Optimal dose and timing in phytohaemagglutinin skin-testing of deer

AIM: To establish the optimal dose of the mitogen phytohaemagglutinin (PHA) and the optimal time for measuring increased skin-fold thickness in red deer following intradermal injection, as an indicator of cell-mediated immune response. METHODS: Three doses (10, 50 and 250 microg) of PHA were injected intradermally in the right side of the neck, and phosphate buffered saline (PBS) was injected at a fourth site as a control, in 20 captive Iberian red deer (Cervus elaphus hispanicus) hinds. Skin-fold thicknesses were measured at 0, 12, 24, 36, 48, 60, 72, 84 and 96 h following injection. RESULTS: The highest dose of PHA tested (250 microg) resulted in a clear and long-lasting cellular response; increases in skin-fold thickness between 48 and 84 h post-injection varied minimally and response correlated positively with liveweight. No correlations with liveweight and no clear increases in skin-fold thickness occurred at the lower doses of PHA or the PBS. CONCLUSIONS AND CLINICAL RELEVANCE: This technique could be applied with minimal training and without specialised equipment in deer, for immunological and ecological research.     

The effects of sex and age on phytohaemagglutinin skin-testing of deer

AIM: To determine if there are sex- or age-related differences in the increase in skinfold thickness in response to the mitogen phytohaemagglutinin (PHA) in red deer. METHODS: One dose of 250 mug PHA was injected intradermally in the right side of the neck, and phosphate buffered saline (PBS) was injected at a second site as a control, in 110 (51 males and 59 females) captive Iberian red deer (Cervus elaphus hispanicus), ranging in age from 21 months to > or =5 years. Skinfold thicknesses were measured immediately before and 72 h following injection. RESULTS: There was a significant effect of gender on the average increase in skinfold thickness; males had greater increases (8.8 (SEM 0.57) mm) than females (4.23 (SEM 0.39) mm) after correcting for other confounding variables. No age-related differences were evident, but differences between sexes were more marked with increasing age. CONCLUSIONS AND CLINICAL RELEVANCE: Effects of gender, probably due to differences in energetic and reproductive constraints in red deer, should be taken into account when interpreting skinfold-test data, both in ecology and in the control of tuberculosis (Tb). Males tend to have a thicker skin than females, so skinfold increase relative to the thickness of the skin, rather than skinfold increase per se, should be used as a more appropriate measure of skinfold increase. This may also have clinical relevance in the interpretation of tuberculin skin testing.     

The testing season affects red deer skinfold increase in response to phytohaemagglutinin

Red deer (Cervus elaphus) have a pronounced seasonality in their physiology. The aim of this study was to assess the effect of the season on red deer responsiveness to skin testing with the phytohaemagglutinin (PHA) mitogen. Study subjects included 270 farmed adult red deer (19 stags and 251 hinds). The skin testing was carried out between January 2009 and August 2010. The animals were injected intradermally with a 0.1 ml volume containing 250 μg of PHA diluted in phosphate buffered saline. The skinfold thickness was measured immediately prior to injection and 72 h after administration, always by the same person and with three repeats per measurement. Single effects of sex and time on skin test responsiveness were significant (p < 0.001) as well as their interaction (p < 0.001). In winter (January), and considering the average of two years, the skinfold increase in response to the intradermal injection of 250 μg PHA was 2.1 times larger in stags and 1.4 times in hinds than in summer (August). While stags had 1.3 times larger responses than hinds in winter, the inverse occurred in summer, with 1.1 times larger responses in hinds. We also evidenced a limited inter-annual variation of skinfold increase in response to PHA in red deer. These findings have important consequences regarding the interpretation of skin test results in the ante-mortem diagnosis of tuberculosis and paratuberculosis, at least in deer.     

