the fetuses were sampled, as were the placentomes. Blood samples were tested using the 'Paralisa' test, a modified immunoglobulin G1 (IgG1) enzyme-linked immunosorbent assay (ELISA). Tissue samples were cultured using the BACTEC system, and fixed samples were sectioned and histological slides examined. RESULTS: All nine hinds and 9/10 fetuses (one hind had twins) were culture-positive for Mycobacterium avium subsp paratuberculosis (M. ptb). Six hinds had gross lesions of Johne's disease, while all hinds had characteristic histopathological lesions affecting the ileum, ileocaecal valve and associated lymph nodes. The only histopathological change observed in the fetuses was some mild inflammation in the lungs of one individual. Acid fast organisms (AFOs) were seen in histological sections of the lymph nodes and ileum of six hinds, and none were seen in tissues from the fetuses. These six hinds were Paralisa-positive, whereas the remaining hinds and fetuses were serologically-negative. CONCLUSIONS: These results confirm that there is a high risk of transmission of M. ptb from clinically affected hinds to their fetuses during pregnancy. CLINICAL RELEVANCE: Johne's disease is an increasingly important disease responsible for deaths in young red deer. Recognising the influence of intra-uterine transmission on the spread of this disease may be an important step towards improved control of Johne's disease.     

Parapoxvirus infections in New Zealand farmed red deer (Cervus elaphus)

Outbreaks of infection due to a parapoxvirus were reported on eight New Zealand deer farms. Scabby lesions were seen variably on the muzzle, lips, face, ears and neck of red deer (Cervus elaphus) with morbidity rates reaching 100%. On three farms multifocal lesions were also present on the velvet. Deaths were reported on two properties where the lesions were extensive and secondary bacterial infections had occurred. On one of these farms multifactorial disease was suspected. Poxvirus particles were seen by negative contrast electron microscopy in scab material from all eight properties. Morphologically the deer virus resembled a parapoxvirus, but restriction endonuclease analysis showed its DNA fragment patterns were distinct from those of orf (contagious ecthyma) virus.     

Blood constituents of farmed red deer (Cervus elaphus). II: biochemical values

Approximately 80 farmed male and female red deer aged three months and over were blood sampled and biochemical parameters measured. The deer were sampled by jugular venepuncture without tranquillization. Mean values recorded were serum gamma-glutamyltransferase (19.5 IU/l), glutamic oxaloacetic transaminase (43.0 IU/l), creatine phosphokinase (197.9 IU/l), plasma pepsinogen (0.91 IU/i), blood urea nitrogen (8.56 mmol/l), serum glucose (6.9 mmol/l), phosphorus (1.75 mmol/l), calcium (2.13 mmol/l), magnesium (0.74 mmol/l), potassium (4.43 mmol/l) and sodium (138.5 mmol/l). There were no significant differences in any parameter between sexes or between age groups 3-8 months, 9-18 months, or older.     

Potential wildlife sources of Yersinia pseudotuberculosis for farmed red deer (Cervus elaphus)

During 1982 and 1983 15 serotype I,6 serotype II, 1 serotype III and 3 untyped strains of Yersinia pseudotuberculosis were isolated from 675 apparently normal small mammals and birds from the Invermay farm and nearby rubbish tip with the following prevalence rates: feral cats 27.8%, Norway rats 8.6%, mice 5.5%, hares 3.8%, rabbits 1.9%, ducks 5.3%, sparrows 2.3%, seagulls 2.3% and starlings 1.7%. For rabbits a significantly higher prevalence of infection was found in the autumn/winter period (4.8%) than the spring/summer period (0%).

Insufficient numbers of other mammals were obtained to demonstrate any seasonal difference in prevalence. All bird isolations were obtained between March and July (8/158) compared with none from August to October (0/144). It appears that a number of free-living species of small mammal and birds may be reservoir hosts for Y. pseudotuberculosis and potential sources of infection for red deer on the Invermay farm.