Evaluation of three enzyme-linked immunosorbent assays (ELISAs) for the detection of serum antibodies in sheep infected with Echinococcus granulosus

The aim of this study was to develop an immunological method for the identification of sheep infected with Echinococcus granulosus which would allow the monitoring of animals imported into countries free from hydatidosis and as an aid to countries where control schemes for the disease are in operation. Three enzyme-linked immunosorbent assays (ELISAs) were developed and validated, using as antigen either a purified 8 kDa hydatid cyst fluid protein (8kDaELISA), a recombinant EG95 oncosphere protein (OncELISA) or a crude protoscolex preparation (ProtELISA). Sera used for the assay validations were obtained from 249 sheep infected either naturally or experimentally with E. granulosus and from 1012 non-infected sheep. The highest diagnostic sensitivity was obtained using the ProtELISA at 62.7 and 51.4%, depending on the cut-off. Assay sensitivities were lower for the 8kDaELISA and the OncELISA. Diagnostic specificities were high, ranging from 95.8 to 99.5%, depending on the ELISA type and cut-off level chosen. A few sera from 39 sheep infected with T. hydatigena and from 19 sheep infected with T. ovis were recorded as positive. Western immunoblot analysis revealed that the dominant antigenic components in the crude protoscolex antigen preparation were macromolecules of about 70-150 kDa, most likely representing polysaccharides. This study demonstrated that the ProtELISA was the most effective immunological method of those assessed for detection of infection with E. granulosus in sheep. Because of its limited diagnostic sensitivity of about 50-60%, it should be useful for the detection of the presence of infected sheep on a flock basis and cannot be used for reliable identification of individual animals infected with E. granulosus.     

Cytological analysis of equine bronchoalveolar lavage fluid. Part 2: Comparison of smear and cytocentrifuged preparations

The aim of this study was to develop a diagnostically useful smear method for preparation of equine bronchoalveolar lavage fluid (BALF) for use by practitioners. A smear method for equine BALF preparation which included the addition of serum was developed, and cell morphology, differential cell counts (DCC) and repeatability of counting DCC compared with those of cytocentrifuged BALF preparations. BALF samples (n = 21) were collected from 5 control horses and 5 heaves-susceptible horses. Smear preparations of BALF produced smaller, darker, staining cells, making cytological identification more difficult than on cytocentrifuged preparations. There was a significantly higher (P<0.01) macrophage DCC and lower lymphocyte DCC on cytocentrifuged compared to smear preparations. Mast cell and eosinophil DCC were significantly higher (P<0.05) on cytocentrifuged compared to smear preparations of BALF. Smear preparations were shown to be reliable for the cytological diagnosis of equine neutrophilic pulmonary disease and offer practitioners an alternative to sending equine BALF to a laboratory for processing and cytological analysis.     

Canine atopic dermatitis: a retrospective study of 169 cases examined at the University of California, Davis, 1992-1998. Part II. Response to hyposensitization

One hundred and sixty-nine dogs were diagnosed with atopic dermatitis, and treated with hyposensitization for at least 1 year based on the results of either intradermal skin tests (IDST) or enzyme-linked immunosorbant serum assays (ELISA). Excellent (i.e. hyposensitization alone controlled clinical signs), good (> 50% improvement), moderate (< 50% improvement) and no (clinical signs were unchanged) responses were seen in 19.5, 32.5, 20.1 and 27.8%, respectively. Age of onset, age when treatment was initiated or the duration of clinical signs had no influence on response to hyposensitization. Dogs having concurrent flea allergy dermatitis were statistically more likely to respond better than dogs with concurrent food allergies. Although not statistically significant, there were trends for Golden Retriever and male dogs to respond better than other breeds and female dogs, respectively. Dogs having more than 21 positive reactions in allergy tests and treated with more than 21 allergens had lower response scores, and a longer time course before achieving beneficial response. Lower response scores were seen in dogs having positive reactions to cultivated plants, grasses, trees or insects. There was no difference in response to hyposensitization whether based on IDST or ELISA results.     

Major outer membrane proteins of Brucella spp.: past,present and future

The major outer membrane proteins (OMPs) of Brucella spp. were initially identified in the early 1980s and characterised as potential immunogenic and protective antigens. They were classified according to their apparent molecular mass as 36–38 kDa OMPs or group 2 porin proteins and 31–34 and 25–27 kDa OMPs which belong to the group 3 proteins. The genes encoding the group 2 porin proteins were identified in the late 1980s and consist of two genes, omp2a and omp2b, which are closely linked in the Brucella genome, and which share a great degree of identity (>85%). In the 1990s, two genes were identified coding for the group 3 proteins and were named omp25 and omp31. The predicted amino acid sequences of omp25 and omp31 share 34% identity. The recent release of the genome sequence of B. melitensis 16 M has revealed the presence of five additional gene products homologous to Omp25 and Omp31. The use of recombinant protein technology and monoclonal antibodies (MAbs) has shown that the major OMPs appear to be of little relevance as antigens in smooth (S) B. abortus or B. melitensis infections i.e. low or no protective activity in the mouse model of infection and low or no immunogenicity during host infection. However, group 3 proteins, in particular Omp31, appear as immunodominant antigen in the course of rough (R) B. ovis infection in rams and as important protective antigen in the B. ovis mouse model of infection. The major OMP genes display diversity and specific markers have been identified for Brucella species, biovars, and strains, including the recent marine mammal Brucella isolates for which new species names have been proposed. Recently, Omp25 has been shown to be involved in virulence of B. melitensis, B. abortus and B. ovis. Mutants lacking Omp25 are indeed attenuated in animal models of infection, and moreover provide levels of protection similar or better than currently used attenuated vaccine strain B. melitensis Rev.1. Therefore, these mutant strains appear interesting vaccine candidates for the future. The other group 3 proteins identified in the genome merit also further investigation related to the development of new vaccines.     

