Use of monoclonal antibodies against human HLA II antigens for the detection of bovine B lymphocytes and macrophages]

The crossreactivity of mouse monoclonal antibodies (MoAbs) (Tab. I) prepared against human HLA-DR and HLA-DP antigens was studied in various bovine cells: lymphocytes from lymph nodes and peripheral blood, adherent (B) and nonadherent (T) lymphocytes, monocytes, granulocytes and platelets. In the immunofluorescence test, MoAbs Bra13, Bra14, Bra20, Bra22, Bra30, Bra70, HL-38 reacted with bovine B lymphocytes and monocytes, but not with other tested cells (Tab. III, IV). These antibodies, except Bra22, were positive with B lymphocytes in the complement dependent cytotoxic test (Tab. II). The similarity of the bovine antigens and HLA-DR antigens determined by used MoAbs was also proved by immunoblotting. Monoclonal antibodies Bra38 and BraFB6 did not react with the bovine cells and separated antigens. The epitope (HLA-DR) recognized by the antibody Bra38 is probably absent in cattle. The presence of HLA-DP analogue determined by the antibody BraFB6 has not been confirmed. The crossreactive MoAbs could be used for the detection of B lymphocytes and macrophages in veterinary immunology.     

Toxicity of Haemophilus pleuropneumoniae to porcine lung macrophages

Viable Haemophilus pleuropneumoniae bacteria were toxic for porcine alveolar macrophages in vitro. This cytotoxic effect proved to be dose-related. A cell-free extract of H. pleuropneumoniae, heat-killed bacteria, and a Pasteurella multocida field strain were nontoxic. When macrophages were cultured with H. pleuropneumoniae bacteria in a ratio of 100 macrophages to six bacteria, ultrastructural signs of cellular degeneration were observed within 1 h. This degeneration was observed in macrophages with or without phagosomes containing H. pleuropneumoniae. A cytotoxic substance was filtered from a H. pleuropneumoniae culture in Eagle's minimal essential medium supplemented with Earle's salts (EMEM) and 10% foetal calf serum that was incubated for 10 h at 37 degrees C. This substance was destroyed by heating at 65 degrees C for 30 min. Macrophages were less susceptible to the toxic effect of H. pleuropneumoniae when serum of convalescent pigs was added.     

Mystery swine disease in The Netherlands: the isolation of Lelystad virus.

In early 1991, the Dutch pig-industry was struck by the so-called mystery swine disease. Large-scale laboratory investigations were undertaken to search for the etiological agent. We focused on isolating viruses and mycoplasmas, and we tested paired sera of affected sows for antibodies against ten known pig viruses. The mycoplasmas M. hyosynoviae, M. hyopneumoniae, and Acholeplasma laidlawii, and the viruses encephalomyocarditis virus and porcine enterovirus types 2 and 7 were isolated from individual pigs. An unknown agent, however, was isolated from 16 of 20 piglets and from 41 of 63 sows. This agent was characterised as a virus and designated Lelystad virus. No relationship between this virus and other viruses has yet been established. Of 165 sows reportedly afflicted by the disease, 123 (75 per cent) seroconverted to Lelystad virus, whereas less than 10 per cent seroconverted to any of the other virus isolates or to the known viral pathogens. Antibodies directed against Lelystad virus were also found in pigs with mystery swine disease in England, Germany, and in the United States. We conclude that infection with Lelystad virus is the likely cause of mystery swine disease.     