Experimental tuberculosis in red deer (Cervus elaphus)

This study was designed to investigate experimental Mycobacterium bovis infection of red deer (Cervus elaphus). Three intravenously inoculated deer (dose 10µg–1000µg) developed miliary tuberculosis of the lungs and all died within 28 days of being infected. No clinical illnesses were observed in four subcutaneously (dose 1 uµg–100uµg) and three intratracheally (dose lµg–100µg) inoculated deer. At the conclusion of the experiment six weeks post inoculation, these seven animals reacted to 2 mg/ml of bovine purified protein derivative. The principal lesions in the intravenously inoculated deer were in the lungs which had multiple foci of necrosis containing very large numbers of acid fast bacilli. A gradation of changes was seen in the subcutaneously inoculated deer. The animal receiving the 1µg dose only had lesions at the injection site and the draining prescapular lymph node. Deer receiving higher doses also had histopathological changes in the lungs and liver. Microscopic changes in the intratracheally infected animals were restricted to the thoracic cavity. The ability of the deer to control infection was related to the route of inoculation.     

Comparison of concentrations of copper in plasma and serum from farmed red deer (Cervus elaphus)

AIM: To determine the relationship between the concentrations of Cu in plasma and serum in red deer, and to compare this relationship with those previously reported in cattle and sheep.

METHODS: Paired serum and heparinised plasma samples from 114 red deer from 10 herds (n=6-20 per herd) were analysed for concentrations of Cu. Samples were collected either at slaughter (n=84; eight herds) or by jugular venepuncture (n=30; two herds). Thirty-nine of the samples taken at slaughter were from adult hinds from four herds, while other samples were taken from 10–14-month-old males, except for one herd (10 samples) where an equal number of 8–9-month-old males and females were sampled. The effect of age, gender and herd on the relationship between concentrations of Cu in plasma and serum was assessed using univariate ANOVA. The individual results for concentrations of Cu in serum were compared with those in plasma, using limits-of-agreement plotting.

RESULTS: The mean concentration of Cu in plasma was not significantly different from that of serum (0.048; 95% CI=-0.14 to 0.24 µmol/L). There was no effect of age, sex or herd on this relationship.

CONCLUSIONS: In deer, there was no significant difference between concentrations of Cu in plasma and serum regardless of age, sex or herd of origin.

CLINICAL RELEVANCE: In contrast to the situation in cattle and sheep, the concentration of Cu in serum can be used interchangeably with that in plasma for the estimation of concentration of Cu in blood of red deer.     

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 long-acting injectable selenium formulations on blood and liver selenium concentrations and liveweights of red deer (Cervus elaphus)

AIM: To evaluate the efficacy of a new long acting injectable selenium ( Se ) formulation to increase the Se status and prevent Se deficiency in red deer. METHODS: Groups of weaned red deer (four stags and six hinds/group) grazing pastures containing <30 mg Se/kg DM were injected subcutaneously with either 0.5, 1.0 or 2.0 mg Se/kg as a new formulation of BaSeO4 (Deposel Multidose), 1.0mgSe/kg of a current formulation (Deposel), or not treated. Blood Se concentrations and liveweight were measured nine times at intervals over 377 and 270 days, respectively. RESULTS: Both formulations of Se elevated blood Se concentrations from 105 nmol/l pre-injection for at least 377 days with peak levels of 1894, 1395 and 818 nmol/l for high, medium and low doses of Deposel Multidose, respectively, at 73141 days, and 1508 nmol/l at 73-141 days for the medium dose of Deposel, which persisted at similar levels for the duration of the study. Deposel Multidose produced fewer and less severe subcutaneous tissue reactions than Deposel. Pastures contained 10 to 30 mg Se/kg DM. There was no significant difference in growth rate between treated and control deer. There was a significant (p<0. 01) linear relationship (y = 1.25x + 71.6, R2=0.86) between blood (x) and liver (y) Se concentrations in the range of 120 - 2100 nmol/l for blood concentrations, and 200 - 3000 nmol/kg for liver concentrations. CONCLUSION: Injections of BaSeO4 in both formulations studied were effective in increasing the Se status of deer but the new formulation produced fewer and less-severe tissue reactions. Young growing red deer appear less sensitive to Se deficiency as measured by weight gain, than sheep and cattle, suggesting that reference ranges for those species are not appropriate for deer. There was a linear correlation between blood and liver selenium concentrations.     