Intravenous administration of levothyroxine for treatment of suspected myxedema coma complicated by severe hypothermia in a dog

A 7-year-old male English Coonhound with suspected myxedema coma complicated by severe hypothermia and metabolic abnormalities was treated with a combination of active external and core rewarming techniques, i.v. and oral administration of levothyroxine, supplemental oxygen, and administration of fluids (0.9% NaCl solution). Myxedema coma develops as a consequence of severe hypothyroidism and is characterized by a hypometabolic, stuporous state. Myxedema coma is associated with a high mortality rate, and most reported cases have involved Doberman Pinschers. Intravenous administration of levothyroxine can be used successfully in combination with oral administration to restore normal metabolic function and assist in warming and thermoregulation, although dosages should be conservative to avoid adverse cardiovascular effects.     

Anticancer vaccines.

With the tools of molecular biology and a greater understanding of mechanisms to harness the immune system, effective tumor immunotherapy is becoming a reality. This new class of therapeutics offers a more targeted, and therefore precise, approach to the treatment of cancer. The recent conditional licensure of a xenogeneic DNA vaccine for advanced canine malignant melanoma strongly suggests that immunotherapy can play an extremely important role alongside the classic cancer treatment triad components of surgery, radiation therapy, and chemotherapy.     

Land Use – Land Cover Conversion, Regeneration and Degradation in the High Elevation Bolivian Andes

Regional land-cover change affects biodiversity, hydrology, and biogeochemical cycles at local, watershed, and landscape scales. Developing countries are experiencing rapid land cover change, but assessment is often restricted by limited financial resources, accessibility, and historical data. The assessment of regional land cover patterns is often the first step in developing conservation and management plans. This study used remotely sensed land cover and topographic data (Landsat and Shuttle Radar Topography Mission), supervised classification techniques, and spectral mixture analysis to characterize current landscape patterns and quantify land cover change from 1985 to 2003 in the Altiplano (2535–4671 m) and Intermediate Valley (Mountain) (1491–4623 m) physiographic zones in the Southeastern Bolivian Andes. Current land cover was mapped into six classes with an overall accuracy of 88% using traditional classification techniques and limited field data. The land cover change analysis showed that extensive deforestation, desertification, and agricultural expansion at a regional scale occurred in the last 20 years (17.3% of the Mountain Zone and 7.2% of the Altiplano). Spectral mixture analysis (SMA) indicated that communal rangeland degradation has also occurred, with increases in soil and non-photosynthetic vegetation fractions in most cover classes. SMA also identified local areas with intensive management activities that are changing differently from the overall region (e.g., localized areas of increased green vegetation). This indicates that actions of local communities, governments, and environmental managers can moderate the potentially severe future changes implied by the results of this study.     

An upper limit for elevated root zone dissolved oxygen concentration for tomato

It is well understood that insufficient oxygen within plant root zones can greatly diminish plant productivity. However, little is known about the effect of elevated root zone oxygen concentrations. Tomato (Lycopersicon lycopersicum Mill., cv. Trust) seedlings were grown in nutrient solutions containing dissolved oxygen (DO) concentration ranging from 5.3 to 40 mg L⁻¹ for 4 weeks. There were no visible symptoms observed on the leaves or stems in any of the treatments. Leaf chlorophyll content was higher in the 40 mg L⁻¹ treatment than with 20 and 30 mg L⁻¹ DO treatments. Two weeks from the start of the experiment, roots in the 40 mg L⁻¹ treatment exhibited stunted growth, became thicker, and had fewer side and fine roots compared to roots in the lower levels of DO treatment. Almost all the measured growth parameters (fresh and dry weights of root, stem, and leaf, leaf area, stem diameter) were significantly reduced in plants grown in the 40 mg L⁻¹ treatment compared to plants in the lower level of DO treatments, except that the plant height increased with the increasing DO concentration. Root respiration increased linearly with increasing DO concentration; however, there was no effect on leaf net CO₂ exchange rate. It is suggested that it was safe to enrich root zone DO to as high as 30 mg L⁻¹, although the growth benefit was minor by increasing DO from ambient air saturated level (∼8.5 mg L⁻¹) to 30 mg L⁻¹. Higher than 30 mg L⁻¹ could cause reduction in tomato plant growth.