Physiological interpretation of a hyperspectral time series in a citrus orchard

Hyperspectral remote sensing for monitoring horticultural production systems requires the understanding of how plant physiology, canopy structure, management and solar elevation affect the retrieved canopy reflectance during different stages of the phenological cycle. Hence, the objective of this study was to set up and to interpret a hyperspectral time series for a mature and healthy citrus orchard in the Western Cape province of South Africa considering these effects. Based on the remotely sensed data, biophysical parameters at the canopy level were derived and related to known observed physiological and phenological changes at the leaf level and to orchard management. Fractions of mature fruit, flowers, and sunburnt leaves were considered, and indices related to canopy structure chlorophyll content and canopy water status were calculated.Results revealed small cover fractions of mature fruit, flowers and sunburnt leaves of respectively 2.1%, 3.1% and 7.0%, but the high spectral contrast between flowers and leaves allowed a successful classification of flowering intensity. Furthermore, it was shown that canopy level time series of vegetation indices were sensitive to changes in solar elevation and soil reflectance which could be reduced by applying an empirical soil line correction for the most affected indices. Most trends in vegetation indices at the canopy level could be explained by a combination of changes at the leaf level (chlorophyll, carotenoids, dry matter), changes in canopy structure (leaf area index and leaf angle distribution) and changes in cover fractions of vegetative flushes, flowers and sunburnt leaves. The transformed chlorophyll absorption ratio index over the optimised soil adjusted vegetation index (MCARI/OSAVI) was best related to leaf level trends in chlorophyll content. Seasonal changes in the photochemical reflectance index (PRI) were linked to inverse changes in the carotenoid-to-chlorophyll ratio. Canopy structure indices (the modified triangular vegetation index or MTVI2 and the standardized leaf area index determining index or sLAIDI) were sensitive to changes in leaf area index, average leaf angle as well to management interactions (pruning and harvest). Canopy water status was highly impacted during the spring flush due to expanding leaves that concealed trends in the underlying mature leaves. Seasonal trends in soil and weeds reflectance were related to changes in volumetric soil water content and to the earlier and reduced growth period of non-irrigated weeds.     

Seasonal variation in canopy reflectance and its application to determine the water status and water use by citrus trees in the Western Cape, South Africa

This study describes the diurnal and seasonal dynamics of the canopy reflectance, water use and water status of Midknight Valencia citrus trees under semi-arid conditions. Hyperspectral canopy reflectance data was collected on 30 trees at monthly intervals over a period of 16 months in a commercial orchard in South Africa. The mean canopy reflectance in the wavelength range 350-2500 nm followed a clear seasonal trend influenced by environmental conditions and tree phenology. Mean monthly reflectance peaked in summer (∼22%) while the lowest value (∼15%) was reached in winter with the seasonal changes in the sun's position accounting for a significant proportion of the variations. A sensitivity analysis of a Penman-Monteith transpiration model showed that water use by individual trees changed by up to 13% when the canopy reflectance was varied over the seasonal range of measured values. This suggested that the seasonal changes in tree water use influenced the seasonal trend of the canopy reflectance. Thus monitoring the canopy reflectance of citrus trees could offer information on the tree water status. To test this, sap flow data of water uptake and loss by the trees were compared with the canopy spectra. Sap flow data showed a heavy reliance by the citrus trees on the internally stored water with up to 25% of the daily total transpiration withdrawn from the trees’ internal water storage pools when soil water was limited. This depletion of internally stored water, and hence the change in tree water status, was detected using spectral indices based on the first order derivatives of the canopy reflectance centered at two and, at most, four spectral bands. We conclude that even if citrus trees are evergreen, their canopy reflectance changes significantly throughout the year with a considerable impact on tree energy balance and water use. In addition, the contribution of the internally stored water to daily transpiration is a possible indicator of drought stress for citrus trees detectable from changes in canopy reflectance and it has potential applications in irrigation scheduling using canopy level spectral information.     