Evaluation of bovine cutaneous delayed-type hypersensitivity (DTH) to various test antigens and a mitogen using several adjuvants

The Bacillus Calmette Guerin (BCG)-induced/purified protein derivative (PPD)-elicited tuberculin skin test is a reliable measure of cell-mediated immune response (CMIR), specifically delayed-type hypersensitivity (DTH); however, its use in livestock may confound diagnosis of Mycobacterium tuberculosis. Therefore, various alternative antigen/adjuvant combinations were evaluated as inducers of DTH that were compared to the BCG/PPD test system with the purpose of finding a skin DTH protocol that does not cross-react with the tuberculin test and allows identification of high and low CMIR responder phenotypes. Specifically, 30 non-lactating cows (five/treatment) were sensitized on day 0 with mycobacteria [BCG, M. tuberculosis or Mycobacterium phlei cell wall extract (MCWE)], and ovalbumin (OVA) emulsified in Freund's complete adjuvant (FCA), non-ulcerative Freund's adjuvant (NUFA), complete NUFA or MCWE. On day 21, cows were injected intradermally with various test antigens including PPD tuberculin, phlein, and OVA. Phosphate buffered saline was included as the negative control and the T-cell mitogen phytohemagglutinin (PHA) was also administered. Double skin-fold thickness was evaluated before and at 6, 24, and 48 h post-injection. Skin biopsies were taken at 24 and 48 h to assess oedema, necrosis, and inflammatory cell infiltration. BCG/PPD and M. phlei/phlein treatments when given with a Freund's adjuvant induced equivalent DTH with peak reactions at 24-48 h after antigen injection. Cows receiving NUFA had fewer injection site granulomas than FCA or CNUFA treatments. The change in skin thickness response to PHA peaked at 6 h. Only cows receiving mycobacteria in NUFA had skin response to OVA, which peaked 6-24 h post-injection. Only sites tested with PPD or phlein had significantly higher lymphocyte infiltration than control, whereas neutrophils were significantly higher at PHA test sites and eosinophils predominated at the PHA test sites. Macrophages were significantly more numerous at the PPD and/or phlein test sites in treatment groups that received killed mycobacteria in a Freund's adjuvant and/or with BCG, and at the PHA test sites in all treatment groups. It was concluded that the M. phlei/phlein system did induce DTH and was similar to the DTH induced by the BCG/PPD system when MCWE was administered with a Freund's adjuvant. Therefore, this protocol is suitable for detecting high/low CMIR responders in research herds. However, cross-reaction to PPD was evident following induction of DTH using M. phlei. Hence, this protocol does not alleviate the problem of artificial induction of DTH cross-reactivity and would not be suitable for commercial herds where tuberculin testing is required.     

A critical evaluation of serum methylmalonic acid and vitamin B12 for the assessment of cobalt deficiency of growing lambs in New Zealand

AIM: To derive reference ranges for serum methylmalonic acid (MMA) for the diagnosis of cobalt/vitamin B₁₂-responsiveness in lambs and critique existing serum vitamin B₁₂ reference ranges.

METHODS: Individual animal data from earlier supplementation trials, involving 225 ewes, 106 suckling lambs, 301 lambs during the suckling and post-weaning periods and 414 weaned lambs, for which weight gain to supplementation was observed, were used to derive relationships between serum vitamin B₁₂ and MMA, and liveweight gain.

RESULTS: Serum MMA concentrations were rarely elevated above the norm of <2 µmol/L when serum vitamin B₁₂ concentrations were >375 pmol/L, and not elevated into the range where a liveweight response to supplementation occurred (>10 µmol/L) unless serum vitamin B₁₂ concentrations were below 200 pmol/L. Suckling lambs were able to maintain high growth rates despite elevated serum MMA concentrations (>20 µmol/L).

