An ELISA for the detection of anergic tuberculous cattle

An enzyme-linked immunosorbent assay (ELISA) for bovine antibody to antigens in unheated Mycobacterium bovis culture filtrate was standardised against a reference serum from an experimentally infected cow. Two Northern Territory herds with a total of 561 cattle were tested. All cattle reacting in the caudal fold tuberculin test, those giving strong reactions in the ELISA and those with visible lesions of tuberculosis were subjected to a detailed bacteriological examination. Of the 19 cattle which yielded isolates of M. bovis, only 4 were positive to the tuberculin test. Serum samples from 5 cattle gave ELISA values greater than 7.0 units. None of these 5 reacted in the tuberculin-test and 2 had no visible lesions. Of the 10 remaining cattle from which M. bovis was isolated, 3 had ELISA values between 6.5 and 7.0 units and were also without visible lesions. The ELISA values for the remaining 7 infected cattle ranged down to 4.6 units. Forty cattle yielded no M. bovis on culture of their tissues. They included 7 which were reactors in the tuberculin test and 23 with ELISA values of 7.0 units or more. The evident low specificity and sensitivity of the ELISA make it of little value as an alternative to the tuberculin test, but it can detect some anergic cattle at the cost of increasing the number of false positive reactors. This may be acceptable in some circumstances and would justify the use of the ELISA as a complement to the tuberculin test or to an in vitro assay of T-cell immunity. In the 2 Northern Territory herds described, the removal of 5 of the anergic cattle would have required a cull of 28 animals of 5% of the total. A cut off value of 6.5 units would have eliminated 3 more, but at the cost of culling 80 animals or nearly 15% of the cattle. Even so, 7 cattle from which M. bovis was isolated would have remained undetected by either test.     

Protection of sheep by vaccination against experimental challenge with Leptospira borgpetersenii serovar Hardjo and L. interrogans serovar Pomona

AIMS: To evaluate a multivalent leptospiral and clostridial vaccine for prevention of renal colonisation and urinary shedding in sheep, following experimental challenge with New Zealand strains of Leptospira borgpetersenii serovar Hardjo type Hardjobovis and L. interrogans serovar Pomona.

METHODS: Two separate but similarly designed studies were conducted. In both studies, Romney-cross lambs, aged 9–11 weeks, were randomly allocated to a vaccinated group and a control group. Vaccinated lambs each received two 1.5-mL S/C doses of a multivalent leptospiral and clostridial vaccine, 4 weeks apart, and animals in the control groups received the same dose of saline. Groups of 12 vaccinated and 12 control lambs were randomly selected in each study for challenge with serovars Hardjo or Pomona. Challenge was initiated 16 weeks following the second vaccination with three daily doses of live leptospires by intranasal and conjunctival routes. Following challenge, urine samples were collected weekly for 6 weeks, for dark field microscopy and leptospiral culture; 6 weeks after challenge the lambs were slaughtered and kidneys collected for leptospiral culture.

RESULTS: In lambs challenged with serovar Hardjo, 8/12 unvaccinated lambs had ≥1 urine or kidney sample that was positive for leptospires following culture, compared with 0/12 lambs in the vaccinated group (p=0.001). In lambs challenged with serovar Pomona, 9/12 unvaccinated lambs had ≥1 urine or kidney sample that was positive following culture, compared with 0/12 lambs in the vaccinated group (p<0.001). Prevention of renal colonisation and urinary shedding, expressed as the prevented fraction, was 100 (95% CI=61.7–100)% and 100 (95% CI=68.3–100)% against challenge with serovars Hardjo and Pomona, respectively, at 4 months after vaccination.

CONCLUSIONS AND CLINICAL RELEVANCE: Use of a multivalent leptospiral and clostridial vaccine demonstrated protection against challenge from New Zealand strains of serovars of Hardjo and Pomona 4 months after vaccination in lambs first vaccinated at 9–11 weeks of age. Further studies are required to assess the duration of immunity against challenge in sheep.     

Detection of Mycobacterium avium subsp. paratuberculosis in skeletal muscle and blood of ewes from a sheep farm in New Zealand

Abstract

AIM: To determine whether viable Mycobacterium avium subsp. paratuberculosis (Map) is present in skeletal muscle and blood in ewes with and without Johne's disease confirmed histologically.

METHODS: A total of 51 mixed-aged ewes in poor body condition from a farm with a history of clinical Johne's disease were culled and examined at necropsy. BACTEC radiometric culture was performed on samples of skeletal muscle from the biceps femoris, mononuclear cells in peripheral blood (hereafter referred to as blood), and ileum. Histological sections and Ziehl-Neelsen (ZN)-stained impression smears of terminal ileum and mesenteric lymph nodes were examined. Ewes were defined as having confirmed Johne's disease if there was histopathological evidence typical of the disease within the ileum and adjacent lymph nodes.

RESULTS: Eighteen of 21 (86%) ewes with confirmed clinical Johne's disease were culture-positive for Map from sites peripheral to the alimentary tract, comprising 15 from skeletal muscle and 13 from blood. Five of 30 (17%) ewes that did not have Johne's disease were culture-positive, with four from skeletal muscle and one from blood. The likelihood that ewes with confirmed Johne's disease had systemic Map infection compared with ewes without was determined as OR=30 (95% CI=6.3–142.0; p<0.001).