Effect of an immunomodulator on the efficacy of an attenuated vaccine against avian pneumovirus in turkeys

Since 1997, avian pneumovirus (APV) has caused estimated annual losses of $15 million to the Minnesota turkey industry. In order to develop an attenuated live vaccine against APV, we serially passaged a Minnesota isolate of APV (APV/MN/turkey/1-a/97) in vitro in cell cultures for 41 passages. Laboratory experiments with this high-passage virus (P41) indicated that the attenuated virus provided immunogenic protection to turkeys against challenge with virulent APV, although some birds showed mild to moderate dinical signs after inoculation. To reduce the residual pathogenicity of P41, while maintaining its immunogenicity, we decided to vaccinate turkeys with P41 in the presence of an immunomodulator, S-28828 (1-n-butyl-2-ethoxymethyl-1H-imidazo[4,5-c]quinolin-4-amine-hydrochloride), which is a potent cytokine inducer. The combined inoculation of S-28828 (5 mg/kg body weight) and P41 resulted in a significant reduction in the incidence of virus-induced clinical signs in comparison with birds that received P41 without immunomodulator (P < 0.05). Only 17% of birds inoculated with S-28828 + APV P41 showed mild respiratory symptoms at 5 days postinoculation as compared with 46% of the vaccinated turkeys that did not receive S-28828. Vaccination with either P41 or with P41 + S-28828 protected turkeys against dinical signs and viral replication after challenge with virulent APV. These results indicate that immunomodulators, such as S-28828, may act as good vaccine adjuvants that can reduce the pathogenicity but maintain the immunogenicity of partially attenuated vaccines.     

Experimental reproduction of malabsorption syndrome with different combinations of reovirus, Escherichia coli, and treated homogenates obtained from broilers

Attempts to reproduce malabsorption syndrome (MAS) by oral inoculation with several different combinations including intestinal homogenate, reovirus, and hemolytic Escherichia coli obtained from MAS-affected chickens and intestinal homogenate from healthy chickens (healthy homogenate) were performed in 1-day-old specific-pathogen-free (SPF) broilers. The MAS homogenate, serving as a positive control, induced weight gain depression and intestinal lesions such as cystic crypts of Lieberkuhn, villus atrophy, and lymphoid and/or granulocytic infiltration. The healthy homogenate, the formalin-treated MAS homogenate, the formalin-treated healthy homogenate, and phosphate-buffered saline caused neither weight gain depression nor intestinal lesions. We were able to reproduce both weight gain depression and intestinal lesions by inoculation of reovirus either combined with the formalin-treated MAS homogenate or combined with healthy homogenate. Surprisingly, when hemolytic E. coli was added to the combination of reovirus with formalin-treated MAS homogenate, this did not cause weight gain depression although this combination caused the described intestinal lesions. Identical results were obtained with the combination of formalin-treated MAS homogenate with hemolytic E coli or the combination of reovirus with hemolytic E. coli. The intestinal lesions were more severe and developed faster by combinations including reovirus and formalin-treated MAS homogenate. This study indicates that a combination of enteropathogenic reovirus with other agents or substances that are present in an intestinal homogenate from MAS-affected and healthy chickens can induce MAS in SPF broilers. Escherichia coli is not essential for induction of weight gain depression but can play a role in development of intestinal lesions. Furthermore, intestinal lesions alone will not always result in weight gain depression.     

Immunological basis of differences in disease resistance in the chicken

Genetic resistance to diseases is a multigenic trait governed mainly by the immune system and its interactions with many physiologic and environmental factors. In the adaptive immunity, T cell and B cell responses, the specific recognition of antigens and interactions between antigen presenting cells, T cells and B cells are crucial. It occurs through a network of mediator proteins such as the molecules of the major histocompatibility complex (MHC), T cell receptors, immunoglobulins and secreted proteins such as the cytokines and antibodies. The diversity of these proteins that mainly is due to an intrinsic polymorphism of the genes causes phenotypic variation in disease resistance. The well-known linkage of MHC polymorphism and Marek's disease resistance difference represents a classic model revealing immunological factors in resistance differences and diversity of mediator molecules. The molecular bases in any resistance variation to infectious pathogens are vaguely understood. This paper presents a review of the major immune mediators involved in resistance and susceptibility to infectious diseases and their functional mechanisms in the chicken. The genetic interaction of disease resistance with production traits and the environment is mentioned.