CONCLUSIONS: The current reference ranges used in New Zealand for serum vitamin B₁₂ are set conservatively high. Serum MMA concentrations appear to allow better differentiation of a responsive condition than vitamin B₁₂ concentrations. Serum MMA concentrations <13 µmol/L indicate responsiveness to supplementation whilst concentrations <7 µmol/L indicate unresponsiveness. In the range 7–13 µmol/L, variation in response was observed and predictability of response is less certain, but supplementation is advisable.

CLINICAL RELEVANCE: The current reference ranges for vitamin B₁₂ responsiveness are conservatively high and lead to over-diagnosis of vitamin B₁₂ deficiency in ill-thriftiness of sheep.     

Bodyweight and serum copper concentrations of farmed red deer stags following oral copper oxide wire administration

A diagnosis of secondary copper deficiency was established on a deer farm with a peat soil type, on the basis of confirmed enzootic ataxia in hinds, liver and serum copper concentrations and pasture and soil element analyses. Seventy-four weaner stags were selected for a trial to investigate a growth response to copper supplementation.

Thirty-seven red and red x wapiti type stags were treated with 4g copper oxide wire particles at four months of age (April). A further 8g copper oxide wire was given in June. Thirty-seven untreated animals acted as controls. Body weights were measured on five occasions, from April 24 to November 26. Serum copper analyses were undertaken on ten deer prior to commencement of the trial, and on seven treated and eight control deer in June, July and October.

Serum copper concentrations ranged from 2.0 to 19.3μmol/l prior to the trial. In June, July and October serum copper ranged from 0.1–6.7, 0.6–5.0, and 1.3–6.3μmol/l respectively, in control deer. In treated deer concentrations ranged from 7.2–14.7, 5.2–10.8, and 6.9–13.7µmoM in June, July and October respectively. The difference between mean copper concentration at each post-treatment sampling date was highly significant, (P<0.001). At the conclusion of the trial (November 26) the treated deer averaged 3.lkg heavier than controls, but this difference was not statistically significant.

In view of these results and the variation in growth response trials in other species, further investigation of the effects of copper on the growth of young deer is warranted.     

Liveweight gain and copper status of young deer treated or untreated with copper oxide wire particles on ten deer farms in Canterbury

AIMS: To measure the liveweight response of young deer to copper supplementation on a range of deer farms in the Canterbury region of the South Island, New Zealand, and relate any response to plasma copper concentration. METHODS: On each of 10 farms throughout north and mid-Canterbury, young deer (4-7 months old) were allocated by liveweight to two groups of 45-50. The deer in one group each received 5 g copper oxide wire particles (COWP) at the start of the project (March-June 2000) and a further 10 g COWP 3 months later (+Cu group), while the other group received no copper supplementation (-Cu group). Liveweight (+/-1 kg) of all deer was recorded monthly. A 10 ml blood sample was taken from a sub-sample of 10 animals in each group at the beginning of the trial and from the same animals 3 and 6 months later for measurement of plasma copper concentration. RESULTS: There was no significant effect (p=0.96) of treatment with COWP on liveweight gain (overall mean liveweight gain = 169 g/day). This result was consistent for all farms and for both periods. Mean plasma copper concentration declined from 12.5 (SD 3.4) to 10.3 (SD 3.0) micromol/l in +Cu groups, and to a significantly lower level (p<0.001) of 8.6 (SD 3.2) in -Cu groups after 3 months, at which time 38% of -Cu animals were considered hypocupraemic (<8 micromol/l). At the end of the second 3-month period there was no significant treatment effect on plasma copper concentration, which averaged 9.4 (SD 2.3) micromol/l. Of the feed samples submitted (n=46), only 17% had a copper concentration <5.0 mg/kg dry matter (DM), 13% had a molybdenum concentration >1.0 mg/kg, and 21% had a sulphur content >3.5 g/kg. CONCLUSIONS: Under the conditions of these trials, no response in liveweight gain to copper supplementation was observed despite evidence of hypocupraemia in 38% of -Cu animals, which gained weight at similar rates to those that had adequate plasma copper levels. The extent of the hypocupraemia was either not sufficiently severe, or not maintained for a long enough period to cause copper deficiency resulting in reduced liveweight gain. No other signs of copper deficiency were evident. There is scope for deer farmers to reassess the need for copper supplementation in young deer.     