CONCLUSION: The prevalence of Map infection of skeletal muscle and blood in ewes with confirmed Johne's disease was 71% and 62% respectively, and in unaffected ewes was 13% for muscle and 3% for blood.

CLINICAL RELEVANCE: Skeletal muscle and blood are potential sources of exposure of humans to Map, and the risk appears higher from sheep with Johne's disease.     

Effect of liveweight and condition score of ewes at mating,and shearing mid-pregnancy,on birthweights and growth rates of twin lambs to weaning

AIMS: To determine the effects of liveweight and condition score of ewes at mating, and shearing mid-pregnancy, on birthweights and growth rates of twin lambs from birth to the time of weaning, and; to determine whether any effect of condition score is independent of liveweight.

METHODS: Romney ewes (n=435) which conceived within a 17-day mating period and successfully gave birth to and reared twin lambs to weaning were used. Ewes were managed during pregnancy so that their total liveweight increased at a level equivalent to the expected growth of the conceptuses. Approximately half of the ewes were shorn 70 days after the mid-point of mating, and the remainder were left unshorn, at which time they had 6-months' wool growth. All lambs were weighed within 12 h of birth and again at weaning.

RESULTS: There was a significant linear relationship (R²=0.99) between mean group liveweight and condition score of ewes at mating (difference of 7.3 kg per 1.0 unit condition score). Liveweight of ewes at mating had small but significant (p<0.05) positive effects on both birthweights and growth rates of twin lambs to weaning. Ewes that had condition scores of 3.5–4.0 (scale 0–5) at mating gave birth to significantly (p<0.05) lighter lambs than ewes that had condition scores <3.5. The condition score of ewes at mating was positively associated with growth rates of lambs, although this effect was largely explained by variation in the liveweight of ewes at mating.

Twin lambs born to ewes shorn mid-pregnancy were significantly (p<0.05) heavier at birth (by 0.4 kg) and at weaning (p<0.05) than those born to unshorn ewes. The heavier weaning weights of lambs born to ewes shorn mid-pregnancy were due to a heavier birthweights rather than increased growth rate.

CONCLUSIONS: Liveweight and condition score of ewes at mating had small positive effects on growth rates to weaning of twin lambs. Shearing mid-pregnancy increased birthweights and weaning weights of twin lambs. To achieve high weaning weights of twin lambs, farmers should ensure ewes attain high liveweights and condition scores at mating and shear mid-pregnancy. It is speculated that the effects of liveweight and condition score of ewes at mating on weaning weights of twin lambs would be greater than those observed in the present study under conditions in which nutrition of the dam is below optimum.     

Reproductive dominance of pasture trees in a fragmented tropical forest mosaic

Tropical forest fragmentation threatens biodiversity, yet basic information on population responses for major groups such as plants is lacking. Hypervariable genetic markers were used to reconstruct a population-level pedigree in fragmented tropical forest for the tree Symphonia globulifera. Though seedlings occurred only in remnant forest, the pedigree showed that most seedlings had been produced by sequentially fewer adults in pasture, creating a genetic bottleneck. The pedigree also implicated shifts in the foraging of animals that disperse pollen and seed in a secondary constriction of the bottleneck. These results suggest that tropical conservation strategies should anticipate complex, cryptic responses to fragmentation.     

Experimental demonstration of guiding and bending of electromagnetic waves in a photonic crystal

The routing and interconnection of optical signals through narrow channels and around sharp corners are important for large-scale all-optical circuit applications. A recent computational result suggests that photonic crystals may offer a novel way of achieving this goal by providing a mechanism for guiding light that is fundamentally different from traditional index guiding. Waveguiding in a photonic crystal and near 100 percent transmission of electromagnetic waves around sharp 90 degree corners were observed experimentally. Bending radii were made smaller than one wavelength.     

Evidence for extreme climatic warmth from late cretaceous arctic vertebrates

A Late Cretaceous (92 to 86 million years ago) vertebrate assemblage from the high Canadian Arctic (Axel Heiberg Island) implies that polar climates were warm (mean annual temperature exceeding 14 degreesC) rather than near freezing. The assemblage includes large (2.4 meters long) champsosaurs, which are extinct crocodilelike reptiles. Magmatism at six large igneous provinces at this time suggests that volcanic carbon dioxide emissions helped cause the global warmth.     

A comparison of the interferon gamma assay with the absorbed ELISA for the diagnosis of Johne''s disease in cattle

Two interferon gamma (IFN-gamma) assays, the IFN-gamma enzyme immunoassay (EIA) and the IFN-gamma bioassay and an absorbed ELISA were used to screen 6 cattle herds for Johne's disease. Each herd had a history of Johne's disease but the majority of infected animals did not show clinical signs. The disease status of the cattle, which were removed from the herds, was confirmed by bacteriological culture of faeces or histopathological examination and culture of tissues collected at necropsy. The sensitivities of the IFN-gamma assays and the absorbed ELISA were determined using test results from infected animals. The sensitivity of the IFN-gamma EIA in detecting subclinical (71.8 to 93.3%) and clinical animals (100%) was not significantly different. However, the IFN-gamma bioassay and the absorbed ELISA were more sensitive in detecting cattle with advanced infections (80%) than those that were subclinically affected (16.7 to 33.3%).