Impact of molybdenum on the copper status of red deer (Cervus elaphus)

AIM: To determine the effect of increasing molybdenum (Mo) intakes on serum and liver copper (Cu) concentrations and growth rates of grazing red deer (Cervus elaphus).

METHODS: Molybdenum- and Cu-amended fertilisers were applied to six 1.1-ha paddocks in a 3 × 2 design. Three levels of Mo were applied on two paddocks at each level in mid April (designated Day 1); levels were: none (control), 0.5 (medium) and 1.0 (high) kg Mo/ha as sodium molybdate. In late May (Day 39), two levels of Cu (none and 3.0 kg Cu/ha, as copper sulphate) were applied to each of the three levels of Mo-treated paddocks. Pasture Mo, Cu and sulphur (S) concentrations were measured at about fortnightly intervals. In late June (Day 74), ten 6-month-old red deer hinds were placed on the six experimental pastures, and serum and liver Cu concentrations were monitored at about monthly intervals for 102 days. The hinds were weighed on four occasions during the trial.

RESULTS: Mean pasture Mo concentrations on Day 56 were 2, 4.6 and 11.3 mg/kg dry matter (DM) for the untreated control, medium and high Mo-treated pastures, respectively. Pasture Cu concentration was 95 mg/kg DM on Day 59, 53 mg/kg DM on Day 90, and 9 mg/kg DM by Day 153. Mean S concentration in pasture was 3.3 (range 3.03–3.45) g/kg DM. Copper application to pasture had no significant effect on serum and liver Cu concentrations in deer so data were pooled within Mo treatment. Mean initial (Day 74) serum Cu concentration was 9.2 µmol/L. In the deer grazing the control Mo pasture, this increased to 10.3 µmol/L on Day 112, before decreasing to 6.4 µmol/L on Day 176. In deer grazing the medium and high Mo-treated pastures, mean serum Cu concentrations were 3.8 and 3.9 µmol/L, respectively, on Day 112, and 2.5 and 3.3 µmol/L, respectively, on Day 176. Mean initial (Day 74) liver Cu concentration was 131 µmol/kg fresh tissue. In the deer grazing the control Mo pasture, this declined to 120 and 52 µmol/kg on Days 112 and Day 176, respectively. In deer grazing the medium and high Motreated pastures, liver Cu concentrations decreased to 55 and 52 µmol/kg fresh tissue, respectively, on Day 112, and 21 and 20 µmol/kg fresh tissue, respectively, on Day 176. Mean serum and liver Cu concentrations were not significantly different between deer grazing the medium and high Mo-treated pastures, and were lower (serum p=0.003, liver p<0.001) in those groups than in deer grazing the untreated control pastures. No clinical signs of Cu deficiency associated with lameness were observed. Deer grazing pastures that had Mo concentrations >10 mg/kg DM had lower (p=0.002) growth rates (100 vs 130 g/day) than those on pastures containing <2.4 mg Mo/kg DM.

CONCLUSION: Increasing pasture Mo concentrations from 2 mg/kg DM to ≥4.6 mg/kg DM significantly reduced serum and liver Cu concentrations in grazing deer. Reduced growth rate was observed at pasture Mo concentrations >10 mg/kg DM.     

Copper supplementation, velvet antler production and growth of rising 2-year-old red deer stags

AIMS: To investigate whether copper supplementation to rising 2-year-old red deer stags (Cervus elaphus) in winter and/or spring improved velvet antler weight, grade and financial value, and liveweight gain. METHODS: Rising 2-year-old red deer stags (n=142) on a commercial deer farm in Hawke's Bay were allocated to two groups on May 14, 1996. One group received 20 g boluses of copper-oxide wire particles. On August 15 each group was reallocated in a crossover design, each sub-group receiving either the same copper-oxide treatment or no treatment, to produce control, winter-only, winter-plus-spring, and spring-only copper treatment groups. Blood samples were collected for serum ferroxidase measurements from seven deer per group and all deer were weighed at 4-6-week intervals. Dates of antler casting and velvet removal, and velvet antler weight and grade were recorded, and the financial value of velvet calculated. Livers from a sample of deer slaughtered at the end of the trial mid-December were analysed to determine copper content. RESULTS: Supplementation with copper did not significantly alter velvet antler weight, daily velvet antler growth rate, days from casting to removal, grade or value, or stag liveweight gain. Serum ferroxidase concentrations averaged 10.0-23.7 IU/l in control deer. Copper supplementation increased mean serum ferroxidase concentrations by approximately 10 IU/l. Mean liver copper concentration in control deer was 99 micromol/kg and ranged from 194 to 386 micromol/kg in the three treated groups. CONCLUSION: Group mean serum ferroxidase concentrations of 10 IU/l and above are adequate for optimum velvet production and liveweight gain in rising 2-year-old stags.     

Dietary phosphorus and an inflammatory challenge affect performance and immune function of weanling pigs

Ninety-six 3-wk-old pigs (6.3+/-0.12 kg initial BW) were allotted to one of eight treatments based on BW and litter origin to determine the effect of dietary phosphorus and an inflammatory challenge on performance and immune function. Four corn-soybean meal-based treatment diets were formulated to contain 0.16, 0.24, 0.32, or 0.40% available P. Monocalcium-dicalcium phosphate was used as the supplemental P source. The Ca:available P ratio was maintained at 2:1. To challenge the pigs, half of the pigs in each dietary treatment were injected i.m. with E. coli lipopolysaccharide (200 microg/kg of BW) on d 7 and 14. This resulted in a 2 x 4 factorial arrangement of treatments. Average daily gain for the 35-d study was increased linearly (P < 0.01) by increasing supplemental P. Average daily gain and ADFI were decreased (P < 0.05) by lipopolysaccharide injection. Serum P concentrations increased linearly (P < 0.01) with increasing supplemental P. Antibody titers to the injection of sheep red blood cells and ovalbumin on d 21 decreased linearly (P < 0.10) by increasing supplemental P. In vitro blastogenic response of lymphocytes to phytohemagglutinin (PHA) on d 25 was increased linearly (P < 0.05) by increasing supplemental P. Blastogenic response of lymphocytes to pokeweed mitogen on d 25 was not affected. On d 31, skinfold thickness 6 h following an intradermal injection of PHA was increased quadratically (P < 0.07) by increasing supplemental P. There were no P x lipopolysaccharide interactions for any immune response measure. In conclusion, increasing supplemental P increased ADG and enhanced cell-mediated immune response but decreased humoral immune response.     

Selenium supplementation of young red deer (Cervus elaphus)

Abstract

Two selenium (Se) supplementation trials were conducted in successive years involving a total of 70 red deer calves 3–15 months of age grazing pasture containing 30–57 ppb of Se on a dry matter basis. The trials compared growth rate, whole blood Se and glutathione peroxidase (GSHpx) concentrations of calves which received periodic doses of oral Se or a single injection of barium selenate (equivalent to 50 mg Se) or no Se supplementation. There were no significant weight gain differences between treated and untreated groups in either trial. Whole blood GSHpx levels were strongly correlated with blood Se levels (r = 0.9278) and produced the following regression equation: GSHpx 0.0155 Se ? 2.292.

In both years the 3 month old calves had GSHpx levels of 6–9 kU/l which probably derived from maternal transfer of Se. The GSHpx levels in unsupplemented calves declined from these levels to a minimum in winter (group means ? 2.6) and then progressively rose the following spring and summer.

Periodical oral dosing with Se or a single injection of barium selenate significantly elevated blood Se and GSHpx levels throughout the trials.     

Feline gingivitis

AIM: To discover whether cross infection between red deer (Cervus elaphus) and cattle is possible with either a bovine isolate of the cattle lungworm, Dictyocaulus viviparus, or with a cervine isolate of the lungworm, Dictyocaulus eckerti.

METHOD: Twelve cattle and 12 red deer were reared parasitefree from birth. At 3–4 months of age, half of each group (n=6) were experimentally infected with D. viviparus and the other half with D. eckerti. The course of infection was monitored for 34 days, after which the animals were slaughtered and the lungs removed to assess levels of infection.

RESULTS: Faecal larval counts demonstrated that patent Dictyocaulus infections occurred in all groups. At necropsy, adult worms were found in the lungs in all groups except the cattle that were infected with D. eckerti. The largest numbers of adult worms were found in the lungs of the red deer infected with D. eckerti.

CONCLUSION: It was demonstrated that both cattle and red deer could be infected with either D. viviparus or D. eckerti. However, D. eckerti larvae that originated from deer established more successfully in deer and D. viviparus larvae that originated from cattle established more successfully in cattle.     

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.     

Effect of forage legumes containing condensed tannins on lungworm (Dictyocaulus sp.) and gastrointestinal parasitism in young red deer (Cervus elaphus)

To investigate the effect of feeding forage legumes containing condensed tannins ( ) on internal parasitism, red deer calves were fed either lucerne (Medicago sativa; 0.1 per cent ), birdsfoot trefoil (Lotus corniculatus; 1.9 per cent ) or sulla (Hedysarum coronarium; 3.5 per cent ) and trickle-infected with deer-origin gastrointestinal nematode and lungworm (Dictyocaulus sp.) larvae for 5 weeks, then slaughtered at 7 weeks. There was a significant negative linear relationship between dietary concentration and abomasal nematode burdens. No significant differences in faecal egg counts, lungworm burdens or voluntary feed intake were found. Deer fed sulla had higher liveweight gain, carcass weight and carcass dressing-out percentage, higher serum total protein and albumin concentration and lower serum gastrin concentration and faecal lungworm larval count, compared with lucerne-fed deer. Inclusion of sulla in diets for young red deer may reduce the impact of internal parasites and/or reduce the dependence on anthelmintic treatment.     

Congenital nutritional myodegeneration (white muscle disease) in a red deer (Cervus elaphus) calf

CASE HISTORY A 5-day-old red deer calf was submitted with tachypnoea and dyspnoea, and was reluctant to move.

CLINICAL FINDINGS: Muscular damage was established via elevated creatinine phosphokinase (CPK) activities (5,000 U/L), while concentrations of Se in whole blood were low (24.8 nmol/L). The animal died despite treatment with penicillin-streptomycin and 0.1 mg/kg Se/vitamin E administered by S/C injection.

DIAGNOSIS: Necropsy and histological examination of cardiac and skeletal muscle confirmed the presumptive diagnosis of congenital white muscle disease (WMD). Prophylactic administration of a Se/vitamin E commercial preparation (as above) to another calf born in the same herd one month later was associated with good health and apparently normal growth and development.

CLINICAL RELEVANCE: Congenital WMD due to Se deficiency can be fatal in red deer calves. However, prophylactic administration of Se and vitamin E to neonatal calves may be beneficial for neonatal red deer calves.     

Intradermal testing of swine to monitor changes in delayed hypersensitivity response

Pigs were tested (intradermal injection) on the abdomen with either phytohemagglutinin (PHA) or PHA and tuberculin. The response to PHA was consistent; double-skin thickness averaged 247% of the preinjection thickness during daily testing. Pigs that were primed with Freund's complete adjuvant had a mean double-skin thickness of 208% for tuberculin and 236% for PHA, indicating that the mitogen induced an antigen-like response. Unexpectedly, injections of dexamethasone (0.1 mg/kg of body weight on 2 days) increased the reactivity to both tuberculin and PHA, indicating that the lymphocyte and macrophage response to dexamethasone in swine may be slightly different than the response reported in other species.